Name
stringlengths 1
127
| ID
stringlengths 1
6
| Flame
stringclasses 4
values | Purpose
stringlengths 0
474
| Depositing Lab
stringlengths 0
42
| Publication
stringlengths 0
220
| GenBank File
stringlengths 0
144
| Sequence Type
stringclasses 2
values | GenBank Raw
stringlengths 0
181k
| Backbone
dict | Growth in Bacteria
dict | Terms and Licenses
dict | References
dict | Gene/Insert 1
dict | Cloning Information for Gene/Insert 1
dict | Gene/Insert 2
dict | Cloning Information for Gene/Insert 2
dict | Gene/Insert 3
dict | Cloning Information for Gene/Insert 3
dict |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
cmv 16 E7 68CVQ | 13691 | Karl Munger | CMV E7 plasmids (unpublished) | https://media.addgene.org/snapgene-media/v2.0.0/sequences/99773/7ff514bd-902f-4529-b6c3-5cb8c0224fac/addgene-plasmid-13691-sequence-99773.gbk | partial | LOCUS sequence-99773-e 1049 bp DNA linear UNA 26-AUG-2024
DEFINITION natural linear DNA
ACCESSION .
VERSION .
KEYWORDS .
SOURCE natural DNA sequence
ORGANISM unspecified
REFERENCE 1 (bases 1 to 1049)
TITLE CMV E7 plasmids
REFERENCE 2 (bases 1 to 1049)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..1049
/mol_type="genomic DNA"
/organism="unspecified"
primer_bind complement(556..575)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_signal 621..676
/label=beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(675..694)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
ORIGIN
1 atgatataca ctgtttgaga tgaggataaa atactctgag tccaaaccgg gcccctctgc
61 taaccatgtt catgccttct tctctttcct acagctcctg ggcaacgtgc tggttgttgt
121 gctgtctcat cattttggca aagaattcct cgacggatcc accatgcatg gagatacacc
181 tacattgcat gaatatatgt tagatttgca accagagaca actgatctct actgttatga
241 gcaattaaat gacagctcag aggaggagga tgaaatagat ggtccagctg gacaagcaga
301 accggacaga gcccattaca atattgtaac cttttgttgc aagtgtgact ctacgcttcg
361 gttggctgct gcaagtactc acgtagacat tcgtactttg gaagacctgt taatgggcac
421 actaggaatt gtgtgcccca tctgttctca gaaaccataa agatccgtcg aggaattcac
481 tcctcaggtg caggctgcct atcagaaggt ggtggctggt gtggccaatg ccctggctca
541 caaataccac tgagatcttt ttccctctgc caaaaattat ggggacatca tgaagcccct
601 tgagcatctg acttctggct aataaaggaa atttattttc attgcaatag tgtgttggaa
661 ttttttgtgt ctctcactcg gaaggacata tgggagggca aatcatttaa aacatcagaa
721 tgagtatttg gtttagagtt tggcaacata tgcccatatg ctggctgcca tgaacaaagg
781 ttggctataa agaggtcatc agtatatgaa acagccccct gctgtccatt ccttattcca
841 tagaaaagcc ttgacttgag gttagatttt ttttatattt tgttttgtgt tatttttttc
901 tttaacatcc ctaaaatttt ccttacatgt tttactagcc agatttttcc tcctctcctg
961 actactccca gtcatagctg tccctcttct cttatggaga tccctcgacg gatcggccgc
1021 aattcgtaat catgtcatag ctgtttcct
//
| {
"Vector backbone": "pCMV bam neo",
"Backbone size w/o insert (bp)": "6550",
"Vector type": "Mammalian Expression",
"Selectable markers": "Neomycin (select with G418)"
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "",
"Publication": "",
"DOI": "",
"Authors": "",
"PubMed Link": "",
"PubMed ID": ""
} | {
"Gene/Insert name": "HPV 16 E7 CVQ6870AAA",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "300",
"Mutation": "aa 68-70 changed from CVQ to AAA",
"Entrez Gene": "E7",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "BamH1 (not destroyed)",
"3′ cloning site": "BamH1 (destroyed during cloning)",
"5′ sequencing primer": "B glob intron-F",
"3′ sequencing primer": "B glob-pA-R"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
||
pGBW-m4252975 | 153873 | Mammalian expression plasmid for SARS-CoV-2 RNA-dependent RNA polymerase | SARS-CoV-2 protein Mammalian expression constructs, tagged, for protein expression and/or purification (unpublished) | https://media.addgene.org/snapgene-media/v2.0.0/sequences/300418/fa0853bc-6096-4c4a-8354-ad39c8e0031b/addgene-plasmid-153873-sequence-300418.gbk | full | LOCUS sequence-300418- 8888 bp DNA circular SYN 26-AUG-2024
DEFINITION Mammalian expression plasmid for SARS-CoV-2 RNA-dependent RNA
polymerase.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8888)
TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for
protein expression and/or purification
REFERENCE 2 (bases 1 to 8888)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..8888
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 85..156
/label=5' beta-globin insulator
/note="insulator upstream of the human beta-globin locus
(Farrell et al., 2002)"
terminator complement(161..207)
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
polyA_signal 225..273
/note="synthetic polyadenylation signal"
enhancer 354..657
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 659..861
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 812..832
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 930..939
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
polyA_signal 3781..3902
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3792..3811)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 3846..3865
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
terminator 4017..4063
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
terminator 4093..4229
/label=rrnG terminator
/note="transcription terminator from the E. coli ribosomal
RNA rrnG operon (Albrechtsen et al., 1991)"
misc_feature 4274..4345
/label=3' beta-globin insulator
/note="insulator downstream of the human beta-globin locus
(Farrell et al., 2002)"
rep_origin complement(4571..5159)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4651..4670)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
promoter 5659..6868
/label=UbC promoter
/note="human ubiquitin C promoter"
primer_bind 6705..6724
/label=hUBCpro-F
/note="Human Ubiquitin C (UbC) promoter, forward primer"
CDS 6872..7471
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind complement(6872..6891)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 7368..7388
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
polyA_signal 7498..7546
/note="synthetic polyadenylation signal"
CDS 7812..8672
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(8030..8049)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
terminator 8685..8821
/label=rrnG terminator
/note="transcription terminator from the E. coli ribosomal
RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
1 gccgtggcgc tcatttgctc gtcgggcatc gaatctcgtc agctatcgtc agcttacctt
61 tttggcagcg atcgcggctc ccgacatctt ggaccattag ctccacaggt atcttcttcc
121 ctctagtggt cataacagca gcttcagcta cctctcaatt caaaaaaccc ctcaagaccc
181 gtttagaggc cccaaggggt tatgctatca atcgttgcgt tacacacaca aaaaaccaac
241 acacatccat cttcgatgga tagcgatttt attatctaac tgctgatcga gtgtagccag
301 atctagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
361 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
421 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
481 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
541 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
601 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatgctg
661 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
721 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
781 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
841 gaggtctata taagcagagc tggtttagtg aaccgtcaga tcagatcttt gtcgatccta
901 ccatccactc gacacacccg ccagcggccg ccaccatggc tgatgcacaa tcgtttttaa
961 accgggtttg cggtgtaagt gcagcccgtc ttacaccgtg cggcacaggc actagtactg
1021 atgtcgtata cagggctttt gacatctaca atgataaagt agctggtttt gctaaattcc
1081 taaaaactaa ttgttgtcgc ttccaagaaa aggacgaaga tgacaattta attgattctt
1141 actttgtagt taagagacac actttctcta actaccaaca tgaagaaaca atttataatt
1201 tacttaagga ttgtccagct gttgctaaac atgacttctt taagtttaga atagacggtg
1261 acatggtacc acatatatca cgtcaacgtc ttactaaata cacaatggca gacctcgtct
1321 atgctttaag gcattttgat gaaggtaatt gtgacacatt aaaagaaata cttgtcacat
1381 acaattgttg tgatgatgat tatttcaata aaaaggactg gtatgatttt gtagaaaacc
1441 cagatatatt acgcgtatac gccaacttag gtgaacgtgt acgccaagct ttgttaaaaa
1501 cagtacaatt ctgtgatgcc atgcgaaatg ctggtattgt tggtgtactg acattagata
1561 atcaagatct caatggtaac tggtatgatt tcggtgattt catacaaacc acgccaggta
1621 gtggagttcc tgttgtagat tcttattatt cattgttaat gcctatatta accttgacca
1681 gggctttaac tgcagagtca catgttgaca ctgacttaac aaagccttac attaagtggg
1741 atttgttaaa atatgacttc acggaagaga ggttaaaact ctttgaccgt tattttaaat
1801 attgggatca gacataccac ccaaattgtg ttaactgttt ggatgacaga tgcattctgc
1861 attgtgcaaa ctttaatgtt ttattctcta cagtgttccc acctacaagt tttggaccac
1921 tagtgagaaa aatatttgtt gatggtgttc catttgtagt ttcaactgga taccacttca
1981 gagagctagg tgttgtacat aatcaggatg taaacttaca tagctctaga cttagtttta
2041 aggaattact tgtgtatgct gctgaccctg ctatgcacgc tgcttctggt aatctattac
2101 tagataaacg cactacgtgc ttttcagtag ctgcacttac taacaatgtt gcttttcaaa
2161 ctgtcaaacc cggtaatttt aacaaagact tctatgactt tgctgtgtct aagggtttct
2221 ttaaggaagg aagttctgtt gaattaaaac acttcttctt tgctcaggat ggtaatgctg
2281 ctatcagcga ttatgactac tatcgttata atctaccaac aatgtgtgat atcagacaac
2341 tactatttgt agttgaagtt gttgataagt actttgattg ttacgatggt ggctgtatta
2401 atgctaacca agtcatcgtc aacaacctag acaaatcagc tggttttcca tttaataaat
2461 ggggtaaggc tagactttat tatgattcaa tgagttatga ggatcaagat gcacttttcg
2521 catatacaaa acgtaatgtc atccctacta taactcaaat gaatcttaag tatgccatta
2581 gtgcaaagaa tagagctcgc accgtagctg gtgtctctat ctgtagtact atgaccaata
2641 gacagtttca tcaaaaatta ttgaaatcaa tagccgccac tagaggagct actgtagtaa
2701 ttggaacaag caaattctat ggtggttggc acaacatgtt aaaaactgtt tatagtgatg
2761 tagaaaaccc tcaccttatg ggttgggatt atcctaaatg tgatagagcc atgcctaaca
2821 tgcttagaat tatggcctca cttgttcttg ctcgcaaaca tacaacgtgt tgtagcttgt
2881 cacaccgttt ctatagatta gctaatgagt gtgctcaagt attgagtgaa atggtcatgt
2941 gtggcggttc actatatgtt aaaccaggtg gaacctcatc aggagatgcc acaactgctt
3001 atgctaatag tgtttttaac atttgtcaag ctgtcacggc caatgttaat gcacttttat
3061 ctactgatgg taacaaaatt gccgataagt atgtccgcaa tttacaacac agactttatg
3121 agtgtctcta tagaaataga gatgttgaca cagactttgt gaatgagttt tacgcatatt
3181 tgcgtaaaca tttctcaatg atgatactct ctgacgatgc tgttgtgtgt ttcaatagca
3241 cttatgcatc tcaaggtcta gtggctagca taaagaactt taagtcagtt ctttattatc
3301 aaaacaatgt ttttatgtct gaagcaaaat gttggactga gactgacctt actaaaggac
3361 ctcatgaatt ttgctctcaa catacaatgc tagttaaaca gggtgatgat tatgtgtacc
3421 ttccttaccc agatccatca agaatcctag gggccggctg ttttgtagat gatatcgtaa
3481 aaacagatgg tacacttatg attgaacggt tcgtgtcttt agctatagat gcttacccac
3541 ttactaaaca tcctaatcag gagtatgctg atgtctttca tttgtactta caatacataa
3601 gaaagctaca tgatgagtta acaggacaca tgttagacat gtattctgtt atgcttacta
3661 atgataacac ttcaaggtat tgggaacctg agttttatga ggctatgtac acaccgcata
3721 cagtcttata agctagcttg actgactgag atacagcgta ccttcagctc acagacatga
3781 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
3841 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
3901 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
3961 tttaaagcaa gtaaaacctc tacaaatgtg gtattggccc atctctatcg gtatcgtagc
4021 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgtgcccct cgggccggat
4081 tgctatctac cggcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac
4141 tcccacatat gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg
4201 tgtcctcctt accagaaatt tatccttaag gtcgtcagct atcctgcagg cgatctctcg
4261 atttcgatca agacattcct ttaatggtct tttctggaca ccactagggg tcagaagtag
4321 ttcatcaaac tttcttccct ccctaatctc attggttacc ttgggctatc gaaacttaat
4381 taaccagtca agtcagctac ttggcgagat cgacttgtct gggtttcgac tacgctcaga
4441 attgcgtcag tcaagttcga tctggtcctt gctattgcac ccgttctccg attacgagtt
4501 tcatttaaat catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
4561 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
4621 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
4681 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
4741 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
4801 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
4861 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
4921 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
4981 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
5041 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
5101 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
5161 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
5221 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
5281 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
5341 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcattta
5401 aatttccgaa ctctccaagg ccctcgtcgg aaaatcttca aacctttcgt ccgatccatc
5461 ttgcaggcta cctctcgaac gaactatcgc aagtctcttg gccggccttg cgccttggct
5521 attgcttggc agcgcctatc gccaggtatt actccaatcc cgaatatccg agatcgggat
5581 cacccgagag aagttcaacc tacatcctca atcccgatct atccgagatc cgaggaatat
5641 cgaaatcggg gcgcgcctgg cctccgcgcc gggttttggc gcctcccgcg ggcgcccccc
5701 tcgtcacggc gagcgctgcc acgtcagacg aagggcgcag gagcgtcctg atccttccgc
5761 ccggacgctc aggacagcgg cccgctgctc ataagactcg gccttagaac cccagtatca
5821 gcagaaggac attttaggac gggacttggg tgactctagg gcactggttt tctttccaga
5881 gagcggaaca ggcgaggaaa agtagtccct tctcggcgat tctgcggagg gatctccgtg
5941 gggcggtgaa cgccgatgat tatataagga cgcgccgggt gtggcacagc tagttccgtc
6001 gcagccggga tttgggtcgc ggttcttgtt tgtggatcgc tgtgatcgtc acttggtgag
6061 tagcgggctg ctgggctggc cggggctttc gtggccgccg ggccgctcgg tgggacggaa
6121 gcgtgtggag agaccgccaa gggctgtagt ctgggtccgc gagcaaggtt gccctgaact
6181 gggggttggg gggagcgcag caaaatggcg gctgttcccg agtcttgaat ggaagacgct
6241 tgtgaggcgg gctgtgaggt cgttgaaaca aggtgggggg catggtgggc ggcaagaacc
6301 caaggtcttg agcccttcgc taatgcggga aagctcttat tcgggtgaga tgggctgggc
6361 accatctggg gaccctgacg tgaagtttgt cactgactgg agaactcggt ttgtcgtctg
6421 ttgcgggggc ggcagttatg gcggtgccgt tgggcagtgc acccgtacct ttgggagcgc
6481 gcgccctcgt cgtgtcgtga cgtcacccgt tctgttggct tataatgcag ggtggggcca
6541 ccagccggta ggtgtgcggt aggcttttct ccgtcgcagg acgcagggtt cgggcctagg
6601 gtaggctctc ctgaatcgac aggcgccgga cctctggtga ggggagggat aagtgaggcg
6661 tcagtttctt tggtcggttt tatgtaccta tcttcttaag tagctgaagc tccggttttg
6721 aactatgcgc tcggggttgg cgagtgtgtt ttgtgaagtt ttttaggcac cttttgaaat
6781 gtaatcattt gggtcaatat gtaattttca gtgttagact tgtaaattgt ccgctaaatt
6841 ctggccgttt ttggcttttt tgttagacaa catgaccgag tacaagccca cggtgcgcct
6901 cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta
6961 ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca
7021 agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg
7081 cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga
7141 gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga
7201 gggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt
7261 ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc
7321 cgccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt
7381 ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac
7441 ctggtgcatg acccgcaagc ccggtgccta ggctagtatg taagcctagt cttagataat
7501 aaaatcgcta tccatcgaag atggatgtgt gttggttttt tgtgtgtgta acgctaggcg
7561 cgcctggtgt accgagaacg atcctctcag tgcgagtctc gacgatccat atcgttgctt
7621 ggcagtcagc cagtcggaat ccagcttggg acccaggaag tccaatcgtc agatattgta
7681 ctcaagcctg gtcacggcag cgtaccgatc tgtttaaacc tagatattga tagtctgatc
7741 ggtcaacgta taatcgagtc ctagcttttg caaacatcta tcaagagaca ggatcagcag
7801 gaggctttcg catgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
7861 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
7921 agttgggtgc gcgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
7981 gttttcgccc cgaagaacgc tttccaatga tgagcacttt taaagttctg ctatgtggcg
8041 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
8101 agaatgactt ggttgagtat tcaccagtca cagaaaagca tcttacggat ggcatgacag
8161 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
8221 tgacaacgat tggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
8281 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
8341 acaccacgat gcctgtagca atggcaacaa ccttgcgtaa actattaact ggcgaactac
8401 ttactctagc ttcccggcaa cagttgatag actggatgga ggcggataaa gttgcaggac
8461 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
8521 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
8581 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
8641 agataggtgc ctcactgatt aagcattggt aaccgattct aggtgcattg gcgcagaaaa
8701 aaatgcctga tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga
8761 tacggcttcc ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta
8821 agatcccgaa tcgtttaaac tcgactctgg ctctatcgaa tctccgtcgt ttcgagctta
8881 cgcgaaca
//
| {
"Vector backbone": "pTwist_CMV_Puro_6 with AarI site removed",
"Backbone size w/o insert (bp)": "6080",
"Vector type": "Mammalian Expression",
"Selectable markers": "Puromycin"
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "OpenMTA",
"Industry Terms": "OpenMTA"
} | {
"Title": "",
"Publication": "",
"DOI": "",
"Authors": "",
"PubMed Link": "",
"PubMed ID": ""
} | {
"Gene/Insert name": "SARS-CoV-2 RNA-dependent RNA polymerase",
"Species": "Severe acute respiratory syndrome coronavirus 2",
"Insert Size (bp)": "2789",
"Mutation": "wt",
"Entrez Gene": "ORF1ab",
"Promoter": "CMV",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Unknown",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
||
Ftz-M3 | 11245 | Robin Reed | Zhou et al Proc Natl Acad Sci U S A. 2002 Sep 17. 99(19):12203-7. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/18117/4d4f22fa-8da5-49ba-804c-7cf938fcbc09/addgene-plasmid-11245-sequence-18117.gbk | partial | LOCUS sequence-18117-e 743 bp DNA linear UNA 25-AUG-2024
DEFINITION natural linear DNA
ACCESSION .
VERSION .
KEYWORDS .
SOURCE natural DNA sequence
ORGANISM unspecified
REFERENCE 1 (bases 1 to 743)
AUTHORS Zhou Z, Sim J, Griffith J, Reed R
TITLE Purification and electron microscopic visualization of functional
human spliceosomes.
JOURNAL Proc Natl Acad Sci U S A. 2002 Sep 17. 99(19):12203-7.
PUBMED 12215496
REFERENCE 2 (bases 1 to 743)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Proc Natl
Acad Sci U S A. 2002 Sep 17. 99(19):12203-7."
FEATURES Location/Qualifiers
source 1..743
/mol_type="genomic DNA"
/organism="unspecified"
promoter complement(542..560)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(543..560)
/label=SP6
/note="SP6 promoter, forward primer"
ORIGIN
1 acgtctactc gccccagtcg cagacgcaga agctgaagaa tggcgacttt gccacccctc
61 cgccaaccac gcccacctct ctgccgcccc tcgaaggcat cagcacgcca ccccaatcgc
121 cgggggagaa atcgtcgtca gctgtcagcc aggagatcaa tcatcgaatt gtgacagccc
181 cgaatggagc cggcgatttc aattggtcgc acatcgagga gactttggca tcaggtaggc
241 atcacacacg attaacaacc cctaaaaata cactttgaaa atattgaaaa tatgtttttg
301 tatacatttt tgatattttc aaacaatacg cagttataaa actcattagc taacccattt
361 tttctttgct tatgcttaca gattgcaaag actcgaaacg cacccgtcag acgtacaccc
421 gctaccagac cctggagctc gatatccgta caccatcagg gtacgagcta gcccatggcg
481 tacaccatca gggtacgact agtagatctc gtacaccatc agggtacgga attccgtgta
541 ttctatagtg tcacctaaat cgtatgtgta tgatacataa ggttatgtat taattgtagc
601 cgcgttctaa cgacaatatg tacaagccta attgtgtagc atctggctta ctgaagcaga
661 ccctatcatc tctctcgtaa actgccgtca gagtcgggtt tggttggacg aaccttctga
721 gtttctgggt acgccgtccc gca
//
| {
"Vector backbone": "pSP65",
"Backbone size w/o insert (bp)": "3000",
"Vector type": "Bacterial Expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Purification and electron microscopic visualization of functional human spliceosomes",
"Publication": "Proc Natl Acad Sci U S A. 2002 Sep 17. 99(19):12203-7.",
"DOI": "10.1073/pnas.182427099",
"Authors": "Zhou Z, Sim J, Griffith J, Reed R",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/12215496",
"PubMed ID": "PubMed 12215496"
} | {
"Gene/Insert name": "Ftz-M3",
"Species": "D. melanogaster (fly)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": "3 MS2 hairpins (C terminal on insert)"
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "",
"3′ cloning site": "EcoRI (not destroyed)",
"5′ sequencing primer": "T7",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
||
Or65a-Gal4 | 63044 | To express Gal4 in the pattern of the Drosophila melanogaster odorant receptor Or65a, using the promoter sequences of the gene cloned 5' of Gal4 | Leslie Vosshall | Fishilevich et al Curr Biol. 2005 Sep 6;15(17):1548-53. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/109048/701db0ae-20be-4d91-a222-ee7c761f144d/addgene-plasmid-63044-sequence-109048.gbk | partial | LOCUS sequence-109048- 980 bp DNA linear UNA 26-AUG-2024
DEFINITION To express Gal4 in the pattern of the Drosophila melanogaster
odorant receptor Or65a, using the promoter sequences of the gene
cloned 5' of Gal4.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE natural DNA sequence
ORGANISM unspecified
REFERENCE 1 (bases 1 to 980)
AUTHORS Fishilevich E, Vosshall LB
TITLE Genetic and functional subdivision of the Drosophila antennal lobe.
JOURNAL Curr Biol. 2005 Sep 6;15(17):1548-53.
PUBMED 16139209
REFERENCE 2 (bases 1 to 980)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Curr
Biol."; date: "2005-09-6"; volume: "15(17)"; pages: "1548-53"
FEATURES Location/Qualifiers
source 1..980
/mol_type="genomic DNA"
/organism="unspecified"
ORIGIN
1 cgccggggat cagatccgcg gccgcgggaa ttcgattagc gttcagcgtt caccatcagg
61 actaacaagt tggcccctgc cagagcggct gtttcggcca ggaaatgcag cacaaataaa
121 tgaggcagca gtcgcttgac cctcgtccgc caatatttta gccatttgta attttatatt
181 ttgtgccacc ggacggacgg acgaacggac ggggcaatgt ttgcatttta gatttgcctt
241 attggaaatt ctatctggag cgggaacaca tgttcccgtt ttcgatacat atgaatgtat
301 ttttatttta ttttaatttt ttattttttt gggccagcac gagactggca gctgttatgg
361 cccggggtgc aagtgtttgg gccaacacct taatggcccg aaatgagcaa gggagttgcc
421 ttaactttgg ctaattgcca ggcgggtcgt agacaagtgt gtctttcgcg ccccgtgggg
481 cagaaatgca gacgcagcac tcagacagac agtccggcat ttgcataaac atgcaacgtg
541 tctaataccc ggagcgacga gatggaccag accgggtcca gacttccggc tcaaggccgc
601 aacaagcgcc aaactactgg caaacttgaa tacactgttt gagtgactaa ttctagtttt
661 tgccataaaa tttcaaaaca acagggattc aattaacagc cgaaacggac atgaaaaagg
721 cgtggtctgg caaataaaac taagctggaa ctgctgattt cggaccaaag ttcctcaaag
781 ttctatcaac caaaagaact aactttccaa ttgaatctcc taccacttgg ccctggccaa
841 acattcgaac ttgaacttgg cagaaaaact acctaccgca tttcgtggcc agcatccgaa
901 aatggacaaa gtcaacgttt cggcggcatc tcgcaagtgg cgttttgtca aatatttccg
961 aacagtttgt gttgcaggcg
//
| {
"Vector backbone": "pCasperAUG-Gal4-X",
"Backbone size w/o insert (bp)": "12000",
"Vector type": "Insect Expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "Unknown"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Genetic and functional subdivision of the Drosophila antennal lobe",
"Publication": "Curr Biol. 2005 Sep 6;15(17):1548-53.",
"DOI": "10.1016/j.cub.2005.07.066",
"Authors": "Fishilevich E, Vosshall LB",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/16139209",
"PubMed ID": "PubMed 16139209"
} | {
"Gene/Insert name": "Drosophila melanogaster Or65a promoter",
"Species": "D. melanogaster (fly)",
"Insert Size (bp)": "7764",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": "Drosophila ADH (aa's 1-18) fused to Yeast Gal4 (aa's 1-881) (C terminal on backbone)"
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "NotI (not destroyed)",
"3′ cloning site": "NotI (not destroyed)",
"5′ sequencing primer": "pCasper-F 5'-gggttttattaacttacat-3'",
"3′ sequencing primer": "GAL4Nterm-R 5'-tcagatgtgccctagtcagc-3'"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
Rab6bb cds in pGem-T Easy | 80521 | Can be used for in situ hybridisation or other purposes | Rob Parton | Hall et al Cell Logist. 2017 Mar 1;7(1):e1301151. doi: 10.1080/21592799.2017.1301151. eCollection 2017. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/176244/efdc1e88-7ea3-4db4-b57b-dc1b86b4c1c8/addgene-plasmid-80521-sequence-176244.gbk | full | LOCUS sequence-176244- 3646 bp DNA circular SYN 26-AUG-2024
DEFINITION Can be used for in situ hybridisation or other purposes.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3646)
AUTHORS Hall TE, Martel N, Lo HP, Xiong Z, Parton RG
TITLE A plasmid library of full-length zebrafish rab proteins for in vivo
cell biology.
JOURNAL Cell Logist. 2017 Mar 1;7(1):e1301151. doi:
10.1080/21592799.2017.1301151. eCollection 2017.
PUBMED 28396820
REFERENCE 2 (bases 1 to 3646)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Cell
Logist."; date: "2017-03-1"; volume: "7(1):e1301151. doi"; pages: "
10.1080/21592799.2017.1301151. eCollection 2017"
FEATURES Location/Qualifiers
source 1..3646
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(2..19)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(173..761)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(253..272)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(932..1792)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 1555..1574
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(1793..1897)
/gene="bla"
/label=AmpR promoter
rep_origin complement(1975..2430)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2112..2133)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2324..2343
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2556..2578
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 2570..2587
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 2571..2587
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 2594..2613
/label=T7
/note="T7 promoter, forward primer"
promoter 2594..2612
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory 2643..2652
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
regulatory 2647..2656
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
promoter complement(3381..3399)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(3382..3399)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(3417..3433)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3417..3433)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3430..3452)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3441..3457
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(3465..3495)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3510..3531
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
ORIGIN
1 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
61 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
121 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
181 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
241 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
301 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
361 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
421 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
481 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
541 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
601 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
661 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
721 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
781 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
841 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
901 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
961 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
1021 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
1081 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
1141 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
1201 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
1261 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
1321 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
1381 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
1441 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
1501 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
1561 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
1621 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
1681 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
1741 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
1801 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
1861 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
1921 ctgatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggaaattg
1981 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
2041 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
2101 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
2161 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
2221 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
2281 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
2341 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
2401 ccgcgcttaa tgcgccgcta cagggcgcgt ccattcgcca ttcaggctgc gcaactgttg
2461 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
2521 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
2581 ggccagtgaa ttgtaatacg actcactata gggcgaattg ggcccgacgt cgcatgctcc
2641 cggccgccat ggcggccgcg ggaattcgat ttcatgtcca tgggaagtga ctttgggaac
2701 cctttacgga aattcaaact tgttttttta ggggagcaaa gcgttgggaa gacatcgctg
2761 ataacaaggt tcatgtatga cagttttgac aacacttatc aggcaactat tggcattgac
2821 tttttatcaa aaaccatgta tctggaggac cgaactgtga ggttacagct atgggacact
2881 gcagggcagg agcgttttag gagtcttatc ccgagctaca ttagagactc cacagtggct
2941 gtggtggtgt atgatatcac aaatatcaat tcattccagc taacctctaa atggattgat
3001 gatgtcagaa cagagagggg aagtgatgtc attatcatgc tggtgggcaa taaaacagat
3061 ctagaggaaa agaggcaaat cactatagag gagggggagc agcgggccaa agagttgaat
3121 gtgatgttta tcgagactag tgcaaagacc ggcagtaatg ttaaacagtt gtttcggcga
3181 gtggccgctg ctttgcctgg tatggagagc ttggatgata ataacaaaga aggaatgatt
3241 gacattaagc tggacaaaca gccggatcct ccggccactg aaagcggctg ctcctgttag
3301 aatcactagt gaattcgcgg ccgcctgcag gtcgaccata tgggagagct cccaacgcgt
3361 tggatgcata gcttgagtat tctatagtgt cacctaaata gcttggcgta atcatggtca
3421 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
3481 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
3541 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
3601 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgctt
//
| {
"Vector backbone": "pGemT easy",
"Backbone size w/o insert (bp)": "",
"Vector type": "",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "A plasmid library of full-length zebrafish rab proteins for in vivo cell biology",
"Publication": "Cell Logist. 2017 Mar 1;7(1):e1301151. doi: 10.1080/21592799.2017.1301151. eCollection 2017.",
"DOI": "10.1080/21592799.2017.1301151.",
"Authors": "Hall TE, Martel N, Lo HP, Xiong Z, Parton RG /",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/28396820",
"PubMed ID": "PubMed 28396820"
} | {
"Gene/Insert name": "rab6bb",
"Species": "D. rerio (zebrafish)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "rab6bb",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pJet1.2-tHe-Tub3 | 201726 | Tardigrade (Hypsibius exemplaris) gamma-tubulin Tub3 | Stanislav Vinopal | Novotna Floriancicova et al Sci Rep. 2023 Mar 30;13(1):5194. doi: 10.1038/s41598-023-31992-z. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/399737/7a2cf93a-ef82-43a2-b7c5-a3c36a5f286d/addgene-plasmid-201726-sequence-399737.gbk | full | LOCUS sequence-399737- 4393 bp DNA circular SYN 27-AUG-2024
DEFINITION Tardigrade (Hypsibius exemplaris) gamma-tubulin Tub3.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4393)
AUTHORS Novotna Floriancicova K, Baltzis A, Smejkal J, Czernekova M,
Kaczmarek L, Maly J, Notredame C, Vinopal S
TITLE Phylogenetic and functional characterization of water bears
(Tardigrada) tubulins.
JOURNAL Sci Rep. 2023 Mar 30;13(1):5194. doi: 10.1038/s41598-023-31992-z.
PUBMED 36997657
REFERENCE 2 (bases 1 to 4393)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 27, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Sci Rep.";
date: "2023-03-30"; volume: "13(1):5194. doi"; pages: "
10.1038/s41598-023-31992-z"
FEATURES Location/Qualifiers
source 1..4393
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 305..324
/label=T7
/note="T7 promoter, forward primer"
promoter 305..323
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter complement(2225..2255)
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
protein_bind 2270..2291
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(2411..2428)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2582..3170)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2662..2681)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3341..4201)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 3964..3983
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(4202..4306)
/gene="bla"
/label=AmpR promoter
primer_bind 4374..4392
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
ORIGIN
1 gcccctgcag ccgaattata ttatttttgc caaataattt ttaacaaaag ctctgaagtc
61 ttcttcattt aaattcttag atgatacttc atctggaaaa ttgtcccaat tagtagcatc
121 acgctgtgag taagttctaa accatttttt tattgttgta ttatctctaa tcttactact
181 cgatgagttt tcggtattat ctctattttt aacttggagc aggttccatt cattgttttt
241 ttcatcatag tgaataaaat caactgcttt aacacttgtg cctgaacacc atatccatcc
301 ggcgtaatac gactcactat agggagagcg gccgccagat cttccggatg gctcgagttt
361 ttcagcaaga tatgcctcgc tccatcatct ccctgcagct cggccagtgc ggcaaccagg
421 tcggcatgga attctggaag aaattgtgca tggagcatgg catcgagagc aacggccagc
481 tgaagaattc cggcgcggag ggcgactttc gggataggaa ggatttcttc ttctaccagg
541 ccgacgacag ccactacatt ccgcgcgcca tcctcatcga tctcgagccg cgtgtgattc
601 agaccatcaa gaattccgac tacggcaagc tgtttaatcc ggaaaacatt tatgtgtcgg
661 agcagggcgg tggtgcggga aacaactggg ctgcaggatt cggaatgggg caagacgtct
721 cggagaaagt gttcgacatg attgagcgtg aagcggaagg ctgcgacgga ttagagggct
781 tcattctcag tcactccatt gccggcggta ccgggtccgg catgggctca tacgtcctgg
841 agcatttgca tgatcgtttc ccgaagaagc tcattcagac cttctccatc ttccccaatc
901 aggacgactc tgccagcgat gtggtggttc agccgtacaa ctcgatcttg acgatgcgcc
961 ggctggcgga gaacgccgac tgtgtggtgg tgctggacaa cacggcgctc aattcgatcg
1021 ctgcagaacg gctgcgaatc gaccaaccga gcttgacaga gatcaacaat atggtctcaa
1081 cggtgatggc cgccagtacg tccacattgc gctatcccgg ttacatgtac aacaactttg
1141 tcggcttgat ggccccgctc attcccatgc cgaagctcca ttttctcatg gccggctatg
1201 ctccactgac gtcgttggac cagacgagcg gtggcatcca gaaaacgacg gtatttgacg
1261 ttatgcgtcg tttacttcag ccgaagaaca tgatgctgtc cccggggaag gatgccctgg
1321 ctgccgacaa tcgctatctc tccattttga acatcattca gggcgaagtg gaatctggcg
1381 agattcacaa aagcctgatg cgcattaaag aacgcaatct ggtcaagttt atggaatggg
1441 gtccggccag catccatgtg gcccagtcga agcgctcgcc gtttgtcgcg tccaaccacc
1501 gcatcagcgg actgatgttg gccaaccaca ccagcgtgca gggactcttc gacagcatcg
1561 tcaagcagta cgacaagatg aagaagcgca atgcctactt ggatcgcttc accaatgaga
1621 agaattgggc cggtgtgtcg gacatggagc agtcgaggga ggtggtggcc aacctgattg
1681 aggagtacaa ggcagccgcg aaaccggatt acatagacta tctgcggggg aaactgcaga
1741 gtaagaagat gtgaagggat ccttggctcc ttcacccttt acatcgtccc atctttctag
1801 aagatctcct acaatattct cagctgccat ggaaaatcga tgttcttctt ttattctctc
1861 aagattttca ggctgtatat taaaacttat attaagaact atgctaacca cctcatcagg
1921 aaccgttgta ggtggcgtgg gttttcttgg caatcgactc tcatgaaaac tacgagctaa
1981 atattcaata tgttcctctt gaccaacttt attctgcatt ttttttgaac gaggtttaga
2041 gcaagcttca ggaaactgag acaggaattt tattaaaaat ttaaattttg aagaaagttc
2101 agggttaata gcatccattt tttgctttgc aagttcctca gcattcttaa caaaagacgt
2161 ctcttttgac atgtttaaag tttaaacctc ctgtgtgaaa ttattatccg ctcataattc
2221 cacacattat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
2281 aactcacatt aattgcgttg cgctcactgc caattgcttt ccagtcggga aacctgtcgt
2341 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
2401 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
2461 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
2521 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
2581 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
2641 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
2701 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
2761 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
2821 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
2881 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
2941 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
3001 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
3061 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
3121 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
3181 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
3241 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
3301 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
3361 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
3421 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
3481 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
3541 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
3601 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
3661 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
3721 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
3781 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
3841 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
3901 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
3961 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
4021 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
4081 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
4141 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
4201 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
4261 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
4321 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
4381 gtatcacgag gcc
//
| {
"Vector backbone": "pJet1.2",
"Backbone size w/o insert (bp)": "2974",
"Vector type": "",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Phylogenetic and functional characterization of water bears (Tardigrada) tubulins",
"Publication": "Sci Rep. 2023 Mar 30;13(1):5194. doi: 10.1038/s41598-023-31992-z.",
"DOI": "10.1038/s41598-023-31992-z.",
"Authors": "Novotna Floriancicova K, Baltzis A, Smejkal J, Czernekova M, Kaczmarek L, Maly J, Notredame C, Vinopal S /s---z",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/36997657",
"PubMed ID": "PubMed 36997657"
} | {
"Gene/Insert name": "Tub3",
"Species": "Hypsibius exemplaris",
"Insert Size (bp)": "1383",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "TOPO Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "pJET1.2 Forward Sequencing Primer (5'-CGACTCACTATAGGGAGAGCGGC-3')",
"3′ sequencing primer": "pJET1.2 Reverse Sequencing Primer (5'-AAGAACATCGATTTTCCATGGCAG-3')"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pSrpR-S2 | 74692 | SrpR-S2 gate | Christopher Voigt | Nielsen et al Science. 2016 Apr 1;352(6281):aac7341. doi: 10.1126/science.aac7341. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/141196/75d96771-a855-40c1-b4f7-6b7006c1da68/addgene-plasmid-74692-sequence-141196.gbk | full | LOCUS sequence-141196- 5814 bp DNA circular SYN 26-AUG-2024
DEFINITION SrpR-S2 gate.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5814)
AUTHORS Nielsen AA, Der BS, Shin J, Vaidyanathan P, Paralanov V, Strychalski
EA, Ross D, Densmore D, Voigt CA
TITLE Genetic circuit design automation.
JOURNAL Science. 2016 Apr 1;352(6281):aac7341. doi: 10.1126/science.aac7341.
PUBMED 27034378
REFERENCE 2 (bases 1 to 5814)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Science.";
date: "2016-04-1"; volume: "352(6281):aac7341. doi"; pages: "
10.1126/science.aac7341"
FEATURES Location/Qualifiers
source 1..5814
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin complement(308..852)
/direction=LEFT
/label=p15A ori
/note="Plasmids containing the medium-copy-number p15A
origin of replication can be propagated in E. coli cells
that contain a second plasmid with the ColE1 origin.
"
primer_bind complement(388..407)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(941..1756)
/codon_start=1
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
CDS complement(941..1756)
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind 1664..1683
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
primer_bind 1748..1767
/label=pENTR-R
/note="pENTR vectors, reverse primer"
promoter 1878..1906
/label=tac promoter
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
protein_bind 1912..1931
/label=lac operator (symmetric)
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG). The
symmetric lac operator was optimized for tight binding of
lac repressor."
misc_RNA 2865..2914
/label=sTRSV HHRz
/note="hammerhead ribozyme from the tobacco ringspot virus
satellite RNA (Khvorova et al., 2003)"
CDS 2964..3683
/codon_start=1
/product="enhanced YFP"
/label=EYFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKLHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(3009..3030)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(3270..3289)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 3617..3638
/label=EGFP-C
/note="EGFP, forward primer"
CDS 3831..4913
/codon_start=1
/gene="lacI"
/product="lac repressor"
/label=lacI
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
primer_bind complement(3850..3869)
/label=LacI-R
/note="LacI, reverse primer"
CDS 4932..5555
/codon_start=1
/gene="tetR from transposon Tn10"
/product="tetracycline repressor TetR"
/label=TetR
/note="TetR binds to the tetracycline operator tetO to
inhibit transcription. This inhibition can be relieved by
adding tetracycline or doxycycline."
/translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
primer_bind complement(5005..5024)
/label=Tet-R
/note="Tetracycline resistance gene, reverse primer"
primer_bind complement(5738..5757)
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
ORIGIN
1 gcttcctcgc tcactgactc gctgcacgag gcagacctca gcgctagcgg agtgtatact
61 ggcttactat gttggcactg atgagggtgt cagtgaagtg cttcatgtgg caggagaaaa
121 aaggctgcac cggtgcgtca gcagaatatg tgatacagga tatattccgc ttcctcgctc
181 actgactcgc tacgctcggt cgttcgactg cggcgagcgg aaatggctta cgaacggggc
241 ggagatttcc tggaagatgc caggaagata cttaacaggg aagtgagagg gccgcggcaa
301 agccgttttt ccataggctc cgcccccctg acaagcatca cgaaatctga cgctcaaatc
361 agtggtggcg aaacccgaca ggactataaa gataccaggc gtttcccctg gcggctccct
421 cgtgcgctct cctgttcctg cctttcggtt taccggtgtc attccgctgt tatggccgcg
481 tttgtctcat tccacgcctg acactcagtt ccgggtaggc agttcgctcc aagctggact
541 gtatgcacga accccccgtt cagtccgacc gctgcgcctt atccggtaac tatcgtcttg
601 agtccaaccc ggaaagacat gcaaaagcac cactggcagc agccactggt aattgattta
661 gaggagttag tcttgaagtc atgcgccggt taaggctaaa ctgaaaggac aagttttggt
721 gactgcgctc ctccaagcca gttacctcgg ttcaaagagt tggtagctca gagaaccttc
781 gaaaaaccgc cctgcaaggc ggttttttcg ttttcagagc aagagattac gcgcagacca
841 aaacgatctc aagaagatca tcttattaag gggtctgacg ctcagtggaa cgaaaaatca
901 atctaaagta tatatgagta aacttggtct gacagttacc ttagaaaaac tcatcgagca
961 tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc
1021 gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt
1081 atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa
1141 aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca
1201 aaagcttatg catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa
1261 aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg agacgaaata
1321 cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca
1381 ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg
1441 ctgttttccc ggggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat
1501 gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg
1561 taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct
1621 tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga gcccatttat
1681 acccatataa atcagcatcc atgttggaat ttaatcgcgg cctcgagcaa gacgtttccc
1741 gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagac agttttattg
1801 ttcatgatga tatattttta tcttgtgcaa tgtacatcag agattttgag acacaaggag
1861 aacgatcgtt ggctgtgttg acaattaatc atcggctcgt ataatgtgtg gaattgtgag
1921 cgctcacaat tagcgctcaa cgggtgtgct tcccgttctg atgagtccgt gaggacgaaa
1981 gcgcctctac aaataatttt gtttaagagt ctatggacta tgttttcaca tatgagatac
2041 caggatggca cgtaaaaccg cagcagaagc agaagaaacc cgtcagcgta ttattgatgc
2101 agcactggaa gtttttgttg cacagggtgt tagtgatgca accctggatc agattgcacg
2161 taaagccggt gttacccgtg gtgcagttta ttggcatttt aatggtaaac tggaagttct
2221 gcaggcagtt ctggcaagcc gtcagcatcc gctggaactg gattttacac cggatctggg
2281 tattgaacgt agctgggaag cagttgttgt tgcaatgctg gatgcagttc atagtccgca
2341 gagcaaacag tttagcgaaa ttctgattta tcagggtctg gatgaaagcg gtctgattca
2401 taatcgtatg gttcaggcaa gcgatcgttt tctgcagtat attcatcagg ttctgcgtca
2461 tgcagttacc cagggtgaac tgccgattaa tctggatctg cagaccagca ttggtgtttt
2521 taaaggtctg attaccggtc tgctgtatga aggtctgcgt agcaaagatc agcaggcaca
2581 gattatcaaa gttgcactgg gtagcttttg ggcactgctg cgtgaaccgc ctcgttttct
2641 gctgtgtgaa gaagcacaga ttaaacaggt gaaatccttc gaataattca gccaaaaaac
2701 ttaagaccgc cggtcttgtc cactaccttg cagtaatgcg gtggacagga tcggcggttt
2761 tcttttctct tctcaatact tctatgattg gtccagattc gttaccaatt gacagctagc
2821 tcagtcctag gtatatacat acatgcttgt ttgtttgtaa acagctgtca ccggatgtgc
2881 tttccggtct gatgagtccg tgaggacgaa acagcctcta caaataattt tgtttaatac
2941 tagagaaaga ggggaaatac tagatggtga gcaagggcga ggagctgttc accggggtgg
3001 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
3061 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accacaggca
3121 agctgcccgt gccctggccc accctcgtga ccaccttcgg ctacggcctg caatgcttcg
3181 cccgctaccc cgaccacatg aagctgcacg acttcttcaa gtccgccatg cccgaaggct
3241 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
3301 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
3361 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
3421 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg
3481 aggacggcag cgtgcagctc gccgaccact accagcagaa caccccaatc ggcgacggcc
3541 ccgtgctgct gcccgacaac cactacctta gctaccagtc cgccctgagc aaagacccca
3601 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
3661 gcatggacga gctgtacaag taactcggta ccaaattcca gaaaagaggc ctcccgaaag
3721 gggggccttt tttcgttttg gtccaatggc ggcgcgccat cgaatggcgc aaaacctttc
3781 gcggtatggc atgatagcgc ccggaagaga gtcaattcag ggtggtgaat atgaaaccag
3841 taacgttata cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg
3901 tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg
3961 agctgaatta cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga
4021 ttggcgttgc cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta
4081 aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg
4141 tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca
4201 ttaactatcc gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc
4261 cggcgttatt tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg
4321 aggacggtac gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc
4381 tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat
4441 atctcactcg caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt
4501 ccggttttca acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg
4561 ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg
4621 ttggtgcgga tatctcggta gtgggatacg acgataccga agatagctca tgttatatcc
4681 cgccgttaac caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct
4741 tgctgcaact ctctcagggc caggcggtga agggcaatca gctgttgcca gtctcactgg
4801 tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
4861 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgataatcca
4921 ggaggaaaaa aatgtccaga ttagataaaa gtaaagtgat taacagcgca ttagagctgc
4981 ttaatgaggt cggaatcgaa ggtttaacaa cccgtaaact cgcccagaag ctaggtgtag
5041 agcagcctac attgtattgg catgtaaaaa ataagcgggc tttgctcgac gccttagcca
5101 ttgagatgtt agataggcac catactcact tttgcccttt agaaggggaa agctggcaag
5161 attttttacg taataacgct aaaagtttta gatgtgcttt actaagtcat cgcgatggag
5221 caaaagtaca tttaggtaca cggcctacag aaaaacagta tgaaactctc gaaaatcaat
5281 tagccttttt atgccaacaa ggtttttcac tagagaatgc attatatgca ctcagcgctg
5341 tggggcattt tactttaggt tgcgtattgg aagatcaaga gcatcaagtc gctaaagaag
5401 aaagggaaac acctactact gatagtatgc cgccattatt acgacaagct atcgaattat
5461 ttgatcacca aggtgcagag ccagccttct tattcggcct tgaattgatc atatgcggat
5521 tagaaaaaca acttaaatgt gaaagtgggt cctaataatt ggtaacgaat cagacaattg
5581 acggctcgag ggagtagcat agggtttgca gaatccctgc ttcgtccatt tgacaggcac
5641 attatgcatc gatgataagc tgtcaaacat gagcagatcc tctacgccgg acgcatcgtg
5701 gccggcatca ccggcgccac aggtgcggtt gctggcgcct atatcgccga catcaccgat
5761 ggggaagatc gggctcgcca cttcgggctc atgagcaaat attttatctg aggt
//
| {
"Vector backbone": "pAN801",
"Backbone size w/o insert (bp)": "3930",
"Vector type": "Bacterial Expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Kanamycin, 50 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "Low Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Genetic circuit design automation",
"Publication": "Science. 2016 Apr 1;352(6281):aac7341. doi: 10.1126/science.aac7341.",
"DOI": "10.1126/science.aac7341.",
"Authors": "Nielsen AA, Der BS, Shin J, Vaidyanathan P, Paralanov V, Strychalski EA, Ross D, Densmore D, Voigt CA /scienceaac",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/27034378",
"PubMed ID": "PubMed 27034378"
} | {
"Gene/Insert name": "PTac-RBSS2-SrpR-PSrpR-YFP",
"Species": "E. coli",
"Insert Size (bp)": "1884",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
CAMK1D gRNA (BRDN0001145707) | 77444 | 3rd generation lentiviral gRNA plasmid targeting human CAMK1D | Doench et al Nat Biotechnol. 2016 Jan 18. doi: 10.1038/nbt.3437. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/187772/a49dfde9-9966-4a7b-b203-9dddea35307e/addgene-plasmid-77444-sequence-187772.gbk | full | LOCUS sequence-187772- 8323 bp DNA circular SYN 26-AUG-2024
DEFINITION 3rd generation lentiviral gRNA plasmid targeting human CAMK1D.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8323)
AUTHORS Doench JG, Fusi N, Sullender M, Hegde M, Vaimberg EW, Donovan KF,
Smith I, Tothova Z, Wilen C, Orchard R, Virgin HW, Listgarten J,
Root DE
TITLE Optimized sgRNA design to maximize activity and minimize off-target
effects of CRISPR-Cas9.
JOURNAL Nat Biotechnol. 2016 Jan 18. doi: 10.1038/nbt.3437.
PUBMED 26780180
REFERENCE 2 (bases 1 to 8323)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat
Biotechnol. 2016 Jan 18. doi: 10.1038/nbt.3437."
FEATURES Location/Qualifiers
source 1..8323
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 87..1265
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 318..1256
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 1213..1233
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
CDS 1278..1877
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind complement(1278..1297)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 1774..1794
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
misc_feature 1893..2481
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(1946..1966)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(2364..2375)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 2553..2786
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 2864..2985
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2901..2920)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2955..2974
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 3025..3160
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(3180..3199)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(3181..3199)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(3209..3226)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(3209..3225)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3218..3240)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 3367..3822
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3454..3473)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3664..3685
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 3848..3952
/gene="bla"
/label=AmpR promoter
CDS 3953..4813
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(4171..4190)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 4984..5572
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5473..5492
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 5726..5743
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 5860..5881
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 5896..5926
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5934..5950
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
primer_bind 5939..5961
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 5958..5974
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 5958..5974
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 5993..6013
/label=T3
/note="T3 promoter, forward primer"
promoter 5995..6013
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 6041..6267
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 6268..6448
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 6495..6620
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 7113..7346
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 7531..7575
/codon_start=1
/product="antigenic peptide corresponding to amino acids
655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
et al., 2013)"
/label=gp41 peptide
/note="recognized by the 2H10 single-chain llama nanobody"
/translation="KNEQELLELDKWASL"
promoter 7753..8001
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA
(Domitrovich & Kunkel, 2003)"
primer_bind 7753..7773
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 7924..7943
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
misc_RNA 8023..8098
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
misc_feature 8154..8271
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
ORIGIN
1 cccgggtgca aagatggata aagttttaaa cagagaggaa tctttgcagc taatggacct
61 tctaggtctt gaaaggagtg ggaattggct ccggtgcccg tcagtgggca gagcgcacat
121 cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgatccggt gcctagagaa
181 ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg
241 gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt
301 ttgccgccag aacacaggta agtgccgtgt gtggttcccg cgggcctggc ctctttacgg
361 gttatggccc ttgcgtgcct tgaattactt ccacctggct gcagtacgtg attcttgatc
421 ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa ggagcccctt
481 cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt
541 ggcaccttcg cgcctgtctc gctgctttcg ataagtctct agccatttaa aatttttgat
601 gacctgctgc gacgcttttt ttctggcaag atagtcttgt aaatgcgggc caagatctgc
661 acactggtat ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca
721 catgttcggc gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc
781 aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg
841 cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga aagatggccg cttcccggcc
901 ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac
961 ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac tccacggagt
1021 accgggcgcc gtccaggcac ctcgattagt tctcgagctt ttggagtacg tcgtctttag
1081 gttgggggga ggggttttat gcgatggagt ttccccacac tgagtgggtg gagactgaag
1141 ttaggccagc ttggcacttg atgtaattct ccttggaatt tgcccttttt gagtttggat
1201 cttggttcat tctcaagcct cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt
1261 cgtgacgtac ggccaccatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg
1321 acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc
1381 acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca
1441 cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gccgtggcgg
1501 tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca
1561 tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc
1621 cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggagtctcg cccgaccacc
1681 agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg
1741 gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg
1801 gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc
1861 gcaagcccgg tgcctgaacg cgttaagtcg acaatcaacc tctggattac aaaatttgtg
1921 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
1981 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
2041 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
2101 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
2161 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
2221 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
2281 cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg
2341 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
2401 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
2461 ccctttgggc cgcctccccg cgtcgacttt aagaccaatg acttacaagg cagctgtaga
2521 tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag
2581 acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga
2641 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
2701 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
2761 ttagtcagtg tggaaaatct ctagcagtac gtatagtagt tcatgtcatc ttattattca
2821 gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc
2881 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
2941 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta
3001 gctatcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
3061 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
3121 gagctattcc agaagtagtg aggaggcttt tttggaggcc tagggacgta cccaattcgc
3181 cctatagtga gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg
3241 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
3301 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
3361 aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
3421 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
3481 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
3541 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
3601 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
3661 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
3721 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
3781 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg
3841 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
3901 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
3961 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
4021 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
4081 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
4141 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
4201 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
4261 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
4321 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
4381 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
4441 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
4501 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
4561 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
4621 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
4681 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
4741 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
4801 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
4861 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
4921 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
4981 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
5041 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
5101 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
5161 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
5221 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
5281 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
5341 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
5401 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
5461 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
5521 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
5581 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
5641 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
5701 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
5761 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
5821 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
5881 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
5941 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa
6001 ccctcactaa agggaacaaa agctggagct gcaagcttaa tgtagtctta tgcaatactc
6061 ttgtagtctt gcaacatggt aacgatgagt tagcaacatg ccttacaagg agagaaaaag
6121 caccgtgcat gccgattggt ggaagtaagg tggtacgatc gtgccttatt aggaaggcaa
6181 cagacgggtc tgacatggat tggacgaacc actgaattgc cgcattgcag agatattgta
6241 tttaagtgcc tagctcgata cataaacggg tctctctggt tagaccagat ctgagcctgg
6301 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
6361 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
6421 ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag
6481 ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag
6541 gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga
6601 gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa
6661 ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca
6721 gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac
6781 aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat
6841 ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg
6901 aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg
6961 ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag tgaattatat
7021 aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc aaagagaaga
7081 gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg gttcttggga
7141 gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc cagacaatta
7201 ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc gcaacagcat
7261 ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct ggctgtggaa
7321 agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa actcatttgc
7381 accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca gatttggaat
7441 cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt aatacactcc
7501 ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt ggaattagat
7561 aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta tataaaatta
7621 ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt actttctata
7681 gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct cccaaccccg
7741 aggggaccca gagagggcct atttcccatg attccttcat atttgcatat acgatacaag
7801 gctgttagag agataattag aattaatttg actgtaaaca caaagatatt agtacaaaat
7861 acgtgacgta gaaagtaata atttcttggg tagtttgcag ttttaaaatt atgttttaaa
7921 atggactatc atatgcttac cgtaacttga aagtatttcg atttcttggc tttatatatc
7981 ttgtggaaag gacgaaacac cggatgtagg caatcactcc gagttttaga gctagaaata
8041 gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt
8101 ttttaagctt ggcgtaacta gatcttgaga caaatggcag tattcatcca caattttaaa
8161 agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca
8221 gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat
8281 tacagggaca gcagagatcc actttggcgc cggctcgagg ggg
//
| {
"Vector backbone": "lentiGuide-Puro",
"Backbone size w/o insert (bp)": "",
"Vector type": "Mammalian Expression, Lentiviral, CRISPR",
"Selectable markers": "Puromycin"
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Optimized sgRNA design to maximize activity and minimize off-target effects of CRISPR-Cas9",
"Publication": "Nat Biotechnol. 2016 Jan 18. doi: 10.1038/nbt.3437.",
"DOI": "10.1038/nbt.3437.",
"Authors": "Doench JG, Fusi N, Sullender M, Hegde M, Vaimberg EW, Donovan KF, Smith I, Tothova Z, Wilen C, Orchard R, Virgin HW, Listgarten J, Root DE /nbt",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/26780180",
"PubMed ID": "PubMed 26780180"
} | {
"Gene/Insert name": "CAMK1D",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "CAMK1D",
"Promoter": "hU6",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Unknown",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "U6-F",
"3′ sequencing primer": "pS2-R"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
||
pJOP-HTT-HR81Q | 92250 | HTT donor DNA with 81Q for homologous recombination-mediated gene modification | Mahmoud Pouladi | Ooi et al Cell Rep. 2019 Feb 26;26(9):2494-2508.e7. doi: 10.1016/j.celrep.2019.02.008. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/315766/4489c536-cc2d-47b4-ac24-ff13b9b1f890/addgene-plasmid-92250-sequence-315766.gbk | full | LOCUS sequence-315766- 12265 bp DNA circular SYN 26-AUG-2024
DEFINITION HTT donor DNA with 81Q for homologous recombination-mediated gene
modification.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 12265)
AUTHORS Ooi J, Langley SR, Xu X, Utami KH, Sim B, Huang Y, Harmston NP, Tay
YL, Ziaei A, Zeng R, Low D, Aminkeng F, Sobota RM, Ginhoux F,
Petretto E, Pouladi MA
TITLE Unbiased Profiling of Isogenic Huntington Disease hPSC-Derived CNS
and Peripheral Cells Reveals Strong Cell-Type Specificity of CAG
Length Effects.
JOURNAL Cell Rep. 2019 Feb 26;26(9):2494-2508.e7. doi:
10.1016/j.celrep.2019.02.008.
PUBMED 30811996
REFERENCE 2 (bases 1 to 12265)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1016/j.celrep.2019.02"; journalName: "Cell Rep"; date:
"2019-02-26- 26"; volume: "26"; issue: "9"; pages: "2494-2508.e7"
FEATURES Location/Qualifiers
source 1..12265
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 1..861
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(219..238)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 1032..1620
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 1521..1540
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 1774..1791
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 1908..1929
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 1944..1974
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 1982..1998
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
primer_bind 1987..2009
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 2006..2022
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 2006..2022
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
repeat_region 4074..4136
/label=piggyBac right (3') inverted repeat
/note="piggyBac transposon-specific inverted terminal
repeat sequence (ITR)"
promoter 4589..4800
/label=EF-1-alpha core promoter
/note="core promoter for human elongation factor
EF-1-alpha"
LTR 4813..5081
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from human
T-cell leukemia virus (HTLV) type 1"
regulatory 5110..5119
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 5140..5802
/codon_start=1
/product="green fluorescent protein 2 from Pontellina
plumata, also known as ppluGFP2 (Shagin et al., 2004)"
/label=CopGFP
/translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT
FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRC
EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG
YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA"
CDS 5872..5925
/codon_start=1
/product="2A peptide from Thosea asigna virus capsid
protein"
/label=T2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="EGRGSLLTCGDVEENPGP"
CDS 5926..6522
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind complement(5926..5945)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 6422..6442
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
CDS 6529..6582
/codon_start=1
/product="2A peptide from Thosea asigna virus capsid
protein"
/label=T2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="EGRGSLLTCGDVEENPGP"
CDS 6583..7710
/codon_start=1
/product="herpes simplex virus thymidine kinase"
/label=HSV TK
/note="confers sensitivity to nucleoside analogs such as
acyclovir or ganciclovir"
/translation="ASYPGHQHASAFDQAARSRGHSNRRTALRPRRQQEATEVRPEQKM
PTLLRVYIDGPHGMGKTTTTQLLVALGSRDDIVYVPEPMTYWRVLGASETIANIYTTQH
RLDQGEISAGDAAVVMTSAQITMGMPYAVTDAVLAPHIGGEAGSSHAPPPALTLIFDRH
PIAALLCYPAARYLMGSMTPQAVLAFVALIPPTLPGTNIVLGALPEDRHIDRLAKRQRP
GERLDLAMLAAIRRVYGLLANTVRYLQCGGSWREDWGQLSGTAVPPQGAEPQSNAGPRP
HIGDTLFTLFRAPELLAPNGDLYNVFAWALDVLAKRLRSMHVFILDYDQSPAGCRDALL
QLTSGMVQTHVTTPGSIPTICDLARTFAREMGEAN"
polyA_signal 7759..7880
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(7796..7815)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 7850..7869
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
repeat_region 8637..8671
/label=piggyBac left (5') inverted repeat
/note="piggyBac transposon-specific inverted terminal
repeat sequence (ITR)"
primer_bind complement(11195..11212)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(11195..11211)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(11204..11226)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 11424..11879
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(11511..11530)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 11721..11742
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(11896..11915)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(12075..12093)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 12161..12265
/gene="bla"
/label=AmpR promoter
ORIGIN
1 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
61 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
121 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
181 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
241 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
301 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
361 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
421 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
481 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
541 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
601 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
661 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtggttct
721 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
781 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
841 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
901 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
961 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
1021 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
1081 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
1141 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
1201 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
1261 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
1321 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
1381 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
1441 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
1501 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
1561 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
1621 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
1681 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
1741 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
1801 cagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
1861 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
1921 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
1981 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagctc
2041 cttcctcttc cagcccttcc tctttcactg actgactgac tgcgtctcaa aacctcggct
2101 gggtcaacgg aacattgtac cacagggagt agtcccgctg ttcgccctcc tattctagag
2161 ctagcgtcac acttggggtc ctcaggtcgt gccgaccacg cgcattctct gcgctctgcg
2221 caggagctcg cccaccctct ccccgtgcag agagccccgc agctggctcc ccgcagggct
2281 gtccgggtga gtatggctct ggccacgggc cagtgtggcg ggagggcaaa ccccaaggcc
2341 acctcggctc agagtccacg gccggctgtc gccccgctcc aggcgtcggc gggggatcct
2401 ttccgcatgg gcctgcgccc gcgctcggcg ccccctccac ggccccgccc cgtccatggc
2461 cccgtccttc atgggcgagc ccctccatgg ccctgcccct ccgcgcccca cccctccctc
2521 gccccacctc tcaccttcct gccccgcccc cagcctcccc acccctcacc ggccagtccc
2581 ctcccctatc ccgctccgcc cctcagccgc cccgcccctc agccggcctg cctaatgtcc
2641 ccgtccccag catcgccccg ccccgccccc gtctcgcccc gcccctcagg cggcctccct
2701 gctgtgcccc gccccggcct cgccacgccc ctacctcacc acgccccccg catcgccacg
2761 ccccccgcat cgccacgcct cccttaccat gcagtcccgc cccgtccctt cctcgtcccg
2821 cctcgccgcg acacttcaca cacagcttcg cctcacccca ttacagtctc accacgcccc
2881 gtcccctctc cgttgagccc cgcgccttcg cccgggtggg gcgctgcgct gtcagcggcc
2941 ttgctgtgtg aggcagaacc tgcgggggca ggggcgggct ggttccctgg ccagccattg
3001 gcagagtccg caggctaggg ctgtcaatca tgctggccgg cgtggccccg cctccgccgg
3061 cgcggccccg cctccgccgg cgcagcgtct gggacgcaag gcgccgtggg ggctgccggg
3121 acgggtccaa gatggacggc cgctcaggtt ctgcttttac ctgcggccca gagccccatt
3181 cattgccccg gtgctgagcg gcgccgcgag tcggcccgag gcctccgggg actgccgtgc
3241 cgggcgggag accgccatgg cgaccctgga aaagctgatg aaggccttcg agtccctcaa
3301 gtccttccag cagcagcagc agcagcagca gcagcagcag cagcagcagc agcagcagca
3361 gcagcagcag cagcagcagc agcagcagca gcagcagcag cagcagcagc agcagcagca
3421 gcagcagcag cagcagcagc agcagcagca gcagcagcag cagcagcagc agcagcagca
3481 gcagcagcag cagcagcagc agcagcagca gcagcagcag cagcagcagc agcagcagca
3541 gcagcaacag ccgccaccgc cgccgccgcc gccgccgcct cctcacctgc cccacccgcc
3601 gccgcaggca cagccgctgc tgcctcagcc gcagccgccc ccgccgccgc ccccgccgcc
3661 acccggcccg gctgtggctg aggagccgct gcaccgaccg tgagtttggg cccgctgcag
3721 ctccctgtcc cggcgggtcc caggctacgg cggggrtggc ggtaaccctg cagcctgcgg
3781 gccggcgaca cgaacccccg gccccgcaga gacagagtga cccagcaacc cagagcccat
3841 gagggacacc cgccccctcc tggggcgagg ccttccccca cttcagcccc gctccctcac
3901 ttgggtcttc ccttgtcctc tcgcgagggg aggcagagcc ttgttggggc ctgtcctgaa
3961 ttcaccgagg ggagtcacgg cctcagccct ctcgcccttc gcaggatgcg aagagttggg
4021 gcgagaactt gtttcttttt atttgcgaga aaccagggcg ggggttcttt taaccctaga
4081 aagataatca tattgtgacg tacgttaaag ataatcatgc gtaaaattga cgcatgtgtt
4141 ttatcggtct gtatatcgag gtttatttat taatttgaat agatattaag ttttattata
4201 tttacactta catactaata ataaattcaa caaacaattt atttatgttt atttatttat
4261 taaaaaaaaa caaaaactca aaatttcttc tataaagtaa caaaactttt atgagggaca
4321 gcccccccca aagcccccag ggatgtaatt acgtccctcc cccgctaggg ggcagcagcg
4381 agccgcccgg ggctccgctc cggtccggcg ctccccccgc atccccgagc cggcagcgtg
4441 cggggacagc ccgggcacgg ggaaggtggc acgggatcgc tttcctctga acgcttctcg
4501 ctgctctttg agcctgcaga cacctggggg gatacgggga aaaggcctcc aaggccaagg
4561 atctgcgatc gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag
4621 aagttggggg gaggggtcgg caattgaacg ggtgcctaga gaaggtggcg cggggtaaac
4681 tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat
4741 ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag
4801 ctgaagcttc gaggggctcg catctctcct tcacgcgccc gccgccctac ctgaggccgc
4861 catccacgcc ggttgagtcg cgttctgccg cctcccgcct gtggtgcctc ctgaactgcg
4921 tccgccgtct aggtaagttt aaagctcagg tcgagacagg gcctttgtcc ggcgctccct
4981 tggagcctac ctagactcag ccggctctcc acgctttgcc tgaccctgct tgctcaactc
5041 tacgtctttg tttcgttttc tgttctgcgc cgttacagat ccaagctgtg accggcgcct
5101 actctagacg ccaccatgga gagcgacgag agcggcctgc ccgccatgga gatcgagtgc
5161 cgcatcaccg gcaccctgaa cggcgtggag ttcgagctgg tgggcggcgg agagggcacc
5221 cccaagcagg gccgcatgac caacaagatg aaaagcacca aaggcgccct gaccttcagc
5281 ccctacctgc tgagccacgt gatgggctac ggcttctacc acttcggcac ctaccccagc
5341 ggctacgaga accccttcct gcacgccatc aacaacggcg gctacaccaa cacccgcatc
5401 gagaagtacg aggacggcgg cgtgctgcac gtgagcttca gctaccgctg cgaggccggc
5461 cgcgtgatcg gcgacttcaa ggtggtgggc accggcttcc ccgaggacag cgtgatcttc
5521 accgacaaga tcatccgcag caacgccacc gtggagcacc tgcaccccat gggcgataac
5581 gtgctggtgg gcagcttcgc ccgcaccttc agcctgcgcg acggcggcta ctacagcttc
5641 gtggtggaca gccacatgca cttcaagagc gccatccacc ccagcatcct gcagaacggg
5701 ggccccatgt tcgccttccg ccgcgtggag gagctgcaca gcaacaccga gctgggcatc
5761 gtggagtacc agcacgcctt caagaccccc atcgccttcg ccagatcccg cgctcagtcg
5821 tccaattctg ccgtggacgg caccgccgga cccggctcca ccggatctcg cgagggcaga
5881 ggaagtcttc taacatgcgg tgacgtggag gagaatcccg gccctatgac cgagtacaag
5941 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc
6001 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg
6061 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
6121 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
6181 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
6241 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
6301 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
6361 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac atccgcgccc
6421 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
6481 gaaggaccgc gcacctggtg catgacccgc aagcccggtg ccaccggtga gggcagagga
6541 agtcttctaa catgcggtga cgtggaggag aatcccggcc ctgcttcgta ccccggccat
6601 caacacgcgt ctgcgttcga ccaggctgcg cgttctcgcg gccatagcaa ccgacgtacg
6661 gcgttgcgcc ctcgccggca gcaagaagcc acggaagtcc gcccggagca gaaaatgccc
6721 acgctactgc gggtttatat agacggtccc cacgggatgg ggaaaaccac caccacgcaa
6781 ctgctggtgg ccctgggttc gcgcgacgat atcgtctacg tacccgagcc gatgacttac
6841 tggcgggtgc tgggggcttc cgagacaatc gcgaacatct acaccacaca acaccgcctc
6901 gaccagggtg agatatcggc cggggacgcg gcggtggtaa tgacaagcgc ccagataaca
6961 atgggcatgc cttatgccgt gaccgacgcc gttctggctc ctcatatcgg gggggaggct
7021 gggagctcac atgccccgcc cccggccctc accctcatct tcgaccgcca tcccatcgcc
7081 gccctcctgt gctacccggc cgcgcggtac cttatgggca gcatgacccc ccaggccgtg
7141 ctggcgttcg tggccctcat cccgccgacc ttgcccggca ccaacatcgt gcttggggcc
7201 cttccggagg acagacacat cgaccgcctg gccaaacgcc agcgccccgg cgagcggctg
7261 gacctggcta tgctggctgc gattcgccgc gtttacgggc tacttgccaa tacggtgcgg
7321 tatctgcagt gcggcgggtc gtggcgggag gactggggac agctttcggg gacggccgtg
7381 ccgccccagg gtgccgagcc ccagagcaac gcgggcccac gaccccatat cggggacacg
7441 ttatttaccc tgtttcgggc ccccgagttg ctggccccca acggcgacct gtataacgtg
7501 tttgcctggg ccttggacgt cttggccaaa cgcctccgtt ccatgcacgt ctttatcctg
7561 gattacgacc aatcgcccgc cggctgccgg gacgccctgc tgcaacttac ctccgggatg
7621 gtccagaccc acgtcaccac ccccggctcc ataccgacga tatgcgacct ggcgcgcacg
7681 tttgcccggg agatggggga ggctaactga ttttcatcga tttctcgagc ttctactgtc
7741 tggttagacc gttaactaaa cttgtttatt gcagcttata atggttacaa ataaagcaat
7801 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
7861 aaactcatca atgtatctta tcatgtctgg aattgactca aatgatgtca attagtctat
7921 cagaagctca tctccttccg ggggacaaga catccctgtt taatatttaa acagcagtgt
7981 tcccaaactg ggttcttata tcccttgctc tggtcaacca ggttgcaggg tttcctgtcc
8041 tcacaggaac gaagtcccta aagaaacagt ggcagccagg tttagccccg gaattgactg
8101 gattcctttt ttagggccca ttggtatggc tttttccccg tatcccccca ggtgtctgca
8161 ggctcaaaga gcagcgagaa gcgttcagag gaaagcgatc ccgtgccacc ttccccgtgc
8221 ccgggctgtc cccgcacgct gccggctcgg ggatgcgggg ggagcgccgg accggagcgg
8281 agccccgggc ggctcgctgc tgccccctag cgggggaggg acgtaattac atccctgggg
8341 gctttggggg ggggctgtcc ctgatatcta taacaagaaa atatatatat aataagttat
8401 cacgtaagta gaacatgaaa taacaatata attatcgtat gagttaaatc ttaaaagtca
8461 cgtaaaagat aatcatgcgt cattttgact cacgcggtcg ttatagttca aaatcagtga
8521 cacttaccgc attgacaagc acgcctcacg ggagctccaa gcggcgactg agatgtccta
8581 aatgcacagc gacggattcg cgctatttag aaagagagag caatatttca agaatgcatg
8641 cgtcaatttt acgcagacta tctttctagg gttaactgcg ttgtgaagag aacttggagg
8701 agccgagatt tgctcagtgc cacttccctc ttctagtctg agagggaaga gggctggggg
8761 cgcgggacac ttcgagagga ggcggggttt ggagctggag agatgtgggg gcagtggatg
8821 acataatgct tttaggacgc ctcggcggga gtggcggggg gggggggggg sggggagtga
8881 gggcgcgtcc aatgggagat ttcttttcct agtggcactt aaaacagcct gagatttgag
8941 gctcttccta cattgtcagg acatttcatt tagttcatga tcacggtggt agtaacacga
9001 ttttaagcac cacctaagag atctgctcat ctaagcctaa gttggtctgc aggcgtttga
9061 atgagttgtg gttgccaagt aaagtggtga acttacgtgg tgattaatga aattatctta
9121 aatattagga agagttgatt gaagtttttt gcctatgtgt gttgggaata aaaccaacac
9181 gttgctgatg gggaggttaa ttgccgaggg atgaatgagg tgtacatttt accagtattc
9241 cagtcaggct tgccagaata cggggggtcc gcagactccg tgggcatctc agatgtgcca
9301 gtgaaagggt ttctgtttgc ttcattgctg acagcttgtt actttttgga agctaggggt
9361 ttctgttgct tgttcttggg gagaattttt gaaacaggaa aagagagacc attaaaacat
9421 ctagcggaac cccaggactt tccctggaag tctgtgtgtc gagtgtacag taggagttag
9481 gaagtactct ggtgcagttc aggcctttct cttacctctc agtattctat ttccgatctg
9541 gatgtgtccc agatggcatt tggtaagaat atctctgtta agactgatta atttttagta
9601 atatttcttg ttctttgttt ctgttatgat ccttgtctcg tcttcaaagt ttaattagaa
9661 aatgattcgg agagcagtgt tagcttattt gttggaataa aatttaggaa taaattattc
9721 taaaggatgg aaaaactttt tggatatttg gagaaatttt aaaacaattt ggcttatctc
9781 ttcagtaagt aatttctcat ccagaaattt actgtagtgc ttttctagga ggtaggtgtc
9841 ataaaagttc acacattgca tgtatcttgt gtaaacacta aacagggctc ctgatgggaa
9901 ggaagacctt tctgctgggc tgcttcagac acttgatcat tctaaaaata tgccttctct
9961 ttcttatgct gatttgacag aacctgcatt tgcttatctt caaaatatgg gtatcaagaa
10021 atttcctttg ctgccttgac aaaggagata gattttgttt cattacttta aggtaatata
10081 tgattacctt atttaaaaaa tttaatcagg actggcaagg tggcttacac ctttaatccg
10141 agcactttgg gaggcctagg tggacgaatc acctgaggtc aggagtttga gaccagcctg
10201 gctaacatgg tgaaaccctg tctctactaa aaatacaaaa attagctggt catggtggca
10261 cgtgcctgta atccaagcta cctgggaggc tgaggcagga aaatcgcttg aacccgggag
10321 gcagagtctg cagtgagttg agatcacgcc actgcactcc agcctgggtg acagagcgag
10381 actctatctc aaaaaaaatt ttttttaatg tattattttt gcataagtaa tacattgaca
10441 tgatacaaat tctgtaatta caaaagggca ataattaaaa tatcttcctt ccaccccttt
10501 cctctgagta cctaactttg tccccaagaa caagcactat ttcagttcct catgtatcct
10561 gccagatata acctgttcat attgtaagat agatttaaaa tgctctaaaa acaaaagtag
10621 tttagaataa tatatatcta tatatttttt gagatgtagt ctcacattgt cacccaggct
10681 ggagtgcagt gatacaatct cggctcactg cagtctctgc ctcccaggtt caaatgcttc
10741 tcctgcctca gccttctgag tagctgggat tacaggcgcc caccaccatg tccagctaat
10801 ttttgtattt ttagtagaga tggggtttca ccatgttggc caggctggtc ttgaactcct
10861 gaccttgtga tctgtccacc tcggcctccc aaagtgctgg gattacaggt gtgagccacc
10921 atgcctggct agaataataa cttttaaagg ttcttagcat gctctgaaat caactgcatt
10981 aggtttattt atagttttat agttatttta aataaaatgc atatttgtca tatttctctg
11041 tattttgctg ttgagaaagg aggtattcac taattttgag taacaaacac tgctcacaaa
11101 gtttggattt tggcagttct gttcacgtga ctagttctga attcactgac tgactgactg
11161 gaaagtcctc tccactgact gtagcctcca attcactggc cgtcgtttta caacgtcgtg
11221 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
11281 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
11341 atggcgaatg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
11401 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
11461 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc
11521 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
11581 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
11641 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
11701 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat
11761 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa
11821 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt
11881 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
11941 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
12001 agctgtgacc gtcaacggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
12061 cgcgacccga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
12121 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
12181 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
12241 tcaataatat tgaaaaagga agagt
//
| {
"Vector backbone": "PBHR100A-1 (SBI)",
"Backbone size w/o insert (bp)": "7925",
"Vector type": "PrecisionX HR targeting vector",
"Selectable markers": "Puromycin"
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Unbiased Profiling of Isogenic Huntington Disease hPSC-Derived CNS and Peripheral Cells Reveals Strong Cell-Type Specificity of CAG Length Effects",
"Publication": "Cell Rep. 2019 Feb 26;26(9):2494-2508.e7. doi: 10.1016/j.celrep.2019.02.008.",
"DOI": "10.1016/j.celrep.2019.02.008.",
"Authors": "Ooi J, Langley SR, Xu X, Utami KH, Sim B, Huang Y, Harmston NP, Tay YL, Ziaei A, Zeng R, Low D, Aminkeng F, Sobota RM, Ginhoux F, Petretto E, Pouladi MA /jcelrep",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/30811996",
"PubMed ID": "PubMed 30811996"
} | {
"Gene/Insert name": "HTT donor DNA",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "Barcode of wobble bases to prevent TALEN cleaving donor DNA: CAC CTG CCC CAC CCG CC",
"Entrez Gene": "HTT",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "NheI/NotI (not destroyed)",
"3′ cloning site": "BspQI (destroyed)/SpeI (destroyed during cloning)",
"5′ sequencing primer": "GTTTCGCCACCTCTGACTTG (-588)",
"3′ sequencing primer": "TCATTTTGACTCACGCGGTC (-196)"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pmiRGLO-TP73-201 | 117333 | Dual luciferase assay for miRNA-targeted UTR | Volker Busskamp | Kutsche et al Cell Syst. 2018 Sep 28. pii: S2405-4712(18)30358-2. doi: 10.1016/j.cels.2018.08.011. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/231339/df1b31ea-9898-4625-a84d-809c88d94616/addgene-plasmid-117333-sequence-231339.gbk | full | LOCUS sequence-231339- 10769 bp DNA circular SYN 26-AUG-2024
DEFINITION Dual luciferase assay for miRNA-targeted UTR.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10769)
AUTHORS Kutsche LK, Gysi DM, Fallmann J, Lenk K, Petri R, Swiersy A, Klapper
SD, Pircs K, Khattak S, Stadler PF, Jakobsson J, Nowick K, Busskamp
V
TITLE Combined Experimental and System-Level Analyses Reveal the Complex
Regulatory Network of miR-124 during Human Neurogenesis.
JOURNAL Cell Syst. 2018 Sep 28. pii: S2405-4712(18)30358-2. doi:
10.1016/j.cels.2018.08.011.
PUBMED 30292704
REFERENCE 2 (bases 1 to 10769)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Cell Syst.
2018 Sep 28. pii: S2405-4712(18)30358-2. doi:
10.1016/j.cels.2018.08.011."
FEATURES Location/Qualifiers
source 1..10769
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 49..548
/label=PGK promoter
/note="mouse phosphoglycerate kinase 1 promoter"
primer_bind complement(84..107)
/label=MSCV-rev
/note="Murine stem cell virus, reverse primer"
primer_bind 481..500
/label=mPGK-F
/note="Mouse PGK promoter, forward primer"
regulatory 589..598
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 595..2247
/codon_start=1
/product="firefly luciferase"
/label=luciferase
/note="synthetic luc2 version of the luciferase gene"
/translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
polyA_signal 5834..5955
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5845..5864)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 5899..5918
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
promoter 6200..6557
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
enhancer 6200..6391
/label=SV40 enhancer
/note="enhancer for the SV40 early promoter (Herr, 1993)"
primer_bind complement(6223..6243)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 6408..6543
/label=SV40 ori
/note="SV40 origin of replication"
regulatory 6602..6611
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 6608..7540
/codon_start=1
/product="Renilla luciferase"
/label=hRluc
/note="human codon-optimized"
/translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
CDS 7592..8383
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="IEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRPV
LFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLSS
HLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQG
LAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIAL
ATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(7643..7662)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 8253..8272
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 8447..8495
/note="synthetic polyadenylation signal"
promoter 8651..8755
/gene="bla"
/label=AmpR promoter
CDS 8756..9616
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(8974..8993)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin complement(9774..10362)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(9854..9873)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
misc_feature 10478..10761
/label=cer region
/note="ColE1-derived recombination site that helps to
maintain plasmids as monomers"
ORIGIN
1 gcagtactcc ggccacgatg cgtccggcgt agaggatcga gatctaccgg gtaggggagg
61 cgcttttccc aaggcagtct ggagcatgcg ctttagcagc cccgctgggc acttggcgct
121 acacaagtgg cctctggcct cgcacacatt ccacatccac cggtaggcgc caaccggctc
181 cgttctttgg tggccccttc gcgccacctt ctactcctcc cctagtcagg aagttccccc
241 ccgccccgca gctcgcgtcg tgcaggacgt gacaaatgga agtagcacgt ctcactagtc
301 tcgtgcagat ggacagcacc gctgagcaat ggaagcgggt aggcctttgg ggcagcggcc
361 aatagcagct ttgctccttc gctttctggg ctcagaggct gggaaggggt gggtccgggg
421 gcgggctcag gggcgggctc aggggcgggg cgggcgcccg aaggtcctcc ggaggcccgg
481 cattctgcac gcttcaaaag cgcacgtctg ccgcgctgtt ctcctcttcc tcatctccgg
541 gcctttcgac ctgcagccca agcttggcaa tccggtactg ttggtaaagc caccatggaa
601 gatgccaaaa acattaagaa gggcccagcg ccattctacc cactcgaaga cgggaccgcc
661 ggcgagcagc tgcacaaagc catgaagcgc tacgccctgg tgcccggcac catcgccttt
721 accgacgcac atatcgaggt ggacattacc tacgccgagt acttcgagat gagcgttcgg
781 ctggcagaag ctatgaagcg ctatgggctg aatacaaacc atcggatcgt ggtgtgcagc
841 gagaatagct tgcagttctt catgcccgtg ttgggtgccc tgttcatcgg tgtggctgtg
901 gccccagcta acgacatcta caacgagcgc gagctgctga acagcatggg catcagccag
961 cccaccgtcg tattcgtgag caagaaaggg ctgcaaaaga tcctcaacgt gcaaaagaag
1021 ctaccgatca tacaaaagat catcatcatg gatagcaaga ccgactacca gggcttccaa
1081 agcatgtaca ccttcgtgac ttcccatttg ccacccggct tcaacgagta cgacttcgtg
1141 cccgagagct tcgaccggga caaaaccatc gccctgatca tgaacagtag tggcagtacc
1201 ggattgccca agggcgtagc cctaccgcac cgcaccgctt gtgtccgatt cagtcatgcc
1261 cgcgacccca tcttcggcaa ccagatcatc cccgacaccg ctatcctcag cgtggtgcca
1321 tttcaccacg gcttcggcat gttcaccacg ctgggctact tgatctgcgg ctttcgggtc
1381 gtgctcatgt accgcttcga ggaggagcta ttcttgcgca gcttgcaaga ctataagatt
1441 caatctgccc tgctggtgcc cacactattt agcttcttcg ctaagagcac tctcatcgac
1501 aagtacgacc taagcaactt gcacgagatc gccagcggcg gggcgccgct cagcaaggag
1561 gtaggtgagg ccgtggccaa acgcttccac ctaccaggca tccgccaggg ctacggcctg
1621 acagaaacaa ccagcgccat tctgatcacc cccgaagggg acgacaagcc tggcgcagta
1681 ggcaaggtgg tgcccttctt cgaggctaag gtggtggact tggacaccgg taagacactg
1741 ggtgtgaacc agcgcggcga gctgtgcgtc cgtggcccca tgatcatgag cggctacgtt
1801 aacaaccccg aggctacaaa cgctctcatc gacaaggacg gctggctgca cagcggcgac
1861 atcgcctact gggacgagga cgagcacttc ttcatcgtgg accggctgaa gagcctgatc
1921 aaatacaagg gctaccaggt agccccagcc gaactggaga gcatcctgct gcaacacccc
1981 aacatcttcg acgccggggt cgccggcctg cccgacgacg atgccggcga gctgcccgcc
2041 gcagtcgtcg tgctggaaca cggtaaaacc atgaccgaga aggagatcgt ggactatgtg
2101 gccagccagg ttacaaccgc caagaagctg cgcggtggtg ttgtgttcgt ggacgaggtg
2161 cctaaaggac tgaccggcaa gttggacgcc cgcaagatcc gcgagattct cattaaggcc
2221 aagaagggcg gcaagatcgc cgtgtaattc tagttgttta aacgagagat ccccgagcag
2281 taccgcatga ccatctggcg gggcctgcag gacctgaagc agggccacga ctacagcacc
2341 gcgcagcagc tgctccgctc tagcaacgcg gccaccatct ccatcggcgg ctcaggggaa
2401 ctgcagcgcc agcgggtcat ggaggccgtg cacttccgcg tgcgccacac catcaccatc
2461 cccaaccgcg gcggcccagg cggcggccct gacgagtggg cggacttcgg cttcgacctg
2521 cccgactgca aggcccgcaa gcagcccatc aaggaggagt tcacggaggc cgagatccac
2581 tgagggcctc gcctggctgc agcctgcgcc accgcccaga gacccaagct gcctcccctc
2641 tccttcctgt gtgtccaaaa ctgcctcagg aggcaggacc ttcgggctgt gcccggggaa
2701 aggcaaggtc cggcccatcc ccaggcacct cacaggcccc aggaaaggcc cagccaccga
2761 agccgcctgt ggacagcctg agtcacctgc agaaccttct ggagctgccc tagtgctggg
2821 cttgtggggc gggggctggc ccactctcag ccctgccact gccccggcgt gctccatggc
2881 aggcgtgggt ggggaccgca gcgtcggctc cgacttccag gcttcatcct agagactgtc
2941 atctcccaac caggcgaggt ccttccaaag gaaaggatcc tctttgctga tggactgcca
3001 aaaagtattt tgcgacatct tttggttctg gatagtagtg agcagccaag tgactgtgtc
3061 tgaaacacca gtgtattttc agggaatgtc cctaactgcg tcttgcccgc gccgggggct
3121 ggggactctc tctgctggac ttgggactgg cctctgcccc cagcacgctg tattctgcag
3181 gaccgcctcc ttcctgcccc taacaacaac cacagtgttg ctgaaattgg agaaaactgg
3241 ggagggcgca acccccccca ggcgcgggga agcatgtggt accgcctcag ccagtgcccc
3301 tcagcctggc cacagtcgcc tctcctcggg gacccctcag cagaaaggga cagcctgtcc
3361 ttagaggact ggaaattgtc aatatttgat aaaatgatac ccttttctac atggtgggtc
3421 agcttttttt tttttttttt taactttctt tctcagcatt ctctttggag ttcaacctag
3481 cgcccatgag ccaggctgag gaagctgagt gagaagccag gtgggcggga cttgttccca
3541 ggaaggccgg gtggggagga agcctagagg gaaccccagg aagggcaaat ccaggcaaat
3601 ctgcaggaat gctctgccat gggagcagct cctcccttgc cacggccacc ttctctagca
3661 ctgcaaggtc cacagggcat tgctttcctt tctaggcggt ggcagtcagg gaacagactg
3721 aggtaggtgt aggggggtct aggccttcgt ggagcacccc agggagttag taggccccgg
3781 ggagacagag tctgcacagg ccctttctgg ggccacctcc atccacgagg agcagcctga
3841 gccttggtgg ccgaaccttg accgtcccgg agcacagctt cagggcaggg aaccggagcc
3901 cctggggggc ctcacgggtg tgacgaggcc cttcattgca ggcaggtggg ccaatgggag
3961 ccctcaccca cgcaagccga gacaccaccc agagtgcagg ctgcctggcc ccttctggca
4021 cggccagctc cacaccctct gcctagggta tgtgtggtcc taaggtctag gagcttcccc
4081 tactaacatc tcccagaaaa agcagttaag cccctcaggg cacagcaagg ttagacacag
4141 cccccatccc cagatcagga ctccatcttg ctaagtggca tcaccgtcac cagcctcccc
4201 ttatttaaaa gcagcgactg gtgttgccgc aggtacctgg tctacgaaga cgcaggcgtc
4261 cctctcccac cgtccacctc cccgggggcc gctgacagca cagtcgcctg ggtgcacgct
4321 tgtgggggca gcaggaacgg ggctgtcggc tctcagggga tctggctgca gccagggcga
4381 gggcctggcc cttccttcca gctccttccg gctccttcca gctgaagggc aggaagctct
4441 ggccgcttag cttctagggt tccatctccc tagaaaggtg cccacgccca gggcatcagt
4501 cagtagcggc agcagcagca gactcggggc tttcccaggg tggcgcagcc accccagctg
4561 catgtcacct cagctctcca tcttattgcc attttgtaga tgaggaagct gagaccagaa
4621 aggctaagac ccatgcccca ggcaccacac ccatctcttg ggggctgggc acctgctacc
4681 cgaggccacc tcctgaagcc cccactcttc ccccatgttc cacttcagga gccgcggggg
4741 cccatcctga cacccggggt tcctcagccc agcgcagatg tgcttcagtt ccagagggct
4801 tgttgatttg tttcttaggt acgttacctg tccaccctga gtccagtgag gctgtcccaa
4861 gagcccctgt agtgtgctcc tgggaagggc tgggggggct gggggggctg ggagaggccc
4921 aggggcagct gtcactggaa ccccagccag atgtccaagg aagccggcca gaacacggag
4981 cagccagatg gccccagctg cacctgtcta gggagcccat gcagcctcct tgcactggag
5041 aagcagctgt gaaagtagac agagttgaga cttcgccgtg gtcaggagaa aatgcaaatt
5101 cccaggaaca agaatccttt aagtgatatg tttttataaa actaaacaaa tcaacaaata
5161 aatcttgaag gcggatggtt ttcccagcag tgcaggggtt ggagggaggc tgctggcact
5221 cctggggcca agggggacag gcagtggtcc tgagtctgct cagagaggca aggcagaagg
5281 agctcgccag gcaggtcagc tcacatctgt ccaagtcgct ctggtcagaa acagcgactc
5341 tcccccattc ccccagcgtt cccaccaggc ctgggctgct gggaagccct tgctgtaccc
5401 aggagcccga cccgcagtat cctggcacag agccacttgt cactcagaac agtcagtgtc
5461 tccaacgcac aaacatccac tcctctgtta ccagttaaag cactttaatg ctttaaggtg
5521 aaaacgaaat cccatccgtg tttttcgtgt aagatcgtgc ttctccgagc agtattaatg
5581 gacgccctcc aatgacataa caactgtttt tggtaatgta atcttgggaa aatgtgttat
5641 ttttttagct gtgtttcagt ggggattttt gtttttgtaa cataataaag tgtatgttcc
5701 aatgaagtcg acctgcaggc atgcaagctg atccggctgc taacaaagcc cgaaaggaag
5761 ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg gcggccgctt
5821 cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga
5881 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
5941 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
6001 atgtgggagg tttttttaag caagtaaaac ctctacaaat gtggtaaaat cgaattttaa
6061 caaaatatta acgcttacaa tttcctgatg cggtattttc tccttacgca tctgtgcggt
6121 atttcacacc gcatacgcgg atctgcgcag caccatggcc tgaaataacc tctgaaagag
6181 gaacttggtt aggtaccttc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
6241 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
6301 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
6361 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
6421 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
6481 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
6541 gcctaggctt ttgcaaaaag cttgattctt ctgacacaac agtctcgaac caaaggctgg
6601 agccaccatg gcttccaagg tgtacgaccc cgagcaacgc aaacgcatga tcactgggcc
6661 tcagtggtgg gctcgctgca agcaaatgaa cgtgctggac tccttcatca actactatga
6721 ttccgagaag cacgccgaga acgccgtgat ttttctgcat ggtaacgctg cctccagcta
6781 cctgtggagg cacgtcgtgc ctcacatcga gcccgtggct agatgcatca tccctgatct
6841 gatcggaatg ggtaagtccg gcaagagcgg gaatggctca tatcgcctcc tggatcacta
6901 caagtacctc accgcttggt tcgagctgct gaaccttcca aagaaaatca tctttgtggg
6961 ccacgactgg ggggcttgtc tggcctttca ctactcctac gagcaccaag acaagatcaa
7021 ggccatcgtc catgctgaga gtgtcgtgga cgtgatcgag tcctgggacg agtggcctga
7081 catcgaggag gatatcgccc tgatcaagag cgaagagggc gagaaaatgg tgcttgagaa
7141 taacttcttc gtcgagacca tgctcccaag caagatcatg cggaaactgg agcctgagga
7201 gttcgctgcc tacctggagc cattcaagga gaagggcgag gttagacggc ctaccctctc
7261 ctggcctcgc gagatccctc tcgttaaggg aggcaagccc gacgtcgtcc agattgtccg
7321 caactacaac gcctaccttc gggccagcga cgatctgcct aagatgttca tcgagtccga
7381 ccctgggttc ttttccaacg ctattgtcga gggagctaag aagttcccta acaccgagtt
7441 cgtgaaggtg aagggcctcc acttcagcca ggaggacgct ccagatgaaa tgggtaagta
7501 catcaagagc ttcgtggagc gcgtgctgaa gaacgagcag accggtggtg ggagcggagg
7561 tggcggatca ggtggcggag gctccggagg gattgaacaa gatggattgc acgcaggttc
7621 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
7681 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
7741 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
7801 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
7861 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
7921 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
7981 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
8041 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
8101 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
8161 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
8221 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
8281 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
8341 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
8401 gaaatgaccg accaagcgac gcccaacctg ccatcacgat ggccgcaata aaatatcttt
8461 attttcatta catctgtgtg ttggtttttt gtgtgaatcg atagcgataa ggatcctctt
8521 tgcgcttgcg ttttcccttg tccagatagc ccagtagctg acattcatcc ggggtcagca
8581 ccgtttctgc ggactggctt tctacgtaat ggtttcttag acgtcaggtg gcacttttcg
8641 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
8701 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
8761 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
8821 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
8881 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
8941 acgttttcca atgatgagca ctttcaaagt tctgctatgt ggcgcggtat tatcccgtat
9001 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
9061 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
9121 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa ctatcggagg
9181 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
9241 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
9301 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
9361 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
9421 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
9481 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
9541 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
9601 gattaagcat tggtaattcg aaatgaccga ccaagcgacg cccaaccggt atcagctcac
9661 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
9721 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
9781 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
9841 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
9901 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
9961 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
10021 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
10081 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
10141 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
10201 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
10261 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
10321 gtttgcaagc agcagattac gcgcagaaaa aaaggatttc aagaagatcc tttgatcttt
10381 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
10441 ttatcaaaaa ggatcttcac ctagatcctt ttatagtccg gaaatacagg aacgcacgct
10501 ggatggccct tcgctgggat ggtgaaacca tgaaaaatgg cagcttcagt ggattaagtg
10561 ggggtaatgt ggcctgtacc ctctggttgc ataggtattc atacggttaa aatttatcag
10621 gcgcgattgc ggcagttttt cgggtggttt gttgccattt ttacctgtct gctgccgtga
10681 tcgcgctgaa cgcgttttag cggtgcgtac aattaaggga ttatggtaaa tccacttact
10741 gtctgccctc gtagccatcg agataaacc
//
| {
"Vector backbone": "pmiRGLO",
"Backbone size w/o insert (bp)": "7334",
"Vector type": "Mammalian Expression, Luciferase",
"Selectable markers": "Neomycin (select with G418)"
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA ; Luciferase Limited Use Label License ; Renilla Luciferase Limited Use Label License",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Combined Experimental and System-Level Analyses Reveal the Complex Regulatory Network of miR-124 during Human Neurogenesis",
"Publication": "Cell Syst. 2018 Sep 28. pii: S2405-4712(18)30358-2. doi: 10.1016/j.cels.2018.08.011.",
"DOI": "10.1016/j.cels.2018.08.011.",
"Authors": "Kutsche LK, Gysi DM, Fallmann J, Lenk K, Petri R, Swiersy A, Klapper SD, Pircs K, Khattak S, Stadler PF, Jakobsson J, Nowick K, Busskamp V /jcels",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/30292704",
"PubMed ID": "PubMed 30292704"
} | {
"Gene/Insert name": "3' UTR TP73-201",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "3439",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "Murine PGK-1",
"Tag / Fusion Protein": "luciferase (N terminal on backbone)"
} | {
"Cloning method": "Gibson Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "Fluc-F1 (AGAAGCTGCGCGGTGGTGTTGTG)",
"3′ sequencing primer": "T7terminal"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pAAV-AVPR2-eLOV-TEVcs-FLAG-GAL4-V5 | 104844 | b2AR-SPARK TF component | Alice Ting | Kim et al Elife. 2017 Nov 30;6. doi: 10.7554/eLife.30233. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/201547/30722db9-ed1f-47a2-bf8f-fe48974f6bcc/addgene-plasmid-104844-sequence-201547.gbk | full | LOCUS sequence-201547- 6272 bp DNA circular SYN 26-AUG-2024
DEFINITION b2AR-SPARK TF component.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6272)
AUTHORS Kim MW, Wang W, Sanchez MI, Coukos R, von Zastrow M, Ting AY
TITLE Time-gated detection of protein-protein interactions with
transcriptional readout.
JOURNAL Elife. 2017 Nov 30;6. doi: 10.7554/eLife.30233.
PUBMED 29189201
REFERENCE 2 (bases 1 to 6272)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Elife.";
date: "2017-11-30"; volume: "6. doi"; pages: " 10.7554/eLife.30233"
FEATURES Location/Qualifiers
source 1..6272
/mol_type="other DNA"
/organism="synthetic DNA construct"
repeat_region 1..130
/label=AAV2 ITR (alternate)
/note="Functional equivalent of wild-type AAV2 ITR"
enhancer 201..504
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 505..708
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 659..679
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 753..772
/label=T7
/note="T7 promoter, forward primer"
promoter 753..771
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 2381..2401
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQM"
CDS 2414..2437
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS 2438..2875
/codon_start=1
/gene="Saccharomyces cerevisiae GAL4 (truncated)"
/product="DNA binding domain of the GAL4 transcriptional
activator"
/label=GAL4 DNA binding domain
/translation="KLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTKR
SPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNKD
AVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
CDS 2876..3217
/codon_start=1
/gene="Saccharomyces cerevisiae GAL4 (truncated)"
/product="activation domain of the GAL4 transcriptional
activator"
/label=GAL4 activation domain
/translation="ANFNQSGNIADSSLSFTFTNSSNGPNLITTQTNSQALSQPIASSN
VHDNFMNNEITASKIDDGNNSKPLSPGWTDQTAYNAFGITTGMFNTTTMDDVYNYLFDD
EDTPPNPKKE"
CDS 3218..3259
/codon_start=1
/product="epitope tag from simian virus 5"
/label=V5 tag
/translation="GKPIPNPLLGLDST"
primer_bind complement(3293..3310)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 3299..3506
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
repeat_region 3535..3675
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 3535..3664
/label=AAV2 ITR
rep_origin 3750..4205
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3837..3856)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4047..4068
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(4222..4241)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 4341..4363
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(4401..4419)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 4487..4591
/gene="bla"
/label=AmpR promoter
CDS 4592..5452
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(4810..4829)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 5623..6211
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6112..6131
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
121 aggggttcct gcggccgcac gcgttgtaca tagtaatcaa ttacggggtc attagttcat
181 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
241 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
301 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
361 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
421 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
481 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
541 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
601 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
661 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact
721 agagaaccca ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa
781 gcttgccacc atgctcatgg cctctaccac aagcgctgtg cccggacatc catctcttcc
841 aagtcttccc tccaacagct cacaggagag gcctctggat acccgggacc cacttcttgc
901 aagagcggag ctcgccttgc tctccatcgt tttcgttgcc gtcgcgttga gcaatggtct
961 ggtgctggcc gccctggcgc gacgaggccg ccgaggccac tgggcgccta ttcatgtctt
1021 tatcggccat ctctgtcttg cagacctggc cgtggcgctg tttcaggtgc tccctcaact
1081 ggcttggaaa gccaccgata gattccgggg gccagacgca ctgtgtcggg ccgtgaaata
1141 cctgcagatg gtgggaatgt atgccagttc atatatgata cttgctatga ccttggacag
1201 acacagggct atctgtcgcc ccatgctggc gtataggcac ggttccggag cccattggaa
1261 caggccagtt ctggttgcgt gggctttttc cctgcttctg agcttgcccc agttgtttat
1321 tttcgcacag agaaatgtgg agggcggatc tggcgtgacc gattgctggg cttgcttcgc
1381 tgagccatgg ggcagaagga cctacgtgac gtggattgct ctcatggtgt tcgttgctcc
1441 caccctgggg atcgccgcat gccaggtcct gatattcaga gaaatacacg cctccttggt
1501 tcctgggcca agcgagcgcc ctggtggaag aagacggggt cgaaggactg gctctccagg
1561 cgaaggcgcc cacgtatcag ctgcggtagc taagaccgta cgcatgacac tggtcatcgt
1621 cgtcgtttac gttctttgct gggccccgtt ttttttggtg cagctctggg cggcctggga
1681 ccccgaagca cccctggaag gtgctccatt cgttctgctc atgcttctcg cttcactgaa
1741 tagctgtacc aatccttgga tttacgcctc tttctcctcc agtgttagct ccgagctgcg
1801 gagtcttctc tgttgcgccc ggggcaggac ccccccaagc ctgggtccac aggacgaatc
1861 ctgcactacc gcctccagtt ccctggccaa agacacctct tctgagctgg ccgagaagct
1921 ggccggcctg gacatcaacg gtggagctag cggcagccgg gcgaccaccc tggaacgtat
1981 tgaaaagagc tttgtgatta ccgatccgcg tctgccggat aacccgatta tttttgtgag
2041 cgatagcttt ctgcagctga ccgaatatag ccgtgaagaa attctgggcc gtaactgccg
2101 ttttctgcag ggcccggaaa ccgatcgtgc gaccgtgcgt aaaattcgtg atgcgattga
2161 taaccagacc gaagtgaccg tgcagctgat taactatacc aaaagcggca aaaaattttg
2221 gaacctgttt catctgcagc cgatgcgtga tcagaaaggc gatgtgcagt attttattgg
2281 cgtgcagctg gatggtaccg aacgtgtgcg tgatgcggcg gaacgtgaag ctgtgatgct
2341 ggtcaagaaa accgcggagg aaatcgacga agccgctaag gaaaatctct acttccagat
2401 gggcggagga tccgactaca aggatgacga cgataagaag ctactgtctt ctatcgaaca
2461 agcatgcgat atttgccgac ttaaaaagct caagtgctcc aaagaaaaac cgaagtgcgc
2521 caagtgtctg aagaacaact gggagtgtcg ctactctccc aaaaccaaaa ggtctccgct
2581 gactagggca catctgacag aagtggaatc aaggctagaa agactggaac agctatttct
2641 actgattttt cctcgagaag accttgacat gattttgaaa atggattctt tacaggatat
2701 aaaagcattg ttaacaggat tatttgtaca agataatgtg aataaagatg ccgtcacaga
2761 tagattggct tcagtggaga ctgatatgcc tctaacattg agacagcata gaataagtgc
2821 gacatcatca tcggaagaga gtagtaacaa aggtcaaaga cagttgactg tatcggccaa
2881 ttttaatcaa agtgggaata ttgctgatag ctcattgtcc ttcactttca ctaacagtag
2941 caacggtccg aacctcataa caactcaaac aaattctcaa gcgctttcac aaccaattgc
3001 ctcctctaac gttcatgata acttcatgaa taatgaaatc acggctagta aaattgatga
3061 tggtaataat tcaaaaccac tgtcacctgg ttggacggac caaactgcgt ataacgcgtt
3121 tggaatcact acagggatgt ttaataccac tacaatggat gatgtatata actatctatt
3181 cgatgatgaa gataccccac caaacccaaa aaaagagggc aagccgatcc ctaaccctct
3241 gctgggcctg gacagcactt aagaattcct cgagctagag ctcgctgatc agcctcgact
3301 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
3361 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
3421 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
3481 gaagacaata gcaggcatgc tggggaggta accacgtgcg gaccgagcgg ccgcaggaac
3541 ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc
3601 gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc
3661 gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
3721 cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc
3781 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc
3841 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
3901 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
3961 tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
4021 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
4081 ctctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg tctattggtt
4141 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac
4201 aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
4261 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
4321 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
4381 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
4441 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
4501 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
4561 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
4621 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
4681 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
4741 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
4801 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
4861 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
4921 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
4981 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
5041 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
5101 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
5161 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
5221 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
5281 tgataaatct ggagccggtg agcgtggaag ccgcggtatc attgcagcac tggggccaga
5341 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
5401 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
5461 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
5521 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
5581 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
5641 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
5701 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
5761 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
5821 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
5881 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
5941 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
6001 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
6061 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
6121 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
6181 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
6241 cctggccttt tgctggcctt ttgctcacat gt
//
| {
"Vector backbone": "pAAV",
"Backbone size w/o insert (bp)": "",
"Vector type": "Mammalian Expression, AAV",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "NEB Stable",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Time-gated detection of protein-protein interactions with transcriptional readout",
"Publication": "Elife. 2017 Nov 30;6. doi: 10.7554/eLife.30233.",
"DOI": "10.7554/eLife.30233.",
"Authors": "Kim MW, Wang W, Sanchez MI, Coukos R, von Zastrow M, Ting AY /eLife",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/29189201",
"PubMed ID": "PubMed 29189201"
} | {
"Gene/Insert name": "AVPR2-eLOV-TEVcs-FLAG-GAL4-V5",
"Species": "Synthetic",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "CMV",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Unknown",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "CMV-F",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
LZRS-Mek2-K101A (KA7) | 21191 | Low | Paul Khavari | Paul Khavari plasmids (unpublished) | https://media.addgene.org/snapgene-media/v2.0.0/sequences/26975/15353ceb-5192-441c-8f74-5ec450134536/addgene-plasmid-21191-sequence-26975.gbk | partial | LOCUS sequence-26975-e 893 bp DNA linear UNA 26-AUG-2024
DEFINITION natural linear DNA
ACCESSION .
VERSION .
KEYWORDS .
SOURCE natural DNA sequence
ORGANISM unspecified
REFERENCE 1 (bases 1 to 893)
TITLE Paul Khavari plasmids
REFERENCE 2 (bases 1 to 893)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..893
/mol_type="genomic DNA"
/organism="unspecified"
protein_bind 58..82
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
ORIGIN
1 gggacacngg gngacatcct ctgactgccg gatcccagnt gtggtggtac gggaatcaca
61 agtttgtaca aaaaagcagg ctttaaagga accaattcag tcgactggat ccatgctggc
121 ccggaggaag ccggtgctgc cggcgctcac catcaaccct accatcgccg agggcccatc
181 ccctaccagc gagggcgcct ccgaggcaaa cctggtggac ctgcagaaga agctggagga
241 gctggaactt gacgagcagc agaagaagcg gctggaagcc tttctcaccc agaaagccaa
301 ggttggcgaa ctcaaagacg atgacttcga aaggatctca gagctgggcg cgggcaacgg
361 cggggtggtc accaaagtcc agcacagacc ctcgggcctc atcatggcca gggcgctgat
421 ccacctcgag atcaagccgg ccatccggaa ccagatcatc cgcgagctgc aggtcctgca
481 cgaatgcaac tcgccgtaca tcgtgggctt ctacggggcc ttctacagtg acggggagat
541 cagcatttgc atggaacaca tggacggcgg ctccctggac caggtgctga aagaggccaa
601 gaggattccc gaggagatcc tggggaaagt cagcatcgcg gttctccggg gcttggcgta
661 cctccgagag aagcaccaga tcatgcaccg agatgtgaag ccctccaaca tcctcgtgaa
721 ctctagaggg gagatcaagc tgtgtgactt cggggtgagc ggccactcat agactccatg
781 gccacntctt cgtggnacgc gctcctacat ggctcggagc gttgcaggca cnattactcg
841 tgcatcgaan tggactggcc tgtcntgtga ctgcctcgaa gaccctcccc gcc
//
| {
"Vector backbone": "LZRS-RfA",
"Backbone size w/o insert (bp)": "11100",
"Vector type": "Retroviral",
"Selectable markers": "Puromycin"
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "Stbl3",
"Copy number": "Low Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "",
"Publication": "",
"DOI": "",
"Authors": "",
"PubMed Link": "",
"PubMed ID": ""
} | {
"Gene/Insert name": "Mek2",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "1200",
"Mutation": "K101A (catalytically inactive)",
"Entrez Gene": "MAP2K2",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "BamHI (not destroyed)",
"3′ cloning site": "NotI (not destroyed)",
"5′ sequencing primer": "TGGATACACGCCGCCCACGTG",
"3′ sequencing primer": "ATCGTCGACCACTGTGCTGG"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pAc5.1A-DmXRN1-P1341AP1342A-V5His6_Q | 147252 | Insect Expression of DmXRN1-P1341AP1342A | Elisa Izaurralde | Izaurralde lab plasmids (unpublished) | https://media.addgene.org/snapgene-media/v2.0.0/sequences/289699/f1d0f6ad-2f2f-4037-9083-804ffd02568a/addgene-plasmid-147252-sequence-289699.gbk | full | LOCUS sequence-289699- 10211 bp DNA circular SYN 26-AUG-2024
DEFINITION Insect Expression of DmXRN1-P1341AP1342A.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10211)
TITLE Izaurralde lab plasmids
REFERENCE 2 (bases 1 to 10211)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..10211
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 56..2568
/label=Ac5 promoter
/note="Drosophila melanogaster actin 5C promoter"
CDS 7496..7537
/codon_start=1
/product="epitope tag from simian virus 5"
/label=V5 tag
/translation="GKPIPNPLLGLDST"
CDS 7547..7564
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(7587..7604)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 7637..7771
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(7661..7680)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 7715..7734
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
rep_origin complement(8393..8981)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(8473..8492)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(9152..10012)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 9775..9794
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(10013..10117)
/gene="bla"
/label=AmpR promoter
primer_bind 10185..10203
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
ORIGIN
1 cttcaaggaa ttaattctat attctaaaaa cacaaatgat acttctaaaa aaaatcatga
61 atggcatcaa ctctgaatca aatctttgca gatgcaccta cttctcattt ccactgtcac
121 atcatttttc cagatctcgc tgcctgttat gtggcccaca aaccaagaca cgttttatgg
181 ccattaaagc tggctgatcg tcgccaaaca ccaaatacat atcaatatgt acattcgaga
241 aagaagcgat caaagaagcg tcttcgggcg agtaggagaa tgcggaggag aaggagaacg
301 agctgatcta gtatctctcc acaatccaat gccaactgac caactggcca tattcggagc
361 aatttgaagc caatttccat cgcctggcga tcgctccatt cttggctata tgtttttcac
421 cgttcccggg gccattttca aagactcgtc ggtaagataa gattgtgtca ctcgctgtct
481 ctcttcattt gtcgaagaat gctgaggaat ttcgcgatga cgtcggcgag tattttgaag
541 aatgagaata atttgtattt atacgaaaat cagttagtgg aattttctac aaaaacatgt
601 tatctataga taattttgtt gcaaaatatg ttgactatga caaagattgt atgtatatac
661 ctttaatgta ttctcatttt cttatgtatt tataatggca atgatgatac tgatgatatt
721 ttaagatgat gccagaccac aggctgattt ctgcgtcttt tgccgaacgc agtgcatgtg
781 cggttgttgt tttttggaat agtttcaatt ttcggactgt ccgctttgat ttcagtttct
841 tggcttattc aaaaagcaaa gtaaagccaa aaaagcgaga tggcaatacc aaatgcggca
901 aaacggtagt ggaaggaaag gggtgcgggg cagcggaagg aagggtgggg cggggcgtgg
961 cggggtctgt ggctgggcgc gacgtcaccg acgttggagc cactcctttg accatgtgtg
1021 cgtgtgtgta ttattcgtgt ctcgccactc gccggttgtt tttttctttt tatctcgctc
1081 tctctagcgc catctcgtac gcatgctcaa cgcaccgcat gttgccgtgt cctttatgcg
1141 tcattttggc tcgaaatagg caattattta aacaaagatt agtcaacgaa aacgctaaaa
1201 taaataagtc tacaatatgg ttacttattg ccatgtgtgt gcagccaacg atagcaacaa
1261 aagcaacaac acagtggctt tccctctttc actttttgtt tgcaagcgcg tgcgagcaag
1321 acggcacgac cggcaaacgc aattacgctg acaaagagca gacgaagttt tggccgaaaa
1381 acatcaaggc gcctgatacg aatgcatttg caataacaat tgcgatattt aatattgttt
1441 atgaagctgt ttgacttcaa aacacacaaa aaaaaaaata aaacaaatta tttgaaagag
1501 aattaggaat cggacagctt atcgttacgg gctaacagca caccgagacg aaatagctta
1561 cctgacgtca cagcctctgg aagaactgcc gccaagcaga gagagagaga aaaagaggga
1621 gagcagctta gaccgcatgt gcttgtgtgt gaggcgtctc tctcttcgtc tcctgtttgc
1681 gcaaacgcat agactgcact gagaaaatcg attacctatt ttttatgaat gaatatttgc
1741 actattacta ttcaaaacta ttaagatagc aatcacattc aatagccaaa tactatacca
1801 cctgagcgat gcaacgaaat gatcaatttg agcaaaaatg ctgcatattt aggacggcat
1861 cattatagaa atgcttcttg ctgtgtactt ttctctcgtc tggcagctgt ttcgccgtta
1921 ttgttaaaac cggcttaagt taggtgtgtt ttctacgact agtgatgccc ctactagaag
1981 atgtgtgttg cacaaatgtc cctgaataac caatttgaag tgcagatagc agtaaacgta
2041 agctaatatg aatattattt aactgtaatg ttttaatatc gctggacatt actaataaac
2101 ccactataaa cacatgtaca tatgtatgtt ttggcataca atgagtagtt ggggaaaaaa
2161 tgtgtaaaag caccgtgacc atcacagcat aaagataacc agctgaagta tcgaatatga
2221 gtaaccccca aattgaatca catgccgcaa ctgataggac ccatggaagt acactcttca
2281 tggcgatata caagacacac acaagcacga acacccagtt gcggaggaaa ttctccgtaa
2341 atgaaaaccc aatcggcgaa caattcatac ccatatatgg taaaagtttt gaacgcgact
2401 tgagagcgga gagcattgcg gctgataagg ttttagcgct aagcgggctt tataaaacgg
2461 gctgcgggac cagttttcat atcactaccg tttgagttct tgtgctgtgt ggatactcct
2521 cccgacacaa agccgctcca tcagccagca gtcgtctaat ccagagaccc cggatcgggg
2581 tacctactag tccagtgtgg tggaattctg cagatatcca gcacagtggc ggccgcccaa
2641 catgggcgtt cccaagttct ttcgctacat cagcgagcga tatccgtgcc tcagcgaatt
2701 ggcgcgggag cactgcatac ccgagttcga caatctgtac ttggacatga acggcattgt
2761 ccacaactgc tcgcatccgg acgacaacaa catacacttt cacctggagg aggagcagat
2821 attccaggaa atcttcaact acgtagacaa gctgttctac ctgatcaagc cgcagcgcct
2881 cttcttcctt tcagttgacg gggtggctcc acgggccaag atgaaccagc agcgcagccg
2941 gcgattccgg acggcgcgcg aggcggagca gcaggaggcc aaggcggccc agcgaggcga
3001 gttgcgagag cacgagcgct tcgacagcaa ctgcattacg ccgggcaccg aattcatggt
3061 gcgtctgcag gagggactgc gcgcctttct caaaacgaag atttcgacgg atccgctgtg
3121 gcagcgttgc accgtaatcc taagtggcca ggaggcgccc ggcgagggcg agcacaagat
3181 catggactac attcggtaca tgaagacgca gccggactat gatcccaata cacgacactg
3241 cctttacggc ctggacgccg atctgatcat tctgggcctc tgcacccacg agctgcactt
3301 cgtggtgctg cgcgaggagg tcaagttcgg gcgcaacgtc aagcgcacct cggtggagga
3361 gacgcgcttc ttcctgctgc acctgggtct gctgcgcgag tacctcgagc tggagttcga
3421 cgccctgaga acagacgagc acaaattgga cattgcgcag cttatcgacg actgggtgct
3481 gatgggcttt ttggtcggca acgacttcat tccccacctg ccgtgcctgc acatttcatc
3541 aaatgcactg ccgctgctct accgcaccta catcggcatt tacccgaccc tcggcggcaa
3601 catcaacgag aacggcaagc tcaacctgcg ccgcttgcag attttcatat cggccctcac
3661 cgaggtggag ctggatcact tcaaggagca cgccgacgac ttgaagtaca tgaacaacaa
3721 atcagaggcg ttcgacatgg acgtcggcga gatcaccgag tctcaaaacc tcgactccga
3781 tctcggagcc ctcatcaaca agagcatgct gctctacgat gacgacagcg aggaggactg
3841 ctccgacgag aacgcggttc ttctgaagga gttccagaac tacaagcgaa acttctaccg
3901 caacaagttc aagcgtgatc cgaatgatga actgatcgag gaactgtgcc accactacgt
3961 caatgcgctg cagtgggtgc tcgactacta ctaccgtggc gtgcagtcct gggactggta
4021 ctatcccttc cactacactc cattcatcag cgacctcaag aacatcgagc aggtggagat
4081 cgccttccac atgggcacgc cctttctgcc attccagcag cttctggcgg tactgccggc
4141 ggccagtgca aagcttctcc ctgtcgccta ccacgacctc atgctgctgc cgaccagccc
4201 gctggccgag ttctatccac tggagttcga gagcgacctc aatggcaaga aacacgattg
4261 ggaggcggtt gtgctgatac ccttcatcga tgagggtcgg ctgctggccg ctatgctgcc
4321 atgcgaggct cagctgtcgc tggaggagcg cgaacgcaac cgacacggtc ccatgtatgt
4381 gtacaaatac agtacggtgg cgcagggtcc catgccagcg tatccaccgc tgagggctct
4441 tcccgtgctc tactgcacgg aggtggccaa gtggtcgcac gaaatcgcag tcaatctgcc
4501 gtacagcgtg tgcatcgagc taccaaatgc ggcgagaacc gttttcttcc cgggatttcc
4561 caccatgcag catctgccct tcgatttcga gctgcgcaac gaccgcgtca aggtgttcga
4621 acaggtcagt cgcaaccaga acattgtgct aaaaccacgc aagcgccagc tggaggacac
4681 actgacggcg gtggcctccc agtacctcgg gaaggtgatc cacgtcggct ggccgcatct
4741 cgtcaaggcg atagtggtgc gcgtggccac gcgagatcag cgcgtcgaca gcgagggaat
4801 cacgctaaac gattcgcggc gtttcgactc ggaatgcaag gcactgcagg aacactttat
4861 caatcgtatg ggaatacagt ttgccaacta tgatgtgctc gtctatgtgc gcacctttgc
4921 cggcaattcg acggagttcc gcgacaaggg ggcattgatg gtgcgcgact cgtggagcag
4981 ctccgtcacc ggctatccag cccagggcgt ggtggccgat ctgactgtgt gggaaagaat
5041 gcgcaagaat ttcctcaacg tggagcacta cttccccgtt ggcagcacga tcttcttgat
5101 cacggatccg tattacggca gcgagggaac tgtgcaggac ccacggctgg cctacaccaa
5161 cggccggatt caagtcagca tcatggtgcg accggaaccg aaagtaaatg cggcacgtca
5221 gctgcaggaa gagcgggacc gggactatct gagcaccttc caggtctgca acttgctccg
5281 aatcagtggc cgcaccctgg gacgcctaag tggcaccgtg tgggtggtgc tcggtccgcg
5341 gcgtcagaag atggaaaacg tgaccaagca caacattggc ctgcagctca agtatccgcg
5401 gcagaacgag gagcgtgccg gatactgctt ccgcacgaac aaccagtggt actactccag
5461 cctggccgtg gatctgatgc gtaactattg ccagcgttac ccagacgtta tcgacttctt
5521 tggcgactcc aacgatcgcg ccgagttcgt attcgaacag gacgtgttcc ccaatgcggt
5581 tggccatcgc cgcgtcgagg aactggccaa ctgggtgcgc cagcagccgc acatgaaagt
5641 ggagcgcatc tcttgcggct ccaagacggt ctgccgcgag accatcgaac tgctgatagc
5701 cgccgtcgac gacctgcgct cactgcccgt caagcacgtt aaactgcagg tgaagccgca
5761 cttgctcatc aagcccaatg tcacactgcc agatgtctac cgatcaaagc gcccggtgcg
5821 tctcttcgat cgcgtcgtaa tcgtacgcac catttacatg gtgcccgtgg gcactaaggg
5881 caccgtgatt ggcattcatc ccgttacgga ccccaacccg gtccgcctgg agtgcgtcca
5941 tgctgtcgac acgttctgca aggtgctgtt cgacagcccc gttccgaact gcaacaacat
6001 ccatggcatc gccgaagacc gagtttacaa ggtgcccgaa attgccctgg tgatcatcaa
6061 gacggatgag gaaggaaaga agcagaacga ctgcgaactg ccggtgaggg atcctcagcc
6121 gaaccaggca caggacgagc cagtccgcgc gaccagcagt cgctatgtga ccgccgcagg
6181 gtccacctcg gtcccgatca cgatgaagac ccaaattagc gacgaattcg tcaagacgcg
6241 tagcgatccg atagcccgta cggattcgta caagccatcg tcggagccaa aaccggtgcc
6301 ggtgccggag cagattacca actggcggga acgcgtcagt acgccaacaa acaaaccgca
6361 gcctgcgcca aataattggc gcataaatcg gtcttcatca cgtcaacagg gcggtagtat
6421 ttttgttgct ccgcccacaa aaactcctga tgcagctgca agcactgctt ccactgcatt
6481 caccgcagcc tcgtccgcaa cgctgactcc tctggatcag accttggcgc tcatgtcggt
6541 tttgggcgtg ggtgaggacc agagctcgcc gccactccag gaggcggtgc agcagcaacg
6601 tccgccacta cttcaacagc aaagagctcc gtttcccgga caaatgccca atctacccaa
6661 ggcagcgctc ttctggcagc aagaggcgca gaagcaggag gcgctgcagc aggaggcgca
6721 gcagcaggag gcgcagaaga agcagcagca ggcgcatgcg caaatggagc ctgaacgtat
6781 caatagccag catttctaca gaagcggcca gacaggagcg gcgttgaatc agccgccatt
6841 gggtgctcca tcaaaaaggc agtggcatga gtgggtgcac ccgcgcatgc agcacgccaa
6901 cgcattccac gctggcgtca acaatggata tcagatgcgt cccaaaaaga atatagctgc
6961 gcagagcacc tttaacaaca acgtgcacat gcacctccag caaccatatt acccgaatca
7021 gcagcagcag cagcagcagc agcagccact ccagctgacg gagatcaaca atgcgccacc
7081 gcgatactca acgatccaag attttgtgcc catacaggcg taccgtccca agaaattaaa
7141 tcgcgtccaa ccagccggtc gtcaggacgt cgatgccacc aagaatccta gtcgttcgcc
7201 tgtgttgcag caaccgacga acgagactat agacacaaag gccagttcta gtcttccggt
7261 acaatcagca ggcgaacagg tgattggact gatgcaaacg cttgaaatta agccggctgc
7321 gtcgcagtcc gaatctgatg gggtctcaac aggcagtgcg aacgcgccca cagcaacgac
7381 atcatcccag gcggtcaacc gaaggaagca tcgagtgcct cgcatcggag ccaagttcga
7441 cttggaatac atactgcccg actccccgca tccaacttct agagggccct tcgaaggtaa
7501 gcctatccct aaccctctcc tcggtctcga ttctacgcgt accggtcatc atcaccatca
7561 ccattgagtt taaacccgct gatcagcctc gactgtgcct tctaaggcct gagctcgctg
7621 atcagcctcg atcgaggatc cagacatgat aagatacatt gatgagtttg gacaaaccac
7681 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
7741 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
7801 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
7861 tatggctgat tatgatcagt cgaccgatgc ccttgagagc cttcaaccca gtcagctcct
7921 tccggtgggc gcggggcatg actatcgtcg ccgcacttat gactgtcttc tttatcatgc
7981 aactcgtagg acaggtgccg gcagcgctct gggtcatttt cggcgaggac cgctttcgct
8041 ggagcgcgac gatgatcggc ctgtcgcttg cggtattcgg aatcttgcac gccctcgctc
8101 aagccttcgt cactggtccc gccaccaaac gtttcggcga gaagcaggcc attatcgccg
8161 gcatggcggc cgacgcgctg ggctacgtct tgctggcgtt cgcgacgcga ggctggatgg
8221 ccttccccat tatgattctt ctcgcttccg gcggcatcgg gatgcccgcg ttgcaggcca
8281 tgctgtccag gcaggtagat gacgaccatc agggacagct tcaaggatcg ctcgcggctc
8341 ttaccagcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
8401 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
8461 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
8521 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
8581 tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
8641 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
8701 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
8761 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
8821 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
8881 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
8941 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
9001 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
9061 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
9121 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
9181 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
9241 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
9301 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
9361 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
9421 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctgca ggcatcgtgg
9481 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
9541 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
9601 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
9661 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
9721 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca cgggataata
9781 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
9841 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
9901 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
9961 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
10021 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
10081 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
10141 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
10201 ggccctttcg t
//
| {
"Vector backbone": "Unknown",
"Backbone size w/o insert (bp)": "",
"Vector type": "Insect Expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "Low Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "",
"Publication": "",
"DOI": "",
"Authors": "",
"PubMed Link": "",
"PubMed ID": ""
} | {
"Gene/Insert name": "DmXRN1-P1341AP1342A",
"Species": "D. melanogaster (fly)",
"Insert Size (bp)": "",
"Mutation": "one silent mtuation compared to the sequence given by NCBI:NM_078684.3 variant A.",
"Entrez Gene": "pcm",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Unknown",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
G417A_pLenti-Tet-BattP(GA)-BFP-2A-iCasp9-2A-Blast_rtTA3 | 200631 | Negative selection blasticidin resistant Tet-inducible Matreyek Bxb1(GA) landing pad | Kenneth Matreyek | Roelle et al ACS Synth Biol. 2023 Nov 3. doi: 10.1021/acssynbio.3c00355. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/394835/c04e26bf-e85f-42d5-88ad-6e339c4df6c2/addgene-plasmid-200631-sequence-394835.gbk | full | LOCUS sequence-394835- 10410 bp DNA circular SYN 26-AUG-2024
DEFINITION Negative selection blasticidin resistant Tet-inducible Matreyek
Bxb1(GA) landing pad.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10410)
AUTHORS Roelle SM, Kamath ND, Matreyek KA
TITLE Mammalian Genomic Manipulation with Orthogonal Bxb1 DNA Recombinase
Sites for the Functional Characterization of Protein Variants.
JOURNAL ACS Synth Biol. 2023 Nov 3. doi: 10.1021/acssynbio.3c00355.
PUBMED 37922210
REFERENCE 2 (bases 1 to 10410)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth
Biol. 2023 Nov 3. doi: 10.1021/acssynbio.3c00355."
FEATURES Location/Qualifiers
source 1..10410
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 1..861
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(219..238)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 1032..1620
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 1521..1540
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 1774..1791
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 1908..1929
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 1944..1974
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 1982..1998
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
primer_bind 1987..2009
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 2006..2022
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 2006..2022
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 2041..2061
/label=T3
/note="T3 promoter, forward primer"
promoter 2043..2061
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 2089..2315
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 2316..2496
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 2543..2668
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 3161..3394
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 3579..3623
/codon_start=1
/product="antigenic peptide corresponding to amino acids
655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
et al., 2013)"
/label=gp41 peptide
/note="recognized by the 2H10 single-chain llama nanobody"
/translation="KNEQELLELDKWASL"
misc_feature 3890..4006
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 4149..4498
/label=TRE3GV promoter
/note="3rd-generation Tet-responsive promoter that can be
activated by binding of Tet-On(R) 3G, optimized for
retroviral and lentiviral vectors"
protein_bind 4157..4175
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 4193..4211
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 4229..4247
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 4265..4283
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 4301..4319
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 4337..4355
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 4373..4391
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 4500..4547
/label=Bxb1 attP
/bound_moiety="Mycobacteriophage Bxb1 integrase"
/note="Minimal attP site for the Mycopbacteriophage Bxb1
integrase (Voutev and Mann, 2017)"
regulatory 4548..4557
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 4554..5264
/codon_start=1
/product="enhanced monomeric blue fluorescent protein
(Subach et al., 2011)"
/label=mTagBFP2
/note="mammalian codon-optimized"
/translation="MVSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTM
RIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
VLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRND
MALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAV
ARYCDLPSKLGHKLN"
CDS 5274..5330
/codon_start=1
/product="2A peptide from porcine teschovirus-1
polyprotein"
/label=P2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="ATNFSLLKQAGDVEENPGP"
CDS 5346..5666
/codon_start=1
/product="F36V mutant of FK506-binding protein FKBP12"
/label=DmrB
/note="binds synthetic ligands that do not bind wild-type
FKBP"
/translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKP
FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
KLE"
CDS complement(6018..6029)
/codon_start=1
/product="WELQut protease recognition and cleavage site"
/label=WELQut site
/translation="WELQ"
CDS 6543..6569
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 6588..6641
/codon_start=1
/product="2A peptide from Thosea asigna virus capsid
protein"
/label=T2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="EGRGSLLTCGDVEENPGP"
CDS 6642..7037
/codon_start=1
/gene="Aspergillus terreus BSD"
/product="blasticidin S deaminase"
/label=BSD
/note="confers resistance to blasticidin"
/translation="AKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
AIVKDSDGQPTAVGIRELLPSGYVWEG"
primer_bind complement(7130..7149)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_signal 7195..7250
/label=beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(7249..7268)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
enhancer 7584..7887
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 7888..8091
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
protein_bind 8196..8220
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
CDS 8223..8930
/codon_start=1
/product="reverse tetracycline transactivator 3"
/label=rtTA3
/note="activates expression from Tet-responsive promoters
in the presence of doxycycline"
/translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
DALDDFDLDMLPG"
LTR 9017..9250
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 9322..9443
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(9359..9378)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 9413..9432
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 9483..9618
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(9638..9657)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(9639..9657)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(9667..9684)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(9667..9683)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(9676..9698)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 9825..10280
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(9912..9931)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 10122..10143
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 10306..10410
/gene="bla"
/label=AmpR promoter
ORIGIN
1 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
61 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
121 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
181 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
241 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
301 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
361 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
421 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
481 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
541 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
601 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
661 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
721 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
781 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
841 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
901 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
961 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
1021 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
1081 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
1141 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
1201 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
1261 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
1321 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
1381 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
1441 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
1501 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
1561 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
1621 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
1681 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
1741 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
1801 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
1861 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
1921 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
1981 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagcgc
2041 gcaattaacc ctcactaaag ggaacaaaag ctggagctgc aagcttaatg tagtcttatg
2101 caatactctt gtagtcttgc aacatggtaa cgatgagtta gcaacatgcc ttacaaggag
2161 agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg gtacgatcgt gccttattag
2221 gaaggcaaca gacgggtctg acatggattg gacgaaccac tgaattgccg cattgcagag
2281 atattgtatt taagtgccta gctcgataca taaacgggtc tctctggtta gaccagatct
2341 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
2401 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
2461 tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa
2521 agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac
2581 ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta
2641 gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg
2701 ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg
2761 ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg
2821 ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag aagaacttag
2881 atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga
2941 caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca
3001 gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat tggagaagtg
3061 aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa
3121 agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt
3181 tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca
3241 gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc
3301 aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg
3361 ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac
3421 tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga
3481 tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac acaagcttaa
3541 tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg
3601 aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata
3661 taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac
3721 tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag acccacctcc
3781 caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga gagagagaca
3841 gagacagatc cattcgatta gtgaacggat ctcgacggta tcggttaact tttaaaagaa
3901 aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca
3961 tacaaactaa agaattacaa aaacaaatta caaaattcaa aattttatcg ataagcttgg
4021 gagttcaggg acagcagaga tccagtttat cggatgttta taaattgcga tgaggccctt
4081 tcgtcttcag cagagatcca gtttatcgat attaaggctt atgcgatgag gccctttcgt
4141 cttcactcga gtttactccc tatcagtgat agagaacgta tgaagagttt actccctatc
4201 agtgatagag aacgtatgca gactttactc cctatcagtg atagagaacg tataaggagt
4261 ttactcccta tcagtgatag agaacgtatg accagtttac tccctatcag tgatagagaa
4321 cgtatctaca gtttactccc tatcagtgat agagaacgta tatccagttt actccctatc
4381 agtgatagag aacgtatgtc gaggtaggcg tgtacggtgg gcgcctataa aagcagagct
4441 cgtttagtga accgtcagat cgcctggagc aattccacaa cacttttgtc ttatacttgg
4501 gtttgtctgg tcaaccaccg cggactcagt ggtgtacggt acaaaccgcc accatggtgt
4561 ctaagggcga agagctgatt aaggagaaca tgcacatgaa gctgtacatg gagggcaccg
4621 tggacaacca tcacttcaag tgcacatccg agggcgaagg caagccctac gagggcaccc
4681 agaccatgag aatcaaggtg gtcgagggcg gccctctccc cttcgccttc gacatcctgg
4741 ctactagctt cctctacggc agcaagacct tcatcaacca cacccagggc atccccgact
4801 tcttcaagca gtccttccct gagggcttca catgggagag agtcaccaca tacgaagacg
4861 ggggcgtgct gaccgctacc caggacacca gcctccagga cggctgcctc atctacaacg
4921 tcaagatcag aggggtgaac ttcacatcca acggccctgt gatgcagaag aaaacactcg
4981 gctgggaggc cttcaccgag acgctgtacc ccgctgacgg cggcctggaa ggcagaaacg
5041 acatggccct gaagctcgtg ggcgggagcc atctgatcgc aaacgccaag accacatata
5101 gatccaagaa acccgctaag aacctcaaga tgcctggcgt ctactatgtg gactacagac
5161 tggaaagaat caaggaggcc aacaacgaga cctacgtcga gcagcacgag gtggcagtgg
5221 ccagatactg cgacctccct agcaaactgg ggcacaagct taatgggagc ggagctacta
5281 acttcagcct gctgaagcag gctggagacg tggaggagaa ccctggacct tccgccaccc
5341 tcgagggagt gcaggtggag actatctccc caggagacgg gcgcaccttc cccaagcgcg
5401 gccagacctg cgtggtgcac tacaccggga tgcttgaaga tggaaagaaa gttgattcct
5461 cccgggacag aaacaagccc tttaagttta tgctaggcaa gcaggaggtg atccgaggct
5521 gggaagaagg ggttgcccag atgagtgtgg gtcagagagc caaactgact atatctccag
5581 attatgccta tggtgccact gggcacccag gcatcatccc accacatgcc actctcgtct
5641 tcgatgtgga gcttctaaaa ctggaatctg gcggtggatc cggagtcgac ggatttggtg
5701 atgtcggtgc tcttgagagt ttgaggggaa atgcagattt ggcttacatc ctgagcatgg
5761 agccctgtgg ccactgcctc attatcaaca atgtgaactt ctgccgtgag tccgggctcc
5821 gcacccgcac tggctccaac atcgactgtg agaagttgcg gcgtcgcttc tcctcgctgc
5881 atttcatggt ggaggtgaag ggcgacctga ctgccaagaa aatggtgctg gctttgctgg
5941 agctggcgcg gcaggaccac ggtgctctgg actgctgcgt ggtggtcatt ctctctcacg
6001 gctgtcaggc cagccacctg cagttcccag gggctgtcta cggcacagat ggatgccctg
6061 tgtcggtcga gaagattgtg aacatcttca atgggaccag ctgccccagc ctgggaggga
6121 agcccaagct ctttttcatc caggcctgtg gtggggagca gaaagaccat gggtttgagg
6181 tggcctccac ttcccctgaa gacgagtccc ctggcagtaa ccccgagcca gatgccaccc
6241 cgttccagga aggtttgagg accttcgacc agctggacgc catatctagt ttgcccacac
6301 ccagtgacat ctttgtgtcc tactctactt tcccaggttt tgtttcctgg agggacccca
6361 agagtggctc ctggtacgtt gagaccctgg acgacatctt tgagcagtgg gctcactctg
6421 aagacctgca gtccctcctg cttagggtcg ctaatgctgt ttcggtgaaa gggatttata
6481 aacagatgcc tggttgcttt aatttcctcc ggaaaaaact tttctttaaa acatcagtcg
6541 actatccgta cgacgtacca gactacgcac tcgactctgg cagcggagag ggcagaggaa
6601 gtcttctaac atgcggtgac gtggaggaga atcccggccc tgccaagcct ttgtctcaag
6661 aagaatccac cctcattgaa agagcaacgg ctacaatcaa cagcatcccc atctctgaag
6721 actacagcgt cgccagcgca gctctctcta gcgacggccg catcttcact ggtgtcaatg
6781 tatatcattt tactggggga ccttgtgcag aactcgtggt gctgggcact gctgctgctg
6841 cggcagctgg caacctgact tgtatcgtcg cgatcggaaa tgagaacagg ggcatcttga
6901 gcccctgcgg acggtgccga caggtgcttc tcgatctgca tcctgggatc aaagccatag
6961 tgaaggacag tgatggacag ccgacggcag ttgggattcg tgaattgctg ccctctggtt
7021 atgtgtggga gggctagtaa acgcgtgaat tcactcctca ggtgcaggct gcctatcaga
7081 aggtggtggc tggtgtggcc aatgccctgg ctcacaaata ccactgagat ctttttccct
7141 ctgccaaaaa ttatggggac atcatgaagc cccttgagca tctgacttct ggctaataaa
7201 ggaaatttat tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga
7261 catatgggag ggcaaatcat ttaaaacatc agaatgagta tttggtttag agtttggcaa
7321 catatgccca tatgctggct gccatgaaca aaggttggct ataaagaggt catcagtata
7381 tgaaacagcc ccctgctgtc cattccttat tccatagaaa agccttgact tgaggttaga
7441 ttttttttat attttgtttt gtgttatttt tttctttaac atccctaaaa ttttccttac
7501 atgttttact agccagattt ttcctcctct cctgactact cccagtcata gctgtccctc
7561 ttctcttatg gagatctaaa ccgcgttaca taacttacgg taaatggccc gcctggctga
7621 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
7681 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
7741 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
7801 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
7861 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
7921 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
7981 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
8041 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg
8101 aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccga
8161 ctctagtcca gtgtggtgga attctgcaga tatcaacaag tttgtacaaa aaagttggca
8221 ccatgtctag actggacaag agcaaagtca taaacggagc tctggaatta ctcaatggtg
8281 tcggtatcga aggcctgacg acaaggaaac tcgctcaaaa gctgggagtt gagcagccta
8341 ccctgtactg gcacgtgaag aacaagcggg ccctgctcga tgccctgcca atcgagatgc
8401 tggacaggca tcatacccac ttctgccccc tggaaggcga gtcatggcaa gactttctgc
8461 ggaacaacgc caagtcatac cgctgtgctc tcctctcaca tcgcgacggg gctaaagtgc
8521 atctcggcac ccgcccaaca gagaaacagt acgaaaccct ggaaaatcag ctcgcgttcc
8581 tgtgtcagca aggcttctcc ctggagaacg cactgtacgc tctgtccgcc gtgggccact
8641 ttacactggg ctgcgtattg gaggaacagg agcatcaagt agcaaaagag gaaagagaga
8701 cacctaccac cgattctatg cccccacttc tgagacaagc aattgagctg ttcgaccggc
8761 agggagccga acctgccttc cttttcggcc tggaactaat catatgtggc ctggagaaac
8821 agctaaagtg cgaaagcggc gggccgaccg acgcccttga cgattttgac ttagacatgc
8881 tcccagccga tgcccttgac gattttgacc ttgacatgct ccccgggtaa gcacaattcg
8941 agctcggtac ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta
9001 aaagaaaagg ggggactgga agggctagct cactcccaac gaagacaaga tctgcttttt
9061 gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
9121 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
9181 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
9241 atctctagca gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa
9301 tgaatatcag agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc
9361 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
9421 tccaaactca tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc
9481 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
9541 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
9601 gaggcttttt tggaggccta gggacgtacc caattcgccc tatagtgagt cgtattacgc
9661 gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
9721 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
9781 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg
9841 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
9901 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
9961 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
10021 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
10081 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
10141 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
10201 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
10261 aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt
10321 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
10381 atgcttcaat aatattgaaa aaggaagagt
//
| {
"Vector backbone": "pLenti",
"Backbone size w/o insert (bp)": "",
"Vector type": "Lentiviral",
"Selectable markers": "Blasticidin ; mTagBFP2"
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "NEB Stable",
"Copy number": "Unknown"
} | {
"Academic/Nonprofit Terms": "UBMTA ; Evrogen Limited Use Label License for FPs ; Tet Systems Limited Use Label License",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Mammalian Genomic Manipulation with Orthogonal Bxb1 DNA Recombinase Sites for the Functional Characterization of Protein Variants",
"Publication": "ACS Synth Biol. 2023 Nov 3. doi: 10.1021/acssynbio.3c00355.",
"DOI": "10.1021/acssynbio.3c00355.",
"Authors": "Roelle SM, Kamath ND, Matreyek KA /acssynbioc",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/37922210",
"PubMed ID": "PubMed 37922210"
} | {
"Gene/Insert name": "iCasp9",
"Species": "Synthetic",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "pTRE3G",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Gibson Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
PROM_7 | 201300 | D7: Promoter of TraesCS3D02G316100 as Golden Gate compatible MoClo PROM part | Benjamin Schwessinger | Schwessinger Lab Plasmids 2023 (unpublished) | https://media.addgene.org/snapgene-media/v2.0.0/sequences/409511/6159e3b1-f86b-44b9-9604-7235545fe5a8/addgene-plasmid-201300-sequence-409511.gbk | full | LOCUS sequence-409511- 2847 bp DNA circular SYN 27-AUG-2024
DEFINITION D7: Promoter of TraesCS3D02G316100 as Golden Gate compatible MoClo
PROM part.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2847)
TITLE Schwessinger Lab Plasmids 2023
REFERENCE 2 (bases 1 to 2847)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 27, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..2847
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(891..907)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(891..907)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(904..926)
/label=M13/pUC Reverse
/note="In lacZ gene"
rep_origin complement(1079..1667)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1159..1178)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
terminator complement(1800..1894)
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
CDS complement(1915..2574)
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind 2489..2508
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
promoter complement(2575..2677)
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
primer_bind 2764..2786
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 2778..2795
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 2779..2795
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
ORIGIN
1 ctctgtggtc tcaggagggt tgagaggacg taggcaaatt acaagttatc ttcccaaaag
61 aagaggccct cccatgtcta cgatgcttcc tgtcacctac tatagctcca ccaagaaata
121 atcaggtgaa tagtatcggt actgcatcaa aacacaccgg aatccgaaaa ctaggcctgc
181 aaaattctct ttaatcctca tcgcaaacca tcaagtgtga tcacaactga cgagcaacgg
241 cgttagcgcg taggactcga acacatatca actgaacgga aatttcctgc gagtcgagtc
301 atttaagatg accacgtcaa aatgccgtcc agcagctaca caaatcggaa ataaatacgc
361 caccttcctc gagatcacct cagtttatac tagttatatg atcagccaga cacattgacc
421 tgactctgcc tattttctta atgactcata aagaaaaaga ggcacacatc atcacatcac
481 atgtgacccc tgcaagcaat ccggcacaag cctccttcct tccgtcgtac gacgtgctgg
541 ctacttcgca gaaaccaacc gaaaaccgcc gggcgcagca acgtcctaga catagctaag
601 ccagccacga cgcctgcaat tacctgtact ttggttggca tccatggaga ggctgcttcc
661 ttcccagaac ctatggactc cttcctcccc cgtcaccacc cccttattta agcacccacc
721 caccattctc ctctcattct ccattcacca ctgttgcaag ctgggccggg tgttcttgat
781 cctttgttct tcgccgtcga cgaggatcgg gcacagcaat gcgagaccac gaagaggcta
841 ggtggaggct cagtgatgat aagtctgcga tggtggatgc atgtgtcatg gtcatagctg
901 tttcctgtgt gaaattgtta tccgctcaga gggcacaatc ctattccgcg ctatccgaca
961 atctccaaga cattaggtgg agttcagttc ggcgtatggc atatgtcgct ggaaagaaca
1021 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
1081 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
1141 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
1201 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
1261 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
1321 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
1381 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
1441 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
1501 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
1561 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
1621 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
1681 tcttttctac ggggtctgac gctctattca acaaagccgc cgtcccgtca agtcagcgta
1741 aatgggtagg gggcttcaaa tcgtcccccc atacgatata agttgttact agtgcttgga
1801 ttctcaccaa taaaaaacgc ccggcggcaa ccgagcgttc tgaacaaatc cagatggagt
1861 tctgaggtca ttactggatc tatcaacagg agtccaagcg agctcgatat caaattacgc
1921 cccgccctgc cactcatcgc agtactgttg taattcatta agcattctgc cgacatggaa
1981 gccatcacaa acggcatgat gaacctgaat cgccagcggc atcagcacct tgtcgccttg
2041 cgtataatat ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt
2101 taaatcaaaa ctggtgaaac tcacccaggg attggctgag acgaaaaaca tattctcaat
2161 aaacccttta gggaaatagg ccaggttttc accgtaacac gccacatctt gcgaatatat
2221 gtgtagaaac tgccggaaat cgtcgtggta ttcactccag agcgatgaaa acgtttcagt
2281 ttgctcatgg aaaacggtgt aacaagggtg aacactatcc catatcacca gctcaccgtc
2341 tttcattgcc atacgaaatt ccggatgagc attcatcagg cgggcaagaa tgtgaataaa
2401 ggccggataa aacttgtgct tatttttctt tacggtcttt aaaaaggccg taatatccag
2461 ctgaacggtc tggttatagg tacattgagc aactgactga aatgcctcaa aatgttcttt
2521 acgatgccat tgggatatat caacggtggt atatccagtg atttttttct ccattttagc
2581 ttccttagct cctgaaaatc tcgataactc aaaaaatacg cccggtagtg atcttatttc
2641 attatggtga aagttggaac ctcttacgtg ccgatcaacg tctcattgat acctgaaaca
2701 aaacccatcg tacggccaag gaagtctcca ataactgtga tccaccacaa gcgccagggt
2761 tttcccagtc acgacgttgt aaaacgacgg ccagtcatgc ataatccgca cgcatctgga
2821 ataaggaagt gccattccgc ctgacct
//
| {
"Vector backbone": "Twist Bioscience Chloramphenicol vector",
"Backbone size w/o insert (bp)": "",
"Vector type": "Plant Expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Chloramphenicol, 25 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "",
"Publication": "",
"DOI": "",
"Authors": "",
"PubMed Link": "",
"PubMed ID": ""
} | {
"Gene/Insert name": "D7: Promoter of TraesCS3D02G316100",
"Species": "Triticum aestivum",
"Insert Size (bp)": "",
"Mutation": "removal of BsaI and BpiI sites",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "unknown (unknown if destroyed)",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pET28a-SENP8 (full length)) | 16361 | Low | Guy Salvesen | Mikolajczyk et al J Biol Chem. 2007 Sep 7. 282(36):26217-24. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/307095/4abc8b6b-da71-45e7-92d0-1e5b38f4541f/addgene-plasmid-16361-sequence-307095.gbk | full | LOCUS sequence-307095- 5928 bp DNA circular SYN 26-AUG-2024
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5928)
AUTHORS Mikolajczyk J, Drag M, Bekes M, Cao JT, Ronai Z, Salvesen GS
TITLE Small ubiquitin-related modifier (SUMO)-specific proteases:
profiling the specificities and activities of human SENPs.
JOURNAL J Biol Chem. 2007 Sep 7. 282(36):26217-24.
PUBMED 17591783
REFERENCE 2 (bases 1 to 5928)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J Biol
Chem. 2007 Sep 7. 282(36):26217-24."
FEATURES Location/Qualifiers
source 1..5928
/mol_type="other DNA"
/organism="synthetic DNA construct"
terminator complement(26..73)
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
primer_bind 69..87
/label=T7 Term
/note="T7 terminator, reverse primer"
CDS complement(140..157)
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS complement(806..823)
/codon_start=1
/product="thrombin recognition and cleavage site"
/label=thrombin site
/translation="LVPRGS"
CDS complement(833..850)
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
RBS 869..891
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
protein_bind 906..930
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
primer_bind complement(930..949)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(931..949)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(1033..1052)
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
promoter 1258..1335
/gene="lacI"
/label=lacI promoter
CDS 1336..2418
/codon_start=1
/gene="lacI"
/product="lac repressor"
/label=lacI
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
primer_bind complement(1355..1374)
/label=LacI-R
/note="LacI, reverse primer"
CDS 3227..3418
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
primer_bind complement(3412..3434)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
misc_feature 3520..3659
/label=bom
/note="basis of mobility region from pBR322"
primer_bind 3575..3594
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(3674..3691)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3845..4433)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3925..3944)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind complement(4544..4563)
/label=pENTR-R
/note="pENTR vectors, reverse primer"
CDS 4555..5370
/codon_start=1
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
CDS 4555..5370
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind complement(4628..4647)
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
rep_origin complement(5462..5917)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5599..5620)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 5811..5830
/label=F1ori-R
/note="F1 origin, reverse primer"
ORIGIN
1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagctacttt gtagcaagtg
181 tggcaatgag atctttccat tctcccctct tctttgtgat gtatgcaggg gtgagtagct
241 gcagcagtga ttctgtctgt tgcctaaaga agttctgaca caaggcctca gtgttacata
301 tcacgtacat cccacagtca tagctgtttt gttgggcagg ggctttctct tccacaaagg
361 ccagtttgtc tccttttctg cctaagaaag cctccagttt ctctgctacc tgctttgcgt
421 gaactgagtt gctcctgcta tgggaatcat aatgaaaaaa gctattttta tcttggaggt
481 agaccaataa actccagtgg gttcctccag ctgcctggtt ggagttatca ttgatggcta
541 aaaatacaac tctcttgttg gggaggtcca gtggttcaag gaacatggca atctctgctg
601 ggttgctagt gcacttgatg aactgggtga cttcagggct gatgaaactg acgtgatcag
661 agcagtcatg aaactgactg ttggcaaagt actcaaacgc aaacccaata atatggtcat
721 tgagccagct tggcggatcc aatagtgaga catctgattg ccgcagtaga ctgtccatgt
781 aactcaagac tacggggtcc atatggctgc cgcgcggcac caggccgctg ctgtgatgat
841 gatgatgatg gctgctgccc atggtatatc tccttcttaa agttaaacaa aattatttct
901 agaggggaat tgttatccgc tcacaattcc cctatagtga gtcgtattaa tttcgcggga
961 tcgagatctc gatcctctac gccggacgca tcgtggccgg catcaccggc gccacaggtg
1021 cggttgctgg cgcctatatc gccgacatca ccgatgggga agatcgggct cgccacttcg
1081 ggctcatgag cgcttgtttc ggcgtgggta tggtggcagg ccccgtggcc gggggactgt
1141 tgggcgccat ctccttgcat gcaccattcc ttgcggcggc ggtgctcaac ggcctcaacc
1201 tactactggg ctgcttccta atgcaggagt cgcataaggg agagcgtcga gatcccggac
1261 accatcgaat ggcgcaaaac ctttcgcggt atggcatgat agcgcccgga agagagtcaa
1321 ttcagggtgg tgaatgtgaa accagtaacg ttatacgatg tcgcagagta tgccggtgtc
1381 tcttatcaga ccgtttcccg cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg
1441 gaaaaagtgg aagcggcgat ggcggagctg aattacattc ccaaccgcgt ggcacaacaa
1501 ctggcgggca aacagtcgtt gctgattggc gttgccacct ccagtctggc cctgcacgcg
1561 ccgtcgcaaa ttgtcgcggc gattaaatct cgcgccgatc aactgggtgc cagcgtggtg
1621 gtgtcgatgg tagaacgaag cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc
1681 gcgcaacgcg tcagtgggct gatcattaac tatccgctgg atgaccagga tgccattgct
1741 gtggaagctg cctgcactaa tgttccggcg ttatttcttg atgtctctga ccagacaccc
1801 atcaacagta ttattttctc ccatgaagac ggtacgcgac tgggcgtgga gcatctggtc
1861 gcattgggtc accagcaaat cgcgctgtta gcgggcccat taagttctgt ctcggcgcgt
1921 ctgcgtctgg ctggctggca taaatatctc actcgcaatc aaattcagcc gatagcggaa
1981 cgggaaggcg actggagtgc catgtccggt tttcaacaaa ccatgcaaat gctgaatgag
2041 ggcatcgttc ccactgcgat gctggttgcc aacgatcaga tggcgctggg cgcaatgcgc
2101 gccattaccg agtccgggct gcgcgttggt gcggatatct cggtagtggg atacgacgat
2161 accgaagaca gctcatgtta tatcccgccg ttaaccacca tcaaacagga ttttcgcctg
2221 ctggggcaaa ccagcgtgga ccgcttgctg caactctctc agggccaggc ggtgaagggc
2281 aatcagctgt tgcccgtctc actggtgaaa agaaaaacca ccctggcgcc caatacgcaa
2341 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
2401 ctggaaagcg ggcagtgagc gcaacgcaat taatgtaagt tagctcactc attaggcacc
2461 gggatctcga ccgatgccct tgagagcctt caacccagtc agctccttcc ggtgggcgcg
2521 gggcatgact atcgtcgccg cacttatgac tgtcttcttt atcatgcaac tcgtaggaca
2581 ggtgccggca gcgctctggg tcattttcgg cgaggaccgc tttcgctgga gcgcgacgat
2641 gatcggcctg tcgcttgcgg tattcggaat cttgcacgcc ctcgctcaag ccttcgtcac
2701 tggtcccgcc accaaacgtt tcggcgagaa gcaggccatt atcgccggca tggcggcccc
2761 acgggtgcgc atgatcgtgc tcctgtcgtt gaggacccgg ctaggctggc ggggttgcct
2821 tactggttag cagaatgaat caccgatacg cgagcgaacg tgaagcgact gctgctgcaa
2881 aacgtctgcg acctgagcaa caacatgaat ggtcttcggt ttccgtgttt cgtaaagtct
2941 ggaaacgcgg aagtcagcgc cctgcaccat tatgttccgg atctgcatcg caggatgctg
3001 ctggctaccc tgtggaacac ctacatctgt attaacgaag cgctggcatt gaccctgagt
3061 gatttttctc tggtcccgcc gcatccatac cgccagttgt ttaccctcac aacgttccag
3121 taaccgggca tgttcatcat cagtaacccg tatcgtgagc atcctctctc gtttcatcgg
3181 tatcattacc cccatgaaca gaaatccccc ttacacggag gcatcagtga ccaaacagga
3241 aaaaaccgcc cttaacatgg cccgctttat cagaagccag acattaacgc ttctggagaa
3301 actcaacgag ctggacgcgg atgaacaggc agacatctgt gaatcgcttc acgaccacgc
3361 tgatgagctt taccgcagct gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca
3421 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
3481 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga cccagtcacg
3541 tagcgatagc ggagtgtata ctggcttaac tatgcggcat cagagcagat tgtactgaga
3601 gtgcaccatt gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc
3661 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
3721 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
3781 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
3841 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
3901 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
3961 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
4021 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
4081 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
4141 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
4201 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
4261 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
4321 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
4381 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
4441 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
4501 tggtcatgaa caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc
4561 catattcaac gggaaacgtc ttgctctagg ccgcgattaa attccaacat ggatgctgat
4621 ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcga
4681 ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc
4741 aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg
4801 accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc
4861 gggaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat
4921 gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac
4981 agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat
5041 gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg
5101 cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat
5161 aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc
5221 gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca
5281 ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag
5341 tttcatttga tgctcgatga gtttttctaa gaattaattc atgagcggat acatatttga
5401 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
5461 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
5521 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
5581 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
5641 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
5701 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
5761 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
5821 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
5881 acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgcca
//
| {
"Vector backbone": "pET28a",
"Backbone size w/o insert (bp)": "5369",
"Vector type": "Bacterial Expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Kanamycin, 50 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "Unknown"
} | {
"Academic/Nonprofit Terms": "UBMTA ; Sanford-Burnham Medical Research Institute Ancillary Agreement",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Small ubiquitin-related modifier (SUMO)-specific proteases: profiling the specificities and activities of human SENPs",
"Publication": "J Biol Chem. 2007 Sep 7. 282(36):26217-24.",
"DOI": "10.1074/jbc.M702444200",
"Authors": "Mikolajczyk J, Drag M, Bekes M, Cao JT, Ronai Z, Salvesen GS",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/17591783",
"PubMed ID": "PubMed 17591783"
} | {
"Gene/Insert name": "human SENP8 full length",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "663",
"Mutation": "human SENP8 full length, see Depositor Comments",
"Entrez Gene": "SENP8",
"Promoter": "",
"Tag / Fusion Protein": "His (N terminal on backbone)"
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "NdeI (not destroyed)",
"3′ cloning site": "XhoI (not destroyed)",
"5′ sequencing primer": "T7",
"3′ sequencing primer": "T7"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
EF1a-HexaPro_Foldon-PDGFR-T2A-mCherry (pZYW079) | 194234 | Lentiviral expression vector expressing Jason McClellan lab's 6-proline Spike extracellular domain on the cell surface attached to PDGFR's transmembrane domain and mCherry transduction marker | Hana El-Samad | Weinberg et al bioRxiv 2021.04.20.440678 | https://media.addgene.org/snapgene-media/v2.0.0/sequences/383907/d52907fa-724b-44a3-9c46-c602b979d1e0/addgene-plasmid-194234-sequence-383907.gbk | full | LOCUS sequence-383907- 14465 bp DNA circular SYN 27-AUG-2024
DEFINITION Lentiviral expression vector expressing Jason McClellan lab's
6-proline Spike extracellular domain on the cell surface attached to
PDGFR's transmembrane domain and mCherry transduction marker.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 14465)
AUTHORS Weinberg ZY, Hilburger CE, Kim M, Cao L, Khalid M, Elmes S, Diwanji
D, Hernandez E, Lopez J, Schaefer K, Smith AM, Zhou F, QCRG
Structural Biology Consortium, Kumar GR, Ott M, Baker D, El-Samad H
TITLE Sentinel cells enable genetic detection of SARS-CoV-2 Spike protein
JOURNAL bioRxiv 2021.04.20.440678
REFERENCE 2 (bases 1 to 14465)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 27, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv
2021.04.20.440678"
FEATURES Location/Qualifiers
source 1..14465
/mol_type="other DNA"
/organism="synthetic DNA construct"
LTR 99..732
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 779..904
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1401..1634
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1819..1863
/codon_start=1
/product="antigenic peptide corresponding to amino acids
655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
et al., 2013)"
/label=gp41 peptide
/note="recognized by the 2H10 single-chain llama nanobody"
/translation="KNEQELLELDKWASL"
misc_feature 2148..2265
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2326..3504
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 2557..3495
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 3452..3472
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
CDS 3574..3603
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 7327..7473
/codon_start=1
/product="transmembrane domain from platelet derived growth
factor receptor beta"
/label=PDGFR-beta TM domain
/translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
KKPR"
CDS 7489..7542
/codon_start=1
/product="2A peptide from Thosea asigna virus capsid
protein"
/label=T2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="EGRGSLLTCGDVEENPGP"
CDS 7552..8262
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
primer_bind complement(7697..7715)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 7965..7984
/label=mCherry-F
/note="mCherry, forward primer"
misc_feature 8277..8865
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(8330..8350)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(8748..8759)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(8887..8903)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(8888..8904)
/label=pBluescriptKS
/note="For pBluescript vector"
LTR 8994..9227
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
promoter complement(9252..9270)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(9253..9270)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(9560..9579)
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind complement(9793..9812)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 9912..9934
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(9972..9990)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
rep_origin complement(10169..10757)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(10249..10268)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
promoter 10860..10964
/gene="bla"
/label=AmpR promoter
CDS 10965..11825
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(11183..11202)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
primer_bind 12067..12084
/label=L4440
/note="L4440 vector, forward primer"
primer_bind complement(12154..12174)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 12159..12488
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 12339..12474
/label=SV40 ori
/note="SV40 origin of replication"
intron 13622..13687
/label=small t intron
/note="SV40 (simian virus 40) small t antigen intron"
CDS 13817..13837
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
polyA_signal 14262..14396
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(14299..14318)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 14353..14372
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
ORIGIN
1 tacctgtggt ttcatttact ctaaacctgt gattcctctg aattattttc attttaaaga
61 aattgtattt gttaaatatg tactacaaac ttagtagttg gaagggctaa ttcactccca
121 aagaagacaa gatatccttg atctgtggat ctaccacaca caaggctact tccctgatta
181 gcagaactac acaccagggc caggggtcag atatccactg acctttggat ggtgctacaa
241 gctagtacca gttgagccag ataaggtaga agaggccaat aaaggagaga acaccagctt
301 gttacaccct gtgagcctgc atgggatgga tgacccggag agagaagtgt tagagtggag
361 gtttgacagc cgcctagcat ttcatcacgt ggcccgagag ctgcatccgg agtacttcaa
421 gaactgctga tatcgagctt gctacaaggg actttccgct ggggactttc cagggaggcg
481 tggcctgggc gggactgggg agtggcgagc cctcagatcc tgcatataag cagctgcttt
541 ttgcctgtac tgggactctc tggttagacc agatctgagc ctgggagctc tctggctaac
601 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
661 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
721 aaatctctag cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac cagaggagct
781 ctctcgacgc aggactcggc ttgctgaagc gcgcacggca agaggcgagg ggcggcgact
841 ggtgagtacg ccaaaaattt tgactagcgg aggctagaag gagagagatg ggtgcgagag
901 cgtcagtatt aagcggggga gaattagatc gcgatgggaa aaaattcggt taaggccagg
961 gggaaagaaa aaatataaat taaaacatat agtatgggca agcagggagc tagaacgatt
1021 cgcagttaat cctggcctgt tagaaacatc agaaggctgt agacaaatac tgggacagct
1081 acaaccatcc cttcagacag gatcagaaga acttagatca ttatataata cagtagcaac
1141 cctctattgt gtgcatcaaa ggatagagat aaaagacacc aaggaagctt tagacaagat
1201 agaggaagag caaaacaaaa gtaagaccac cgcacagcaa gcggccggcc gctgatcttc
1261 agacctggag gaggagatat gagggacaat tggagaagtg aattatataa atataaagta
1321 gtaaaaattg aaccattagg agtagcaccc accaaggcaa agagaagagt ggtgcagaga
1381 gaaaaaagag cagtgggaat aggagctttg ttccttgggt tcttgggagc agcaggaagc
1441 actatgggcg cagcgtcaat gacgctgacg gtacaggcca gacaattatt gtctggtata
1501 gtgcagcagc agaacaattt gctgagggct attgaggcgc aacagcatct gttgcaactc
1561 acagtctggg gcatcaagca gctccaggca agaatcctgg ctgtggaaag atacctaaag
1621 gatcaacagc tcctggggat ttggggttgc tctggaaaac tcatttgcac cactgctgtg
1681 ccttggaatg ctagttggag taataaatct ctggaacaga tttggaatca cacgacctgg
1741 atggagtggg acagagaaat taacaattac acaagcttaa tacactcctt aattgaagaa
1801 tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg aattagataa atgggcaagt
1861 ttgtggaatt ggtttaacat aacaaattgg ctgtggtata taaaattatt cataatgata
1921 gtaggaggct tggtaggttt aagaatagtt tttgctgtac tttctatagt gaatagagtt
1981 aggcagggat attcaccatt atcgtttcag acccacctcc caaccccgag gggacccgac
2041 aggcccgaag gaatagaaga agaaggtgga gagagagaca gagacagatc cattcgatta
2101 gtgaacggat ctcgacggta tcgccaaatg gcagtattca tccacaattt taaaagaaaa
2161 ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc aacagacata
2221 caaactaaag aattacaaaa acaaattaca aaaattcaaa attttcgggt ttattacagg
2281 gacagcagag atccagtttg gccctctgat gagacgaacg cgtgaggctc cggtgcccgt
2341 cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat
2401 tgaaccggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg
2461 ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac
2521 gttctttttc gcaacgggtt tgccgccaga acacaggtaa gtgccgtgtg tggttcccgc
2581 gggcctggcc tctttacggg ttatggccct tgcgtgcctt gaattacttc cacctggctg
2641 cagtacgtga ttcttgatcc cgagcttcgg gttggaagtg ggtgggagag ttcgaggcct
2701 tgcgcttaag gagccccttc gcctcgtgct tgagttgagg cctggcctgg gcgctggggc
2761 cgccgcgtgc gaatctggtg gcaccttcgc gcctgtctcg ctgctttcga taagtctcta
2821 gccatttaaa atttttgatg acctgctgcg acgctttttt tctggcaaga tagtcttgta
2881 aatgcgggcc aagatctgca cactggtatt tcggtttttg gggccgcggg cggcgacggg
2941 gcccgtgcgt cccagcgcac atgttcggcg aggcggggcc tgcgagcgcg gccaccgaga
3001 atcggacggg ggtagtctca agctggccgg cctgctctgg tgcctggcct cgcgccgccg
3061 tgtatcgccc cgccctgggc ggcaaggctg gcccggtcgg caccagttgc gtgagcggaa
3121 agatggccgc ttcccggccc tgctgcaggg agctcaaaat ggaggacgcg gcgctcggga
3181 gagcgggcgg gtgagtcacc cacacaaagg aaaagggcct ttccgtcctc agccgtcgct
3241 tcatgtgact ccacggagta ccgggcgccg tccaggcacc tcgattagtt ctcgagcttt
3301 tggagtacgt cgtctttagg ttggggggag gggttttatg cgatggagtt tccccacact
3361 gagtgggtgg agactgaagt taggccagct tggcacttga tgtaattctc cttggaattt
3421 gccctttttg agtttggatc ttggttcatt ctcaagcctc agacagtggt tcaaagtttt
3481 tttcttccat ttcaggtgtc gtgagccact atggccttgc ccgttacggc acttctgctc
3541 ccactcgcat tgctgcttca cgccgcccga ccagaacaga aactcataag tgaagaagac
3601 cttgggggag gcggttccgg gggaggcggt tctagccagt gcgtgaacct gaccacccga
3661 acccagctcc caccagccta caccaacagc tttacacggg gcgtgtacta ccctgacaag
3721 gtgttcagat ctagcgtcct gcacagcact caggacctct tcctgccgtt cttcagcaac
3781 gtgacatggt tccacgccat ccacgtgagc ggcacaaacg gaaccaagcg gtttgataac
3841 cccgtcctgc cattcaatga tggagtttac ttcgccagta ccgagaagag taacatcatc
3901 cggggctgga tcttcggcac caccctggat agcaaaacac agagcctcct gatcgtgaac
3961 aatgccacga acgtcgtgat caaggtgtgc gagttccagt tttgcaatga tcctttcctg
4021 ggtgtgtact accacaagaa caacaagagc tggatggaaa gcgagttcag agtctacagc
4081 agcgccaaca actgcacatt cgagtatgtc tctcagcctt ttctgatgga ccttgagggg
4141 aaacaaggca acttcaagaa cctgagagaa ttcgtgttca agaacatcga cggctacttc
4201 aaaatctact ccaagcacac acccatcaac ctggtccggg acctccctca gggcttcagc
4261 gccctggaac ccctggtcga cctgcccata ggcatcaaca taacgcggtt ccaaaccctg
4321 ctggccctgc atagatccta cctgactcct ggcgacagca gcagcggatg gaccgccgga
4381 gctgcagcct actatgtggg ctacctgcaa cctagaacct tcctgctgaa gtacaacgag
4441 aacggcacaa tcacagacgc cgtcgactgc gccctggacc ctctctctga gacaaagtgc
4501 accctgaagt ccttcaccgt ggaaaagggc atctaccaga ccagcaactt ccgggtgcag
4561 cctacagaga gcatcgtgcg atttccaaac attaccaacc tctgcccctt cggcgaggtg
4621 tttaacgcca caagatttgc ctccgtttac gcctggaata gaaagagaat cagcaattgt
4681 gtggccgact actccgtgct gtataacagc gcctctttca gcaccttcaa gtgctacggc
4741 gtttccccaa caaagctgaa tgacctgtgc ttcaccaacg tgtacgccga ctccttcgta
4801 attagaggcg atgaggtgcg gcagatcgca ccaggccaga ccggtaagat cgctgactac
4861 aactataagc tgcctgatga ttttacaggc tgcgtgatcg cctggaactc taacaacctg
4921 gatagcaagg tgggcggcaa ctacaactac ctgtaccggc tgtttcgcaa gtctaacctg
4981 aaacctttcg agagagacat ctccacagag atctaccagg ccggttctac accttgtaac
5041 ggggtggaag gcttcaactg ttacttccct ctgcaaagct acggcttcca gcctaccaat
5101 ggagtcggct accagccata ccgggtggtc gtgctgtcct tcgagttact ccacgccccc
5161 gccaccgtct gcggtcctaa gaagtccacc aatctggtta agaacaaatg cgtgaacttc
5221 aacttcaacg gcctgaccgg gaccggcgtg ctgaccgaaa gcaacaaaaa gttcctcccc
5281 ttccagcagt tcggccgtga tatcgctgac accacagatg ccgtcagaga tccacagacc
5341 ctggaaatcc tggatattac accctgctcc ttcggaggag tttctgtgat cacccccggg
5401 accaatacca gcaaccaggt ggctgtgctg taccaagatg ttaactgcac cgaggttcct
5461 gtggccatcc acgccgatca gctgacacct acttggagag tgtactccac tggctccaat
5521 gtgttccaga ccagggccgg atgtctgatc ggcgccgagc acgtgaataa cagttacgag
5581 tgcgacatcc ctatcggcgc cggcatctgt gccagctacc agacccagac aaacagccct
5641 gggtctgctt cctctgtagc tagccagagc atcatcgcct acaccatgag cctgggcgca
5701 gagaacagcg tggcctattc caacaactct atcgccattc ccaccaactt tacaattagc
5761 gtcacaacag agatcctgcc cgtgagcatg accaagacca gcgtggactg tacaatgtac
5821 atctgtggcg acagcactga atgcagcaac ctgctgctgc aatacggctc cttttgcacc
5881 caactgaacc gggcgctgac cggaatcgcc gtggaacagg acaaaaatac ccaggaggtg
5941 ttcgcccaag tgaagcagat ctacaagacc ccacctatca aggacttcgg cggctttaac
6001 tttagccaga ttctccctga tccttctaag cctagcaagc ggagccctat cgaggatctg
6061 ctgttcaaca aggtcaccct ggccgatgcc ggctttatca aacagtatgg cgattgcctg
6121 ggcgacatag ccgccagaga tctgatctgc gcccagaaat tcaacggcct gacagttctc
6181 ccacctctgc tgaccgacga gatgatcgct cagtacacct ctgccctgct ggctggcacc
6241 atcacatctg ggtggacatt tggcgccggc cccgccctgc agatcccctt tcccatgcag
6301 atggcctata gattcaacgg aatcggcgtg acccagaacg tgctgtatga aaaccagaag
6361 ctgatcgcta accagttcaa ttctgccatc ggcaagatcc aggactccct ctcctctacc
6421 cccagcgccc tgggcaaact gcaggacgtg gtgaatcaga acgcccaagc cctgaacacc
6481 ctggtgaagc agctcagcag caattttggc gccatcagct ctgtgctgaa cgatatcctg
6541 tctagactgg accctccaga agccgaagtc cagatcgata gactgatcac aggcagactg
6601 cagtccctgc aaacctacgt gacccaacag ctgatcaggg ccgctgaaat aagagccagc
6661 gccaatctcg ccgctaccaa gatgtccgag tgtgtgctgg gacagtctaa acgcgttgac
6721 ttctgcggca aaggctatca cctgatgagc ttcccccaga gcgcgccgca cggcgtggtg
6781 ttcctgcatg tgacatacgt gcctgcccaa gagaagaatt tcacaaccgc ccctgccatc
6841 tgccacgacg gcaaggccca cttcccaaga gagggcgttt tcgtttccaa tggcacacac
6901 tggttcgtga cacaaagaaa cttctacgaa ccccagatta tcaccaccga caacaccttc
6961 gtgagtggca attgtgacgt ggtcatcgga atcgtgaaca acacagtgta cgaccctctg
7021 caacctgagc tggactcttt taaggaagag ctggacaagt actttaaaaa ccacaccagc
7081 cccgatgtgg acctgggcga catcagtggc attaacgcca gcgtggtgaa catccaaaag
7141 gaaatcgaca gactgaacga ggtggccaag aacctgaacg agtccctgat cgacctgcag
7201 gagctcggca aatacgagca gggatccgga tacatccccg aggcccccag agatggccag
7261 gcctacgtgc ggaaggacgg cgagtgggta ctgctgagca cattcctggg cagatccggt
7321 tctaatgctg tgggccagga cacgcaggag gtcatcgtgg tgccacactc cttgcccttt
7381 aaggtggtgg tgatctcagc catcctggcc ctggtggtgc tcaccatcat ctcccttatc
7441 atcctcatca tgctttggca gaagaagcca cgtggatccg ggagtggcga aggcaggggc
7501 tcccttttga cgtgcgggga tgttgaagag aaccctgggc ccggatctgg aatggtgagc
7561 aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg
7621 gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac
7681 gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc cttcgcctgg
7741 gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca ccccgccgac
7801 atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac
7861 ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggcgagttc
7921 atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt aatgcagaag
7981 aagaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg cgccctgaag
8041 ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc tgaggtcaag
8101 accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt caacatcaag
8161 ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga acgcgccgag
8221 ggccgccact ccaccggcgg catggacgag ctgtacaagt aatggcgctt atcgataatc
8281 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
8341 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
8401 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
8461 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
8521 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
8581 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
8641 gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct
8701 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
8761 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
8821 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatgct gcgtctcaag
8881 catacacgat accgtcgacc tcgagggaat taattcgagc tcggtacctt taagaccaat
8941 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg
9001 gctaattcac tcccaacgaa gacaagatct gctttttgct tgtactgggt ttctctggtt
9061 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
9121 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
9181 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagca tctagaatta
9241 attccgtgta ttctatagtg tcacctaaat cgtatgtgta tgatacataa ggttatgtat
9301 taattgtagc cgcgttctaa cgacaatatg tacaagccta attgtgtagc atctggctta
9361 ctgaagcaga ccctatcatc tctctcgtaa actgccgtca gagtcggttt ggttggacga
9421 accttctgag tttctggtaa cgccgtcccg cacccggaaa tggtcagcga accaatcagc
9481 agggtcatcg ctagccagat cctctacgcc ggacgcatcg tggccggcat caccggcgcc
9541 acaggtgcgg ttgctggcgc ctatatcgcc gacatcaccg atggggaaga tcgggctcgc
9601 cacttcgggc tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc cgtggccggg
9661 ggactgttgg gcgccatctc cttgcatgca ccattccttg cggcggcggt gctcaacggc
9721 ctcaacctac tactgggctg cttcctaatg caggagtcgc ataagggaga gcgtcgaatg
9781 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
9841 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
9901 tgtgaccatc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
9961 gagatgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
10021 ttcttagacg tcaggtggca cttttcgggg aaatgtgccg agcggccgca cggttatcca
10081 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
10141 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
10201 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
10261 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
10321 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
10381 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
10441 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
10501 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
10561 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
10621 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
10681 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
10741 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
10801 acgaaaactc acgttaaggg attttggtca tgacaggtgg cacttttcgg ggaaatgtgc
10861 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
10921 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
10981 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
11041 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
11101 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
11161 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
11221 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
11281 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
11341 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
11401 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
11461 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
11521 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
11581 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
11641 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtcccgcggt atcattgcag
11701 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
11761 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
11821 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
11881 aatttaaaag gatctaggtg aagatccttt ttgataatcg cggccgccaa ttttttacgg
11941 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
12001 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
12061 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
12121 cccgcgcgtt ggccgattca ttaatgcagc tgtggaatgt gtgtcagtta gggtgtggaa
12181 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
12241 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
12301 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
12361 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
12421 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
12481 tttgcaaaaa gcttggacac aagacaggct tgcgagatat gtttgagaat accactttat
12541 cccgcgtcag ggagaggcag tgcgtaaaaa gacgcggact catgtgaaat actggttttt
12601 agtgcgccag atctctataa tctcgcgcaa cctattttcc cctcgaacac tttttaagcc
12661 gtagataaac aggctgggac acttcacatg agcgaaaaat acatcgtcac ctgggacatg
12721 ttgcagatcc atgcacgtaa actcgcaagc cgactgatgc cttctgaaca atggaaaggc
12781 attattgccg taagccgtgg cggtctgtac cgggtgcgtt actggcgcgt gaactgggta
12841 ttcgtcatgt cgataccgtt tgtatttcca gctacgatca cgacaaccag cgcgagctta
12901 aagtgctgaa acgcgcagaa ggcgatggcg aaggcttcat cgttattgat gacctggtgg
12961 ataccggtgg tactgcggtt gcgattcgtg aaatgtatcc aaaagcgcac tttgtcacca
13021 tcttcgcaaa accggctggt cgtccgctgg ttgatgacta tgttgttgat atcccgcaag
13081 atacctggat tgaacagccg tgggatatgg gcgtcgtatt cgtcccgcca atctccggtc
13141 gctaatcttt tcaacgcctg gcactgccgg gcgttgttct ttttaacttc aggcgggtta
13201 caatagtttc cagtaagtat tctggaggct gcatccatga cacaggcaaa cctgagcgaa
13261 accctgttca aaccccgctt taaacatcct gaaacctcga cgctagtccg ccgctttaat
13321 cacggcgcac aaccgcctgt gcagtcggcc cttgatggta aaaccatccc tcactggtat
13381 cgcatgatta accgtctgat gtggatctgg cgcggcattg acccacgcga aatcctcgac
13441 gtccaggcac gtattgtgat gagcgatgcc gaacgtaccg acgatgattt atacgatacg
13501 gtgattggct accgtggcgg caactggatt tatgagtggg ccccggatct ttgtgaagga
13561 accttacttc tgtggtgtga cataattgga caaactacct acagagattt aaagctctaa
13621 ggtaaatata aaatttttaa gtgtataatg tgttaaacta ctgattctaa ttgtttgtgt
13681 attttagatt ccaacctatg gaactgatga atgggagcag tggtggaatg cctttaatga
13741 ggaaaacctg ttttgctcag aagaaatgcc atctagtgat gatgaggcta ctgctgactc
13801 tcaacattct actcctccaa aaaagaagag aaaggtagaa gaccccaagg actttccttc
13861 agaattgcta agttttttga gtcatgctgt gtttagtaat agaactcttg cttgctttgc
13921 tatttacacc acaaaggaaa aagctgcact gctatacaag aaaattatgg aaaaatattc
13981 tgtaaccttt ataagtaggc ataacagtta taatcataac atactgtttt ttcttactcc
14041 acacaggcat agagtgtctg ctattaataa ctatgctcaa aaattgtgta cctttagctt
14101 tttaatttgt aaaggggtta ataaggaata tttgatgtat agtgccttga ctagagatca
14161 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
14221 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
14281 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
14341 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatcaact
14401 ggataactca agctaaccaa aatcatccca aacttcccac cccataccct attaccactg
14461 ccaat
//
| {
"Vector backbone": "pAN2414 Lentiviral Destination",
"Backbone size w/o insert (bp)": "",
"Vector type": "Lentiviral",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "NEB Stable",
"Copy number": "Low Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA ; Takara Bio Limited Use Label License (formerly Clontech)",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Sentinel cells enable genetic detection of SARS-CoV-2 Spike protein",
"Publication": "bioRxiv 2021.04.20.440678",
"DOI": "10.1101/2021.04.20.440678",
"Authors": "Weinberg ZY, Hilburger CE, Kim M, Cao L, Khalid M, Elmes S, Diwanji D, Hernandez E, Lopez J, Schaefer K, Smith AM, Zhou F, QCRG Structural Biology Consortium, Kumar GR, Ott M, Baker D, El-Samad H",
"PubMed Link": "",
"PubMed ID": ""
} | {
"Gene/Insert name": "EF1a-Spike ECD-PDGFR TMD-T2A-mCherry",
"Species": "Synthetic",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "S",
"Promoter": "EF1a",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "unknown (unknown if destroyed)",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pFGC5941-PacI-ath-STTM163 | 84089 | target miRNA163 for destruction in Arabidopsis thaliana | Guiliang Tang | Tang et al Methods. 2012 Oct;58(2):118-25. doi: 10.1016/j.ymeth.2012.10.006. Epub 2012 Oct 23. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/166823/ec38137f-b94e-4765-9f8b-c401abcc5bc6/addgene-plasmid-84089-sequence-166823.gbk | full | LOCUS sequence-166823- 11477 bp DNA circular SYN 26-AUG-2024
DEFINITION target miRNA163 for destruction in Arabidopsis thaliana.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11477)
AUTHORS Tang G, Yan J, Gu Y, Qiao M, Fan R, Mao Y, Tang X
TITLE Construction of short tandem target mimic (STTM) to block the
functions of plant and animal microRNAs.
JOURNAL Methods. 2012 Oct;58(2):118-25. doi: 10.1016/j.ymeth.2012.10.006.
Epub 2012 Oct 23.
PUBMED 23098881
REFERENCE 2 (bases 1 to 11477)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1016/j.ymeth.2012.10.006"; journalName: "Methods"; date:
"2012-10"; volume: "58"; issue: "2"; pages: "118-25"
FEATURES Location/Qualifiers
source 1..11477
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 1..346
/label=CaMV 35S promoter
/note="strong constitutive promoter from cauliflower mosaic
virus"
primer_bind 289..309
/label=35S promoter
/note="CaMV 35S promoter, forward primer"
polyA_signal 513..689
/label=CaMV poly(A) signal
/note="cauliflower mosaic virus polyadenylation signal"
promoter complement(1251..1269)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(1252..1269)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind 1908..1927
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
promoter complement(2030..2058)
/label=tet promoter
/note="E. coli promoter for tetracycline efflux protein
gene"
promoter 2333..2713
/label=MAS promoter
/note="mannopine synthase promoter (Velten et al., 1984)"
CDS 2719..3270
/codon_start=1
/gene="Streptomyces hygroscopicus bar"
/product="phosphinothricin acetyltransferase"
/label=BlpR
/note="confers resistance to bialophos or phosphinothricin"
/translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
WQLDFSLPVPPRPVLPVTEI"
terminator 3280..3532
/label=MAS terminator
/note="mannopine synthase terminator"
misc_feature 3618..3642
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
CDS 4067..4861
/codon_start=1
/gene="aphA-3"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin"
/translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
rep_origin 4948..5536
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5690..5707
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 5722..5862
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(5788..5807)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 5948..5970
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
rep_origin 6206..6400
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
CDS complement(6466..7539)
/codon_start=1
/product="replication protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/label=pVS1 RepA
/translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
LVMRYRNLIEGEASAGS"
CDS complement(7968..8597)
/codon_start=1
/product="stability protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/label=pVS1 StaA
/translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
misc_feature 9898..9922
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
primer_bind 10110..10132
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 10124..10141
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 10125..10141
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
terminator complement(10166..10873)
/label=OCS terminator
/note="octopine synthase terminator"
primer_bind 11052..11071
/label=T7
/note="T7 promoter, forward primer"
promoter 11052..11070
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
1 tgagactttt caacaaaggg taatttcggg aaacctcctc ggattccatt gcccagctat
61 ctgtcacctc atcgaaagga cagtagaaaa ggaaggtggc tcctacaaat gccatcattg
121 cgataaagga aaggctatca ttcaagatgc ctctgccgac agtggtccca aagatggacc
181 cccacccacg aggagcatcg tggaaaaaga agacgttcca accacgtctt caaagcaagt
241 ggattgatgt gacatctcca ctgacataag ggatgacgca caaccccact atccttcgca
301 agacccttcc tctatataag gaagttcatt tcatttggag aggacagccc aagcttatcg
361 aagttccaag ctatcctctt caagttgttg ttgttatggt ctaatttaaa tatggtctaa
421 agaagaagaa tatcgaagtt ccaagctatc ctcttcaaga attcggtacg ctgaaatcac
481 cagtctctct ctacaaatct atctctctct attttctcca taaataatgt gtgagtagtt
541 tcccgataag ggaaattagg gttcttatag ggtttcgctc atgtgttgag catataagaa
601 acccttagta tgtatttgta tttgtaaaat acttctatca ataaaatttc taattcctaa
661 aaccaaaatc cagtactaaa atccagatct cctaaagtcc ctatagatct ttgtcgtgaa
721 tataaaccag acacgagacg actagacctg gagcccagac gccgttcgaa gctagaagta
781 ccgcttaggc aggaggccgt tagggaaaag atgctaaggc agggttggtt acgttgactc
841 ccccgtaggt ttggtttaaa tatgatgaag tggacggaag gaaggaggaa gacaaggaag
901 gataaggttg caggccctgt gcaaggtaag aagatggaaa tttgatagag gtacgctact
961 atacttatac tatacgctaa gggaatgatt gtatttatac cctatacccc ctaataaccc
1021 cttatcaatt taagaaataa tccgcataag cccccgctta aaaattggta tcagagccat
1081 gaataggtct atgaccaaaa ctcaagagga taaaacctca ccaaaatacg aaagagtcct
1141 taactctaaa gataaaagat ctttcaagat caaaactagt tccctcacac cggtgacggg
1201 gatcgcatgc gacctctaga ccatcacatt ggagcttcag ctgctcgagt tctatagtgt
1261 cacctaaatc gtatgtgtat gatacataag gttatgtatt aattgtagcc gcgttcttac
1321 gtcgacgtac caattacgcc ccgccctgtc actcatcgca gtactgttgt aattcattaa
1381 gcattctgcc gacatggaag ccatcacaaa cggcatgatg aacctgaatc gccagcggca
1441 tcagcacctt gtcgccttgc gtataatatt tgcccatggt gaaaacgggg gcgaagaagt
1501 tgtccatatt ggccacgttt aaatcaaaac tggtgaaact cacccaggga ttggctgaga
1561 cgaaaaacat attctcaata aaccctttag ggaaataggc caggttttca ccgtaacacg
1621 ccacatcttg cgaatatatg tgtagaaact gccggaaatc gtcgtggtat tcactccaga
1681 gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta acaagggtga acactatccc
1741 atattaccag ctcaccgtct ttcattgcca tacggaactc cggatgagca ttcatcaggc
1801 gggcaagaat gtgaataaag gccggataaa acttgtgctt atttttcttt acggtcttta
1861 aaaaggccgt aatatccagc tgaacggtct ggttataggt acattgagca actgactgaa
1921 atgcctcaaa atgttcttta cgatgccatt ggggtatatc aacggtggta tatccagtga
1981 tttctttctc cattttagct tccttagctc ctgaaaatct cgataactca ttaaagctta
2041 tcgatgataa gctgtcaaac atgagaattt agcccgccta atgagcgggc tttttgttgt
2101 gatccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
2161 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
2221 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
2281 gcctttctta attaactctc tagactcacc taggatccat gaattctgag atttttcaaa
2341 tcagtgcgca agacgtgacg taagtatccg agtcagtttt tatttttcta ctaatttggt
2401 cgtttatttc ggcgtgtagg acatggcaac cgggcctgaa tttcgcgggt attctgtttc
2461 tattccaact ttttcttgat ccgcagccat taacgacttt tgaatagata cgctgacacg
2521 ccaagcctcg ctagtcaaaa gtgtaccaaa caacgcttta cagcaagaac ggaatgcgcg
2581 tgacgctcgc ggtgacgcca tttcgccttt tcagaaatgg ataaatagcc ttgcttccta
2641 ttatatcttc ccaaattacc aatacattac actagcatct gaatttcata accaatctcg
2701 atacaccaaa tcgaatcgat gagcccagaa cgacgcccgg ccgacatccg ccgtgccacc
2761 gaggcggaca tgccggcggt ctgcaccatc gtcaaccact acatcgagac aagcacggtc
2821 aacttccgta ccgagccgca ggaaccgcag gagtggacgg acgacctcgt ccgtctgcgg
2881 gagcgctatc cctggctcgt cgccgaggtg gacggcgagg tcgccggcat cgcctacgcg
2941 ggcccctgga aggcacgcaa cgcctacgac tggacggccg agtcgaccgt gtacgtctcc
3001 ccccgccacc agcggacggg actgggctcc acgctctaca cccacctgct gaagtccctg
3061 gaggcacagg gcttcaagag cgtggtcgct gtcatcgggc tgcccaacga cccgagcgtg
3121 cgcatgcacg aggcgctcgg atatgccccc cgcggcatgc tgcgggcggc cggcttcaag
3181 cacgggaact ggcatgacgt gggtttctgg cagctggact tcagcctgcc ggtaccgccc
3241 cgtccggtcc tgcccgtcac cgagatttga ctgcagaatc ttggactccc atgttggcaa
3301 aggcaaccaa acaaacaatg aatgatccgc tcctgcatat ggggcggttt gagtatttca
3361 actgccattt gggctgaatt gaagacatgc tcctgtcaga aattccgtga tcttactcaa
3421 tattcagtaa tctcggccaa tatcctaaat gtgcgtggct ttatctgtct ttgtattgtt
3481 tcatcaattc atgtaacgtt tgcttttctt atgaattttc aaataaatta tcagatccga
3541 gctcgaatta attcggcgtt aattcagtac attaaaaacg tccgcaatgt gttattaagt
3601 tgtctaagcg tcaatttgtt tacaccacaa tatatcctgc caccagccag ccaacagctc
3661 cccgaccggc agctcggcac aaaatcacca ctcgatacag gcagcccatc agtccgggac
3721 ggcgtcagcg ggagagccgt tgtaaggcgg cagactttgc tcatgttacc gatgctattc
3781 ggaagaacgg caactaagct gccgggtttg aaacacggat gatctcgcgg agggtagcat
3841 gttgattgta acgatgacag agcgttgctg cctgtgatca ccgcggtttc aaaatcggct
3901 ccgtcgatac tatgttatac gccaactttg aaaacaactt tgaaaaagct gttttctggt
3961 atttaaggtt ttagaatgca aggaacagtg aattggagtt cgtcttgtta taattagctt
4021 cttggggtat ctttaaatac tgtagaaaag aggaaggaaa taataaatgg ctaaaatgag
4081 aatatcaccg gaattgaaaa aactgatcga aaaataccgc tgcgtaaaag atacggaagg
4141 aatgtctcct gctaaggtat ataagctggt gggagaaaat gaaaacctat atttaaaaat
4201 gacggacagc cggtataaag ggaccaccta tgatgtggaa cgggaaaagg acatgatgct
4261 atggctggaa ggaaagctgc ctgttccaaa ggtcctgcac tttgaacggc atgatggctg
4321 gagcaatctg ctcatgagtg aggccgatgg cgtcctttgc tcggaagagt atgaagatga
4381 acaaagccct gaaaagatta tcgagctgta tgcggagtgc atcaggctct ttcactccat
4441 cgacatatcg gattgtccct atacgaatag cttagacagc cgcttagccg aattggatta
4501 cttactgaat aacgatctgg ccgatgtgga ttgcgaaaac tgggaagaag acactccatt
4561 taaagatccg cgcgagctgt atgatttttt aaagacggaa aagcccgaag aggaacttgt
4621 cttttcccac ggcgacctgg gagacagcaa catctttgtg aaagatggca aagtaagtgg
4681 ctttattgat cttgggagaa gcggcagggc ggacaagtgg tatgacattg ccttctgcgt
4741 ccggtcgatc agggaggata tcggggaaga acagtatgtc gagctatttt ttgacttact
4801 ggggatcaag cctgattggg agaaaataaa atattatatt ttactggatg aattgtttta
4861 gtacctagaa tgcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
4921 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
4981 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
5041 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
5101 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
5161 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
5221 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
5281 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
5341 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
5401 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
5461 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
5521 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
5581 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
5641 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
5701 gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
5761 ttcacaccgc atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
5821 gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca
5881 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
5941 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg
6001 cagggtgcct tgatgtgggc gccggcggtc gagtggcgac ggcgcggctt gtccgcgccc
6061 tggtagattg cctggccgta ggccagccat ttttgagcgg ccagcggccg cgataggccg
6121 acgcgaagcg gcggggcgta gggagcgcag cgaccgaagg gtaggcgctt tttgcagctc
6181 ttcggctgtg cgctggccag acagttatgc acaggccagg cgggttttaa gagttttaat
6241 aagttttaaa gagttttagg cggaaaaatc gccttttttc tcttttatat cagtcactta
6301 catgtgtgac cggttcccaa tgtacggctt tgggttccca atgtacgggt tccggttccc
6361 aatgtacggc tttgggttcc caatgtacgt gctatccaca ggaaagagac cttttcgacc
6421 tttttcccct gctagggcaa tttgccctag catctgctcc gtacattagg aaccggcgga
6481 tgcttcgccc tcgatcaggt tgcggtagcg catgactagg atcgggccag cctgccccgc
6541 ctcctccttc aaatcgtact ccggcaggtc atttgacccg atcagcttgc gcacggtgaa
6601 acagaacttc ttgaactctc cggcgctgcc actgcgttcg tagatcgtct tgaacaacca
6661 tctggcttct gccttgcctg cggcgcggcg tgccaggcgg tagagaaaac ggccgatgcc
6721 gggatcgatc aaaaagtaat cggggtgaac cgtcagcacg tccgggttct tgccttctgt
6781 gatctcgcgg tacatccaat cagctagctc gatctcgatg tactccggcc gcccggtttc
6841 gctctttacg atcttgtagc ggctaatcaa ggcttcaccc tcggataccg tcaccaggcg
6901 gccgttcttg gccttcttcg tacgctgcat ggcaacgtgc gtggtgttta accgaatgca
6961 ggtttctacc aggtcgtctt tctgctttcc gccatcggct cgccggcaga acttgagtac
7021 gtccgcaacg tgtggacgga acacgcggcc gggcttgtct cccttccctt cccggtatcg
7081 gttcatggat tcggttagat gggaaaccgc catcagtacc aggtcgtaat cccacacact
7141 ggccatgccg gccggccctg cggaaacctc tacgtgcccg tctggaagct cgtagcggat
7201 cacctcgcca gctcgtcggt cacgcttcga cagacggaaa acggccacgt ccatgatgct
7261 gcgactatcg cgggtgccca cgtcatagag catcggaacg aaaaaatctg gttgctcgtc
7321 gcccttgggc ggcttcctaa tcgacggcgc accggctgcc ggcggttgcc gggattcttt
7381 gcggattcga tcagcggccg cttgccacga ttcaccgggg cgtgcttctg cctcgatgcg
7441 ttgccgctgg gcggcctgcg cggccttcaa cttctccacc aggtcatcac ccagcgccgc
7501 gccgatttgt accgggccgg atggtttgcg accgctcacg ccgattcctc gggcttgggg
7561 gttccagtgc cattgcaggg ccggcaggca acccagccgc ttacgcctgg ccaaccgccc
7621 gttcctccac acatggggca ttccacggcg tcggtgcctg gttgttcttg attttccatg
7681 ccgcctcctt tagccgctaa aattcatcta ctcatttatt catttgctca tttactctgg
7741 tagctgcgcg atgtattcag atagcagctc ggtaatggtc ttgccttggc gtaccgcgta
7801 catcttcagc ttggtgtgat cctccgccgg caactgaaag ttgacccgct tcatggctgg
7861 cgtgtctgcc aggctggcca acgttgcagc cttgctgctg cgtgcgctcg gacggccggc
7921 acttagcgtg tttgtgcttt tgctcatttt ctctttacct cattaactca aatgagtttt
7981 gatttaattt cagcggccag cgcctggacc tcgcgggcag cgtcgccctc gggttctgat
8041 tcaagaacgg ttgtgccggc ggcggcagtg cctgggtagc tcacgcgctg cgtgatacgg
8101 gactcaagaa tgggcagctc gtacccggcc agcgcctcgg caacctcacc gccgatgcgc
8161 gtgcctttga tcgcccgcga cacgacaaag gccgcttgta gccttccatc cgtgacctca
8221 atgcgctgct taaccagctc caccaggtcg gcggtggccc atatgtcgta agggcttggc
8281 tgcaccggaa tcagcacgaa gtcggctgcc ttgatcgcgg acacagccaa gtccgccgcc
8341 tggggcgctc cgtcgatcac tacgaagtcg cgccggccga tggccttcac gtcgcggtca
8401 atcgtcgggc ggtcgatgcc gacaacggtt agcggttgat cttcccgcac ggccgcccaa
8461 tcgcgggcac tgccctgggg atcggaatcg actaacagaa catcggcccc ggcgagttgc
8521 agggcgcggg ctagatgggt tgcgatggtc gtcttgcctg acccgccttt ctggttaagt
8581 acagcgataa ccttcatgcg ttccccttgc gtatttgttt atttactcat cgcatcatat
8641 acgcagcgac cgcatgacgc aagctgtttt actcaaatac acatcacctt tttagacggc
8701 ggcgctcggt ttcttcagcg gccaagctgg ccggccaggc cgccagcttg gcatcagaca
8761 aaccggccag gatttcatgc agccgcacgg ttgagacgtg cgcgggcggc tcgaacacgt
8821 acccggccgc gatcatctcc gcctcgatct cttcggtaat gaaaaacggt tcgtcctggc
8881 cgtcctggtg cggtttcatg cttgttcctc ttggcgttca ttctcggcgg ccgccagggc
8941 gtcggcctcg gtcaatgcgt cctcacggaa ggcaccgcgc cgcctggcct cggtgggcgt
9001 cacttcctcg ctgcgctcaa gtgcgcggta cagggtcgag cgatgcacgc caagcagtgc
9061 agccgcctct ttcacggtgc ggccttcctg gtcgatcagc tcgcgggcgt gcgcgatctg
9121 tgccggggtg agggtagggc gggggccaaa cttcacgcct cgggccttgg cggcctcgcg
9181 cccgctccgg gtgcggtcga tgattaggga acgctcgaac tcggcaatgc cggcgaacac
9241 ggtcaacacc atgcggccgg ccggcgtggt ggtgtcggcc cacggctctg ccaggctacg
9301 caggcccgcg ccggcctcct ggatgcgctc ggcaatgtcc agtaggtcgc gggtgctgcg
9361 ggccaggcgg tctagcctgg tcactgtcac aacgtcgcca gggcgtaggt ggtcaagcat
9421 cctggccagc tccgggcggt cgcgcctggt gccggtgatc ttctcggaaa acagcttggt
9481 gcagccggcc gcgtgcagtt cggcccgttg gttggtcaag tcctggtcgt cggtgctgac
9541 gcgggcatag cccagcaggc cagcggcggc gctcttgttc atggcgtaat gtctccggtt
9601 ctagtcgcaa gtattctact ttatgcgact aaaacacgcg acaagaaaac gccaggaaaa
9661 gggcagggcg gcagcctgtc gcgtaactta ggacttgtgc gacatgtcgt tttcagaaga
9721 cggctgcact gaacgtcaga agccgactgc actatagcag cggaggggtt ggatcaaagt
9781 actttgatcc cgaggggaac cctgtggttg gcatgcacat acaaatggac gaacggataa
9841 accttttcac gcccttttaa atatccgatt attctaataa acgctctttt ctcttaggtt
9901 tacccgccaa tatatcctgt caaacactga tagtttaaac tgaaggcggg aaacgacaat
9961 ctgatccaag ctcaagctgc tctagcattc gccattcagg ctgcgcaact gttgggaagg
10021 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
10081 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
10141 tgccaagctt gcatgcctgc aggtcctgct gagcctcgac atgttgtcgc aaaattcgcc
10201 ctggacccgc ccaacgattt gtcgtcactg tcaaggtttg acctgcactt catttggggc
10261 ccacatacac caaaaaaatg ctgcataatt ctcggggcag caagtcggtt acccggccgc
10321 cgtgctggac cgggttgaat ggtgcccgta actttcggta gagcggacgg ccaatactca
10381 acttcaagga atctcaccca tgcgcgccgg cggggaaccg gagttccctt cagtgagcgt
10441 tattagttcg ccgctcggtg tgtcgtagat actagcccct ggggcacttt tgaaatttga
10501 ataagattta tgtaatcagt cttttaggtt tgaccggttc tgccgctttt tttaaaattg
10561 gatttgtaat aataaaacgc aattgtttgt tattgtggcg ctctatcata gatgtcgcta
10621 taaacctatt cagcacaata tattgttttc attttaatat tgtacatata agtagtaggg
10681 tacaatcagt aaattgaacg gagaatatta ttcataaaaa tacgatagta acgggtgata
10741 tattcattag aatgaaccga aaccggcggt aaggatctga gctacacatg ctcaggtttt
10801 ttacaacgtg cacaacagaa ttgaaagcaa atatcatgcg atcataggcg tctcgcatat
10861 ctcattaaag caggactcta gggactagtc ccgggtctta attaagtttg caagcagcag
10921 attacgcgca gaaaaaaagg atctcaagga gatcctttgg atcacaaaaa aaaagcccgc
10981 tcattaggcg ggttaaattc ccgggtactg cagccgattc attaatgcag gttaacctgg
11041 cttatcgaaa ttaatacgac tcactatagg gagaccggca gatctgatcc ctactccaaa
11101 aatgtcaaag atacagtctc agaagaccaa agggctattg agacttttca acaaagggta
11161 atttcgggaa acctcctcgg attccattgc ccagctatct gtcacttcat cgaaaggaca
11221 gtggaaaagg aaggtggctc ctacaaatgc catcattgcg ataaaggaaa ggctatcatt
11281 caagatgcct ctgccgacag tggtcccaaa gatggacccc cacccacgag gagcatcgtg
11341 ggaaaagaag gcgttccaac cacgtcttca aagcaagtgg attgatgtga catctccact
11401 gacgtaaggg atgacgcaca atcccacccc tactccaaaa atgtcaaaga tactgtctca
11461 gaagaccaag gggctat
//
| {
"Vector backbone": "pFGC5941-PacI",
"Backbone size w/o insert (bp)": "8608",
"Vector type": "Plant Expression",
"Selectable markers": "Basta"
} | {
"Bacterial Resistance(s)": "Chloramphenicol and Kanamycin, 25 & 50 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Construction of short tandem target mimic (STTM) to block the functions of plant and animal microRNAs",
"Publication": "Methods. 2012 Oct;58(2):118-25. doi: 10.1016/j.ymeth.2012.10.006. Epub 2012 Oct 23.",
"DOI": "10.1016/j.ymeth.2012.10.006.",
"Authors": "Tang G, Yan J, Gu Y, Qiao M, Fan R, Mao Y, Tang X /jymeth",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/23098881",
"PubMed ID": "PubMed 23098881"
} | {
"Gene/Insert name": "STTM163",
"Species": "A. thaliana (mustard weed)",
"Insert Size (bp)": "2846",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "2x35S",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "PacI (not destroyed)",
"3′ cloning site": "PacI (not destroyed)",
"5′ sequencing primer": "CACTATCCTTCGCAAGACCCTTCC",
"3′ sequencing primer": "CAACACATGAGCGAAACCCTATAA"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pCW57-GFP-P2A-MCS (Neo) | 89181 | Medium | (Empty Backbone)All-in-one doxycycline inducible lentiviral vector for expression of one gene in combination with turbo GFP using the P2A self-cleaving peptide. | Adam Karpf | Barger et al Elife. 2021 Apr 23;10. pii: 55070. doi: 10.7554/eLife.55070. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/176294/f8db23f0-e26a-4e53-aab3-0f3fb09c158a/addgene-plasmid-89181-sequence-176294.gbk | full | LOCUS sequence-176294- 8639 bp DNA circular SYN 25-AUG-2024
DEFINITION All-in-one doxycycline inducible lentiviral vector for expression of
one gene in combination with turbo GFP using the P2A self-cleaving
peptide..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8639)
AUTHORS Barger CJ, Chee L, Albahrani M, Munoz-Trujillo C, Boghean L, Branick
C, Odunsi K, Drapkin R, Zou L, Karpf AR
TITLE Co-regulation and function of FOXM1/RHNO1 bidirectional genes in
cancer.
JOURNAL Elife. 2021 Apr 23;10. pii: 55070. doi: 10.7554/eLife.55070.
PUBMED 33890574
REFERENCE 2 (bases 1 to 8639)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Elife.";
date: "2021-04-23"; volume: "10. pii: 55070. doi"; pages: "
10.7554/eLife.55070"
FEATURES Location/Qualifiers
source 1..8639
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind 58..76
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 94..112
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 129..147
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 165..183
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 201..219
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 236..254
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 272..290
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
misc_feature 352..469
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
CDS complement(682..726)
/codon_start=1
/product="antigenic peptide corresponding to amino acids
655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
et al., 2013)"
/label=gp41 peptide
/note="recognized by the 2H10 single-chain llama nanobody"
/translation="KNEQELLELDKWASL"
misc_feature 911..1144
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
misc_feature 1637..1762
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
LTR 1809..1989
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
promoter complement(1990..2216)
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
primer_bind complement(2244..2264)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(2244..2262)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(2283..2299)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2283..2299)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(2296..2318)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 2307..2323
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(2331..2361)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2376..2397
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(2514..2531)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2685..3273)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2765..2784)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3444..4304)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 4067..4086
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(4305..4376)
/gene="bla"
/label=AmpR promoter
rep_origin 4376..4511
/label=SV40 ori
/note="SV40 origin of replication"
polyA_signal 4551..4672
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4562..4581)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 4616..4635
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
LTR 4744..4977
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
CDS 5182..5193
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind 5591..5611
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(5652..6455)
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MSRMIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(5763..5782)
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind 6373..6392
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
CDS complement(6456..6509)
/codon_start=1
/product="2A peptide from Thosea asigna virus capsid
protein"
/label=T2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="EGRGSLLTCGDVEENPGP"
CDS complement(6525..7268)
/codon_start=1
/product="improved tetracycline-controlled transactivator"
/label=rtTA-Advanced
/note="In the Tet-On(R) system, rtTA-Advanced binds to the
Tet-responsive element and stimulates transcription only in
the presence of tetracycline or doxycycline."
/translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
DALDDFDLDMLPADALDDFDLDMLPG"
promoter complement(7292..7802)
/label=hPGK promoter
/note="human phosphoglycerate kinase 1 promoter"
primer_bind complement(7355..7373)
/label=hPGK-F
/note="Human PGK promoter, forward primer"
CDS complement(7830..7886)
/codon_start=1
/product="2A peptide from porcine teschovirus-1
polyprotein"
/label=P2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="ATNFSLLKQAGDVEENPGP"
CDS complement(7899..8594)
/codon_start=1
/product="improved green fluorescent protein from
Pontellina plumata (Evdokimov et al., 2006)"
/label=TurboGFP
/note="mammalian codon-optimized"
/translation="MESDESGLPAMEIECRITGTLNGVEFELVGGGEGTPEQGRMTNKM
KSTKGALTFSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVL
HVSFSYRYEAGRVIGDFKVMGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNDLDGSFTR
TFSLRDGGYYSSVVDSHMHFKSAIHPSILQNGGPMFAFRRVEEDHSNTELGIVEYQHAF
KTPDADAGEE"
regulatory 8591..8600
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
promoter complement(join(8623..8639,1..298))
/label=tight TRE promoter
/note="Tet-responsive promoter PTight, consisting of seven
tet operator sequences followed by the minimal CMV
promoter"
primer_bind complement(join(8626..8639,1..11))
/label=LNCX
/note="Human CMV promoter, forward primer"
ORIGIN
1 ctaaacgagc tctgcttata taggcctccc accgtacacg cctacctcga catacgttct
61 ctatcactga tagggagtaa actcgacata cgttctctat cactgatagg gataaactcg
121 acatacgttc tctatcactg atagggagta aactcgacat acgttctcta tcactgatag
181 ggagtaaact cgacatacgt tctctatcac tgatagggag taaactcgac atcgttctct
241 atcactgata gggagtaaac tcgacatacg ttctctatca ctgataggga gtaaactcga
301 catatcgatt cgcggccaaa gtggatctct gctgtccctg taataaaccc gaaaattttg
361 aatttttgta atttgttttt gtaattcttt agtttgtatg tctgttgcta ttatgtctac
421 tattctttcc cctgcactgt accccccaat cccccctttt cttttaaaat tgtggatgaa
481 tactgccatt tgtctcgagg tcgagaattg tcccctcggg gttgggaggt gggtctgaaa
541 cgataatggt gaatatccct gcctaactct attcactata gaaagtacag caaaaactat
601 tcttaaacct accaagcctc ctactatcat tatgaataat tttatatacc acagccaatt
661 tgttatgtta aaccaattcc acaaacttgc ccatttatct aattccaata attcttgttc
721 attcttttct tgctggtttt gcgattcttc aattaaggag tgtattaagc ttgtgtaatt
781 gttaatttct ctgtcccact ccatccaggt cgtgtgattc caaatctgtt ccagagattt
841 attactccaa ctagcattcc aaggcacagc agtggtgcaa atgagttttc cagagcaacc
901 ccaaatcccc aggagctgtt gatcctttag gtatctttcc acagccagga ttcttgcctg
961 gagctgcttg atgccccaga ctgtgagttg caacagatgc tgttgcgcct caatagccct
1021 cagcaaattg ttctgctgct gcactatacc agacaataat tgtctggcct gtaccgtcag
1081 cgtcattgac gctgcgccca tagtgcttcc tgctgctccc aagaacccaa ggaacaaagc
1141 tcctattccc actgctcttt tttctctctg caccactctt ctctttgcct tggtgggtgc
1201 tactcctaat ggttcaattt ttactacttt atatttatat aattcacttc tccaattgtc
1261 cctcatatct cctcctccag gtctgaagat cagcggccgc ttgctgtgcg gtggtcttac
1321 ttttgttttg ctcttcctct atcttgtcta aagcttcctt ggtgtctttt atctctatcc
1381 tttgatgcac acaatagagg gttgctactg tattatataa tgatctaagt tcttctgatc
1441 ctgtctgaag ggatggttgt agctgtccca gtatttgtct acagccttct gatgtttcta
1501 acaggccagg attaactgcg aatcgttcta gctccctgct tgcccatact atatgtttta
1561 atttatattt tttctttccc cctggcctta accgaatttt ttcccatcgc gatctaattc
1621 tcccccgctt aatactgacg ctctcgcacc catctctctc cttctagcct ccgctagtca
1681 aaatttttgg cgtactcacc agtcgccgcc cctcgcctct tgccgtgcgc gcttcagcaa
1741 gccgagtcct gcgtcgagag agctcctctg gtttcccttt cgctttcaag tccctgttcg
1801 ggcgccactg ctagagattt tccacactga ctaaaagggt ctgagggatc tctagttacc
1861 agagtcacac aacagacggg cacacactac ttgaagcact caaggcaagc tttattgagg
1921 cttaagcagt gggttcccta gttagccaga gagctcccag gctcagatct ggtctaacca
1981 gagagacccg tttatgtatc gagctaggca cttaaataca atatctctgc aatgcggcaa
2041 ttcagtggtt cgtccaatcc atgtcagacc cgtctgttgc cttcctaata aggcacgatc
2101 gtaccacctt acttccacca atcggcatgc acggtgcttt ttctctcctt gtaaggcatg
2161 ttgctaactc atcgttacca tgttgcaaga ctacaagagt attgcataag actacattaa
2221 gcttgcagct ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca
2281 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
2341 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
2401 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
2461 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
2521 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
2581 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
2641 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
2701 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
2761 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
2821 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
2881 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
2941 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
3001 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
3061 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
3121 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
3181 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
3241 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
3301 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
3361 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
3421 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
3481 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
3541 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
3601 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
3661 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
3721 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
3781 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
3841 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
3901 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
3961 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
4021 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
4081 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
4141 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
4201 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
4261 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
4321 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaaaggcct
4381 ccaaaaaagc ctcctcacta cttctggaat agctcagagg ccgaggcggc ctcggcctct
4441 gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact gggcggagtt
4501 aggggcggga tgggcggagt taggggcggg atagctagag ccagacatga taagatacat
4561 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
4621 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttcctctcac
4681 tctctgatat tcatttcttt gcaagttata aatactgaat aataagatga catgaactac
4741 tactgctaga gattttccac actgactaaa agggtctgag ggatctctag ttaccagagt
4801 cacacaacag acgggcacac actacttgaa gcactcaagg caagctttat tgaggcttaa
4861 gcagtgggtt ccctagttag ccagagagct cccaggctca gatctggtct aaccagagag
4921 acccagtaca agcaaaaagc agatcttgtc ttcgttggga gtgaattagc ccttccagtc
4981 cccccttttc ttttaaaaag tggctaagat ctacagctgc cttgtaagtc attggtctta
5041 aaggtacctc tagtccggac gccgaggcga aacaggcggg gaggcggccc aaagggagat
5101 ccgactcgtc tgagggcgaa ggcgaagacg cggaagaggc cgcagagccg gcagcaggcc
5161 gcgggaagga aggtccgctg gattgagggc cgaagggacg tagcagaagg acgtcccgcg
5221 cagaatccag gtggcaacac aggcgagcag ccatggaaag gacgtcagct tccccgacaa
5281 caccacggaa ttgtcagtgc ccaacagccg agcccctgtc cagcagcggg caaggcaggc
5341 ggcgatgagt tccgccgtgg caatagggag ggggaaagcg aaagtcccgg aaaggagctg
5401 acaggtggtg gcaatgcccc aaccagtggg ggttgcgtca gcaaacacag tgcacaccac
5461 gccacgttgc ctgacaacgg gccacaactc ctcataaaga gacagcaacc aggatttata
5521 caaggaggag aaaatgaaag ccatacggga agcaatagca tgatacaaag gcattaaagc
5581 agcgtatcca catagcgtaa aaggagcaac atagttaaga atacgatatc ttggatcgat
5641 cctcaatgca ttcagaagaa ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg
5701 ggagcggcga taccgtaaag cacgaggaag cggtcagccc attcgccgcc aagctcttca
5761 gcaatatcac gggtagccaa cgctatgtcc tgatagcggt ccgccacacc cagccggcca
5821 cagtcgatga atccagaaaa gcggccattt tccaccatga tattcggcaa gcaggcatcg
5881 ccatgggtca cgacgagatc ctcgccgtcg ggcatgcgcg ccttgagcct ggcgaacagt
5941 tcggctggcg cgagcccctg atgctcttcg tccagatcat cctgatcgac aagaccggct
6001 tccatccgag tacgtgctcg ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta
6061 gccggatcaa gcgtatgcag ccgccgcatt gcatcagcca tgatggatac tttctcggca
6121 ggagcaaggt gagatgacag gagatcctgc cccggcactt cgcccaatag cagccagtcc
6181 cttcccgctt cagtgacaac gtcgagcaca gctgcgcaag gaacgcccgt cgtggccagc
6241 cacgatagcc gcgctgcctc gtcctgcagt tcattcaggg caccggacag gtcggtcttg
6301 acaaaaagaa ccgggcgccc ctgcgctgac agccggaaca cggcggcatc agagcagccg
6361 attgtctgtt gtgcccagtc atagccgaat agcctctcca cccaagcggc cggagaacct
6421 gcgtgcaatc catcttgttc aatcattcta gacattggac cagggttttc ttcaacatca
6481 ccacaagtga ggagagaacc tctaccttct ccgcttcctt cgaacccggg gagcatgtca
6541 aggtcaaaat cgtcaagagc gtcagcaggc agcatatcaa ggtcaaagtc gtcaagggca
6601 tcggctggga gcatgtctaa gtcaaaatcg tcaagggcgt cggtcggccc gccgctttcg
6661 cactttagct gtttctccag gccacatatg attagttcca ggccgaaaag gaaggcaggt
6721 tcggctccct gccggtcgaa cagctcaatt gcttgtctca gaagtggggg catagaatcg
6781 gtggtaggtg tctctctttc ctcttttgct acttgatgct cctgttcctc caatacgcag
6841 cccagtgtaa agtggcccac ggcggacaga gcgtacagtg cgttctccag ggagaagcct
6901 tgctgacaca ggaacgcgag ctgattttcc agggtttcgt actgtttctc tgttgggcgg
6961 gtgccgagat gcactttagc cccgtcgcga tgtgagagga gagcacagcg gtatgacttg
7021 gcgttgttcc gcagaaagtc ttgccatgac tcgccttcca gggggcagaa gtgggtatga
7081 tgcctgtcca gcatctcgat tggcagggca tcgagcaggg cccgcttgtt cttcacgtgc
7141 cagtacaggg taggctgctc aactcccagc ttttgagcga gtttccttgt cgtcaggcct
7201 tcgataccga caccattgag taattccaga gctccgttta tgactttgct cttgtccagt
7261 ctagacatgg tggccgcgcg cgggtgaatt gctggggaga gaggtcggtg attcggtcaa
7321 cgagggagcc gactgccgac gtgcgctccg gaggcttgca gaatgcggaa caccgcgcgg
7381 gcaggaacag ggcccacact accgccccac accccgcctc ccgcaccgcc ccttcccggc
7441 cgctgctctc ggcgcgccct gctgagcagc cgctattggc cacagcccat cgcggtcggc
7501 gcgctgccat tgctccctgg cgctgtccgt ctgcgagggt actagtgaga cgtgcggctt
7561 ccgtttgtca cgtccggcac gccgcgaacc gcaaggaacc ttcccgactt aggggcggag
7621 caggaagcgt cgccgggggg cccacaaggg tagcggcgaa gatccgggtg acgctgcgaa
7681 cggacgtgaa gaatgtgcga gacccagggt cggcgccgct gcgtttcccg gaaccacgcc
7741 cagagcagcc gcgtccctgc gcaaacccag ggctgccttg gaaaaggcgc aaccccaacc
7801 ccggatcctg tacaacgcgt ctgcagccta ggaccggggt tttcttccac gtctcctgct
7861 tgctttaaca gagagaagtt cgtggctccg gaaccggttt cttcaccggc atctgcatcc
7921 ggggtcttga aggcgtgctg gtactccacg atgcccagct cggtgttgct gtgatcctcc
7981 tccacgcggc ggaaggcgaa catggggccc ccgttctgca ggatgctggg gtggatggcg
8041 ctcttgaagt gcatgtggct gtccaccacg gagctgtagt agccgccgtc gcgcaggctg
8101 aaggtgcggg tgaagctgcc atccagatcg ttatcgccca tggggtgcag gtgctccacg
8161 gtggcgttgc tgcggatgat cttgtcggtg aagatcacgc tgtcctcggg gaagccggtg
8221 cccatcacct tgaagtcgcc gatcacgcgg ccggcctcgt agcggtagct gaagctcacg
8281 tgcagcacgc cgccgtcctc gtacttctcg atgcgggtgt tggtgtagcc gccgttgttg
8341 atggcgtgca ggaaggggtt ctcgtagccg ctggggtagg tgccgaagtg gtagaagccg
8401 tagcccatca cgtggctcag caggtagggg ctgaaggtca gggcgccttt ggtgctcttc
8461 atcttgttgg tcatgcggcc ctgctcgggg gtgccctctc cgccgcccac cagctcgaac
8521 tccacgccgt tcagggtgcc ggtgatgcgg cactcgatct ccatggcggg caggccgctc
8581 tcgtcgctct ccatggtggc gaattcgcta gccaattctc caggcgatct gacggttca
//
| {
"Vector backbone": "pCW57.1",
"Backbone size w/o insert (bp)": "",
"Vector type": "Mammalian Expression, Lentiviral ; Doxycycline inducible; P2A Self Cleaving Peptide",
"Selectable markers": "Neomycin (select with G418) ; Turbo GFP"
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "NEB Stable",
"Copy number": "Unknown"
} | {
"Academic/Nonprofit Terms": "UBMTA ; Evrogen Limited Use Label License for FPs",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Co-regulation and function of FOXM1/RHNO1 bidirectional genes in cancer",
"Publication": "Elife. 2021 Apr 23;10. pii: 55070. doi: 10.7554/eLife.55070.",
"DOI": "10.7554/eLife.55070.",
"Authors": "Barger CJ, Chee L, Albahrani M, Munoz-Trujillo C, Boghean L, Branick C, Odunsi K, Drapkin R, Zou L, Karpf AR /eLife",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/33890574",
"PubMed ID": "PubMed 33890574"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "pCW57.MCS_Seq_F CGTATGTCGAGGTAGGCGTG",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
TFORF3455 | 144931 | Lentiviral vector for overexpressing transcription factor ORFs with unique 24-bp barcodes. Barcodes facilitate identification of transcription factors in pooled screens. | Feng Zhang | Joung et al Cell. 2023 Jan 5;186(1):209-229.e26. doi: 10.1016/j.cell.2022.11.026. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/404505/68b411fe-7e40-4559-81d0-ab16e05458b8/addgene-plasmid-144931-sequence-404505.gbk | full | LOCUS sequence-404505- 10093 bp DNA circular SYN 26-AUG-2024
DEFINITION Lentiviral vector for overexpressing transcription factor ORFs with
unique 24-bp barcodes. Barcodes facilitate identification of
transcription factors in pooled screens..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10093)
AUTHORS Joung J, Ma S, Tay T, Geiger-Schuller KR, Kirchgatterer PC, Verdine
VK, Guo B, Arias-Garcia MA, Allen WE, Singh A, Kuksenko O, Abudayyeh
OO, Gootenberg JS, Fu Z, Macrae RK, Buenrostro JD, Regev A, Zhang F
TITLE A transcription factor atlas of directed differentiation.
JOURNAL Cell. 2023 Jan 5;186(1):209-229.e26. doi:
10.1016/j.cell.2022.11.026.
PUBMED 36608654
REFERENCE 2 (bases 1 to 10093)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1016/j.cell.2022.11"; journalName: "Cell"; date: "2023-01-5- 5";
volume: "186"; issue: "1"; pages: "209-229.e26"
FEATURES Location/Qualifiers
source 1..10093
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 5..231
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 232..412
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 459..584
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1077..1310
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1495..1539
/codon_start=1
/product="antigenic peptide corresponding to amino acids
655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
et al., 2013)"
/label=gp41 peptide
/note="recognized by the 2H10 single-chain llama nanobody"
/translation="KNEQELLELDKWASL"
promoter 1770..1980
/label=SV40 promoter
/note="SV40 early promoter"
promoter 1792..1988
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 1839..1974
/label=SV40 ori
/note="SV40 origin of replication"
CDS 2020..2622
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGAS"
primer_bind complement(2020..2039)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 2516..2536
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
misc_feature 2665..2782
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2920..4098
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 3151..4089
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 4046..4066
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
protein_bind 4118..4142
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
protein_bind complement(5766..5790)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
CDS 5798..5839
/codon_start=1
/product="epitope tag from simian virus 5"
/label=V5 tag
/translation="GKPIPNPLLGLDST"
misc_feature 5950..6538
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(6003..6023)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(6421..6432)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 6610..6843
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 6921..7042
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(6958..6977)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 7012..7031
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 7082..7217
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(7237..7256)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(7238..7256)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(7266..7283)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(7266..7282)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(7275..7297)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 7424..7879
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(7511..7530)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 7721..7742
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 7905..8009
/gene="bla"
/label=AmpR promoter
CDS 8010..8870
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(8228..8247)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 9041..9629
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 9530..9549
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 9783..9800
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 9917..9938
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 9953..9983
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 9991..10007
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
primer_bind 9996..10018
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 10015..10031
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 10015..10031
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 10050..10070
/label=T3
/note="T3 promoter, forward primer"
promoter 10052..10070
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
1 ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat gagttagcaa
61 catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt aaggtggtac
121 gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg aaccactgaa
181 ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa cgggtctctc
241 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
301 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
361 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
421 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
481 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
541 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
601 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
661 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
721 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
781 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
841 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
901 gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg
961 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta
1021 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga
1081 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg
1141 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg
1201 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc
1261 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg
1321 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat
1381 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac
1441 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat
1501 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca
1561 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga
1621 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg
1681 tttcagaccc acctcccaac cccgagggga cccatgcatt gcatctcaat tagtcagcaa
1741 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcgtctca
1801 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
1861 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
1921 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
1981 tttgcaaaaa gctttctaga ggtaccaccg gagcttacca tgaccgagta caagcccacg
2041 gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc
2101 gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc
2161 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg
2221 gacgacggcg ccgccgtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg
2281 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa
2341 cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc
2401 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga
2461 gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac
2521 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga
2581 ccgcgcacct ggtgcatgac ccgcaagccc ggtgccagtt aacgaattct agatcttgag
2641 acaaatggca gtattcatcc acaattttaa aagaaaaggg gggattgggg ggtacagtgc
2701 aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat tacaaaaaca
2761 aattacaaaa attcaaaatt ttcgggttta ttacagggac agcagagatc cactttggcg
2821 ccggctcgag gcctgcaggt gcaaagatgg ataaagtttt aaacagagag gaatctttgc
2881 agctaatgga ccttctaggt cttgaaagga gtgggaattg gctccggtgc ccgtcagtgg
2941 gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacc
3001 ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc
3061 ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt
3121 tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc ccgcgggcct
3181 ggcctcttta cgggttatgg cccttgcgtg ccttgaatta cttccacctg gctgcagtac
3241 gtgattcttg atcccgagct tcgggttgga agtgggtggg agagttcgag gccttgcgct
3301 taaggagccc cttcgcctcg tgcttgagtt gaggcctggc ctgggcgctg gggccgccgc
3361 gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc tctagccatt
3421 taaaattttt gatgacctgc tgcgacgctt tttttctggc aagatagtct tgtaaatgcg
3481 ggccaagatc tgcacactgg tatttcggtt tttggggccg cgggcggcga cggggcccgt
3541 gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc gagaatcgga
3601 cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg gcctcgcgcc gccgtgtatc
3661 gccccgccct gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc ggaaagatgg
3721 ccgcttcccg gccctgctgc agggagctca aaatggagga cgcggcgctc gggagagcgg
3781 gcgggtgagt cacccacaca aaggaaaagg gcctttccgt cctcagccgt cgcttcatgt
3841 gactccacgg agtaccgggc gccgtccagg cacctcgatt agttctcgag cttttggagt
3901 acgtcgtctt taggttgggg ggaggggttt tatgcgatgg agtttcccca cactgagtgg
3961 gtggagactg aagttaggcc agcttggcac ttgatgtaat tctccttgga atttgccctt
4021 tttgagtttg gatcttggtt cattctcaag cctcagacag tggttcaaag tttttttctt
4081 ccatttcagg tgtcgtgagg ctagcatcga ttgatcaaca agtttgtaca aaaaagttgg
4141 catgccccga aggaagcaaa gccacccgca gcccgtgaaa tgcgaggggg tcaaagtgga
4201 tactgaagac tccctcgacg aaggacccgg ggccctggta ttggagagtg atttgctact
4261 aggccaggat ctggagtttg aggaggaaga ggaagaggag gaaggcgacg gcaacagtga
4321 ccagctcatg ggcttcgaga gagactcgga aggagactct ctgggggcca ggcctgggct
4381 tccctatggg ctgagcgacg atgagtctgg gggcggccgg gcactaagtg cggagagtga
4441 agttgaggag ccagccaggg gtccagggga ggccaggggt gagaggccag gcccagcctg
4501 ccagctgtgt ggggggccga caggtgaggg gccgtgttgt ggggcaggag ggccgggtgg
4561 ggggcccctg ctgcccccac ggctactgta ctcatgccgc ctctgcacct tcgtgtccca
4621 ctactcgagc cacctgaagc ggcacatgca gacacacagc ggagagaagc cgttccgctg
4681 tggccgctgc ccctacgcct cagcccagct cgtcaacctg acacgacata cccgcaccca
4741 cactggcgag aagccctacc gctgtcccca ctgccccttt gcctgcagca gcctgggcaa
4801 cctgaggcgg catcagcgta cccacgcagg gccccccact cctccctgcc cgacctgtgg
4861 cttccgctgc tgtactccac gaccagcccg gcctcccagt cccacagagc aggagggggc
4921 ggtgccccgg cgacctgaag atgctctgct ccttccagat ttgagcctcc atgtgccacc
4981 aggtggtgcc agtttcctgc cagactgtgg gcagctgcgg ggtgaagggg agggcctctg
5041 cgggactgga tcagaaccac tgccagagct gctattccct tggacctgcc ggggctgtgg
5101 acaagagctg gaggagggtg agggtagtcg gctgggagct gccatgtgtg ggcgctgcat
5161 gcgaggagag gctggagggg gtgccagtgg ggggccccag ggccccagtg acaaaggctt
5221 tgcctgtagc ctctgcccct ttgccactca ctatcccaac cacctggccc ggcacatgaa
5281 gacacacagt ggtgagaagc ccttccgctg cgcccgctgt ccttatgcct ctgctcatct
5341 ggataacctg aaacggcacc agcgcgtcca tacaggagag aagccctaca agtgccccct
5401 ctgcccttat gcctgtggca atctggccaa cctcaagcgt catggtcgca tccactctgg
5461 tgacaaacct tttcggtgta gcctttgcaa ctacagctgc aaccagagca tgaacctcaa
5521 acgtcacatg ctgcggcaca caggcgagaa gcccttccgc tgtgccacct gcgcctatac
5581 cacgggccac tgggacaact acaagcgcca ccagaaggtg catggccacg gtggggcagg
5641 agggcctggt ctctctgcct ctgagggctg ggccccacct catagcccac cctctgtttt
5701 gagctctcgg ggcccaccag ccctggggac tgctggcagc cgggctgtcc acacagactc
5761 atcctgccca actttcttgt acaaagtggt tgatatcggt aagcctatcc ctaaccctct
5821 cctcggtctc gattctacgt agtaatgaac tagtccgtaa cttgaaagta tttcgatttc
5881 ttggctttat atatcttgtg gaaaggacga ctgttctcag cttgaacaga tattacgcgt
5941 taagtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
6001 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
6061 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
6121 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
6181 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
6241 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
6301 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct
6361 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
6421 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
6481 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcgt
6541 cgactttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa
6601 aggggggact ggaagggcta attcactccc aacgaagaca agatctgctt tttgcttgta
6661 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
6721 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
6781 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
6841 gcagtacgta tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat
6901 gaatatcaga gagtgagagg aacttgttta ttgcagctta taatggttac aaataaagca
6961 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
7021 ccaaactcat caatgtatct tatcatgtct ggctctagct atcccgcccc taactccgcc
7081 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
7141 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
7201 aggctttttt ggaggcctag ggacgtaccc aattcgccct atagtgagtc gtattacgcg
7261 cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
7321 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
7381 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc
7441 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
7501 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
7561 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
7621 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
7681 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
7741 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
7801 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
7861 atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
7921 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
7981 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
8041 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
8101 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
8161 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
8221 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
8281 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
8341 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
8401 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
8461 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
8521 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
8581 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
8641 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
8701 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
8761 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
8821 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
8881 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
8941 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
9001 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
9061 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
9121 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
9181 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
9241 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
9301 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
9361 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
9421 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
9481 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
9541 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
9601 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
9661 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
9721 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
9781 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
9841 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
9901 gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact
9961 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
10021 acagctatga ccatgattac gccaagcgcg caattaaccc tcactaaagg gaacaaaagc
10081 tggagctgca agc
//
| {
"Vector backbone": "pLX_TRC317",
"Backbone size w/o insert (bp)": "8264",
"Vector type": "Mammalian Expression, Lentiviral",
"Selectable markers": "Puromycin"
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "NEB Stable",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "A transcription factor atlas of directed differentiation",
"Publication": "Cell. 2023 Jan 5;186(1):209-229.e26. doi: 10.1016/j.cell.2022.11.026.",
"DOI": "10.1016/j.cell.2022.11.026.",
"Authors": "Joung J, Ma S, Tay T, Geiger-Schuller KR, Kirchgatterer PC, Verdine VK, Guo B, Arias-Garcia MA, Allen WE, Singh A, Kuksenko O, Abudayyeh OO, Gootenberg JS, Fu Z, Macrae RK, Buenrostro JD, Regev A, Zhang F /jcell",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/36608654",
"PubMed ID": "PubMed 36608654"
} | {
"Gene/Insert name": "ZNF513",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "1829",
"Mutation": "",
"Entrez Gene": "ZNF513",
"Promoter": "EF1a",
"Tag / Fusion Protein": "V5 (C terminal on insert)"
} | {
"Cloning method": "Gateway Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "GGGTGGAGACTGAAGTTAGGCCAG",
"3′ sequencing primer": "CACATAGCGTAAAAGGAGCAACATAG"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pJK688 | 71641 | Produces Treponema denticola 5-aminoimidazole ribonucleotide carboxylase, Lys41>Arg mutant (TdPurE2-K41R) | T. J. Kappock | Serine mutants of Treponema denticola PurE2 (unpublished) | https://media.addgene.org/snapgene-media/v2.0.0/sequences/131662/09115418-8509-4790-9352-b06a15af1301/addgene-plasmid-71641-sequence-131662.gbk | full | LOCUS sequence-131662- 4137 bp DNA circular SYN 26-AUG-2024
DEFINITION Produces Treponema denticola 5-aminoimidazole ribonucleotide
carboxylase, Lys41>Arg mutant (TdPurE2-K41R).
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4137)
AUTHORS Kappock
TITLE Serine mutants of Treponema denticola PurE2
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 4137)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
FEATURES Location/Qualifiers
source 1..4137
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin 12..467
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(99..118)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 309..330
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 494..598
/gene="bla"
/label=AmpR promoter
CDS 599..1459
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(817..836)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 1630..2218
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2119..2138
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 2372..2389
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 2404..2546
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(2472..2491)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 2632..2654
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(2648..2839)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
primer_bind 3348..3367
/label=T7
/note="T7 promoter, forward primer"
promoter 3348..3366
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
RBS 3398..3420
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 3981..3998
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(4051..4069)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 4065..4112
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
ORIGIN
1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
421 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttcag gtggcacttt
481 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
541 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
601 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
661 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
721 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
781 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
841 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
901 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
961 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
1021 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
1081 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
1141 tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
1201 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
1261 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
1321 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
1381 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
1441 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
1501 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
1561 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
1621 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
1681 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
1741 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
1801 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
1861 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
1921 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
1981 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
2041 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
2101 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
2161 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
2221 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
2281 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
2341 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
2401 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatatgg
2461 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat
2521 cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct
2581 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
2641 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct
2701 catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt
2761 tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg
2821 ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa
2881 tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc
2941 ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa
3001 aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta
3061 gccagcagca tcctgcgatg cagatccgga acataatggt gcagggcgct gacttccgcg
3121 tttccagact ttacgaaaca cggaaaccga agaccattca tgttgttgct caggtcgcag
3181 acgttttgca gcagcagtcg cttcacgttc gctcgcgtat cggtgattca ttctgctaac
3241 cagtaaggca accccgccag cctagccggg tcctcaacga caggagcacg atcatgcgca
3301 cccgtggcca ggacccaacg ctgcccgaga tctcgatccc gcgaaattaa tacgactcac
3361 tatagggaga ccacaacggt ttccctctag aaataatttt gtttaacttt aagaaggaga
3421 tatacatatg cgtccgctgg tgattatcct tatgggcagc agttcagata tgggtcacgc
3481 cgagaaaatt gcgtctgaat tgaaaacttt tggtattgag tatgccattc gcatcggctc
3541 cgcgcataga acagctgagc atgtagtgtc tatgctgaaa gaatatgaag ctttggaccg
3601 tcctaaactg tacatcacga tcgctgggcg tagtaacgct ttatcaggct ttgttgatgg
3661 cttcgtcaaa ggtgcaacca ttgcctgccc accgccaagc gatagcttcg ccggcgccga
3721 catttattct tcgttacgga tgccgagcgg gattagtccg gcactggttc ttgaaccgaa
3781 aaatgcggca ctcctggcgg ctcgtatttt ttcactgtac gataaagaaa tcgcagattc
3841 ggtcaaatcc tatatggaat ccaatgcgca gaaaattatc gaagacgatt cgaaactgaa
3901 acgctgaatt cagcgtttca gtttcttgtc gacggagctc gaattcgagc tccgtcgaca
3961 agcttgcggc cgcactcgag caccaccacc accaccactg agatccggct gctaacaaag
4021 cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg
4081 gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata tccggat
//
| {
"Vector backbone": "pET23a",
"Backbone size w/o insert (bp)": "3666",
"Vector type": "Bacterial Expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "",
"Publication": "",
"DOI": "",
"Authors": "",
"PubMed Link": "",
"PubMed ID": ""
} | {
"Gene/Insert name": "phosphoribosylaminoimidazole carboxylase",
"Species": "Treponema denticola",
"Insert Size (bp)": "",
"Mutation": "changes lysine-41 to arginine",
"Entrez Gene": "",
"Promoter": "T7",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "NdeI (not destroyed)",
"3′ cloning site": "EcoRI (not destroyed)",
"5′ sequencing primer": "T7 promoter",
"3′ sequencing primer": "T7 terminator"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pBABE-NLS-HA-TurboID | 215073 | Expresses NLS-HA-TurboID in mammalian cells from a retroviral vector. | Michele Pagano | Rona et al Mol Cell. 2024 Feb 29:S1097-2765(24)00129-1. doi: 10.1016/j.molcel.2024.02.010. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/429287/fcba2dee-ace9-4e5b-89f8-48f7ce8c120a/addgene-plasmid-215073-sequence-429287.gbk | full | LOCUS sequence-429287- 6155 bp DNA circular SYN 27-AUG-2024
DEFINITION Expresses NLS-HA-TurboID in mammalian cells from a retroviral
vector..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6155)
AUTHORS Rona G, Miwatani-Minter B, Zhang Q, Goldberg HV, Kerzhnerman MA,
Howard JB, Simoneschi D, Lane E, Hobbs JW, Sassani E, Wang AA,
Keegan S, Laverty DJ, Piett CG, Pongor LS, Xu ML, Andrade J, Thomas
A, Sicinski P, Askenazi M, Ueberheide B, Fenyo D, Nagel ZD, Pagano M
TITLE CDK-independent role of D-type cyclins in regulating DNA mismatch
repair.
JOURNAL Mol Cell. 2024 Feb 29:S1097-2765(24)00129-1. doi:
10.1016/j.molcel.2024.02.010.
PUBMED 38458201
REFERENCE 2 (bases 1 to 6155)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 27, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Mol Cell.
2024 Feb 29:S1097-2765(24)00129-1. doi:
10.1016/j.molcel.2024.02.010."
FEATURES Location/Qualifiers
source 1..6155
/mol_type="other DNA"
/organism="synthetic DNA construct"
LTR 8..477
/note="long terminal repeat from Moloney murine leukemia
virus"
misc_feature 542..741
/label=MMLV Psi
/note="packaging signal of Moloney murine leukemia virus
(MMLV)"
CDS 942..1358
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
primer_bind 1288..1310
/label=pLXSN 5'
/note="Murine stem cell virus, forward primer. Also called
MSCV"
primer_bind 1326..1342
/label=pBABE 5'
/note="Psi packaging signal, forward primer for pBABE
vectors"
regulatory 1370..1379
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 1388..1408
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 1421..1447
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 1451..1477
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 1481..1507
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 1514..2470
/codon_start=1
/product="mutant of the E. coli biotin ligase BioID
engineered for improved catalytic efficiency (Branon et
al., 2018)"
/label=TurboID
/translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW
GVDVFTVPGKGYSLPEPIPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKSGD
ACIAEYQQAGRGSRGRKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRKLG
ADKVRVKWPNDLYLQDRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQGWI
TLQEAGINLDRNTLAATLIRELRAALELFEQEGLAPYLPRWEKLDNFINRPVKLIIGDK
EIFGISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEK"
primer_bind complement(2485..2505)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2490..2819
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2670..2805
/label=SV40 ori
/note="SV40 origin of replication"
CDS 2829..3428
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind complement(2829..2848)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 3325..3345
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
LTR 3649..4118
/note="long terminal repeat from Moloney murine leukemia
virus"
primer_bind complement(4248..4265)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4419..5007)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4499..4518)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5167..6027)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(6028..6132)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 ctgcagcctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
61 gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt
121 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag
181 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc
241 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga
301 gctcaataaa agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc
361 gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc
421 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt
481 gggggctcgt ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg
541 caagctggcc agcaacttat ctgtgtctgt ccgattgtct agtgtctatg actgatttta
601 tgcgcctgcg tcggtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa
661 ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc
721 gtttttgtgg cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg
781 tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt
841 cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt
901 gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc
961 ccttaagttt gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg
1021 tagatgtcaa gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg
1081 tcggatggcc gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg
1141 tcttttcacc tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag
1201 ccttggcttt tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc
1261 ctcttcctcc atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat
1321 cctcccttta tccagccctc actccttctc taggcgccgg ccggatcccg ccaccatggg
1381 agcatcacca aaaaagaaga gaaaggtacc ggtggctacc tacccgtatg atgttccgga
1441 ttacgctggc tatccctacg acgtgcccga ctatgccggg tacccctatg acgtcccaga
1501 ctacgcagct agcaaagaca atactgtgcc tctgaagctg atcgctctcc tggctaatgg
1561 cgagttccat agtggcgaac agctgggaga aaccctgggc atgtccaggg ccgctatcaa
1621 caagcacatt cagactctgc gcgactgggg cgtggacgtg ttcaccgtgc ccggaaaggg
1681 ctactctctg cccgagccta tcccgctgct gaacgctaaa cagattctgg gacagctgga
1741 cggcgggagc gtggcagtcc tgcctgtggt cgactccacc aatcagtacc tgctggatcg
1801 aatcggcgag ctgaagagtg gggatgcttg cattgcagaa tatcagcagg cagggagagg
1861 aagcagaggg aggaaatggt tctctccttt tggagctaac ctgtacctga gtatgttttg
1921 gcgcctgaag cggggaccag cagcaatcgg cctgggcccg gtcatcggaa ttgtcatggc
1981 agaagcgctg cgaaagctgg gagcagacaa ggtgcgagtc aaatggccca atgacctgta
2041 tctgcaggat agaaagctgg caggcatcct ggtggagctg gccggaataa caggcgatgc
2101 tgcacagatc gtcattggcg ccgggattaa cgtggctatg aggcgcgtgg aggaaagcgt
2161 ggtcaatcag ggctggatca cactgcagga agcagggatt aacctggaca ggaatactct
2221 ggccgctacg ctgatccgag agctgcgggc agccctggaa ctgttcgagc aggaaggcct
2281 ggctccatat ctgccacggt gggagaagct ggataacttc atcaatagac ccgtgaagct
2341 gatcattggg gacaaagaga ttttcgggat tagccggggg attgataaac agggagccct
2401 gctgctggaa caggacggag ttatcaaacc ctggatgggc ggagaaatca gtctgcggtc
2461 tgccgaaaag taactcgacc ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
2521 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
2581 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
2641 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
2701 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
2761 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
2821 agcttaccat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca
2881 gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg
2941 atccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg
3001 ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca
3061 cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt
3121 tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc
3181 ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg
3241 gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg
3301 ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg
3361 tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg
3421 gtgcctgacg cccgccccac gacccgcagc gcccgaccga aaggagcgca cgaccccatg
3481 catcgataaa ataaaagatt ttatttagtc tccagaaaaa ggggggaatg aaagacccca
3541 cctgtaggtt tggcaagcta gcttaagtaa cgccattttg caaggcatgg aaaaatacat
3601 aactgagaat agagaagttc agatcaaggt caggaacaga tggaacagct gaatatgggc
3661 caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga acagatggaa
3721 cagctgaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc
3781 caagaacaga tggtccccag atgcggtcca gccctcagca gtttctagag aaccatcaga
3841 tgtttccagg gtgccccaag gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc
3901 agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa aagagcccac
3961 aacccctcac tcggggcgcc agtcctccga ttgactgagt cgcccgggta cccgtgtatc
4021 caataaaccc tcttgcagtt gcatccgact tgtggtctcg ctgttccttg ggagggtctc
4081 ctctgagtga ttgactaccc gtcagcgggg gtctttcaca tgcagcatgt atcaaaatta
4141 atttggtttt ttttcttaag tatttacatt aaatggccat agttgcatta atgaatcggc
4201 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
4261 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
4321 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
4381 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
4441 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
4501 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
4561 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
4621 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
4681 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
4741 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
4801 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
4861 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
4921 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
4981 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
5041 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
5101 ttcacctaga tccttttgcg gccgcaaatc aatctaaagt atatatgagt aaacttggtc
5161 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
5221 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
5281 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
5341 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
5401 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
5461 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
5521 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
5581 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
5641 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
5701 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
5761 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
5821 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
5881 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
5941 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
6001 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
6061 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
6121 aggggttccg cgcacatttc cccgaaaagt gccac
//
| {
"Vector backbone": "pBABE",
"Backbone size w/o insert (bp)": "",
"Vector type": "Mammalian Expression, Retroviral",
"Selectable markers": "Puromycin"
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "NEB Stable",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "CDK-independent role of D-type cyclins in regulating DNA mismatch repair",
"Publication": "Mol Cell. 2024 Feb 29:S1097-2765(24)00129-1. doi: 10.1016/j.molcel.2024.02.010.",
"DOI": "10.1016/j.molcel.2024.02.010.",
"Authors": "Rona G, Miwatani-Minter B, Zhang Q, Goldberg HV, Kerzhnerman MA, Howard JB, Simoneschi D, Lane E, Hobbs JW, Sassani E, Wang AA, Keegan S, Laverty DJ, Piett CG, Pongor LS, Xu ML, Andrade J, Thomas A, Sicinski P, Askenazi M, Ueberheide B, Fenyo D, Nagel ZD, Pagano M /jmolcel",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/38458201",
"PubMed ID": "PubMed 38458201"
} | {
"Gene/Insert name": "TurboID",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "birA",
"Promoter": "LTR",
"Tag / Fusion Protein": "NLS-HA (N terminal on insert)"
} | {
"Cloning method": "Unknown",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "pBABE 5'",
"3′ sequencing primer": "pBABE 3'"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pT7-EGFP-C1-HsNot1iso1_1-1840-IKGRQtoDSYAA-shRNAres_W | 147890 | Mammalian Expression of HsNot1iso1_1-1840-IKGRQtoDSYAA-shRNAres | Elisa Izaurralde | Chen et al Mol Cell. 2014 Jun 5;54(5):737-50. doi: 10.1016/j.molcel.2014.03.034. Epub 2014 Apr 24. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/290337/bb1e99ed-f6ec-43b5-b58a-f63fdcf68839/addgene-plasmid-147890-sequence-290337.gbk | full | LOCUS sequence-290337- 10278 bp DNA circular SYN 26-AUG-2024
DEFINITION Mammalian Expression of HsNot1iso1_1-1840-IKGRQtoDSYAA-shRNAres.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10278)
AUTHORS Chen Y, Boland A, Kuzuoglu-Ozturk D, Bawankar P, Loh B, Chang CT,
Weichenrieder O, Izaurralde E
TITLE A DDX6-CNOT1 complex and W-binding pockets in CNOT9 reveal direct
links between miRNA target recognition and silencing.
JOURNAL Mol Cell. 2014 Jun 5;54(5):737-50. doi:
10.1016/j.molcel.2014.03.034. Epub 2014 Apr 24.
PUBMED 24768540
REFERENCE 2 (bases 1 to 10278)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1016/j.molcel.2014.03.034"; journalName: "Mol Cell"; date:
"2014-06-5- 5"; volume: "54"; issue: "5"; pages: "737-50"
FEATURES Location/Qualifiers
source 1..10278
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 61..364
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 365..568
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 519..539
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 613..632
/label=T7
/note="T7 promoter, forward primer"
promoter 613..631
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory 659..668
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 665..1381
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 665..1381
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(710..731)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(971..990)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1318..1339
/label=EGFP-C
/note="EGFP, forward primer"
polyA_signal 7066..7187
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(7103..7122)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 7157..7176
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(7194..7649)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(7331..7352)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 7543..7562
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 7676..7780
/gene="bla"
/label=AmpR promoter
promoter 7782..8139
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
enhancer 7782..7973
/label=SV40 enhancer
/note="enhancer for the SV40 early promoter (Herr, 1993)"
primer_bind complement(7805..7825)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 7990..8125
/label=SV40 ori
/note="SV40 origin of replication"
CDS 8174..8968
/codon_start=1
/product="aminoglycoside phosphotransferase"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
CDS 8174..8968
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(8228..8247)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 8838..8857
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(9156..9175)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 9200..9247
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 9576..10164
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 10065..10084
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
541 acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg
601 gcttatcgaa attaatacga ctcactatag ggagacccaa gctctagcgc taccggtcgc
661 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
721 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
781 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
841 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
901 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
961 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
1021 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
1081 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
1141 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
1201 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
1261 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
1321 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
1381 gtccggactc agatctcgag ctatgaatct tgactcgctc tcgctggcct tgtctcaaat
1441 cagctacctg gtggacaatt taaccaagaa aaattaccga gccagccagc aggaaataca
1501 gcatattgtg aatcggcacg gtcctgaggc agacaggcat ttattacgct gcctattttc
1561 gcatgtggat ttcagtggcg atggtaaaag cagtggcaaa gatttccatc agactcagtt
1621 tctgattcag gagtgtgcgt tgctgattac aaagccaaat tttatctcga cgctgtccta
1681 tgccattgat aatccattgc actatcagaa gagtttaaag cctgcacccc acttatttgc
1741 ccagctgagt aaagtgctca aattaagcaa agtacaagag gtaatttttg gccttgccct
1801 gttgaattct tccagctcag atcttagagg tttcgctgcc cagtttatca aacagaagct
1861 tccagatctt ctgcgttctt acattgacgc agacgtcagt ggaaatcaag aaggtggctt
1921 ccaagatata gcaatagagg tcctacacct cctcctctcc catctcctct ttgggcagaa
1981 gggagccttt ggagttggac aagaacagat agacgctttt cttaagacgc tgcgcagaga
2041 ttttccccaa gaacgctgtc ccgtggtgct cgcaccactt ttataccctg aaaaacggga
2101 cattctaatg gacaggatcc tgcctgattc cggaggggta gctaaaacca tgatggagag
2161 ctctttggct gatttcatgc aagaagtagg ctatggcttt tgtgcaagta ttgaagaatg
2221 tcgcaatata atcgtgcagt ttggtgttcg ggaggtcaca gctgcccagg ttgcaagggt
2281 tttgggaatg atggctcgaa ctcattcagg attaacagat ggcattccat tacagagtat
2341 ttctgctccg ggcagtggga tctggagtga tgggaaagat aaaagtgatg gagcacaggc
2401 acacacatgg aatgtagaag tcttgattga cgttcttaaa gaactgaatc caagtttgaa
2461 tttcaaggaa gtaacttatg aactggacca tcctggattt caaattcgtg acagtaaagg
2521 acttcataat gtggtttatg gcattcagag gggtttgggt atggaagtgt tcccagtaga
2581 cctcatatat agaccttgga aacatgctga aggccagctc tccttcatcc agcactcact
2641 aatcaatcca gagatcttct gttttgctga ctatccctgt catactgttg ccactgatat
2701 tctgaaagca ccaccagagg atgacaatcg agaaattgcc acatggaaga gcttggattt
2761 gattgaatct ctgctgaggc ttgcagaggt tgggcagtat gagcaagtca aacagctctt
2821 cagcttccct atcaaacact gtccagacat gctggtattg gccttactac aaattaacac
2881 ctcttggcat accttgcgcc atgaacttat ctccactctg atgccaattt tccttggaaa
2941 ccatcctaac tcagctatta ttttgcacta tgcatggcat gggcagggac agtctccctc
3001 aattcgccaa cttatcatgc atgcaatggc agaatggtac atgagagggg agcagtatga
3061 tcaggccaaa ttgtctcgaa tacttgatgt ggcccaggac ttgaaggcct tgtcaatgct
3121 gctaaatggt actccatttg cctttgttat tgaccttgct gcacttgctt cacgtcgtga
3181 atacctcaaa cttgataagt ggctcacaga taaaattcga gagcatgggg agccttttat
3241 ccaggcgtgt atgacttttt taaagagacg gtgtccttct attttgggcg gacttgcccc
3301 agaaaaagac cagcccaaaa gtgctcaact tcctccagaa actttggcga caatgttggc
3361 ctgtctgcaa gcttgtgcag ggagtgtttc tcaggagcta tcagaaacta tcctcaccat
3421 ggtagccaat tgcagtaatg ttatgaataa ggccagacaa ccaccacctg gagttatgcc
3481 aaaaggacgt cctcctagtg ctagcagctt agatgccatt tctcctgttc agattgaccc
3541 tcttgctgga atgacatctc ttagtatagg tggttcagct gcccctcaca cccagagtat
3601 gcagggtttt cctccaaatt tgggttctgc attcagtacc cctcagtcac cagcaaaagc
3661 atttccaccc ctttcaaccc ccaatcagac cactgcattc agtggtattg gaggactttc
3721 atcacagctt ccagtaggtg gtcttggcac aggcagcctg actggtatag gaactggtgc
3781 tcttggactc cctgcagtga ataacgaccc ttttgtacag aggaaactgg gcacctctgg
3841 actgaatcag cctacattcc agcagagtaa gatgaaacct tcggacttgt ctcaggtgtg
3901 gccagaggca aaccagcact ttagtaaaga gatagatgat gaagcaaaca gctatttcca
3961 gcgaatatat aatcatccac cacatccaac catgtctgtt gatgaggtat tagaaatgct
4021 gcagagattt aaagactcta ctataaagag ggaacgagaa gtatttaact gtatgctaag
4081 gaacttgttt gaagaatatc gtttttttcc ccagtatcct gataaagagt tacatataac
4141 agcctgccta tttggtggta taattgagaa aggactggtc acttacatgg cactaggtct
4201 ggctctacga tatgttcttg aagccttacg caagcctttt ggatccaaaa tgtattattt
4261 cgggattgct gcactagata gatttaaaaa cagattgaag gactatcccc agtattgtca
4321 gcatttggct tctatcagtc actttatgca atttccacat catttacagg agtatattga
4381 gtatggacag cagtctagag atcctcctgt gaaaatgcaa ggctctatca caacccctgg
4441 aagtattgca ctggctcagg cccaggctca ggcccaggtt ccagcaaaag ctcctcttgc
4501 tggtcaagtt agcactatgg taaccacctc aacaactacc actgttgcta aaacggttac
4561 ggtcaccagg ccaactggag tcagctttaa gaaagatgtg ccaccttcta ttaatactac
4621 aaatatagat acgttgcttg tggccacaga tcaaactgag agaattgtgg agcccccaga
4681 aaatatccag gagaaaattg cttttatttt caataatctc tcacagtcaa atatgacaca
4741 aaaggttgaa gagctaaagg aaacggtgaa agaagaattt atgccttggg tttcacagta
4801 tctggttatg aagagagtca gtattgagcc aaactttcat agcctgtatt caaacttcct
4861 tgacacgctg aagaatcctg aatttaacaa gatggttctg aatgagacct acagaaacat
4921 taaagtgctc ctgacctctg ataaagctgc agccaatttc tcagatcgtt ctttgctgaa
4981 gaacttggga cattggctag gaatgatcac attagctaaa aacaaaccca tcttacacac
5041 tgacttggat gtgaaatcat tgctgctaga ggcttatgtt aaaggacaac aagaattgct
5101 ctatgtagtg ccctttgttg ccaaagtctt agaatctagc attcgtagtg tggtttttag
5161 gccaccaaac ccttggacaa tggcaattat gaatgtatta gctgagctac atcaggagca
5221 tgacttaaag ttaaacttga agtttgaaat cgaggttctc tgcaagaacc ttgcattaga
5281 catcaatgag ctaaaacctg gaaacctcct aaaggataaa gatcgcctga agaatttaga
5341 tgagcaactc tctgctccaa agaaagatgt caagcagcca gaagaactcc ctcccatcac
5401 aaccacaaca acttctacta caccagctac caacaccact tgtacagcca cggttccacc
5461 acagccacag tacagctacc acgacatcaa tgtctattcc cttgcgggct tggcaccaca
5521 cattactcta aatccaacaa ttcccttgtt tcaggcccat ccacagttga agcagtgtgt
5581 gcgtcaggca attgaacggg ctgtccagga gctggtccat cctgtggtgg atcgatcaat
5641 taagattgcc atgactactt gtgagcaaga tgtcaggtcg gattttgccc tggattcgga
5701 ggaatctcga atgcgaatag cagctcatca catgatgcgt aacttgacag cttacatggc
5761 tatgattaca tgcgctgaac ctttgctcat gagcatatct accaacttaa aaaacagttt
5821 tgcctcagcc cttcgtactg cttccccaca acaaagagaa atgatggatc aggcagctgc
5881 tcaattagct caggacaatt gtgagttggc ttgctgtttt attcagaaga ctgcagtaga
5941 aaaagcaggc cctgagatgg acaagagatt agcaactgaa tttgagctga gaaaacatgc
6001 taggcaagaa ggacgcagat actgtgatcc tgttgtttta acatatgctg ctgaacggat
6061 gccagagcaa atcaggctga aagttggtgg tgtggaccca aagcagttgg ctgtttacga
6121 agagtttgca cgcaatgttc ctggcttctt acctacaaat gacttaagtc agcccacggg
6181 atttttagcc cagcccatga agcaagcttg ggcaacagat gatgtagctc agatttatga
6241 taagtgtatt acagaactgg agcaacatct acatgccatc ccaccaactt tggccatgaa
6301 ccctcaagct caggctcttc gaagtctctt ggaggttgta gttttatctc gaaactctcg
6361 ggatgccata gctgctcttg gattgctcca aaaggctgta gagggcttac tagatgccac
6421 aagtggtgct gatgctgacc ttctgctgcg ctacagggaa tgccacctct tggtcctaaa
6481 agctctgcag gatggccggg catatgggtc tccatggtgc aacaaacaga tcacaaggtg
6541 cctaattgaa tgtcgagatg aatataaata taatgtggag gctgtggagc tgctaattcg
6601 caatcatttg gttaatatgc agcagtatga tcttcaccta gcgcagtcaa tggagaatgg
6661 cttaaactac atggctgtgg catttgctat gcagttagta aaaatcctgc tggtggatga
6721 aaggagtgtt gctcatgtta ctgaggcaga tctgttccac accattgaaa ccctcatgag
6781 gattaatgct cattccagag gcaatgctcc agaaggattg ccccagctga tggaagtagt
6841 gcgatccaac tatgaagcaa tgattgatcg tgctcatgga ggcccaaact ttatgatgca
6901 ttctgggatc tctcaagcct catagccgcg ggcccgggat ccaccggatc tagataactg
6961 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
7021 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
7081 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
7141 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt
7201 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
7261 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
7321 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
7381 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
7441 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
7501 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
7561 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
7621 cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg
7681 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
7741 accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag
7801 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
7861 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
7921 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
7981 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
8041 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
8101 tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg
8161 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
8221 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
8281 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
8341 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
8401 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
8461 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
8521 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
8581 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
8641 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
8701 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
8761 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
8821 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
8881 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
8941 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
9001 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
9061 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
9121 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg
9181 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg
9241 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
9301 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
9361 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt
9421 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
9481 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
9541 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
9601 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
9661 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
9721 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
9781 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
9841 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
9901 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
9961 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
10021 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
10081 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
10141 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
10201 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
10261 cgtattaccg ccatgcat
//
| {
"Vector backbone": "Unknown",
"Backbone size w/o insert (bp)": "",
"Vector type": "Mammalian Expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Kanamycin, 50 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "A DDX6-CNOT1 complex and W-binding pockets in CNOT9 reveal direct links between miRNA target recognition and silencing",
"Publication": "Mol Cell. 2014 Jun 5;54(5):737-50. doi: 10.1016/j.molcel.2014.03.034. Epub 2014 Apr 24.",
"DOI": "10.1016/j.molcel.2014.03.034.",
"Authors": "Chen Y, Boland A, Kuzuoglu-Ozturk D, Bawankar P, Loh B, Chang CT, Weichenrieder O, Izaurralde E /jmolcel",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/24768540",
"PubMed ID": "PubMed 24768540"
} | {
"Gene/Insert name": "HsNot1iso1_1-1840-IKGRQtoDSYAA-shRNAres",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "1 silent mutation compared to the sequence of isoform 1 given by NCBI: NM_016284.4.",
"Entrez Gene": "CNOT1",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Unknown",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
P[acman]-Omega2 | 165861 | (Empty Backbone)Omega2 Basic cloning vector GB2.0 compatible P[acman]-derived backbone. Allows for copy induction from low to high using AutoFOS or similar. Requires EPI300 bacterial strain (DH10B Derived) or similar. uses blue-white screening | Koen Venken | Matinyan et al Cell Rep. 2021 Sep 14;36(11):109700. doi: 10.1016/j.celrep.2021.109700. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/336788/ef23f1ff-d79f-4b80-be57-71defb40dffa/addgene-plasmid-165861-sequence-336788.gbk | full | LOCUS sequence-336788- 8793 bp DNA circular SYN 26-AUG-2024
DEFINITION Omega2 Basic cloning vector GB2.0 compatible P[acman]-derived
backbone. Allows for copy induction from low to high using AutoFOS
or similar. Requires EPI300 bacterial strain (DH10B Derived) or
similar. uses blue-white screening.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8793)
AUTHORS Matinyan N, Karkhanis MS, Gonzalez Y, Jain A, Saltzman A,
Malovannaya A, Sarrion-Perdigones A, Dierick HA, Venken KJT
TITLE Multiplexed drug-based selection and counterselection genetic
manipulations in Drosophila.
JOURNAL Cell Rep. 2021 Sep 14;36(11):109700. doi:
10.1016/j.celrep.2021.109700.
PUBMED 34525356
REFERENCE 2 (bases 1 to 8793)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Cell Rep.";
date: "2021-09-14"; volume: "36(11):109700. doi"; pages: "
10.1016/j.celrep.2021.109700"
FEATURES Location/Qualifiers
source 1..8793
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS complement(392..1363)
/codon_start=1
/gene="sopB"
/product="partitioning protein for the bacterial F plasmid"
/label=sopB
/translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
CDS complement(1363..2538)
/codon_start=1
/gene="sopA"
/product="partitioning protein for the bacterial F plasmid"
/label=sopA
/translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
misc_feature 2864..3114
/gene="incC"
/label=incC
/note="incompatibility region of the bacterial F plasmid"
CDS complement(3117..3872)
/codon_start=1
/gene="repE"
/product="replication initiation protein for the bacterial
F plasmid"
/label=repE
/translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
rep_origin 3963..4182
/label=ori2
/note="secondary origin of replication for the bacterial F
plasmid; also known as oriS"
rep_origin 4269..4904
/label=oriV
/note="incP origin of replication"
promoter 5839..5941
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
CDS 5942..6601
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind complement(6008..6027)
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
protein_bind 6741..6810
/label=attB
/bound_moiety="phage phi-C31 integrase"
/note="attB site for the phi-C31 integrase (Groth et al.,
2000)"
misc_feature 7025..7257
/label=P element 3' end
/note="P element 3' end"
promoter 7291..7338
/label=EM7 promoter
/note="synthetic bacterial promoter "
promoter 7392..7410
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 7392..7409
/label=SP6
/note="SP6 promoter, forward primer"
misc_feature 7417..7473
/label=MCS
/note="pUC18/19 multiple cloning site"
primer_bind complement(7475..7494)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(7476..7494)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(7501..7518)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(7501..7517)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(7510..7532)
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind complement(7726..7745)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
misc_feature 7778..8363
/label=P element 5' end
misc_feature join(8639..8793,1..319)
/gene="sopC"
/label=sopC
/note="centromere-like partitioning region of the bacterial
F plasmid"
ORIGIN
1 agactaataa tcagaccgac gatacgagtg ggaccgtggt cccagactaa taatcagacc
61 gacgatacga gtgggaccat ggtcccagac taataatcag accgacgata cgagtgggac
121 cgtggtccca gtctgattat cagaccgacg atacgagtgg gaccgtggtc ccagactaat
181 aatcagaccg acgatacgag tgggaccgtg gtcccagact aataatcaga ccgacgatac
241 gagtgggacc gtggtcccag tctgattatc agaccgacga tacaagtgga acagtgggcc
301 cagagagaat attcaggcca gttatgcttt ctggcctgta acaaaggaca ttaagtaaag
361 acagataaac gtagactaaa acgtggtcgc atcagggtgc tggcttttca agttccttaa
421 gaatggcctc aattttctct atacactcag ttggaacacg cgacctgtcc aggttaagca
481 ccattttatc gcccttatac aatactgtcg ctccaggagc aaactgatgt cgtgagctta
541 aactagttct tgatgcagat gacgttttaa gcacagaagt taaaagagtg ataacttctt
601 cagcttcaaa tatcacccca gcttttttct gctcatgaag gttagatgcc tgctgcttaa
661 gtaattcctc tttatctgta aaggcttttt gaagtgcatc acctgaccgg gcagatagtt
721 caccggggtg agaaaaaaga gcaacaactg atttaggcaa tttggcggtg ttgatacagc
781 gggtaataat cttacgtgaa atattttccg catcagccag cgcagaaata tttccagcaa
841 attcattctg caatcggctt gcataacgct gaccacgttc ataagcactt gttgggcgat
901 aatcgttacc caatctggat aatgcagcca tctgctcatc atccagctcg ccaaccagaa
961 cacgataatc actttcggta agtgcagcag ctttacgacg gcgactccca tcggcaattt
1021 ctatgacacc agatactctt cgaccgaacg ccggtgtctg ttgaccagtc agtagaaaag
1081 aagggatgag atcatccagt gcgtcctcag taagcagctc ctggtcacgt tcattacctg
1141 accatacccg agaggtcttt tcaacactat caccccggag cacttcaaga gtaaacttca
1201 catcccgacc acatacaggc aaagtaatgg cattaccgcg agccattact cctacgcgcg
1261 caattaacga atccaccatc ggggcagctg gtgtcgataa cgaagtatct tcaaccggtt
1321 gagtattgag cgtatgtttt ggaataacag gcgcacgctt cattatctaa tctcccagcg
1381 tggtttaatc agacgatcga aaatttcatt gcagacaggt tcccaaatag aaagagcatt
1441 tctccaggca ccagttgaag agcgttgatc aatggcctgt tcaaaaacag ttctcatccg
1501 gatctgacct ttaccaactt catccgtttc acgtacaaca ttttttagaa ccatgcttcc
1561 ccaggcatcc cgaatttgct cctccatcca cggggactga gagccattac tattgctgta
1621 tttggtaagc aaaatacgta catcaggctc gaacccttta agatcaacgt tcttgagcag
1681 atcacgaagc atatcgaaaa actgcagtgc ggaggtgtag tcaaacaact cagcaggcgt
1741 gggaacaatc agcacatcag cagcacatac gacattaatc gtgccgatac ccaggttagg
1801 cgcgctgtca ataactatga catcatagtc atgagcaaca gtttcaatgg ccagtcggag
1861 catcaggtgt ggatcggtgg gcagtttacc ttcatcaaat ttgcccatta actcagtttc
1921 aatacggtgc agagccagac aggaaggaat aatgtcaagc cccggccagc aagtgggctt
1981 tattgcataa gtgacatcgt ccttttcccc aagatagaaa ggcaggagag tatcttctgc
2041 atgaatatga agatctggta cccatccgtg atacattgag gctgttccct gggggtcgtt
2101 accttccacg agcaaaacac gtagcccctt cagagccaga tcctgagcaa gatgaacaga
2161 aactgaggtt ttgtaaacgc cacctttatg ggcagcaacc ccgatcaccg gtggaaatac
2221 atcttcagca cgtcgcaatc gcgtaccaaa cacatcacgc atatgattaa tttgttcaat
2281 tgtataacca acacgttgct caacccgtcc tcgaatttcc atatccgggt gcggtagtcg
2341 ccctgctttc tcggcatctc tgatagcctg agaagaaacc ccaactaaat ccgctgcttc
2401 acctattctc cagcgccggg ttattttcct cgcttccggg ctgtcatcat taaactgtgc
2461 aatggcgata gccttcgtca tttcatgacc agcgtttatg cactggttaa gtgtttccat
2521 gagtttcatt ctgaacatcc tttaatcatt gctttgcgtt tttttattaa atcttgcaat
2581 ttactgcaaa gcaacaacaa aatcgcaaag tcatcaaaaa accgcaaagt tgtttaaaat
2641 aagagcaaca ctacaaaagg agataagaag agcacatacc tcagtcactt attatcacta
2701 gcgctcgccg cagccgtgta accgagcata gcgagcgaac tggcgaggaa gcaaagaaga
2761 actgttctgt cagatagctc ttacgctcag cgcaagaaga aatatccacc gtgggaaaaa
2821 ctccaggtag aggtacacac gcggatagcc aattcagagt aataaactgt gataatcaac
2881 cctcatcaat gatgacgaac taacccccga tatcaggtca catgacgaag ggaaagagaa
2941 ggaaatcaac tgtgacaaac tgccctcaaa tttggcttcc ttaaaaatta cagttcaaaa
3001 agtatgagaa aatccatgca ggctgaagga aacagcaaaa ctgtgacaaa ttaccctcag
3061 taggtcagaa caaatgtgac gaaccaccct caaatctgtg acagataacc ctcagactat
3121 cctgtcgtca tggaagtgat atcgcggaag gaaaatacga tatgagtcgt ctggcggcct
3181 ttctttttct caatgtatga gaggcgcatt ggagttctgc tgttgatctc attaacacag
3241 acctgcagga agcggcggcg gaagtcaggc atacgctggt aactttgagg cagctggtaa
3301 cgctctatga tccagtcgat tttcagagac acgatgcctg agccatccgg cttacgatac
3361 tgacacaggg attcgtataa acgcatggca tacggattgg tgatttcttt tgtttcacta
3421 agccgaaact gcgtaaaccg gttctgtaac ccgataaaga agggaatgag atatgggttg
3481 atatgtacac tgtaaagccc tctggatgga ctgtgcgcac gtttgataaa ccaaggaaaa
3541 gattcatagc ctttttcatc gccggcatcc tcttcagggc gataaaaaac cacttccttc
3601 cccgcgaaac tcttcaatgc ctgccgtata tccttactgg cttccgcaga ggtcaatccg
3661 aatatttcag catatttagc aacatggatc tcgcagatac cgtcatgttc ctgtagggtg
3721 ccatcagatt ttctgatctg gtcaacgaac agatacagca tacgtttttg atcccgggag
3781 agactatatg ccgcctcagt gaggtcgttt gactggacga ttcgcgggct atttttacgt
3841 ttcttgtgat tgataaccgc tgtttccgcc atgacagatc catgtgaagt gtgacaagtt
3901 tttagattgt cacactaaat aaaaaagagt caataagcag ggataacttt gtgaaaaaac
3961 agcttcttct gagggcaatt tgtcacaggg ttaagggcaa tttgtcacag acaggactgt
4021 catttgaggg tgatttgtca cactgaaagg gcaatttgtc acaacacctt ctctagaacc
4081 agcatggata aaggcctaca aggcgctcta aaaaagaaga tctaaaaact ataaaaaaaa
4141 taattataaa aatatccccg tggataagtg gataacccca agggaagttt tttcaggcat
4201 cgtgtgtaag cagaatatat aagtgctgtt ccctggtgct tcctcgctca ctcgaaattc
4261 ccctttccga cgctcaccgg gctggttgcc ctcgccgctg ggctggcggc cgtctatggc
4321 cctgcaaacg cgccagaaac gccgtcgaag ccgtgtgcga gacaccgcgg ccggccgccg
4381 gcgttgtgga tacctcgcgg aaaacttggc cctcactgac agatgagggg cggacgttga
4441 cacttgaggg gccgactcac ccggcgcggc gttgacagat gaggggcagg ctcgatttcg
4501 gccggcgacg tggagctggc cagcctcgca aatcggcgaa aacgcctgat tttacgcgag
4561 tttcccacag atgatgtgga caagcctggg gataagtgcc ctgcggtatt gacacttgag
4621 gggcgcgact actgacagat gaggggcgcg atccttgaca cttgaggggc agagtgctga
4681 cagatgaggg gcgcacctat tgacatttga ggggctgtcc acaggcagaa aatccagcat
4741 ttgcaagggt ttccgcccgt ttttcggcca ccgctaacct gtcttttaac ctgcttttaa
4801 accaatattt ataaaccttg tttttaacca gggctgcgcc ctgtgcgcgt gaccgcgcac
4861 gccgaagggg ggtgcccccc cttctcgaac cctcccggcc cgctaacgct ggcctcccat
4921 ccccccaggg gctgcgcccc tcggccgcga acggcctcac cccaaaaatg gcaggggatc
4981 tcgagggctt cgccctgtcg ctcgactgcg gcgagcacta ctggctgtaa aaggacagac
5041 cacatcatgg ttctgtgttc attaggttgt tctgtccatt gctgacataa tccgctccac
5101 ttcaacgtaa caccgcacga agatttctat tgttcctgaa ggcatattca aatcgttttc
5161 gttaccgctt gcaggcatca tgacagaaca ctacttccta taaacgctac acaggctcct
5221 gagattaata atgcggatct ctacgataat gggagatttt cccgactgtt tcgttcgctt
5281 ctcagtggat aacagccagc ttctctgttt aacagacaaa aacagcatat ccactcagtt
5341 ccacatttcc atataaaggc caaggcattt attctcagga taattgtttc agcatcgcaa
5401 ccgcatcaga ctccggcatc gcaaactgca cccggtgccg ggcagccaca tccagcgcaa
5461 aaaccttcgt gtagacttcc gttgaactga tggacttatg tcccatcagg ctttgcagaa
5521 ctttcagcgg tataccggca tacagcatgt gcatcgcata ggaatggcgg aacgtatgtg
5581 gtgtgaccgg aacagagaac gtcacaccgt cagcagcagc ggcggcaacc gcctccccaa
5641 tccaggtcct gaccgttctg tccgtcactt cccagatccg cgctttctct gtccttcctg
5701 tgcgacggtt acgccgctcc atgagcttat cgcgaataaa tacctgtgac ggaagatcac
5761 ttcgcagaat aaataaatcc tggtgtccct gttgataccg ggaagccctg ggccaacttt
5821 tggcgaaaat gagagtgatg atcggcacgt aagaggttcc aactttcacc ataatgaaat
5881 aagatcacta ccgggcgtat tttttgagtt atcgagattt tcaggagcta aggaagctaa
5941 aatggagaaa aaaatcactg gatataccac cgttgatata tcccaatggc atcgtaaaga
6001 acattttgag gcatttcagt cagttgctca atgtacctat aaccagaccg ttcagctgga
6061 tattacggcc tttttaaaga ccgtaaagaa aaataagcac aagttttatc cggcctttat
6121 tcacattctt gcccgcctga tgaatgctca tccggaattc cgtatggcaa tgaaagacgg
6181 tgagctggtg atatgggata gtgttcaccc ttgttacacc gttttccatg agcaaactga
6241 aacgttttca tcgctctgga gtgaatacca cgacgatttc cggcagtttc tacacatata
6301 ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag ggtttattga
6361 gaatatgttt ttcgtgtcag ccaatccctg ggtgagtttc accagttttg atttaaacgt
6421 ggccaatatg gacaacttct tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg
6481 cgacaaggtg ctgatgccgc tggcgattca ggttcatcat gccgtttgtg atggcttcca
6541 tgtcggcaga atgcttaatg aattacaaca gtactgcgat gagtggcagg gcggggcgta
6601 atttttttaa ggcagttatt ggtgccctta aacgcctggt tgctacgcct gaataagtga
6661 taataagcgg atgaatggca gaaattcgat gataagctgt caaacatgat gttgaattgg
6721 tcgacgatgt aggtcactgt ctcgaagccg cggtgcgggt gccagggcgt gcccttgggc
6781 tccccgggcg cgtactccac ctcacccatc tggtccatca tgatgaacgg gtcgaggtgg
6841 cggtagttga tcccggcgaa cgcgcggcgc accgggaagc cctcgccctc gaaaccgctg
6901 ggcgcggtgg tcacggtgag cacgggacgt gcgacggcgt cggcgggtgc ggatacgcgg
6961 ggcagcgtca gcgggttctc gacggtcacg gcgggcatgt cgatcacgat gagaatggcc
7021 agaccatgat gaaataacat aaggtggtcc cgtcggcaag agacatccac ttaacgtatg
7081 cttgcaataa gtgcgagtga aaggaatagt attctgagtg tcgtattgag tctgagtgag
7141 acagcgatat gattgttgat taacccttag catgtccgtg gggtttgaat taactcataa
7201 tattaattag acgaaattat ttttaaagtt ttatttttaa taatttgcga gtacgcataa
7261 ctgcaggtct cagtcaggag agagacgcct gttgacaatt aatcatcggc atagtatatc
7321 ggcatagtat aatacgacaa ggtgaggaac taaacccagg aggcagatca tgaccatgat
7381 tacgccaagc tatttaggtg acactataga atactcaagc ttgcatgcct gcaggtcgac
7441 tctagaggat ccccgggtac cgagctcgaa ttcgccctat agtgagtcgt attacaattc
7501 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
7561 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
7621 cccttcccaa cagttgcgca gcctgaatgg cgaatgacgc ctgatgcggt attttctcct
7681 tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga
7741 tgccgcatag cgtctctcgc ttgagacctg cagtgacgct gctgctctaa acgacgcatt
7801 tcgtactcca aagtacgaat tttttccctc aagctcttat tttcattaaa caatgaacag
7861 gacctaacgc acagtcacgt tattgtttac ataaatgatt ttttttacta ttcaaactta
7921 ctctgtttgt gtactcccac tggtatagcc ttcttttatc ttttctggtt caggctctat
7981 cactttacta ggtacggcat ctgcgttgag tcgcctcctt ttaaatgtct gaccttttgc
8041 aggtgcagcc ttccactgcg aatcattaaa gtgggtatca caaatttggg agttttcacc
8101 aaggctgcac ccaaggctct gctcccacaa ttttctctta atagcacact tcggcacgtg
8161 aattaatttt actccagtca cagctttgca gcaaaatttg caatatttca ttttttttta
8221 ttccacgtaa gggttaatgt tttcaaaaaa aaattcgtcc gcacacaacc tttcctctca
8281 acaagcaaac gtgcactgaa tttaagtgta tacttcggta agcttcggct atcgacggga
8341 ccaccttatg ttatttcatc atgggccaga cccacgtagt ccagactagt ggatctcgac
8401 agcgacacac ttgcatcgga tgcagcccgg ttaacgtgcc ggcacggcct gggtaaccag
8461 gtattttgtc cacataaccg tgcgcaaaat gttgtggata agcaggacac agcagcaatc
8521 cacagcaggc atacaaccgc acaccgaggt tactccgttc tacaggttac gacgacatgt
8581 caatacttgc ccttgacagg cattgatgga atcgtagtct cacgctgata gtctgatcga
8641 caatacaagt gggaccgtgg tcccagaccg ataatcagac cgacaacacg agtgggatcg
8701 tggtcccaga ctaataatca gaccgacgat acgagtggga ccgtggtccc agactaataa
8761 tcagaccgac gatacgagtg ggaccgtggt tcc
//
| {
"Vector backbone": "P[acman]",
"Backbone size w/o insert (bp)": "",
"Vector type": "Synthetic Biology",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Chloramphenicol, 25 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "EPI300",
"Copy number": "Low Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Multiplexed drug-based selection and counterselection genetic manipulations in Drosophila",
"Publication": "Cell Rep. 2021 Sep 14;36(11):109700. doi: 10.1016/j.celrep.2021.109700.",
"DOI": "10.1016/j.celrep.2021.109700.",
"Authors": "Matinyan N, Karkhanis MS, Gonzalez Y, Jain A, Saltzman A, Malovannaya A, Sarrion-Perdigones A, Dierick HA, Venken KJT /jcelrep",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/34525356",
"PubMed ID": "PubMed 34525356"
} | {
"Gene/Insert name": "None",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pEMJ_042 | 214061 | T7 inducible LanTERN (pRSET) | Pamela Silver | Jones et al ACS Synth Biol. 2024 Feb 20. doi: 10.1021/acssynbio.3c00600. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/424313/6547e049-012f-4525-ae4a-7ce836a49a59/addgene-plasmid-214061-sequence-424313.gbk | full | LOCUS sequence-424313- 4212 bp DNA circular SYN 25-AUG-2024
DEFINITION T7 inducible LanTERN (pRSET).
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4212)
AUTHORS Jones EM, Su Y, Sander C, Justman QA, Springer M, Silver PA
TITLE LanTERN: A Fluorescent Sensor That Specifically Responds to
Lanthanides.
JOURNAL ACS Synth Biol. 2024 Feb 20. doi: 10.1021/acssynbio.3c00600.
PUBMED 38377571
REFERENCE 2 (bases 1 to 4212)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth
Biol. 2024 Feb 20. doi: 10.1021/acssynbio.3c00600."
FEATURES Location/Qualifiers
source 1..4212
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 220..237
/label=L4440
/note="L4440 vector, forward primer"
primer_bind 322..341
/label=T7
/note="T7 promoter, forward primer"
promoter 322..340
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
RBS 372..394
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 414..431
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 435..467
/codon_start=1
/product="leader peptide from bacteriophage T7 gene 10"
/label=T7 tag (gene 10 leader)
/note="promotes efficient translation in E. coli"
/translation="MASMTGGQQMG"
CDS 471..494
/codon_start=1
/product="Xpress(TM) epitope tag, including an enterokinase
recognition and cleavage site"
/label=Xpress(TM) tag
/translation="DLYDDDDK"
regulatory 501..510
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 522..578
/codon_start=1
/product="calmodulin-binding peptide from avian smooth
muscle myosin light chain kinase"
/label=calmodulin-binding peptide
/translation="RRKWNKTGHAVRAIGRLSS"
primer_bind complement(1915..1933)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 1929..1976
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
rep_origin 2073..2528
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2160..2179)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2370..2391
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 2554..2658
/gene="bla"
/label=AmpR promoter
CDS 2659..3519
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(2877..2896)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin join(3690..4212,1..66)
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4179..4198
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
61 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
121 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
181 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
241 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
301 aggatctcga tcccgcgaaa ttaatacgac tcactatagg gagaccacaa cggtttccct
361 ctagaaataa ttttgtttaa ctttaagaag gagatataca tatgcggggt tctcatcatc
421 atcatcatca tggtatggct agcatgactg gtggacagca aatgggtcgg gatctgtacg
481 acgatgacga taaggatctc gccaccatgg tcgactcatc acgtcgtaag tggaataaga
541 caggtcacgc agtcagagct ataggtcggc tgagctcact cgagaacgtg tatattaccg
601 cggataaaca gaaaaacggc attaaagcga actttcacgt gcgccataac gtggaagatg
661 gcagcgtgca gctggcggat cattatcagc agaacacccc gattggcgat ggcccggtgc
721 tgctgccgga taaccattat ctgagcaccc agaccaagct gagcaaagat ccgaacgaaa
781 aacgcgatca catggtgctg ctggaatttg tgaccgcagc gggcattaca cacggcatgg
841 atgaactgta tggcggcacc ggcggcagcg cgagccaggg cgaagaactg tttaccggcg
901 tggtgccgat tctggtggaa ctggatggcg atgtgaacgg ccataaattt agcgtgcgcg
961 gcgaaggcga aggcgatgcg accattggca aactgaccct gaaatttatt tccaccaccg
1021 gcaaactacc ggtgccgtgg ccgaccctgg tgaccacctt aacctatggc gtgcagtgct
1081 ttagccgcta tccggatcat atgaaacgcc atgatttttt taaaagcgcg atgccggaag
1141 gctatgtgca ggaacgcacc attagcttta aagatgatgg caaatataaa acccgcgcgg
1201 tggtgaaatt tgaaggcgat accctggtga accgcattga actgaaaggc accgatttta
1261 aagaagatgg caacattctg gggcataaac tggaatataa ccttcctgac caactgactg
1321 aagagcagat cgcagaattt aaagaggctt tctccctatt tgacccggac ggggatggga
1381 caatagatac caaggagctg gggacggtga tgcggtctct ggggcagaac cccacagaag
1441 cagagctgca ggacatgatc aatgaagtag atccggacgg tgatggcaca atcgacttcc
1501 ctgagttcct gacaatgatg gcaagaaaaa tgaaagacac ggactcggaa gaagaaatta
1561 gagaagcgtt cagggtgttt gatccggatg gcgatggcta catcgacgca gcagagcttc
1621 gccacgtgat gacaaacctt ggagagaagt taacagatga agaggttgat gaaatgatca
1681 gggaagcaga cccggatggg gatggtcagg tagattacga agagtttgta caaatgatga
1741 cagcgaagta agcgactcta gatcataatc agccatacca catttgtaga ggttttactt
1801 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattcga
1861 agcttgatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg
1921 agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga
1981 aaggaggaac tatatccgga tctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
2041 ccaacagttg cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc
2101 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
2161 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
2221 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
2281 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
2341 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
2401 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
2461 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
2521 tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
2581 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
2641 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
2701 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
2761 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
2821 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
2881 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
2941 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
3001 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
3061 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
3121 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
3181 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
3241 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
3301 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
3361 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
3421 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
3481 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
3541 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
3601 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
3661 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
3721 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
3781 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
3841 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
3901 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
3961 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
4021 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
4081 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
4141 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
4201 agtcctgtcg gg
//
| {
"Vector backbone": "pRSET",
"Backbone size w/o insert (bp)": "2955",
"Vector type": "Bacterial Expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "LanTERN: A Fluorescent Sensor That Specifically Responds to Lanthanides",
"Publication": "ACS Synth Biol. 2024 Feb 20. doi: 10.1021/acssynbio.3c00600.",
"DOI": "10.1021/acssynbio.3c00600.",
"Authors": "Jones EM, Su Y, Sander C, Justman QA, Springer M, Silver PA /acssynbioc",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/38377571",
"PubMed ID": "PubMed 38377571"
} | {
"Gene/Insert name": "LanTERN",
"Species": "Synthetic",
"Insert Size (bp)": "1257",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "T7",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Gibson Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "T7",
"3′ sequencing primer": "ggctgattatgatctagagtcgc"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pMT-DREDD-V5-His | 63749 | Expresses drosophila DREDD tagged with V5 and 6xHis epitopes, under control of metallothionein promoter | Cyril Gueydan | pMT-IMD-V5-His (unpublished) | https://media.addgene.org/snapgene-media/v2.0.0/sequences/108845/3f03c015-410d-4816-bc97-3a4eff7de14f/addgene-plasmid-63749-sequence-108845.gbk | full | LOCUS sequence-108845- 5030 bp DNA circular SYN 26-AUG-2024
DEFINITION Expresses drosophila DREDD tagged with V5 and 6xHis epitopes, under
control of metallothionein promoter.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5030)
TITLE pMT-IMD-V5-His
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 5030)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
FEATURES Location/Qualifiers
source 1..5030
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(29..51)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 151..170
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 364..386
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 378..395
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 379..395
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 407..833
/label=MT promoter
/note="Drosophila metallothionein promoter"
CDS 2422..2463
/codon_start=1
/product="epitope tag from simian virus 5"
/label=V5 tag
/translation="GKPIPNPLLGLDST"
CDS 2473..2490
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(2513..2530)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2532..2666
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2556..2575)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 2610..2629
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind complement(2810..2826)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2810..2826)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(2823..2845)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 2834..2850
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(2858..2888)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2903..2924
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(3041..3058)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3212..3800)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3292..3311)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3971..4831)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 4594..4613
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(4832..4936)
/gene="bla"
/label=AmpR promoter
primer_bind 5004..5022
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
ORIGIN
1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
361 tttcccagtc acgacgttgt aaaacgacgg ccagtgccag tgaattaatt cgttgcagga
421 caggatgtgg tgcccgatgt gactagctct ttgctgcagg ccgtcctatc ctctggttcc
481 gataagagac ccagaactcc ggccccccac cgcccaccgc cacccccata catatgtggt
541 acgcaagtaa gagtgcctgc gcatgcccca tgtgccccac caagagtttt gcatcccata
601 caagtcccca aagtggagaa ccgaaccaat tcttcgcggg cagaacaaaa gcttctgcac
661 acgtctccac tcgaatttgg agccggccgg cgtgtgcaaa agaggtgaat cgaacgaaag
721 acccgtgtgt aaagccgcgt ttccaaaatg tataaaaccg agagcatctg gccaatgtgc
781 atcagttgtg gtcagcagca aaatcaagtg aatcatctca gtgcaactaa aggggggatc
841 tagatcgggg tacctactag tccagtgtgg tggaattcca ccatggccgg atcaaacctg
901 ttgattcatc tagacaccat cgatcagaac gatctgatct acgtggaacg tgacatgaac
961 tttgcccaaa aggtgggcct ctgctttctg ctttatggcg acgaccactc ggatgccacc
1021 tacattctgc agaaactttt ggccatgaca cgatcagact tcccgcaaag tgatctactc
1081 ataaagttcg ccaagtcccg gccagaaacc tggagaagac atctcgtgga ggccctgtgc
1141 attattgggg cccgcaaggt gctccggaga ctgggtttct gctggcagga gctgcgaatg
1201 cactacttgc cgcatatcgc tgggatcacg ctgcatgtcc atcctctgtt gaagagcctc
1261 tacaggatgt gtgaggagtt gtcgttggtg cagagtggcc gcttgctcct agatgttcgc
1321 gaaaaggtgg agagccagca ggcaggagat ccacttcgct tctacgatcc cgcgtacctg
1381 gagatctttt tgctggactg gctgaccaga aggagcataa agttagggga cataaatgcc
1441 gcaggcagcg atgttcagct gctggtcggc catttgaagt ccaacggtct gcaggcgcaa
1501 gcaaatttgc tcaaagacac catcattagt aacgctccgg agccagatgc ggctggcact
1561 gctgcgatgg cagtcaagca agagattgaa tcggataacc agcagtcgta ctgttccaca
1621 caaatagatg ctcttaagtt gacccgggag aacgcaggaa tcgccttgat tatcaaccag
1681 cagaagtttc accggaatgt tagcagggat aatatgaaat ttctgtcgcc cgacccactg
1741 cgtagacggg atggcacgga tgtggataaa gaacgactaa tcgaggtgtt ttcctcgatg
1801 ggatacaatg tggaggcgta cgacaacgtg gatcacatgg gcatcataga gcgcattcgc
1861 agtgcgtgcg atagatcgct cgtgcgcgat tccctagtgg tcttcatcct gagccacggc
1921 ttcgaggagg ccgtctacgc atccaacagc atcgccatga agatcacgga tatcgaggac
1981 ttgctctgca gctacgacac cctgtactac aaaccgaagt tgctaatcat acaagcctgc
2041 caggagaagc ttgtccataa aaagaagcca aatgagctct ttaggataga cgtgaccacc
2101 gtgtcgccgg accagcacat cgacatgctg cgggccatgt ccacggtaaa tggatatgct
2161 gctctgcgcc acacccaaac aggtagctgg tttatcggga gcctgtgcga tgccatcgat
2221 cgccgttccg ccagtgaaca catcgccgat atcctgacga ttgtcaccaa cgaagtgtct
2281 aaaaagcgag gcagtaatga cgagtccatg gtgccgaatg ttaaaagcac atttcgccag
2341 cacgtgtact ttccacctcg tctggaattc tgcagatatc cagcacagtg gcggccgctc
2401 gagtctagag ggcccttcga aggtaagcct atccctaacc ctctcctcgg tctcgattct
2461 acgcgtaccg gtcatcatca ccatcaccat tgagtttaaa cccgctgatc agcctcgact
2521 gtgccttcta agatccagac atgataagat acattgatga gtttggacaa accacaacta
2581 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
2641 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
2701 ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtatgg
2761 ctgattatga tcagtcgacc tgcaggcatg caagcttggc gtaatcatgg tcatagctgt
2821 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
2881 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
2941 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
3001 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
3061 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
3121 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
3181 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
3241 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
3301 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
3361 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
3421 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
3481 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
3541 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
3601 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
3661 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
3721 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
3781 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
3841 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
3901 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
3961 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
4021 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
4081 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
4141 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
4201 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
4261 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
4321 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
4381 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
4441 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
4501 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
4561 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
4621 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
4681 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
4741 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
4801 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
4861 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
4921 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
4981 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
//
| {
"Vector backbone": "pMT-V5-His",
"Backbone size w/o insert (bp)": "3500",
"Vector type": "Insect Expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "Unknown"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "",
"Publication": "",
"DOI": "",
"Authors": "",
"PubMed Link": "",
"PubMed ID": ""
} | {
"Gene/Insert name": "Death related ced-3/Nedd2-like caspase isoform E",
"Species": "D. melanogaster (fly)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "Dredd",
"Promoter": "pMT",
"Tag / Fusion Protein": "V5-6XHis (C terminal on backbone)"
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "EcoRI (not destroyed)",
"3′ cloning site": "EcoRI (not destroyed)",
"5′ sequencing primer": "CATCTCAGTGCAACTAAA",
"3′ sequencing primer": "TAGAAGGCACAGTCGAGG"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pEGFP-C1-AR V7 | 86856 | High | Fluorescent human androgen-receptor splice variant 7, lacking the ligand-binding domain (fused to EGFP) | Szafran et al Prostate. 2017 Jan;77(1):82-93. doi: 10.1002/pros.23251. Epub 2016 Oct 4. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/386709/6f4f3de3-67b5-482c-a8c1-46b891106bfc/addgene-plasmid-86856-sequence-386709.gbk | full | LOCUS sequence-386709- 6633 bp DNA circular SYN 25-AUG-2024
DEFINITION Fluorescent human androgen-receptor splice variant 7, lacking the
ligand-binding domain (fused to EGFP).
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6633)
AUTHORS Szafran AT, Stephan C, Bolt M, Mancini MG, Marcelli M, Mancini MA
TITLE High-Content Screening Identifies Src Family Kinases as Potential
Regulators of AR-V7 Expression and Androgen-Independent Cell Growth.
JOURNAL Prostate. 2017 Jan;77(1):82-93. doi: 10.1002/pros.23251. Epub 2016
Oct 4.
PUBMED 27699828
REFERENCE 2 (bases 1 to 6633)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1002/pros.23251"; journalName: "Prostate"; date: "2017-01";
volume: "77"; issue: "1"; pages: "82-93"
FEATURES Location/Qualifiers
source 1..6633
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 61..364
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 365..568
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 519..539
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 607..616
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 613..1329
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 613..1329
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(658..679)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(919..938)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1266..1287
/label=EGFP-C
/note="EGFP, forward primer"
polyA_signal 3421..3542
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3458..3477)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3512..3531
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(3549..4004)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3686..3707)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 3898..3917
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 4031..4135
/gene="bla"
/label=AmpR promoter
promoter 4137..4494
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
enhancer 4137..4328
/label=SV40 enhancer
/note="enhancer for the SV40 early promoter (Herr, 1993)"
primer_bind complement(4160..4180)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 4345..4480
/label=SV40 ori
/note="SV40 origin of replication"
CDS 4529..5323
/codon_start=1
/product="aminoglycoside phosphotransferase"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
CDS 4529..5323
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(4583..4602)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 5193..5212
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(5511..5530)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 5555..5602
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 5931..6519
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6420..6439
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
1321 ctgtacaagt ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc
1381 atggaagtgc agttagggct gggaagggtc taccctcggc cgccgtccaa gacctaccga
1441 ggagctttcc agaatctgtt ccagagcgtg cgcgaagtga tccagaaccc gggccccagg
1501 cacccagagg ccgcgagcgc agcacctccc ggcgccagtt tgctgctgct gcagcagcag
1561 cagcagcagc agcagcagca gcagcagcag cagcagcagc agcagcaaga gactagcccc
1621 aggcagcagc agcagcagca gggtgaggat ggttctcccc aagcccatcg tagaggcccc
1681 acaggctacc tggtcctgga tgaggaacag caaccttcac agccgcagtc ggccctggag
1741 tgccaccccg agagaggttg cgtcccagag cctggagccg ccgtggccgc cagcaagggg
1801 ctgccgcagc agctgccagc acctccggac gaggatgact cagctgcccc atccacgttg
1861 tccctgctgg gccccacttt ccccggctta agcagctgct ccgctgacct taaagacatc
1921 ctgagcgagg ccagcaccat gcaactcctt cagcaacagc agcaggaagc agtatccgaa
1981 ggcagcagca gcgggagagc gagggaggcc tcgggggctc ccacttcctc caaggacaat
2041 tacttagggg gcacttcgac catttctgac aacgccaagg agttgtgtaa ggcagtgtcg
2101 gtgtccatgg gcctgggtgt ggaggcgttg gagcatctga gtccagggga acagcttcgg
2161 ggggattgca tgtacgcccc acttttggga gttccacccg ctgtgcgtcc cactccttgt
2221 gccccattgg ccgaatgcaa aggttctctg ctagacgaca gcgcaggcaa gagcactgaa
2281 gatactgctg agtattcccc tttcaaggga ggttacacca aagggctaga aggcgagagc
2341 ctaggctgct ctggcagcgc tgcagcaggg agctccggga cacttgaact gccgtctacc
2401 ctgtctctct acaagtccgg agcactggac gaggcagctg cgtaccagag tcgcgactac
2461 tacaactttc cactggctct ggccggaccg ccgccccctc cgccgcctcc ccatccccac
2521 gctcgcatca agctggagaa cccgctggac tacggcagcg cctgggcggc tgcggcggcg
2581 cagtgccgct atggggacct ggcgagcctg catggcgcgg gtgcagcggg acccggttct
2641 gggtcaccct cagccgccgc ttcctcatcc tggcacactc tcttcacagc cgaagaaggc
2701 cagttgtatg gaccgtgtgg tggtggtggg ggtggtggcg gcggcggcgg cggcggcggc
2761 ggcggcggcg gcggcggcgg cggcggcgag gcgggagctg tagcccccta cggctacact
2821 cggccccctc aggggctggc gggccaggaa agcgacttca ccgcacctga tgtgtggtac
2881 cctggcggca tggtgagcag agtgccctat cccagtccca cttgtgtcaa aagcgaaatg
2941 ggcccctgga tggatagcta ctccggacct tacggggaca tgcgtttgga gactgccagg
3001 gaccatgttt tgcccattga ctattacttt ccaccccaga agacctgcct gatctgtgga
3061 gatgaagctt ctgggtgtca ctatggagct ctcacatgtg gaagctgcaa ggtcttcttc
3121 aaaagagccg ctgaagggaa acagaagtac ctgtgcgcca gcagaaatga ttgcactatt
3181 gataaattcc gaaggaaaaa ttgtccatct tgtcgtcttc ggaaatgtta tgaagcaggg
3241 atgactctgg gagaaaaatt ccgggttggc aattgcaagc atctcaaaat gaccagaccc
3301 tgatctagat aactgatcat aatcagccat accacatttg tagaggtttt acttgcttta
3361 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
3421 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
3481 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
3541 taacgcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
3601 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
3661 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
3721 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
3781 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
3841 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
3901 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
3961 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg
4021 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
4081 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga
4141 ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
4201 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
4261 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
4321 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
4381 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
4441 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga
4501 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
4561 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
4621 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
4681 cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc
4741 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
4801 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
4861 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
4921 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
4981 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
5041 cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
5101 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
5161 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
5221 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
5281 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
5341 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
5401 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
5461 cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg
5521 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
5581 cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
5641 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
5701 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
5761 gccatagcct caggttactc atatatactt tagattgatt taaaacttca tttttaattt
5821 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
5881 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
5941 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
6001 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
6061 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
6121 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
6181 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
6241 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
6301 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
6361 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
6421 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
6481 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
6541 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
6601 gattctgtgg ataaccgtat taccgccatg cat
//
| {
"Vector backbone": "pEGFP-C1",
"Backbone size w/o insert (bp)": "4731",
"Vector type": "Mammalian Expression",
"Selectable markers": "Neomycin (select with G418)"
} | {
"Bacterial Resistance(s)": "Kanamycin, 50 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "High-Content Screening Identifies Src Family Kinases as Potential Regulators of AR-V7 Expression and Androgen-Independent Cell Growth",
"Publication": "Prostate. 2017 Jan;77(1):82-93. doi: 10.1002/pros.23251. Epub 2016 Oct 4.",
"DOI": "10.1002/pros.23251.",
"Authors": "Szafran AT, Stephan C, Bolt M, Mancini MG, Marcelli M, Mancini MA /pros",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/27699828",
"PubMed ID": "PubMed 27699828"
} | {
"Gene/Insert name": "Androgen receptor (AR)",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "1932",
"Mutation": "Alternative splice variant 7 (alteration/deletion of the C-terminus)",
"Entrez Gene": "AR",
"Promoter": "CMV",
"Tag / Fusion Protein": "EGFP (N terminal on backbone)"
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "Xma I (not destroyed)",
"3′ cloning site": "Xba I (not destroyed)",
"5′ sequencing primer": "EGFP-C",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
RLK X023_pECIA14 | 114821 | Prey vector RLK X023_pECIA14 should be used with bait vector RLK X023_pECIA2. | Youssef Belkhadir | Smakowska-Luzan et al Nature. 2018 Jan 18;553(7688):342-346. doi: 10.1038/nature25184. Epub 2018 Jan 10. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/270426/ec54cc88-7988-4bf4-b28d-dba1a13d4ca8/addgene-plasmid-114821-sequence-270426.gbk | full | LOCUS sequence-270426- 6134 bp DNA circular SYN 26-AUG-2024
DEFINITION Prey vector RLK X023_pECIA14 should be used with bait vector RLK
X023_pECIA2..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6134)
AUTHORS Smakowska-Luzan E, Mott GA, Parys K, Stegmann M, Howton TC,
Layeghifard M, Neuhold J, Lehner A, Kong J, Grunwald K, Weinberger
N, Satbhai SB, Mayer D, Busch W, Madalinski M, Stolt-Bergner P,
Provart NJ, Mukhtar MS, Zipfel C, Desveaux D, Guttman DS, Belkhadir
Y
TITLE An extracellular network of Arabidopsis leucine-rich repeat receptor
kinases.
JOURNAL Nature. 2018 Jan 18;553(7688):342-346. doi: 10.1038/nature25184.
Epub 2018 Jan 10.
PUBMED 29320478
REFERENCE 2 (bases 1 to 6134)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1038/nature25184"; journalName: "Nature"; date: "2018-01-18-
18"; volume: "553"; issue: "7688"; pages: "342-346"
FEATURES Location/Qualifiers
source 1..6134
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 1..17
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 23..449
/label=MT promoter
/note="Drosophila metallothionein promoter"
sig_peptide 467..520
/codon_start=1
/product="Drosophila BiP signal sequence"
/label=BiP signal sequence
/translation="MKLCILLAVVAFVGLSLG"
CDS 1430..1453
/codon_start=1
/product="recognition and cleavage site for human
rhinovirus 3C and PreScission proteases"
/label=HRV 3C site
/translation="LEVLFQGP"
CDS 3128..3151
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS 3167..3184
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(3207..3224)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 3257..3391
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3281..3300)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 3335..3354
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind complement(3535..3551)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3535..3551)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3548..3570)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3559..3575
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(3583..3613)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3628..3649
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(3766..3783)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3937..4525)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4017..4036)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4696..5556)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 5319..5338
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5557..5661)
/gene="bla"
/label=AmpR promoter
primer_bind 5729..5747
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(5785..5807)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 5907..5926
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind join(6120..6134,1..8)
/label=M13/pUC Forward
/note="In lacZ gene"
ORIGIN
1 gtaaaacgac ggccagtgaa ttaattcgtt gcaggacagg atgtggtgcc cgatgtgact
61 agctctttgc tgcaggccgt cctatcctct ggttccgata agagacccag aactccggcc
121 ccccaccgcc caccgccacc cccatacata tgtggtacgc aagtaagagt gcctgcgcat
181 gccccatgtg ccccaccaag agctttgcat cccatacaag tccccaaagt ggagaaccga
241 accaattctt cgcgggcaga acaaaagctt ctgcacacgt ctccactcga atttggagcc
301 ggccggcgtg tgcaaaagag gtgaatcgaa cgaaagaccc gtgtgtaaag ccgcgtttcc
361 aaaatgtata aaaccgagag catctggcca atgtgcatca gttgtggtca gcagcaaaat
421 caagtgaatc atctcagtgc aactaaaggg gggatccgat ctcaatatga agttatgcat
481 attactggcc gtcgtggcct ttgttggcct ctcgctcggg accacagacg gtgttcttct
541 tctctctttc cgttactcaa tcgttgacga tcctctttac gtttttcgga gctggagatt
601 cgacgacgag actccttgct cttggcgtgg tgtcacgtgc gatgcatctt cccggcacgt
661 gactgttctg tcacttccaa gctcgaacct taccggcaca ctaccttcaa atttgggttc
721 actcaattca cttcaaagac ttgatctttc caacaattcc atcaatgggt ctttcccggt
781 ttcgcttctc aacgcgacgg agcttcgatt tcttgatctg tccgataatc acatctccgg
841 tgcactaccg gcgagttttg gcgcgctttc gaacctccaa gtgttgaatc tctccgataa
901 ttccttcgtc ggcgaattac cgaacacatt aggatggaac cggaacttaa cggagatttc
961 acttcagaaa aactatttat ccggcgggat tccgggaggt tttaagtcga cggagtatct
1021 tgatctctcg tcaaatttga tcaaaggctc gttgccgtca catttcagag ggaatcgtct
1081 acgctatttc aacgcttcgt acaacagaat ctccggcgag attccgtcag gtttcgccga
1141 cgaaatcccg gaagacgcca ccgttgatct ctcattcaac caacttacag gtcaaatccc
1201 gggttttcgg gttctcgata accaagaatc caactctttc tccggtaacc cgggtctctg
1261 cggatccgac catgcaaaac acccttgtcg tgacggtgaa gcaacctctc cacctccatc
1321 gccgactcca aattctcctc ctgcattagc tgctatacca aatactattg gcttaaccaa
1381 tcacccaatt agctccaaaa ccggtccgaa atcaaaatgg gatcataaac tggaggtgct
1441 gttccaggga ccaagcggca atcccggcgg cgatctggcc ccccagatgc tgcgcgagct
1501 gcaggagacc aatgccgccc tgcaggacgt gcgcgagctg ctgcgccacc gcgtgaagga
1561 gatcaccttc ctgaagaaca ccgtgatgga gtgcgacgcc tgcggcatgc agccctctag
1621 aggctccggc tccatcatcc cagttgagga ggagaacccg gacttctgga accgcgaggc
1681 agccgaggcc ctgggtgccg ccaagaagct gcagcctgca cagacagccg ccaagaacct
1741 catcatcttc ctgggcgatg ggatgggggt gtctacggtg acagctgcca ggatcctaaa
1801 agggcagaag aaggacaaac tggggcctga gatacccctg gccatggacc gcttcccata
1861 tgtggctctg tccaagacat acaatgtaga caaacatgtg ccagacagtg gagccacagc
1921 cacggcctac ctgtgcgggg tcaagggcaa cttccagacc attggcttga gtgcagccgc
1981 ccgctttaac cagtgcaaca cgacacgcgg caacgaggtc atctccgtga tgaatcgggc
2041 caagaaagca gggaagtcag tgggagtggt aaccaccaca cgagtgcagc acgcctcgcc
2101 agccggcacc tacgcccaca cggtgaaccg caactggtac tcggacgccg acgtgcctgc
2161 ctcggcccgc caggaggggt gccaggacat cgctacgcag ctcatctcca acatggacat
2221 tgacgtgatc ctaggtggag gccgaaagta catgtttcgc atgggaaccc cagaccctga
2281 gtacccagat gactacagcc aaggtgggac caggctggac gggaagaatc tggtgcagga
2341 atggctggcg aagcgccagg gtgcccggta tgtgtggaac cgcactgagc tcatgcaggc
2401 ttccctggac ccgtctgtga cccatctcat gggtctcttt gagcctggag acatgaaata
2461 cgagatccac cgagactcca cactggaccc ctccctgatg gagatgacag aggctgccct
2521 gcgcctgctg agcaggaacc cccgcggctt cttcctcttc gtggagggtg gtcgcatcga
2581 ccatggtcat catgaaagca gggcttaccg ggcactgact gagacgatca tgttcgacga
2641 cgccattgag agggcgggcc agctcaccag cgaggaggac acgctgagcc tcgtcactgc
2701 cgaccactcc cacgtcttct ccttcggagg ctaccccctg cgagggagct ccatcttcgg
2761 gctggcccct ggcaaggccc gggacaggaa ggcctacacg gtcctcctat acggaaacgg
2821 tccaggctat gtgctcaagg acggcgcccg gccggatgtt accgagagcg agagcgggag
2881 ccccgagtat cggcagcagt cagcagtgcc cctggacgaa gagacccacg caggcgagga
2941 cgtggcggtg ttcgcgcgcg gcccgcaggc gcacctggtt cacggcgtgc aggagcagac
3001 cttcatagcg cacgtcatgg ccttcgccgc ctgcctggag ccctacaccg cctgcgacct
3061 ggcgcccccc gccggcacca ccgacgccgc gcacccgggt tctagaggaa gcacctccgg
3121 cgataacgat tacaaggatg atgatgataa ggccagcggc accggtcatc atcaccatca
3181 ccattgagtt taaacccgct gatcagcctc gactgtgcct tctaaggcct gagctcgctg
3241 atcagcctcg atcgaggatc cagacatgat aagatacatt gatgagtttg gacaaaccac
3301 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
3361 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
3421 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
3481 tatggctgat tatgatcagt cgacctgcag gcatgcaagc ttggcgtaat catggtcata
3541 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
3601 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
3661 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
3721 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
3781 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
3841 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
3901 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
3961 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
4021 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
4081 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
4141 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
4201 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
4261 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
4321 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
4381 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
4441 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
4501 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
4561 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
4621 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
4681 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
4741 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
4801 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
4861 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
4921 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
4981 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
5041 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
5101 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
5161 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
5221 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
5281 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
5341 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
5401 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
5461 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
5521 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
5581 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
5641 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
5701 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
5761 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
5821 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
5881 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
5941 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
6001 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
6061 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
6121 ccagtcacga cgtt
//
| {
"Vector backbone": "pECIA14",
"Backbone size w/o insert (bp)": "",
"Vector type": "Insect Expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "An extracellular network of Arabidopsis leucine-rich repeat receptor kinases",
"Publication": "Nature. 2018 Jan 18;553(7688):342-346. doi: 10.1038/nature25184. Epub 2018 Jan 10.",
"DOI": "10.1038/nature25184.",
"Authors": "Smakowska-Luzan E, Mott GA, Parys K, Stegmann M, Howton TC, Layeghifard M, Neuhold J, Lehner A, Kong J, Grunwald K, Weinberger N, Satbhai SB, Mayer D, Busch W, Madalinski M, Stolt-Bergner P, Provart NJ, Mukhtar MS, Zipfel C, Desveaux D, Guttman DS, Belkhadir Y /nature",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/29320478",
"PubMed ID": "PubMed 29320478"
} | {
"Gene/Insert name": "AT5G67280",
"Species": "A. thaliana (mustard weed)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "Metallothionein (Copper-Inducible)",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Ligation Independent Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pcDNA3.4-GFP-TEV-B55 | 217378 | expresses GFP tagged B55 in mammalian cells; tev cleavable | Wolfgang Peti | Padi et al Nature. 2024 Jan;625(7993):195-203. doi: 10.1038/s41586-023-06870-3. Epub 2023 Dec 20. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/430725/21bc6c57-d5aa-4b0c-9568-827db3e38d01/addgene-plasmid-217378-sequence-430725.gbk | full | LOCUS sequence-430725- 8190 bp DNA circular SYN 25-AUG-2024
DEFINITION expresses GFP tagged B55 in mammalian cells; tev cleavable.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8190)
AUTHORS Padi SKR, Vos MR, Godek RJ, Fuller JR, Kruse T, Hein JB, Nilsson J,
Kelker MS, Page R, Peti W
TITLE Cryo-EM structures of PP2A:B55-FAM122A and PP2A:B55-ARPP19.
JOURNAL Nature. 2024 Jan;625(7993):195-203. doi: 10.1038/s41586-023-06870-3.
Epub 2023 Dec 20.
PUBMED 38123684
REFERENCE 2 (bases 1 to 8190)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1038/s41586-023-06870-3"; journalName: "Nature"; date:
"2024-01"; volume: "625"; issue: "7993"; pages: "195-203"
FEATURES Location/Qualifiers
source 1..8190
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 50..429
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 430..633
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 584..604
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 630..654
/label=LNCX
/note="Human CMV promoter, forward primer"
CDS 775..1491
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 775..1491
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(820..841)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1081..1100)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1428..1449
/label=EGFP-C
/note="EGFP, forward primer"
CDS 1507..1527
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQG"
CDS 2887..2904
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
misc_feature 2967..3555
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(3020..3040)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(3438..3449)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(3576..3595)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 3620..3668
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 3870..4298
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3957..3976)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4167..4188
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(4307..4327)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 4312..4641
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 4492..4627
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4554..4573
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 4708..5502
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind 5372..5391
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 5678..5799
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5715..5734)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 5769..5788
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(5848..5864)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5848..5864)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(5861..5883)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 5872..5888
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(5896..5926)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5941..5962
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(6079..6096)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(6250..6838)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6330..6349)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(7009..7869)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 7632..7651
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7870..7974)
/gene="bla"
/label=AmpR promoter
primer_bind complement(8049..8068)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
601 cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg
661 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgg
721 actctagagg atcgaaccct tgccaccatg tataccatgg gctctgataa aattatggtg
781 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
841 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
901 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
961 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
1021 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
1081 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
1141 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
1201 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
1261 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
1321 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
1381 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
1441 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gggagcaggt
1501 gctggagaaa atctttattt tcaaggtcat atgggatccg caggagctgg aggagggaat
1561 gatattcagt ggtgtttttc tcaggtgaaa ggagcagtag atgatgatgt agcagaagca
1621 gatataattt ctacagtaga atttaatcat tctggagaat tactagcaac aggagataaa
1681 ggtggtagag ttgtcatctt tcaacaggag caggagaaca aaatccagtc tcatagcaga
1741 ggagaatata atgtttacag caccttccag agccatgaac cagagtttga ctacttgaaa
1801 agtttagaaa tagaagaaaa gatcaacaaa attaggtggt taccccagaa aaatgctgct
1861 cagtttttat tgtctaccaa tgataaaaca ataaaattat ggaaaatcag tgaaagggac
1921 aaaagaccag aagggtataa cttgaaagag gaggatggaa ggtatagaga tcctactaca
1981 gttactacac tacgagtgcc agtctttagg cctatggatc taatggttga ggccagtcca
2041 cgaagaatat ttgccaatgc tcatacatat cacatcaact caatttctat taatagtgat
2101 tatgaaacat atttatctgc agatgatttg cggattaatc tttggcatct ggaaattaca
2161 gacaggagtt ttaacattgt ggatatcaag cctgccaata tggaagagct aacagaggtg
2221 attacagcag cagaatttca tccaaacagc tgtaacacat ttgtatacag cagcagtaaa
2281 ggaactattc ggctatgtga catgagggca tctgccctct gtgatagaca ttctaaattg
2341 tttgaagaac ctgaagatcc cagtaacagg tcattttttt ccgaaatcat ctcctctatt
2401 tcggatgtaa aattcagcca tagtggtcga tatatgatga ctagagacta tttgtcagtc
2461 aaaatttggg acttaaatat ggaaaacagg cctgtggaaa cataccaggt gcatgaatac
2521 ctcagaagta aactctgttc actgtatgaa aatgactgca tatttgacaa atttgaatgt
2581 tgttggaatg gatctgacag tgttgtcatg actggatctt acaataattt cttcagaatg
2641 tttgacagaa acacaaagcg agacataacc ctagaagcat cgcgggaaaa caataagcct
2701 cgcacagttc tgaagcctcg caaagtctgt gcaagtggca agcgaaagaa agatgaaata
2761 agtgttgaca gcctagactt caataagaaa atccttcaca cagcctggca ccccaaggaa
2821 aatatcattg ccgtagctac tacaaacaat ctgtatatat ttcaagacaa agtgaattga
2881 ctcgagcacc accaccacca ccactgagat ccggctgcta aaagggttcg atccctaccg
2941 gttagtaatg agtttgatat ctcgacaatc aacctctgga ttacaaaatt tgtgaaagat
3001 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
3061 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
3121 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
3181 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
3241 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
3301 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
3361 agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
3421 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
3481 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
3541 gggccgcctc cccgcctgga acgggggagg ctaactgaaa cacggaagga gacaataccg
3601 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg gtgttgggtc
3661 gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag
3721 accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc
3781 gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcagatct
3841 gcgcagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
3901 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
3961 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
4021 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
4081 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
4141 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
4201 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
4261 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
4321 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
4381 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
4441 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
4501 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
4561 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
4621 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
4681 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
4741 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
4801 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
4861 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
4921 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
4981 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
5041 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
5101 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
5161 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
5221 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
5281 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
5341 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
5401 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
5461 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
5521 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
5581 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
5641 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
5701 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
5761 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
5821 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
5881 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
5941 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
6001 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
6061 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
6121 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
6181 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
6241 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
6301 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
6361 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
6421 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
6481 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
6541 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
6601 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
6661 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
6721 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
6781 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
6841 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
6901 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
6961 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
7021 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
7081 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
7141 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
7201 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
7261 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
7321 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
7381 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
7441 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
7501 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
7561 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
7621 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
7681 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
7741 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
7801 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
7861 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
7921 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
7981 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc
8041 actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt
8101 gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg
8161 accgacaatt gcatgaagaa tctgcttagg
//
| {
"Vector backbone": "pcDNA3.4",
"Backbone size w/o insert (bp)": "",
"Vector type": "Mammalian Expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "Unknown"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Cryo-EM structures of PP2A:B55-FAM122A and PP2A:B55-ARPP19",
"Publication": "Nature. 2024 Jan;625(7993):195-203. doi: 10.1038/s41586-023-06870-3. Epub 2023 Dec 20.",
"DOI": "10.1038/s41586-023-06870-3.",
"Authors": "Padi SKR, Vos MR, Godek RJ, Fuller JR, Kruse T, Hein JB, Nilsson J, Kelker MS, Page R, Peti W /s---",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/38123684",
"PubMed ID": "PubMed 38123684"
} | {
"Gene/Insert name": "B55",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "1341",
"Mutation": "",
"Entrez Gene": "PPP2R2A",
"Promoter": "",
"Tag / Fusion Protein": "EGFP TEV (N terminal on insert)"
} | {
"Cloning method": "Gibson Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "CMV forward",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pCMV-EPN-37 (deltaL) | 80117 | Myr1-12GS-I3-01-myc-Late3(delta) | Neil King | Votteler et al Nature. 2016 Dec 8;540(7632):292-295. doi: 10.1038/nature20607. Epub 2016 Nov 30. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/169491/de0cbcaf-e8c8-4705-af53-8217e31f081f/addgene-plasmid-80117-sequence-169491.gbk | full | LOCUS sequence-169491- 6294 bp DNA circular SYN 26-AUG-2024
DEFINITION Myr1-12GS-I3-01-myc-Late3(delta).
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6294)
AUTHORS Votteler J, Ogohara C, Yi S, Hsia Y, Nattermann U, Belnap DM, King
NP, Sundquist WI
TITLE Designed proteins induce the formation of nanocage-containing
extracellular vesicles.
JOURNAL Nature. 2016 Dec 8;540(7632):292-295. doi: 10.1038/nature20607. Epub
2016 Nov 30.
PUBMED 27919066
REFERENCE 2 (bases 1 to 6294)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1038/nature20607"; journalName: "Nature"; date: "2016-12-8- 8";
volume: "540"; issue: "7632"; pages: "292-295"
FEATURES Location/Qualifiers
source 1..6294
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 1..380
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 381..584
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 535..555
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 629..648
/label=T7
/note="T7 promoter, forward primer"
promoter 629..647
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 1370..1399
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
primer_bind complement(1577..1596)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
primer_bind complement(1651..1668)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 1657..1881
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 1927..2355
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2014..2033)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2224..2245
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2364..2384)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2369..2698
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2549..2684
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2611..2630
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2765..3559
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(2819..2838)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3429..3448
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 3733..3854
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3770..3789)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3824..3843
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(3903..3919)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3903..3919)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3916..3938)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3927..3943
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(3951..3981)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3996..4017
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4134..4151)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4305..4893)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4385..4404)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5064..5924)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 5687..5706
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5925..6029)
/gene="bla"
/label=AmpR promoter
primer_bind complement(6104..6123)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
661 gctagcgttt aaacgggccc tctagaatta tcgatccgga ggtaccatgg gtgcgcgtgc
721 gtctggttct aaatctggct ccggctctga ctctggtagc aagatcgaag agctgttcaa
781 gaaacacaag atcgttgccg tgctgcgtgc caatagtgtg gaagaagcga aaaagaaagc
841 gctggcggtt ttcctgggcg gcgttcatct gattgaaatt acctttaccg tgccggatgc
901 ggataccgtg attaaggaac tgagctttct gaaggaaatg ggcgcgatta ttggtgcggg
961 caccgtgacc agcgtggagc agtgccgtaa agcggtggaa agtggcgccg aattcattgt
1021 gagtccgcac ctggacgagg aaattagcca attttgcaag gagaagggtg tgttctatat
1081 gccaggcgtt atgaccccga ccgaactggt gaaagccatg aaactgggcc ataccatctt
1141 aaaactgttt ccgggtgagg tggtgggtcc gcagtttgtt aaagcgatga aaggtccgtt
1201 tccgaatgtg aaatttgtgc caaccggcgg tgttaatctg gacaatgtgt gcgaatggtt
1261 caaagcgggc gtgctggccg tgggcgtggg cagcgcgtta gtgaaaggca ccccggtgga
1321 agtggcggaa aaggccaagg cgttcgttga gaagattcgt ggctgcaccg aacagaaact
1381 gattagcgaa gaagatctgc caattcagca gaagtcccag cacaataaga gcgttgttca
1441 ggagacccca caaactcaga acctggccgc agctgcgtcc gagatcaaga aggaatataa
1501 tgttaaggaa aaggaccagg ttgaggatct gaacttggac tccttgtggg agtgactcga
1561 gaagcttgct agcagatctt tttccctctg ccaaaaatta tggggacatc atgaagcccc
1621 ttgagcactt aagtttaaac cgctgatcag cctcgactgt gccttctagt tgccagccat
1681 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
1741 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
1801 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
1861 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt
1921 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
1981 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
2041 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
2101 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
2161 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
2221 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
2281 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
2341 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
2401 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
2461 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
2521 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
2581 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
2641 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
2701 gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt
2761 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
2821 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
2881 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
2941 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
3001 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
3061 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
3121 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
3181 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
3241 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
3301 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
3361 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
3421 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
3481 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
3541 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
3601 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
3661 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
3721 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
3781 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
3841 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
3901 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
3961 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
4021 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
4081 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
4141 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
4201 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
4261 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
4321 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
4381 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
4441 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
4501 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
4561 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
4621 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
4681 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
4741 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
4801 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
4861 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
4921 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
4981 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
5041 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
5101 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
5161 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
5221 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
5281 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
5341 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
5401 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
5461 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
5521 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
5581 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
5641 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
5701 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
5761 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
5821 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
5881 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
5941 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
6001 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
6061 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
6121 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
6181 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
6241 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgtt
//
| {
"Vector backbone": "pCMV",
"Backbone size w/o insert (bp)": "5443",
"Vector type": "Mammalian Expression, Bacterial Expression",
"Selectable markers": "Neomycin (select with G418)"
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Designed proteins induce the formation of nanocage-containing extracellular vesicles",
"Publication": "Nature. 2016 Dec 8;540(7632):292-295. doi: 10.1038/nature20607. Epub 2016 Nov 30.",
"DOI": "10.1038/nature20607.",
"Authors": "Votteler J, Ogohara C, Yi S, Hsia Y, Nattermann U, Belnap DM, King NP, Sundquist WI /nature",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/27919066",
"PubMed ID": "PubMed 27919066"
} | {
"Gene/Insert name": "EPN-37 (L2)",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pMXs-Sox1 | 32931 | Low | Kevin Eggan | Son et al Cell Stem Cell. 2011 Sep 2;9(3):205-18. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/39839/4d114c5b-1c9b-4b93-bd97-1c39d6723378/addgene-plasmid-32931-sequence-39839.gbk | partial | LOCUS sequence-39839-e 883 bp DNA linear UNA 26-AUG-2024
DEFINITION natural linear DNA
ACCESSION .
VERSION .
KEYWORDS .
SOURCE natural DNA sequence
ORGANISM unspecified
REFERENCE 1 (bases 1 to 883)
AUTHORS Son EY, Ichida JK, Wainger BJ, Toma JS, Rafuse VF, Woolf CJ, Eggan K
TITLE Conversion of mouse and human fibroblasts into functional spinal
motor neurons.
JOURNAL Cell Stem Cell. 2011 Sep 2;9(3):205-18.
PUBMED 21852222
REFERENCE 2 (bases 1 to 883)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Cell Stem
Cell."; date: "2011-09-2"; volume: "9(3)"; pages: "205-18"
FEATURES Location/Qualifiers
source 1..883
/mol_type="genomic DNA"
/organism="unspecified"
protein_bind 45..69
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
ORIGIN
1 gggggtggac catcctctag actgccggat ctagctagtt aatcacaagt ttgtacaaaa
61 aagcaggctt gatgtacagc atgatgatgg agaccgacct gcactcgccc ggcggcgccc
121 aggcgcccac gaacctctcg ggcccggccg gggcgggcgg gggcggcggt gggggcgggg
181 gcggcggcgg cggcgggggc accaaggcca accaggatcg ggtcaagcgg cccatgaacg
241 ccttcatggt gtggtcccgc ggacagcggc gcaagatggc ccaggaaaac cccaagatgc
301 acaactcgga gatcagcaag cgcctcgggg ccgagtggaa ggtcatgtcc gaggccgaga
361 agcggccgtt catcgacgag gccaagagac tgcgcgcgct gcacatgaag gaacacccgg
421 attacaagta ccggccgcgc cgcaagacca agacgctgct caagaaggac aagtactcgc
481 tggccggcgg gctgctagcg gccggcgcgg gtggcggcgg cgcggccgtg gccatgggtg
541 tgggcgtggg cgtcggggcg gcggcggtgg gccagcgctt ggagagccca ggcggcgcgg
601 cgggaggagg ctacgcgcat gtcaacggct gggctaacgg cgcctacccc ggctcggtgg
661 ccgcggcggc tgcggccgcg gccatgatgc aggaggcaca gctggcctac gggcagcacc
721 caggcgcggg cggcgcgcac ccgcacgcac acccggcgca cccgcacccg caccatccgc
781 acgcgcatcc tcacaacccg cagcccatgc accgctacga catgggcgcg ctgcagtaca
841 gccccatctc caactctcag ggctacatga gcgcgtcgcc ttc
//
| {
"Vector backbone": "pMXs",
"Backbone size w/o insert (bp)": "4800",
"Vector type": "Mammalian Expression, Retroviral",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "Stbl3",
"Copy number": "Unknown"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Conversion of mouse and human fibroblasts into functional spinal motor neurons",
"Publication": "Cell Stem Cell. 2011 Sep 2;9(3):205-18.",
"DOI": "10.1016/j.stem.2011.07.014",
"Authors": "Son EY, Ichida JK, Wainger BJ, Toma JS, Rafuse VF, Woolf CJ, Eggan K",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/21852222",
"PubMed ID": "PubMed 21852222"
} | {
"Gene/Insert name": "Sox1",
"Species": "M. musculus (mouse)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "Sox1",
"Promoter": "5'LTR",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Gateway Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "pLXSN 5'",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pDONR223_OR2T33_WT | 81851 | Gateway Donor vector containing OR2T33 , part of the Target Accelerator Plasmid Collection. | Kim et al Cancer Discov. 2016 Jul;6(7):714-26. doi: 10.1158/2159-8290.CD-16-0160. Epub 2016 May 4. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/168749/1b50ae88-55d2-4eb9-b9cf-e7296c600296/addgene-plasmid-81851-sequence-168749.gbk | full | LOCUS sequence-168749- 3756 bp DNA circular SYN 26-AUG-2024
DEFINITION Gateway Donor vector containing OR2T33 , part of the Target
Accelerator Plasmid Collection..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3756)
AUTHORS Kim E, Ilic N, Shrestha Y, Zou L, Kamburov A, Zhu C, Yang X, Lubonja
R, Tran N, Nguyen C, Lawrence MS, Piccioni F, Bagul M, Doench JG,
Chouinard CR, Wu X, Hogstrom L, Natoli T, Tamayo P, Horn H, Corsello
SM, Lage K, Root DE, Subramanian A, Golub TR, Getz G, Boehm JS, Hahn
WC
TITLE Systematic Functional Interrogation of Rare Cancer Variants
Identifies Oncogenic Alleles.
JOURNAL Cancer Discov. 2016 Jul;6(7):714-26. doi:
10.1158/2159-8290.CD-16-0160. Epub 2016 May 4.
PUBMED 27147599
REFERENCE 2 (bases 1 to 3756)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1158/2159-8290.CD-16-0160"; journalName: "Cancer Discov"; date:
"2016-07"; volume: "6"; issue: "7"; pages: "714-26"
FEATURES Location/Qualifiers
source 1..3756
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 43..65
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 57..74
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 58..74
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 90..189
/gene="mutant version of attL"
/label=attL1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
protein_bind complement(1156..1255)
/gene="mutant version of attL"
/label=attL2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
primer_bind complement(1272..1291)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(1273..1291)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(1296..1312)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1296..1312)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
CDS 1743..2534
/codon_start=1
/gene="aadA"
/product="aminoglycoside adenylyltransferase (Murphy,
1985)"
/label=SmR
/note="confers resistance to spectinomycin and
streptomycin"
/translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
rep_origin 2627..3215
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3116..3135
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 3369..3386
/label=L4440
/note="L4440 vector, forward primer"
terminator complement(3545..3572)
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator complement(3664..3750)
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
ORIGIN
1 ggcagttccc tactctcgcg ttaacgctag catggatgtt ttcccagtca cgacgttgta
61 aaacgacggc cagtcttaag ctcgggcccc aaataatgat tttattttga ctgatagtga
121 cctgttcgtt gcaacaaatt gatgagcaat gcttttttat aatgccaact ttgtacaaaa
181 aagttggcac catggagatg agaaatacta ccccagattt tattctccta ggactcttta
241 accacaccag agcccaccaa gtcctcttca tgatggttct gagtatcgtt ttgacctccc
301 tgtttggcaa ttccctcatg attctcctga ttcactggga ccaccggctc cacacgccca
361 tgtacttcct cctgagccaa ctttccctca tggacatgat gctggtttcc accactgtgc
421 ccaaaatggc ggctgactac ttgaccggaa gtaaggccat ctcccgcgct ggctgtggtg
481 tgcagatctt cttcctcccc acactgggtg gtggagagtg cttcctctta gcagccatgg
541 cctatgaccg ctatgcggct gtctgccacc cactccgata tcccactctc atgagctggc
601 agctgtgcct gaggatgacc atgtcgtgtt ggctcctggg tgcagctgac gggctcctgc
661 aggctgttgt taccctgagc ttcccatatt gtggtgcaca cgagatcgat cacttcttct
721 gcgagacccc cgtgctggtg cgtttggctt gtgctgacac ttcagtcttc gaaaacgcca
781 tgtacatctg ctgtgtgtta atgctcctgg tccccttttc cctcatcctg tcctcctatg
841 gtctcatcct cgctgctgtt ctgcacatgc gctctacaga agcccgcaag aaggcctttg
901 ccacctgctc ttcacatgtg gctgtggtgg gactctttta tggagctgcc atttttacct
961 atatgagacc caaatcccat aggtccacta accatgacaa ggttgtgtca gccttctata
1021 ctatgttcac ccctttacta aaccccctca tctacagtgt gaagaacagt gaggtgaagg
1081 gagccctgaa acggtggctg gggacgtgtg taaacataaa acaccagcaa aatgaggccc
1141 acaggtcaag atgattgcca actttcttgt acaaagttgg cattataaga aagcattgct
1201 tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc
1261 cagctgatat cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct
1321 ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgcc
1381 tcctctagac cagccaggac agaaatgcct cgacttcgct gctacccaag gttgccgggt
1441 gacgcacacc gtggaaacgg atgaaggcac gaacccagtg gacataagcc tgttcggttc
1501 gtaagctgta atgcaagtag cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac
1561 gcagcggtgg taacggcgca gtggcggttt tcatggcttg ttatgactgt ttttttgggg
1621 tacagtctat gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga
1681 tgttatggag cagcaacgat gttacgcagc agggcagtcg ccctaaaaca aagttaaaca
1741 ttatgaggga agcggtgatc gccgaagtat cgactcaact atcagaggta gttggcgtca
1801 tcgagcgcca tctcgaaccg acgttgctgg ccgtacattt gtacggctcc gcagtggatg
1861 gcggcctgaa gccacacagt gatattgatt tgctggttac ggtgaccgta aggcttgatg
1921 aaacaacgcg gcgagctttg atcaacgacc ttttggaaac ttcggcttcc cctggagaga
1981 gcgagattct ccgcgctgta gaagtcacca ttgttgtgca cgacgacatc attccgtggc
2041 gttatccagc taagcgcgaa ctgcaatttg gagaatggca gcgcaatgac attcttgcag
2101 gtatcttcga gccagccacg atcgacattg atctggctat cttgctgaca aaagcaagag
2161 aacatagcgt tgccttggta ggtccagcgg cggaggaact ctttgatccg gttcctgaac
2221 aggatctatt tgaggcgcta aatgaaacct taacgctatg gaactcgccg cccgactggg
2281 ctggcgatga gcgaaatgta gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg
2341 gcaaaatcgc gccgaaggat gtcgctgccg actgggcaat ggagcgcctg ccggcccagt
2401 atcagcccgt catacttgaa gctagacagg cttatcttgg acaagaagaa gatcgcttgg
2461 cctcgcgcgc agatcagttg gaagaatttg tccactacgt gaaaggcgag atcaccaagg
2521 tagtcggcaa ataaccctcg agccacccat gaccaaaatc ccttaacgtg agttacgcgt
2581 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
2641 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
2701 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
2761 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
2821 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
2881 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
2941 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
3001 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
3061 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
3121 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
3181 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
3241 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
3301 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
3361 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
3421 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
3481 cgggcagtga gcgcaacgca attaatacgc gtaccgctag ccaggaagag tttgtagaaa
3541 cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg
3601 cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg
3661 cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca
3721 gtcttccgac tgagcctttc gttttatttg atgcct
//
| {
"Vector backbone": "pDONR223",
"Backbone size w/o insert (bp)": "2790",
"Vector type": "Gateway Entry vector",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Spectinomycin, 50 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Systematic Functional Interrogation of Rare Cancer Variants Identifies Oncogenic Alleles",
"Publication": "Cancer Discov. 2016 Jul;6(7):714-26. doi: 10.1158/2159-8290.CD-16-0160. Epub 2016 May 4.",
"DOI": "10.1158/2159-8290.CD-16-0160.",
"Authors": "Kim E, Ilic N, Shrestha Y, Zou L, Kamburov A, Zhu C, Yang X, Lubonja R, Tran N, Nguyen C, Lawrence MS, Piccioni F, Bagul M, Doench JG, Chouinard CR, Wu X, Hogstrom L, Natoli T, Tamayo P, Horn H, Corsello SM, Lage K, Root DE, Subramanian A, Golub TR, Getz G, Boehm JS, Hahn WC /-CD--",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/27147599",
"PubMed ID": "PubMed 27147599"
} | {
"Gene/Insert name": "OR2T33",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "WT",
"Entrez Gene": "OR2T33",
"Promoter": "None",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Gateway Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "M13pUC-fwd",
"3′ sequencing primer": "M13-Rev"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
||
shNg pA_RC3J1_CAGW | 92155 | Bicistronic AAV construct encoding GFP control and shRNA targeting Ng | Weifeng Xu | Han et al Cell Rep. 2017 Jan 24;18(4):892-904. doi: 10.1016/j.celrep.2016.12.084. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/177275/6185a33c-6143-4cc1-9f8d-f67d4901f01a/addgene-plasmid-92155-sequence-177275.gbk | full | LOCUS sequence-177275- 5934 bp DNA circular SYN 26-AUG-2024
DEFINITION Bicistronic AAV construct encoding GFP control and shRNA targeting
Ng.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5934)
AUTHORS Han KS, Cooke SF, Xu W
TITLE Experience-Dependent Equilibration of AMPAR-Mediated Synaptic
Transmission during the Critical Period.
JOURNAL Cell Rep. 2017 Jan 24;18(4):892-904. doi:
10.1016/j.celrep.2016.12.084.
PUBMED 28122240
REFERENCE 2 (bases 1 to 5934)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1016/j.celrep.2016.12"; journalName: "Cell Rep"; date:
"2017-01-24- 24"; volume: "18"; issue: "4"; pages: "892-904"
FEATURES Location/Qualifiers
source 1..5934
/mol_type="other DNA"
/organism="synthetic DNA construct"
repeat_region 1..130
/label=AAV2 ITR
promoter 196..410
/label=H1 promoter
/note="human H1 RNA promoter"
primer_bind 331..350
/label=H1
/note="Human H1 promoter, forward primer"
promoter 743..1020
/label=chicken beta-actin promoter
primer_bind 1300..1322
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 1384..1403
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
regulatory 1455..1464
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 1461..2180
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 1461..2177
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(1506..1527)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1767..1786)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 2114..2135
/label=EGFP-C
/note="EGFP, forward primer"
intron 2206..2338
/label=chimeric intron
/note="chimera between introns from human beta-globin and
immunoglobulin heavy chain genes"
primer_bind 2321..2340
/label=pMT2-F
/note="Synthetic intron, forward primer"
misc_feature 2378..2966
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(2431..2451)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(2849..2860)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
polyA_signal 3009..3130
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3020..3039)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 3074..3093
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
repeat_region 3292..3421
/label=AAV2 ITR
rep_origin complement(3658..4246)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3738..3757)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4417..5277)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 5040..5059
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5278..5382)
/gene="bla"
/label=AmpR promoter
rep_origin complement(5408..5863)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5545..5566)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 5757..5776
/label=F1ori-R
/note="F1 origin, reverse primer"
ORIGIN
1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct
181 aggaagatcg aattcgaacg ctgacgtcat caacccgctc caaggaatcg cgggcccagt
241 gtcactaggc gggaacaccc agcgcgcgtg cgccctggca ggaagatggc tgtgagggac
301 aggggagtgg cgccctgcaa tatttgcatg tcgctatgtg ttctgggaaa tcaccataaa
361 cgtgaaatgt ctttggattt gggaatctta taagttctgt atgagaccac agatgcccgt
421 gacaagactt ccctactgtt tcaagagaac agtagggaag tcttgtcact ttttggaaag
481 cttatcgaag taccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
541 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
601 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
661 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
721 ttagtcatcg ctattaccat ggtcgaggtg agccccacgt tctgcttcac tctccccatc
781 tcccccccct ccccaccccc aattttgtat ttatttattt tttaattatt ttgtgcagcg
841 atgggggcgg gggggggggg ggggcgcgcg ccaggcgggg cggggcgggg cgaggggcgg
901 ggcggggcga ggcggagagg tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt
961 ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg
1021 ggagtcgctg cgacgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
1081 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
1141 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
1201 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggctgtccgc
1261 ggggggacgg ctgccttcgg gggggacggg gcagggcggg gttcggcttc tggcgtgtga
1321 ccggcggctc tagagcctct gctaaccatg ttcatgcctt cttctttttc ctacagctcc
1381 tgggcaacgt gctggttatt gtgctgtctc atcattttgg caaagaattg gatcgggatc
1441 cccgggtacc ggtcgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc
1501 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
1561 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
1621 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
1681 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
1741 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
1801 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
1861 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
1921 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
1981 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
2041 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
2101 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
2161 tggacgagct gtacaagtaa agcggccccg gactcgaggc cgcaggtaag tatcaaggtt
2221 acaagacagg tttaaggaga ccaatagaaa ctgggcttgt cgagacagag aagactcttg
2281 cgtttctgat aggcacctat tggtcttact gacatccact ttgcctttct ctccacaggt
2341 gtcgagtgga gctcgcgact agatcaagct tatcgataat caacctctgg attacaaaat
2401 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
2461 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
2521 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
2581 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
2641 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
2701 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
2761 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
2821 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
2881 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
2941 gatctccctt tgggccgcct ccccgcatcg ataccgtcga cccgggcggc cgcttcgagc
3001 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
3061 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
3121 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggagatgtg
3181 ggaggttttt taaagcaagt aaaacctcta caaatgtggt aaaatcgata aggatcttcc
3241 tagagcatgg ctacgtagat aagtagcatg gcgggttaat cattaactac aaggaacccc
3301 tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac
3361 caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca
3421 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
3481 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
3541 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
3601 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
3661 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
3721 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
3781 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
3841 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
3901 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
3961 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
4021 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
4081 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
4141 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
4201 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
4261 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
4321 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
4381 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
4441 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
4501 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
4561 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
4621 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
4681 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
4741 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
4801 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
4861 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
4921 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
4981 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
5041 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
5101 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
5161 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
5221 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
5281 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
5341 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
5401 gccacctaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
5461 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
5521 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
5581 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
5641 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
5701 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
5761 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
5821 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcccattc gccattcagg
5881 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccag
//
| {
"Vector backbone": "unknown",
"Backbone size w/o insert (bp)": "",
"Vector type": "Mammalian Expression, AAV",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "NEB Stable",
"Copy number": "Unknown"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Experience-Dependent Equilibration of AMPAR-Mediated Synaptic Transmission during the Critical Period",
"Publication": "Cell Rep. 2017 Jan 24;18(4):892-904. doi: 10.1016/j.celrep.2016.12.084.",
"DOI": "10.1016/j.celrep.2016.12.084.",
"Authors": "Han KS, Cooke SF, Xu W /jcelrep",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/28122240",
"PubMed ID": "PubMed 28122240"
} | {
"Gene/Insert name": "shRNA targeting Ng",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "NRGN",
"Promoter": "H1",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Unknown",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "H1",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
mKate2-P2A-APEX2-TAPP1-PH | 67662 | Low | Mammalian expression vector driving APEX2-TAPP1 (Human TAPP1-PH domain, [marks PI(3,4)P2] fused to EM peroxidase marker). Also contains bicistronially expressed cytoplasmic red reporter. | Rob Parton | Ariotti et al Dev Cell. 2015 Nov 23;35(4):513-25. doi: 10.1016/j.devcel.2015.10.016. Epub 2015 Nov 12. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/117187/50521cfd-fae1-4298-b03a-cc764246c8af/addgene-plasmid-67662-sequence-117187.gbk | full | LOCUS sequence-117187- 6082 bp DNA circular SYN 25-AUG-2024
DEFINITION Mammalian expression vector driving APEX2-TAPP1 (Human TAPP1-PH
domain, [marks PI(3,4)P2] fused to EM peroxidase marker). Also
contains bicistronially expressed cytoplasmic red reporter..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6082)
AUTHORS Ariotti N, Hall TE, Rae J, Ferguson C, McMahon KA, Martel N, Webb
RE, Webb RI, Teasdale RD, Parton RG
TITLE Modular Detection of GFP-Labeled Proteins for Rapid Screening by
Electron Microscopy in Cells and Organisms.
JOURNAL Dev Cell. 2015 Nov 23;35(4):513-25. doi:
10.1016/j.devcel.2015.10.016. Epub 2015 Nov 12.
PUBMED 26585296
REFERENCE 2 (bases 1 to 6082)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1016/j.devcel.2015.10.016"; journalName: "Dev Cell"; date:
"2015-11-23- 23"; volume: "35"; issue: "4"; pages: "513-25"
FEATURES Location/Qualifiers
source 1..6082
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 35..53
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 35..52
/label=SP6
/note="SP6 promoter, forward primer"
protein_bind 108..132
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
regulatory 133..142
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 139..834
/codon_start=1
/product="monomeric far-red fluorescent protein (Shcherbo
et al., 2009)"
/label=mKate2
/note="mammalian codon-optimized"
/translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL
KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY
CDLPSKLGHR"
CDS 850..906
/codon_start=1
/product="2A peptide from porcine teschovirus-1
polyprotein"
/label=P2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="ATNFSLLKQAGDVEENPGP"
CDS 925..1671
/codon_start=1
/product="soybean ascorbate peroxidase, engineered to serve
as a monomeric tag for electron microscopy and live-cell
proteomics (Lam et al., 2015)"
/label=APEX2
/note="more active than the original APEX due to the A134P
mutation"
/translation="GKSYPTVSADYQDAVEKAKKKLRGFIAEKRCAPLMLRLAFHSAGT
FDKGTKTGGPFGTIKHPAELAHSANNGLDIAVRLLEPLKAEFPILSYADFYQLAGVVAV
EVTGGPKVPFHPGREDKPEPPPEGRLPDPTKGSDHLRDVFGKAMGLTDQDIVALSGGHT
IGAAHKERSGFEGPWTSNPLIFDNSYFTELLSGEKEGLLQLPSDKALLSDPVFRPLVDK
YAADEDAFFADYAEAHQKLSELGFADA"
protein_bind complement(1676..1700)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
protein_bind complement(2071..2091)
/gene="mutant version of attB"
/label=attB3
/bound_moiety="BP Clonase(TM)"
/note="core recombination site for the Gateway(R) BP
reaction"
polyA_signal 2131..2265
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2155..2174)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 2209..2228
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind complement(2378..2398)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(2378..2396)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(2417..2433)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2417..2433)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(2430..2452)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 2441..2457
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(2465..2495)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2510..2531
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(2648..2665)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2819..3407)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2899..2918)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3578..4438)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 4201..4220
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(4439..4543)
/gene="bla"
/label=AmpR promoter
rep_origin complement(4569..5024)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4706..4727)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 4918..4937
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 5098..6082
/label=CMV IE94 promoter
/note="enhancer/promoter region of simian cytomegalovirus
major immediate early transcription unit IE94"
ORIGIN
1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
61 ctacttgttc tttttgcagg atcccatcga ttcgaattca aggatcaaca agtttgtaca
121 aaaaagcagg ctgccaccat ggtgagcgag ctgattaagg agaacatgca catgaagctg
181 tacatggagg gcaccgtgaa caaccaccac ttcaagtgca catccgaggg cgaaggcaag
241 ccctacgagg gcacccagac catgagaatc aaggcggtcg agggcggccc tctccccttc
301 gccttcgaca tcctggctac cagcttcatg tacggcagca aaaccttcat caaccacacc
361 cagggcatcc ccgacttctt taagcagtcc ttccccgagg gcttcacatg ggagagagtc
421 accacatacg aagacggggg cgtgctgacc gctacccagg acaccagcct ccaggacggc
481 tgcctcatct acaacgtcaa gatcagaggg gtgaacttcc catccaacgg ccctgtgatg
541 cagaagaaaa cactcggctg ggaggcctcc accgagaccc tgtaccccgc tgacggcggc
601 ctggaaggca gagccgacat ggccctgaag ctcgtgggcg ggggccacct gatctgcaac
661 ttgaagacca catacagatc caagaaaccc gctaagaacc tcaagatgcc cggcgtctac
721 tatgtggaca gaagactgga aagaatcaag gaggccgaca aagagaccta cgtcgagcag
781 cacgaggtgg ctgtggccag atactgcgac ctccctagca aactggggca cagacctagg
841 ggcagcggcg ccaccaactt cagcctgctg aagcaggccg gcgacgtgga ggagaacccc
901 ggccccgccc ccggcagcct cgagggaaag tcttacccaa ctgtgagtgc tgattaccag
961 gacgccgttg agaaggcgaa gaagaagctc agaggcttca tcgctgagaa gagatgcgct
1021 cctctaatgc tccgtttggc attccactct gctggaacct ttgacaaggg cacgaagacc
1081 ggtggaccct tcggaaccat caagcaccct gccgaactgg ctcacagcgc taacaacggt
1141 cttgacatcg ctgttaggct tttggagcca ctcaaggcgg agttccctat tttgagctac
1201 gccgatttct accagttggc tggcgttgtt gccgttgagg tcacgggtgg acctaaggtt
1261 ccattccacc ctggaagaga ggacaagcct gagccaccac cagagggtcg cttgcccgat
1321 cccactaagg gttctgacca tttgagagat gtgtttggca aagctatggg gcttactgac
1381 caagatatcg ttgctctatc tgggggtcac actattggag ctgcacacaa ggagcgttct
1441 ggatttgagg gtccctggac ctctaatcct cttattttcg acaactcata cttcacggag
1501 ttgttgagtg gtgagaagga aggtctcctt cagctacctt ctgacaaggc tcttttgtct
1561 gaccctgtat tccgccctct cgttgacaaa tatgcagcgg acgaagatgc cttctttgct
1621 gattacgctg aggctcacca aaagctttcc gagcttgggt ttgctgatgc cgtcgaccca
1681 gctttcttgt acaaagtggt gtttactcct aaaccacctc aagatagtgc ggttatcaaa
1741 gctggatatt gtgtaaaaca aggagcagtg atgaaaaact ggaagagaag atattttcaa
1801 ttggatgaaa acacaatagg ctacttcaaa tctgaactgg aaaaggaacc tcttcgcgta
1861 ataccactta aagaggttca taaagtccag gaatgtaagc aaagcgacat aatgatgagg
1921 gacaacctct ttgaaattgt aacaacgtct cgaactttct atgtgcaggc tgatagccct
1981 gaagagatgc acagttggat taaagcagtc tctggcgcca ttgtagcaca gcggggtccc
2041 ggcagatctg cgtcttctga gcatccctga caactttatt atacatagtt gcactcgagc
2101 ctctagaact atagtgagtc gtattacgta gatccagaca tgataagata cattgatgag
2161 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
2221 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
2281 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaatt cgcggccgcg
2341 gcgccaatgc attgggcccg gtacccagct tttgttccct ttagtgaggg ttaattgcgc
2401 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
2461 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
2521 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
2581 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
2641 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
2701 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
2761 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
2821 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
2881 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
2941 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
3001 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
3061 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
3121 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
3181 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
3241 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
3301 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
3361 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
3421 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
3481 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
3541 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
3601 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
3661 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
3721 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
3781 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
3841 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
3901 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
3961 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
4021 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
4081 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
4141 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
4201 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
4261 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
4321 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
4381 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
4441 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
4501 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
4561 tgccacctaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
4621 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
4681 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
4741 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
4801 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
4861 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
4921 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
4981 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt cgccattcag
5041 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagtcgac
5101 catagccaat tcaatatggc gtatatggac tcatgccaat tcaatatggt ggatctggac
5161 ctgtgccaat tcaatatggc gtatatggac tcgtgccaat tcaatatggt ggatctggac
5221 cccagccaat tcaatatggc ggacttggca ccatgccaat tcaatatggc ggacttggca
5281 ctgtgccaac tggggagggg tctacttggc acggtgccaa gtttgaggag gggtcttggc
5341 cctgtgccaa gtccgccata ttgaattggc atggtgccaa taatggcggc catattggct
5401 atatgccagg atcaatatat aggcaatatc caatatggcc ctatgccaat atggctattg
5461 gccaggttca atactatgta ttggccctat gccatatagt attccatata tgggttttcc
5521 tattgacgta gatagcccct cccaatgggc ggtcccatat accatatatg gggcttccta
5581 ataccgccca tagccactcc cccattgacg tcaatggtct ctatatatgg tctttcctat
5641 tgacgtcata tgggcggtcc tattgacgta tatggcgcct cccccattga cgtcaattac
5701 ggtaaatggc ccgcctggct caatgcccat tgacgtcaat aggaccaccc accattgacg
5761 tcaatgggat ggctcattgc ccattcatat ccgttctcac gccccctatt gacgtcaatg
5821 acggtaaatg gcccacttgg cagtacatca atatctatta atagtaactt ggcaagtaca
5881 ttactattgg aaggacgcca gggtacattg gcagtactcc cattgacgtc aatggcggta
5941 aatggcccgc gatggctgcc aagtacatcc ccattgacgt caatggggag gggcaatgac
6001 gcaaatgggc gttccattga cgtaaatggg cggtaggcgt gcctaatggg aggtctatat
6061 aagcaatgct cgtttaggga ac
//
| {
"Vector backbone": "pCSDEST2",
"Backbone size w/o insert (bp)": "",
"Vector type": "Mammalian Expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA ; Evrogen Limited Use Label License for FPs",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Modular Detection of GFP-Labeled Proteins for Rapid Screening by Electron Microscopy in Cells and Organisms",
"Publication": "Dev Cell. 2015 Nov 23;35(4):513-25. doi: 10.1016/j.devcel.2015.10.016. Epub 2015 Nov 12.",
"DOI": "10.1016/j.devcel.2015.10.016.",
"Authors": "Ariotti N, Hall TE, Rae J, Ferguson C, McMahon KA, Martel N, Webb RE, Webb RI, Teasdale RD, Parton RG /jdevcel",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/26585296",
"PubMed ID": "PubMed 26585296"
} | {
"Gene/Insert name": "mKate2-P2A-APEX2-TAPP1-PH",
"Species": "Synthetic",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "CMV",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Gateway Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "SP6",
"3′ sequencing primer": "T3"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
pENTRY4_ WRKY28_2 | 126900 | Gateway pENTR-plasmid to generate a binary construct for genome editing of the wheat homolog of a susceptibility gene of O. sativa. Encodes wheat optimized Cas9 and gene specific sgRNA modules. | Sophien Kamoun | Genome editing in wheat (unpublished) | https://media.addgene.org/snapgene-media/v2.0.0/sequences/249965/4d87bfe1-9e25-42d5-80d0-95d7aba38a14/addgene-plasmid-126900-sequence-249965.gbk | full | LOCUS sequence-249965- 12097 bp DNA circular SYN 26-AUG-2024
DEFINITION Gateway pENTR-plasmid to generate a binary construct for genome
editing of the wheat homolog of a susceptibility gene of O. sativa.
Encodes wheat optimized Cas9 and gene specific sgRNA modules..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 12097)
TITLE Genome editing in wheat
REFERENCE 2 (bases 1 to 12097)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..12097
/mol_type="other DNA"
/organism="synthetic DNA construct"
terminator 103..189
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 281..308
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
primer_bind 316..339
/label=pENTR-F
/note="pENTR vectors, forward primer"
protein_bind 358..457
/gene="mutant version of attL"
/label=attL1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
regulatory 2327..2336
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 2339..2359
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
regulatory 2408..2417
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 2414..6517
/codon_start=1
/product="Cas9 (Csn1) endonuclease from the Streptococcus
pyogenes Type II CRISPR/Cas system"
/label=Cas9
/note="generates RNA-guided double strand breaks in DNA"
/translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
DATLIHQSITGLYETRIDLSQLGGD"
CDS 2414..6517
/codon_start=1
/product="Cas9 (Csn1) endonuclease from the Streptococcus
pyogenes Type II CRISPR/Cas system"
/label=Cas9
/note="generates RNA-guided double strand breaks in DNA"
/translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
DATLIHQSITGLYETRIDLSQLGGD"
CDS 6521..6547
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 6527..6553
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/note="This is a variant of the standard HA tag, which has
sequence YPYDVPDYA. Some anti-HA antibodies may not
recognize this variant."
/translation="YDVPDYASL"
CDS 6566..6586
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 6593..6613
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 6629..6685
/codon_start=1
/product="2A peptide from porcine teschovirus-1
polyprotein"
/label=P2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="ATNFSLLKQAGDVEENPGP"
CDS 7157..7876
/codon_start=1
/product="GFP variant that folds robustly even when fused
to poorly folded proteins (Pedelacq et al., 2006)"
/label=superfolder GFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLL
EFVTAAGITHGMDELYK"
misc_RNA 8502..8577
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
protein_bind complement(8589..8613)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
misc_RNA 9026..9101
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
protein_bind complement(9113..9137)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
misc_RNA 9550..9625
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
protein_bind complement(9637..9661)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
misc_RNA 10074..10149
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
protein_bind complement(10161..10185)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
primer_bind complement(10289..10308)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(10290..10308)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind complement(10326..10425)
/gene="mutant version of attL"
/label=attL2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
primer_bind complement(10537..10556)
/label=pENTR-R
/note="pENTR vectors, reverse primer"
CDS 10548..11357
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPGDAWLLTTAIPGKTA
FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind complement(10615..10634)
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
rep_origin 11447..12035
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 11936..11955
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgctagca tggatctcgg
61 ggacgtctaa ctactaagcg agagtaggga actgccaggc atcaaataaa acgaaaggct
121 cagtcggagg actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
181 aggacaaatc cgccgggagc ggatttgaac gttgtgaagc aacggcccgg agggtggcgg
241 gcaggacgcc cgccataaac tgccaggcat caaactaagc agaaggccat cctgacggat
301 ggcctttttg cgtttctaca aactcttcct gttagttagt tacttaagct cgggccccaa
361 ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga taagcaatgc
421 ttttttataa tgccaacttt gtacaaaaaa gcaggctcca ccatggtaca aagtggtgaa
481 gcttgcatgc ctgcagtgca gcgtgacccg gtcgtgcccc tctctagaga taatgagcat
541 tgcatgtcta agttataaaa aattaccaca tatttttttt gtcacacttg tttgaagtgc
601 agtttatcta tctttataca tatatttaaa ctttactcta cgaataatat aatctatagt
661 actacaataa tatcagtgtt ttagagaatc atataaatga acagttagac atggtctaaa
721 ggacaattga gtattttgac aacaggactc tacagtttta tctttttagt gtgcatgtgt
781 tctccttttt ttttgcaaat agcttcacct atataatact tcatccattt tattagtaca
841 tccatttagg gtttagggtt aatggttttt atagactaat ttttttagta catctatttt
901 attctatttt agcctctaaa ttaagaaaac taaaactcta ttttagtttt tttatttaat
961 aatttagata taaaatagaa taaaataaag tgactaaaaa ttaaacaaat accctttaag
1021 aaattaaaaa aactaaggaa acatttttct tgtttcgagt agataatgcc agcctgttaa
1081 acgccgtcga cgagtctaac ggacaccaac cagcgaacca gcagcgtcgc gtcgggccaa
1141 gcgaagcaga cggcacggca tctctgtcgc tgcctctgga cccctctcga gagttccgct
1201 ccaccgttgg acttgctccg ctgtcggcat ccagaaattg cgtggcggag cggcagacgt
1261 gagccggcac ggcaggcggc ctcctcctcc tctcacggca ccggcagcta cgggggattc
1321 ctttcccacc gctccttcgc tttcccttcc tcgcccgccg taataaatag acaccccctc
1381 cacaccctct ttccccaacc tcgtgttgtt cggagcgcac acacacacaa ccagatctcc
1441 cccaaatcca cccgtcggca cctccgcttc aaggtacgcc gctcgtcctc cccccccccc
1501 cctctctacc ttctctagat cggcgttccg gtccatggtt agggcccggt agttctactt
1561 ctgttcatgt ttgtgttaga tccgtgtttg tgttagatcc gtgctgctag cgttcgtaca
1621 cggatgcgac ctgtacgtca gacacgttct gattgctaac ttgccagtgt ttctctttgg
1681 ggaatcctgg gatggctcta gccgttccgc agacgggatc gatttcatga ttttttttgt
1741 ttcgttgcat agggtttggt ttgccctttt cctttatttc aatatatgcc gtgcacttgt
1801 ttgtcgggtc atcttttcat gctttttttt gtcttggttg tgatgatgtg gtgtggttgg
1861 gcggtcgttc attcgttcta gatcggagta gaatactgtt tcaaactacc tggtgtattt
1921 attaattttg gaactgtatg tgtgtgtcat acatcttcat agttacgagt ttaagatgga
1981 tggaaatatc gatctaggat aggtatacat gttgatgtgg gttttactga tgcatataca
2041 tgatggcata tgcagcatct attcatatgc tctaaccttg agtacctatc tattataata
2101 aacaagtatg ttttataatt attttgatct tgatatactt ggatgatggc atatgcagca
2161 gctatatgtg gattttttta gccctgcctt catacgctat ttatttgctt ggtactgttt
2221 cttttgtcga tgctcaccct gttgtttggt gttacttctg caggtcgact ctagaggatc
2281 tggaattccc ggggagaatc tggagctctc gagatttctc acgcgtgcca ccatggggcc
2341 gaagaagaag aggaaggtgg gtagggtgtg tcggattagc agccttcggt acaggggtcc
2401 tggcattgcc accatggaca agaagtactc tatcgggttg gatataggaa cgaatagtgt
2461 gggttgggct gtcatcactg acgagtacaa ggtgccatcg aagaaattta aagtgctggg
2521 caacaccgac aggcactcca tcaagaagaa cctaataggt gcgttgctct ttgattccgg
2581 ggagacagcc gaagcgaccc gcctcaagcg cacagcaaga aggcgctata cgcgtagaaa
2641 gaacaggatc tgctacctac aagagatctt ttccaacgaa atggcgaaag tggatgactc
2701 cttcttccat cggctcgagg aaagcttcct cgtagaagag gacaagaagc acgaacgcca
2761 tccgatattc gggaacattg tcgacgaagt cgcataccac gagaagtatc ctaccatcta
2821 tcacctgagg aagaagctcg tagatagcac tgacaaggct gatttgcgcc tcatctattt
2881 ggccctggcc catatgatta agttccgtgg gcacttcctc atcgaaggcg accttaatcc
2941 ggacaattcc gatgttgaca aactcttcat ccaactggtc cagacataca atcagctctt
3001 tgaagagaac cccatcaacg cgtcaggagt tgatgcgaag gcaatcttga gtgctcggct
3061 gtcgaaaagc cggaggctcg aaaatctgat agcccaactg ccaggcgaaa agaagaacgg
3121 attgtttggc aaccttattg cccttagtct agggctcacc cctaacttca agtctaactt
3181 tgacctggcc gaggatgcca agctccaact gtccaaagac acgtacgacg acgatctcga
3241 caatctgctt gcccaaatcg gcgaccagta tgcggacctt ttccttgcag cgaagaactt
3301 gtcagatgca atcctcttgt ccgatattct gcgcgttaac actgagatta ccaaggctcc
3361 tctttcggca tccatgatca aacggtacga cgaacatcac caggatctca cgctcctcaa
3421 ggctctcgtc agacaacagc tgccggagaa gtacaaagag atcttcttcg atcaatcaaa
3481 gaacgggtat gctgggtaca tcgacggtgg cgcgtcacag gaggagttct acaagtttat
3541 caagcccatt ctcgagaaga tggatggcac tgaagagctc ctggttaagc tcaatcgcga
3601 ggatctactc cgcaagcagc gtaccttcga caatggaagc attccacacc aaatccatct
3661 gggcgagctt cacgccatac ttcgtaggca agaggacttt tacccattcc tcaaagacaa
3721 ccgggagaaa atcgagaaga tcctcacatt cagaatcccc tactatgtgg gcccattggc
3781 aaggggcaac tctaggttcg cgtggatgac tagaaaatcg gaggaaacga ttaccccatg
3841 gaacttcgag gaggtggtgg acaagggcgc ttctgcccaa agcttcatcg agaggatgac
3901 caacttcgac aagaatctgc caaacgagaa ggtcctcccc aaacattccc tactgtacga
3961 gtacttcacc gtctacaacg agctgaccaa ggtcaagtac gttactgagg gtatgaggaa
4021 accggccttt ctgtcaggag agcagaagaa ggccatagtg gatctccttt tcaagaccaa
4081 tcgcaaggtc acagtcaagc agctaaagga ggactacttt aagaaaatcg agtgctttga
4141 ctctgtggag ataagcggcg ttgaggatcg cttcaacgcc tcgctgggaa cgtaccacga
4201 cttgctcaag atcattaagg ataaagactt cctcgataac gaagagaacg aggacatctt
4261 ggaggacatc gttctgacgc ttactctctt cgaggaccgc gagatgatcg aagagaggct
4321 caagacgtat gcgcacctct ttgacgacaa agtgatgaag cagctgaagc gcaggcgcta
4381 cacaggttgg ggcaggctga gcaggaagct gatcaatggc attcgcgata agcagtcggg
4441 caagaccatc ctcgacttcc tcaagtccga tgggttcgca aacaggaact ttatgcagct
4501 aatccacgat gactccttga ccttcaagga ggacattcag aaggctcagg tttcaggcca
4561 aggcgattcg ctccacgagc atatcgcgaa tttggccgga tctccagcca tcaagaaggg
4621 tatacttcaa acagtgaagg ttgtcgatga gctggtcaaa gtgatgggta gacataagcc
4681 cgagaatatc gtgattgaga tggctcggga gaaccaaacc acacagaagg ggcagaagaa
4741 ctccagggaa aggatgaaga gaatcgaaga aggcatcaag gagctcggat cacaaattct
4801 caaggagcat ccggtcgaaa acacccaact gcaaaacgag aagctctacc tctactacct
4861 ccaaaacgga cgcgatatgt acgtggacca ggagctcgac atcaatcgcc tatctgacta
4921 cgatgtcgat catatagtac cacaaagctt cctcaaagac gactctatcg acaacaaggt
4981 gctgaccagg tccgacaaga accgcgggaa gtcagacaac gtgccttctg aggaagtcgt
5041 caagaagatg aagaactact ggaggcaact cctcaatgca aagctcatta cacaaaggaa
5101 gttcgacaac cttaccaagg cggagagagg cggcctcagt gagctagata aggccggatt
5161 tatcaagcgc caactggttg agacacggca aatcaccaag cacgttgcac agatcctgga
5221 ctcacgcatg aacacaaaat acgatgagaa tgacaagcta atccgggaag tcaaagttat
5281 tactctgaaa tctaaactgg tgtccgactt tcggaaggac ttccagttct ataaggtccg
5341 tgagatcaac aactaccatc acgcccatga tgcctatctc aatgctgtgg ttggcactgc
5401 actcatcaag aaatatccca agcttgagtc ggagttcgtc tacggtgact acaaggttta
5461 tgacgtaagg aagatgattg cgaagagtga gcaggaaatc ggcaaggcca ccgccaagta
5521 cttcttttac agcaacataa tgaacttctt caagactgag atcaccctgg ccaatggaga
5581 gatcagaaaa cgtccgctga tagagactaa cggagaaact ggtgagatag tgtgggacaa
5641 ggggagagac tttgctactg tccggaaggt gctaagcatg ccacaagtca acatcgtcaa
5701 gaaaacggaa gtccaaaccg gcggcttctc gaaagagtca atccttccga agcgcaatag
5761 cgataagctt atagcccgta agaaagactg ggatccaaag aagtacggtg gctttgacag
5821 tcctacagtg gcgtactccg tgctggttgt tgccaaggtg gaaaagggga agtcgaagaa
5881 gctgaagagc gtaaaggagt tgctcgggat taccattatg gagcgttcca gctttgagaa
5941 gaatcccatt gatttcttgg aggcgaaagg atacaaggaa gtcaagaaag atctcatcat
6001 aaagttgccg aaatacagct tgttcgagct ggagaatggg agaaagagga tgcttgccag
6061 cgcaggcgag ttgcaaaagg ggaacgagct ggcactccca tccaagtacg tcaacttcct
6121 gtatctcgcc tcacactatg agaaacttaa gggttcccca gaggacaatg agcaaaagca
6181 gctgtttgtg gagcaacaca agcactacct cgatgagatc atcgaacaga tcagtgagtt
6241 ctcgaagagg gtgattcttg ctgacgctaa ccttgacaag gtgttgtccg cctacaacaa
6301 gcatcgggac aaacccatcc gggagcaggc agaaaacatc atacacttgt tcacgttgac
6361 gaaccttggc gctcctgcgg cgttcaaata ttttgacacg acgattgata ggaagcgcta
6421 taccagcacc aaagaagtac tcgatgccac cctaattcac cagtctatta caggcctgta
6481 cgaaacgagg atcgacttgt cgcaactagg aggagatgcc tatccgtatg acgtgccaga
6541 ttacgcctcc cttgggtctg gttcccctaa gaagaagcgg aaagtggagg atcccaagaa
6601 gaagcgcaaa gtcgacggcg gatccggagc cacgaacttt tccctgctca agcaagctgg
6661 tgatgtggag gagaatcctg gcccagctcc aggctcaacg tccatggcga aggcagataa
6721 gaaaccagcg gagaagaaac cggcagagaa aactccggca gtcgaaccag cagcagcggc
6781 agagaagaaa ccaaaagccg gaaagaaact ccccaaggaa ccagccgtcg ccggagacaa
6841 gaagaagaag agatcaaaga agaacgttga gacatacaag atctacatct tcaaggtgtt
6901 gaagcaagtt catccagaca tcggaatctc aagcaaagcg atgggaatta tgaacagttt
6961 catcaacgat atttttgaga agctcgctgg tgaatcttcg aagctggcga ggtacaacaa
7021 gaagccgacg attacttcca gggagattca gactgcggtt agacttgtgt tgcccggtga
7081 gttggcgaaa catgctgtgt ctgaagggac taaggcggtt acgaagttta cgagttctgc
7141 tcagggcgcc tctttcatgg tgtccaaagg agaggagctc tttacaggag ttgtgcccat
7201 tttggtcgag ttggatggcg acgtaaatgg ccacaagttc agcgtgagag gcgaaggtga
7261 aggtgatgcc accaatggca agcttaccct caagttcatc tgtaccactg ggaaactgcc
7321 ggttccatgg ccaacgttgg tcaccactct cacatatggt gttcagtgct tttcgaggta
7381 tccggaccac atgaaacgcc atgacttctt caagtcagcc atgcctgagg ggtatgtcca
7441 agaacgcacc atcagcttta aggacgacgg aacgtacaag acacgtgcgg aggtcaagtt
7501 cgagggagat acactggtga acaggatcga actcaaaggg atcgacttca aagaggacgg
7561 caacatactg gggcataagc tcgagtacaa cttcaattct cacaacgtct acattactgc
7621 cgacaagcag aagaacggca ttaaggcgaa cttcaaaatc cggcataatg tggaggatgg
7681 tagtgtccaa ctggctgacc actaccagca gaacacgccg ataggcgatg gcccagtgct
7741 tctgcccgac aatcactacc tgtcaaccca atccgttctg tctaaggacc ctaacgaaaa
7801 gagggatcac atggtgcttc tggagtttgt aacggctgca ggcatcactc atgggatgga
7861 tgagctctac aagtgataac tgcggccgca gatatcacta gtgttcaaac atttggcaat
7921 aaagtttctt aagattgaat cctgttgccg gtcttgcgat gattatcata taatttctgt
7981 tgaattacgt taagcatgta ataattaaca tgtaatgcat gacgttattt atgagatggg
8041 tttttatgat tagagtcccg caattataca tttaatacgc gatagaaaac aaaatataac
8101 tagaattcga gctcggagga ccaagcccgt tattctgaca gttctggtgc tcaacacatt
8161 tatatttatc aaggagcaca ttgttactca ctgctaggag ggaatcgaac taggaatatt
8221 gatcagagga actacgagag agctgaagat aactgccctc tagctctcac tgatctgggt
8281 cgcatagtga gatgcagccc acgtgagttc agcaacggtc tagcgctggg cttttaggcc
8341 cgcatgatcg ggcttttgtc gggtggtcga cgtgttcacg attggggaga gcaacgcagc
8401 agttcctctt agtttagtcc cacctcgcct gtccagcaga gttctgaccg gtttataaac
8461 tcgcttgctg catcagactt ggggcaaggt cagcggcatg agttttagag ctagaaatag
8521 caagttaaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt
8581 ttttctagac ccagctttct tgtacaaagt tggcattacg ctttacgaat tcccatgggg
8641 aggaccaagc ccgttattct gacagttctg gtgctcaaca catttatatt tatcaaggag
8701 cacattgtta ctcactgcta ggagggaatc gaactaggaa tattgatcag aggaactacg
8761 agagagctga agataactgc cctctagctc tcactgatct gggtcgcata gtgagatgca
8821 gcccacgtga gttcagcaac ggtctagcgc tgggctttta ggcccgcatg atcgggcttt
8881 tgtcgggtgg tcgacgtgtt cacgattggg gagagcaacg cagcagttcc tcttagttta
8941 gtcccacctc gcctgtccag cagagttctg accggtttat aaactcgctt gctgcatcag
9001 acttggggca gacttccctc agcctgtttt agagctagaa atagcaagtt aaaataaggc
9061 tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg ctttttttct agacccagct
9121 ttcttgtaca aagttggcat tacgctcaga gaattcgcat gcggaggacc aagcccgtta
9181 ttctgacagt tctggtgctc aacacattta tatttatcaa ggagcacatt gttactcact
9241 gctaggaggg aatcgaacta ggaatattga tcagaggaac tacgagagag ctgaagataa
9301 ctgccctcta gctctcactg atctgggtcg catagtgaga tgcagcccac gtgagttcag
9361 caacggtcta gcgctgggct tttaggcccg catgatcggg cttttgtcgg gtggtcgacg
9421 tgttcacgat tggggagagc aacgcagcag ttcctcttag tttagtccca cctcgcctgt
9481 ccagcagagt tctgaccggt ttataaactc gcttgctgca tcagacttgc cgcagcgaca
9541 gcctcgacag ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact
9601 tgaaaaagtg gcaccgagtc ggtgcttttt ttctagaccc agctttcttg tacaaagttg
9661 gcattacgct tgtggaattc ctcgagggag gaccaagccc gttattctga cagttctggt
9721 gctcaacaca tttatattta tcaaggagca cattgttact cactgctagg agggaatcga
9781 actaggaata ttgatcagag gaactacgag agagctgaag ataactgccc tctagctctc
9841 actgatctgg gtcgcatagt gagatgcagc ccacgtgagt tcagcaacgg tctagcgctg
9901 ggcttttagg cccgcatgat cgggcttttg tcgggtggtc gacgtgttca cgattgggga
9961 gagcaacgca gcagttcctc ttagtttagt cccacctcgc ctgtccagca gagttctgac
10021 cggtttataa actcgcttgc tgcatcagac ttgggcgcgt gggcgaggag aacgttttag
10081 agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg
10141 agtcggtgct ttttttctag acccagcttt cttgtacaaa gttggcatta cgctgagcga
10201 ggatgcacat gtgaccgagg gaggtgtcat ctatgttact gctagtcgat ttaaatcctt
10261 aattaaccct gcaggcatct ccaattcgcc ctatagtgag tcgtattaca ccggtgagga
10321 tctagaccca gctttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg
10381 ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgcagc
10441 tctggcccgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat
10501 gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc
10561 aacgggaaac gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata
10621 aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc
10681 ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag
10741 atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt
10801 ttatccgtac tcctggtgat gcatggttac tcaccactgc gatccccgga aaaacagcat
10861 tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt
10921 tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat
10981 ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg
11041 atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc
11101 cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg
11161 acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc
11221 aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc
11281 tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc
11341 tcgatgagtt tttctaatca gaattggtta attggttgta acattattca gattgggccc
11401 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
11461 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
11521 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
11581 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
11641 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
11701 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
11761 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
11821 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
11881 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
11941 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
12001 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
12061 ggttcctggc cttttgctgg ccttttgctc acatgtt
//
| {
"Vector backbone": "pENTR4",
"Backbone size w/o insert (bp)": "",
"Vector type": "CRISPR",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Kanamycin, 50 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "",
"Publication": "",
"DOI": "",
"Authors": "",
"PubMed Link": "",
"PubMed ID": ""
} | {
"Gene/Insert name": "Wheat_live_Cas9",
"Species": "Synthetic",
"Insert Size (bp)": "8726",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "Unknown (unknown if destroyed)",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pBLV B4 | 68057 | expresses BLV (NC001414) B4 pre-miRNA and miRNAs | Christopher Sullivan | Burke et al Nucleic Acids Res. 2014 Dec 16;42(22):13949-62. doi: 10.1093/nar/gku1247. Epub 2014 Nov 26. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/121454/0c976dde-88c7-49de-a3a9-34243dd1c100/addgene-plasmid-68057-sequence-121454.gbk | partial | LOCUS sequence-121454- 954 bp DNA linear UNA 26-AUG-2024
DEFINITION expresses BLV (NC001414) B4 pre-miRNA and miRNAs.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE natural DNA sequence
ORGANISM unspecified
REFERENCE 1 (bases 1 to 954)
AUTHORS Burke JM, Bass CR, Kincaid RP, Sullivan CS
TITLE Identification of tri-phosphatase activity in the biogenesis of
retroviral microRNAs and RNAP III-generated shRNAs.
JOURNAL Nucleic Acids Res. 2014 Dec 16;42(22):13949-62. doi:
10.1093/nar/gku1247. Epub 2014 Nov 26.
PUBMED 25428356
REFERENCE 2 (bases 1 to 954)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1093/nar/gku1247"; journalName: "Nucleic Acids Res"; date:
"2014-12-16- 16"; volume: "42"; issue: "22"; pages: "13949-62"
FEATURES Location/Qualifiers
source 1..954
/mol_type="genomic DNA"
/organism="unspecified"
primer_bind complement(253..269)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(253..269)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
terminator 317..348
/label=tonB terminator
/note="bidirectional E. coli tonB-P14 transcription
terminator"
promoter 349..451
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
primer_bind complement(525..544)
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
ORIGIN
1 aggtgagcca gtgagttgat tgcagtccag ttacgctgga gtctgaggct cgtcctgaat
61 gatatgcggc ctcctcgagc tgcagctgtg ctaagcgaga ggctctggtg ctggggataa
121 gatgcggccc ctagcaccac agtctctgcg ccttttgggt tcgaatcttc cccatgcctc
181 tagagcgcgt gatcttacgg cattatacgt atgatcggtc cacgatcagc tagattatct
241 agtcagcttg atgtcatagc tgtttcctga ggctcaatac tgaccattta aatcatacct
301 gacctccata gcagaaagtc aaaagcctcc gaccggaggc ttttgacttg atcggcacgt
361 aagaggttcc aactttcacc ataatgaaat aagatcacta ccgggcgtat tttttgagtt
421 atcgagattt tcaggagcta aggaagctaa aatgagccat attcaacggg aaacgtcttg
481 cttgaagccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
541 cgataatgtc gggcaatcag gtgcgacaat ctatcgattg tatgggaagc ccgatgcgcc
601 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
661 caggctaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
721 tcctgatgat gcatggttac tcaccactgc gatcccaggg aaaacagcat tccaggtatt
781 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
841 gttgcattcg attcctgttt gtaattgtcc ttttaacggc gatcgcgtat ttcgtctcgc
901 tcaggcgcaa tcacgaatga ataacggttt ggttggtgcg agtgatttga tgac
//
| {
"Vector backbone": "pIDTSmart-Kan",
"Backbone size w/o insert (bp)": "1962",
"Vector type": "Synthetic Biology",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Kanamycin, 50 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "Unknown"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Identification of tri-phosphatase activity in the biogenesis of retroviral microRNAs and RNAP III-generated shRNAs",
"Publication": "Nucleic Acids Res. 2014 Dec 16;42(22):13949-62. doi: 10.1093/nar/gku1247. Epub 2014 Nov 26.",
"DOI": "10.1093/nar/gku1247.",
"Authors": "Burke JM, Bass CR, Kincaid RP, Sullivan CS /nar/gku",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/25428356",
"PubMed ID": "PubMed 25428356"
} | {
"Gene/Insert name": "BLV-miR-B4",
"Species": "Bovine leukemia virus",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "Xho1 (not destroyed)",
"3′ cloning site": "Xba1 (not destroyed)",
"5′ sequencing primer": "M13 Foward",
"3′ sequencing primer": "M13 reverse"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
His-TEV-GB1 Val54Met | 128691 | Expresses GB1 Val54Met mutant | Sharon Rozovsky | Chen et al J Phys Chem B. 2020 Jan 30;124(4):601-616. doi: 10.1021/acs.jpcb.9b07466. Epub 2020 Jan 7. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/252992/fd7791b0-e55f-4114-9d33-2141d475d9f3/addgene-plasmid-128691-sequence-252992.gbk | full | LOCUS sequence-252992- 4226 bp DNA circular SYN 26-AUG-2024
DEFINITION Expresses GB1 Val54Met mutant.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4226)
AUTHORS Chen Q, Xu S, Lu X, Boeri MV, Pepelyayeva Y, Diaz EL, Soni SD,
Allaire M, Forstner MB, Bahnson BJ, Rozovsky S
TITLE (77)Se NMR Probes the Protein Environment of Selenomethionine.
JOURNAL J Phys Chem B. 2020 Jan 30;124(4):601-616. doi:
10.1021/acs.jpcb.9b07466. Epub 2020 Jan 7.
PUBMED 31846581
REFERENCE 2 (bases 1 to 4226)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1021/acs.jpcb.9b07466"; journalName: "J Phys Chem B"; date:
"2020-01-30- 30"; volume: "124"; issue: "4"; pages: "601-616"
FEATURES Location/Qualifiers
source 1..4226
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin 86..674
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 575..594
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 828..845
/label=L4440
/note="L4440 vector, forward primer"
primer_bind 943..960
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 944..960
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 1423..1442
/label=T7
/note="T7 promoter, forward primer"
promoter 1423..1441
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 1442..1466
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
CDS 1515..1532
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 1554..1574
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQG"
terminator 1753..1800
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
primer_bind complement(2023..2042)
/label=pENTR-R
/note="pENTR vectors, reverse primer"
primer_bind complement(2048..2067)
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
promoter 2109..2200
/gene="bla"
/label=AmpR promoter
CDS 2201..3061
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(2419..2438)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
CDS 3144..4226
/codon_start=1
/gene="lacI"
/product="lac repressor"
/label=lacI
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
primer_bind complement(3163..3182)
/label=LacI-R
/note="LacI, reverse primer"
ORIGIN
1 ctcatgacca aaatccctta acgtgagtta cgcgcgcgtc gttccactga gcgtcagacc
61 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
121 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
181 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
241 tgtagccgta gttagcccac cacttcaaga actctgtagc accgcctaca tacctcgctc
301 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
361 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
421 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
481 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
541 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
601 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
661 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
721 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
781 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
841 gcgaggaagc ggaaggcgag agtagggaac tgccaggcat caaactaagc agaaggcccc
901 tgacggatgg cctttttgcg tttctacaaa ctctttctgt gttgtaaaac gacggccagt
961 cttaagctcg ggccccctgg gcggttctga taacgagtaa tcgttaatcc gcaaataacg
1021 taaaaacccg cttcggcggg tttttttatg gggggagttt agggaaagag catttgtcag
1081 aatatttaag ggcgcctgtc actttgcttg atatatgaga attatttaac cttataaatg
1141 agaaaaaagc aacgcacttt aaataagata cgttgctttt tcgattgatg aacacctata
1201 attaaactat tcatctatta tttatgattt tttgtatata caatatttct agtttgttaa
1261 agagaattaa gaaaataaat ctcgaaaata ataaagggaa aatcagtttt tgatatcaaa
1321 attatacatg tcaacgataa tacaaaatat aatacaaact ataagatgtt atcagtattt
1381 attatcattt agaataaatt ttgtgtcgcc cttccgcgaa attaatacga ctcactatag
1441 gggaattgtg agcggataac aattcccctc tagaaataat tttgtttaac ttttaggagg
1501 taaaacatat gagccatcac caccaccacc acgactacga cattccgacg acggagaact
1561 tgtatttcca gggtcagtac aaactgatcc tgaatggcaa gaccctgaag ggtgaaacca
1621 ccaccgaagc ggtcgatgca gcgacggctg aaaaagtttt caagcaatat gcgaacgata
1681 atggtgttga cggcgagtgg acttacgatg acgccacgaa aacctttacc atgaccgagt
1741 aactcgagcc ccctagcata accccttggg gcctctaaac gggtcttgag gggttttttg
1801 cccctgagac gcgtcaatcg agttcgtacc taagggcgac accccataat tagcccgggc
1861 gaaaggccca gtctttcgac tgagcctttc gttttatttg atgcctggca gttccctact
1921 ctcgcatggg gagtccccac actaccatcg gcgctacggc gtttcacttc tgagttcggc
1981 atggggtcag gtgggaccac cgcgctactg ccgccaggca aacaaggggt gttatgagcc
2041 atattcaggt ataaatgggc tcgcgataat gttcagaatt ggttaattgg ttgtaacact
2101 gacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
2161 accctgataa atgcttcaat aatattgaaa aaggaagaat atgagtattc aacatttccg
2221 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
2281 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
2341 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
2401 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
2461 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
2521 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
2581 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
2641 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
2701 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcga tggcaacaac
2761 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
2821 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
2881 gtttattgct gataaatccg gagccggtga gcgtggttct cgcggtatca tcgcagcgct
2941 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
3001 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
3061 agcggcgcgc catcgaatgg cgcaaaacct ttcgcggtat ggcatgatag cgcccggaag
3121 agagtcaatt cagggtggtg aatatgaaac cagtaacgtt atacgatgtc gcagagtatg
3181 ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca ggccagccac gtttctgcga
3241 aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg
3301 cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc agtctggccc
3361 tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca
3421 gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca
3481 atcttctcgc gcaacgcgtc agtgggctga tcattaacta tccgctggat gaccaggatg
3541 ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt atttcttgat gtctctgacc
3601 agacacccat caacagtatt attttctccc atgaggacgg tacgcgactg ggcgtggagc
3661 atctggtcgc attgggtcac cagcaaatcg cgctgttagc gggcccatta agttctgtct
3721 cggcgcgtct gcgtctggct ggctggcata aatatctcac tcgcaatcaa attcagccga
3781 tagcggaacg ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc
3841 tgaatgaggg catcgttccc actgcgatgc tggttgccaa cgatcagatg gcgctgggcg
3901 caatgcgcgc cattaccgag tccgggctgc gcgttggtgc ggatatctcg gtagtgggat
3961 acgacgatac cgaagatagc tcatgttata tcccgccgtt aaccaccatc aaacaggatt
4021 ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca actctctcag ggccaggcgg
4081 tgaagggcaa tcagctgttg ccagtctcac tggtgaaaag aaaaaccacc ctggcgccca
4141 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
4201 tttcccgact ggaaagcggg cagtga
//
| {
"Vector backbone": "pJexpress414",
"Backbone size w/o insert (bp)": "3973",
"Vector type": "Bacterial Expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Industry MTA"
} | {
"Title": "(77)Se NMR Probes the Protein Environment of Selenomethionine",
"Publication": "J Phys Chem B. 2020 Jan 30;124(4):601-616. doi: 10.1021/acs.jpcb.9b07466. Epub 2020 Jan 7.",
"DOI": "10.1021/acs.jpcb.9b07466.",
"Authors": "Chen Q, Xu S, Lu X, Boeri MV, Pepelyayeva Y, Diaz EL, Soni SD, Allaire M, Forstner MB, Bahnson BJ, Rozovsky S /acsjpcbb",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/31846581",
"PubMed ID": "PubMed 31846581"
} | {
"Gene/Insert name": "spg",
"Species": "Streptococcus sp. group G",
"Insert Size (bp)": "235",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "T7",
"Tag / Fusion Protein": "His (N terminal on insert)"
} | {
"Cloning method": "Unknown",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "T7",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
PATL2_G288-Y543_BV_bio | 194761 | Cheryl Arrowsmith | SGC Toronto 2022-Q1 to Q2 (unpublished) | https://media.addgene.org/snapgene-media/v2.0.0/sequences/383917/685a8f9f-acb4-422a-a563-33a30b640da9/addgene-plasmid-194761-sequence-383917.gbk | full | LOCUS sequence-383917- 6945 bp DNA circular SYN 27-AUG-2024
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6945)
TITLE SGC Toronto 2022-Q1 to Q2
REFERENCE 2 (bases 1 to 6945)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 27, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6945
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter complement(176..204)
/gene="intI1 (promoter lies within the coding sequence)"
/label=Pc promoter
/note="class 1 integron promoter"
polyA_signal 712..760
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
primer_bind 785..804
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
CDS complement(831..1796)
/codon_start=1
/gene="birA"
/product="E. coli biotin protein ligase"
/label=BirA
/translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
CDS complement(834..1796)
/codon_start=1
/product="E. coli biotin protein ligase"
/label=BirA
/translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
promoter complement(1833..1942)
/label=p10 promoter
/note="baculovirus promoter for expression in insect cells"
promoter 1966..2057
/gene="polh from Autographa californica"
/label=polyhedrin promoter
/note="promoter for the baculovirus polyhedrin gene"
primer_bind 1982..1999
/label=Polyhedrin-F
/note="Polyhedrin promoter, forward primer"
CDS 2108..2152
/codon_start=1
/product="peptide tag that allows for enzymatic
biotinylation"
/label=AviTag(TM)
/translation="GLNDIFEAQKIEWHE"
CDS 2954..2971
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
polyA_signal 3106..3240
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3143..3162)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3197..3216
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
mobile_element 3269..3434
/mobile_element_type="transposon:Tn7"
/label=Tn7L
/note="mini-Tn7 element (left end of the Tn7 transposon)"
rep_origin 3618..4072
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3705..3724)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3914..3935
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 4098..4202
/gene="bla"
/label=AmpR promoter
CDS 4203..5063
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(4421..4440)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 5234..5822
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5723..5742
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 5976..5993
/label=L4440
/note="L4440 vector, forward primer"
mobile_element 6127..6351
/mobile_element_type="transposon:Tn7"
/label=Tn7R
/note="mini-Tn7 element (right end of the Tn7 transposon)"
ORIGIN
1 taacatcaaa catcgaccca cggcgtaacg cgcttgctgc ttggatgccc gaggcataga
61 ctgtacaaaa aaacagtcat aacaagccat gaaaaccgcc actgcgccgt taccaccgct
121 gcgttcggtc aaggttctgg accagttgcg tgagcgcata cgctacttgc attacagttt
181 acgaaccgaa caggcttatg tcaactgggt tcgtgccttc atccgtttcc acggtgtgcg
241 tcacccggca accttgggca gcagcgaagt cgaggcattt ctgtcctggc tggcgaacga
301 gcgcaaggtt tcggtctcca cgcatcgtca ggcattggcg gccttgctgt tcttctacgg
361 caaggtgctg tgcacggatc tgcccttgct tcaggagatc ggtagacctc ggccgtcgcg
421 gcgcttgccg gtggtgctga ccccggatga agtggttcgc atcctcggtt ttctggaagg
481 cgagcatcgt ttgttcgccc aggactctag ctatagttct agtggttggc tacgtacccg
541 tagtggctat ggcagggctt gccgccccga cgttggctgc gagccctggg ccttcacccg
601 aacttggggg ttggggtggg gaaaaggaag aaacgcgggc gtattggtcc caatggggtc
661 tcggtggggt atcgacagag tgccagccct gggaccgaac cccgcgttta tgaacaaacg
721 acccaacacc cgtgcgtttt attctgtctt tttattgccg tcatagcgcg ggttccttcc
781 ggtattgtct ccttccgtgt ttcagttagc ctcccccatc tcccggtact tcatttttct
841 gcactacgca gggatatttc accgcccatc cagggtttta ttattccatc ctgctcaagt
901 aataaagccc cctgtttgtc tattccgcgt gaaatgccaa atatttcttt atcaccaatg
961 ataagtttca ctgggcgatt aataaaatta tccagctttt cccagcgcga cagataaggt
1021 gccaatcctt cttgttcgaa gagttccaac gcagcacgta attcacgtat tagcatggcc
1081 gccaacgtat tacgatcgag attgatcccc gcttcctgca gcgtgatcca cccctgatta
1141 acgacactct cttcaacacg gcgcattgcc atgttgatcc cggctccaat gactatttgc
1201 gccgcatcgc cagttttgcc agtcagctcc accagaatgc ctgccagctt gcgatcctgc
1261 agatagaggt cattaggcca tttaacacga actttatctg cacccagctt gcgtaatact
1321 tccgccatca cgataccgat aaccagactt aaaccaatcg ccgccgccgg gccttgttcc
1381 agacgccaga acatcgacaa atataagttt gcgccaaaag gcgaaaacca tttccgaccc
1441 cggcgaccac ggccagcctg ctggtattct gcaatgcaag catcgcccga tttaagctct
1501 ccgatacgat caagaaggta ctgattcgtg gagtcaatca ctggcagcac ggctacacta
1561 ccgccatcca gctgacccaa tatctgttta gcattaagta actggatagg ctcaggcagg
1621 ctgtatcctt tacccggaac ggtaaagaca tcaacgcccc agtcacgcag tgtctgaatg
1681 tgtttattaa tagccgcccg gctcattccc agcgtttcac ccaactgctc gccagagtga
1741 aattcaccgt tcgctaacag ggcaatcaat ttcagtggca cggtgttatc cttcatggct
1801 cgagatcccg ggtgatcaag tcttcgtcga gtgattgtaa ataaaatgta atttacagta
1861 tagtatttta attaatatac aaatgatttg ataataattc ttatttaact ataatatatt
1921 gtgttgggtt gaattaaagg tccgtatact ccggaatatt aatagatcat ggagataatt
1981 aaaatgataa ccatctcgca aataaataag tattttactg ttttcgtaac agttttgtaa
2041 taaaaaaacc tataaatatt ccggattatt cataccgtcc caccatcggg cgcggatccc
2101 gatgagcggc ctgaacgata tttttgaagc gcagaaaatt gaatggcatg aaggcagcgc
2161 tggaggttca ggtgggactc aagagcagga tattgaagcc gccagttcgc agcggttaag
2221 agtgctctat agaattgaaa agatgtttct ccagctttta gagattgagg agggctggaa
2281 ataccgccct cccccaccgt gttttagcga acaacagtcc aaccaagtcg aaaaactttt
2341 ccaaacccta aaaacacagg agcagaataa cctggaggaa gcggctgacg gtttcttaca
2401 ggttctaagt gtcaggaaag gtaaggcgct cgtagcgcgg ctattgccgt ttcttccaca
2461 agaccaagca gtcacaatac tcttagctat cactcaccac ctcccactat tggtgcggag
2521 ggacgttgca gaccaagctc tgcaaatgtt gtttaagccc cttggtaaat gcatttccca
2581 cctcacgctc catgagcttc tgcaaggact ccaaggattg acacttttgc cccctggatc
2641 atctgaacga cccgtgacag tcgttttgca gaatcagttt ggaattagtc tactgtacgc
2701 acttttatca catggggagc aattggtgag cttgcactct tcactggaag agcccaacag
2761 cgaccacacg gcatggaccg atatggtagt actgatcgct tgggagatag cgcaaatgcc
2821 tacagcttcg cttgctgagc ctctggcatt tcctagtaac ctgttgcccc tattctgtca
2881 ccatgtcgat aaacaattag tccagcagct agaggcacga atggagtttg cttggatcta
2941 tggcggaagt ggacatcacc accatcatca ctgatgaagc ttgtcgagaa gtactagagg
3001 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
3061 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
3121 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
3181 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc
3241 tgatcactgc ttgagcctag gagatccgaa ccagataagt gaaatctagt tccaaactat
3301 tttgtcattt ttaattttcg tattagctta cgacgctaca cccagttccc atctattttg
3361 tcactcttcc ctaaataatc cttaaaaact ccatttccac ccctcccagt tcccaactat
3421 tttgtccgcc cacagcgggg catttttctt cctgttatgt ttttaatcaa acatcctgcc
3481 aactccatgt gacaaaccgt catcttcggc tactttttct ctgtcacaga atgaaaattt
3541 ttctgtcatc tcttcgttat taatgtttgt aattgactga atatcaacgc ttatttgcag
3601 cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
3661 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
3721 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
3781 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
3841 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
3901 cacgttctta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
3961 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
4021 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca
4081 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
4141 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
4201 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
4261 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
4321 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
4381 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
4441 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
4501 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
4561 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
4621 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
4681 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
4741 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
4801 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
4861 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
4921 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
4981 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
5041 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
5101 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
5161 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
5221 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
5281 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
5341 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
5401 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
5461 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
5521 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
5581 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
5641 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
5701 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
5761 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
5821 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
5881 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
5941 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
6001 aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatag
6061 accagccgcg taacctggca aaatcggtta cggttgagta ataaatggat gccctgcgta
6121 agcgggtgtg ggcggacaat aaagtcttaa actgaacaaa atagatctaa actatgacaa
6181 taaagtctta aactagacag aatagttgta aactgaaatc agtccagtta tgctgtgaaa
6241 aagcatactg gacttttgtt atggctaaag caaactcttc attttctgaa gtgcaaattg
6301 cccgtcgtat taaagagggg cgtggccaag ggcatggtaa agactatatt cgcggcgttg
6361 tgacaattta ccgaacaact ccgcggccgg gaagccgatc tcggcttgaa cgaattgtta
6421 ggtggcggta cttgggtcga tatcaaagtg catcacttct tcccgtatgc ccaactttgt
6481 atagagagcc actgcgggat cgtcaccgta atctgcttgc acgtagatca cataagcacc
6541 aagcgcgttg gcctcatgct tgaggagatt gatgagcgcg gtggcaatgc cctgcctccg
6601 gtgctcgccg gagactgcga gatcatagat atagatctca ctacgcggct gctcaaactt
6661 gggcagaacg taagccgcga gagcgccaac aaccgcttct tggtcgaagg cagcaagcgc
6721 gatgaatgtc ttactacgga gcaagttccc gaggtaatcg gagtccggct gatgttggga
6781 gtaggtggct acgtctccga actcacgacc gaaaagatca agagcagccc gcatggattt
6841 gacttggtca gggccgagcc tacatgtgcg aatgatgccc atacttgagc cacctaactt
6901 tgttttaggg cgactgccct gctgcgtaac atcgttgctg ctcca
//
| {
"Vector backbone": "pFBD-BirA",
"Backbone size w/o insert (bp)": "",
"Vector type": "Baculovirus Expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "",
"Publication": "",
"DOI": "",
"Authors": "",
"PubMed Link": "",
"PubMed ID": ""
} | {
"Gene/Insert name": "PATL2",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "PATL2",
"Promoter": "Polyhedrin and p10",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Ligation Independent Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "pFBOH-FWD:CCGGATTATTCATACCGTCCCACCA",
"3′ sequencing primer": "pFBOH-REV:CTGATTATGATCCTCTAGTACTTCT"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
||
pICH41295(pro5U)-pAT5G42470 | 205385 | MoClo lvl0 Promoter + 5'UTR part | Jennifer Nemhauser | Yang et al Quant Plant Biol. 2023 Oct 19;4:e12. doi: 10.1017/qpb.2023.10. eCollection 2023. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/404836/0279ba5c-d814-4b69-b06b-42e243d15b62/addgene-plasmid-205385-sequence-404836.gbk | full | LOCUS sequence-404836- 3421 bp DNA circular SYN 27-AUG-2024
DEFINITION MoClo lvl0 Promoter + 5'UTR part.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3421)
AUTHORS Yang EJY, Nemhauser JL
TITLE Building a pipeline to identify and engineer constitutive and
repressible promoters.
JOURNAL Quant Plant Biol. 2023 Oct 19;4:e12. doi: 10.1017/qpb.2023.10.
eCollection 2023.
PUBMED 37901686
REFERENCE 2 (bases 1 to 3421)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 27, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Quant Plant
Biol."; date: "2023-10-19"; volume: "4:e12. doi"; pages: "
10.1017/qpb.2023.10. eCollection 2023"
FEATURES Location/Qualifiers
source 1..3421
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin 932..1520
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 1421..1440
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 1674..1691
/label=L4440
/note="L4440 vector, forward primer"
primer_bind complement(2979..2997)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 3065..3169
/gene="bla"
/label=AmpR promoter
CDS join(3391..3421,1..761)
/codon_start=1
/gene="aadA"
/product="aminoglycoside adenylyltransferase (Murphy,
1985)"
/label=SmR
/note="confers resistance to spectinomycin and
streptomycin"
/translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
ORIGIN
1 ctcaactatc agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg ttgctggccg
61 tacatttgta cggctccgca gtggatggcg gcctgaagcc acacagcgat attgatttgc
121 tggttacggt gaccgtaagg cttgatgaaa caacgcggcg agctttgatc aacgaccttt
181 tggaaacttc ggcttcccct ggagagagcg agattctccg cgctgtagaa gtcaccattg
241 ttgtgcacga cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg caatttggag
301 aatggcagcg caatgacatt cttgcaggta tcttcgagcc agccacgatc gacattgatc
361 tggctatctt gctgacaaaa gcaagagaac atagcgttgc cttggtaggt ccagcggcgg
421 aggaactctt tgatccggtt cctgaacagg atctatttga ggcgctaaat gaaaccttaa
481 cgctatggaa ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg cttacgttgt
541 cccgcatttg gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc gctgccgact
601 gggcaatgga gcgcctgccg gcccagtatc agcccgtcat acttgaagct agacaggctt
661 atcttggaca agaagaagat cgcttggcct cgcgcgcaga tcagttggaa gaatttgtcc
721 attacgtaaa aggcgagatc accaaggtag tcggcaaata actgtcagac caagtttact
781 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
841 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
901 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
961 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
1021 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
1081 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
1141 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
1201 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
1261 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
1321 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
1381 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
1441 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
1501 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
1561 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
1621 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
1681 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
1741 cgattcatta atcactctgt ggtctcagga gtctcatatt tcttcatcac gcacatactt
1801 tttttatttc ttttattgta aagggtaaaa tcacatgttc actattaact taccactaaa
1861 atgcggtagt tttggataga tattgtagaa tgtagtagat ttagaaaggg ttttccaaaa
1921 tgtggtagat tcaacaaaat ttcttttttt ttaaaaaaaa aaatctacca cattcgagag
1981 ttgatacttt gttagctttg ctttgtattg ataatttttt tttttttttt aattgataaa
2041 aagaaaacta gatattgaca ttcggatttc ggatcggatt tgaatcgaag tttttcggat
2101 tcagataatt tggatgggaa agtttattat ccattcggat tttatgacta ttcggtttgg
2161 atgatgttgg attcagatcg gttttaggaa aaaaaattgg ttaaatttta caaaaatttg
2221 gttaaatttt acaaaaaatt gtataattgt aacaaatatg actaaattgt tttggatatc
2281 taggttattt cggatatttt ttggtataaa attataacta aatataacta atatttttta
2341 aaattataac taaatatcat aatattttta gaggtataat tttattttgg gttatcgaat
2401 atccgtttgg tttttggttc gaatcagatt cggttcgaat aattcggata gaagaagtca
2461 ttacccattt ggagttttgt gattattcga tttggtttca aaaccggatt tttggatctg
2521 tttcatattg atttttggat ttcggatttt atgtctactc ctaaaaaaac ccatgtagtt
2581 ttcgttttaa gtgtagaatt tatcttttgg ttgtaagtta tatctagtct atcttttggg
2641 acgtttggtt tcccatgttt ggattgtttg ttgtaagtcc tatgtacaac tccatcttct
2701 atcgagaatc ttttagaagg aaaaaaaaaa ctcgtattta cgaaatttga cagtagaaaa
2761 caaaaaccta agtacaattc aaattgacaa aaaaaaaaaa aaagtaaaaa ttcttaaaag
2821 gggactacag taaaaattca tgttaaatct aactgctcga acgttccagt ttccaccacc
2881 cggcgcgtga gtagtaccac cgtcgtcact ttcacaatct cccaacagag atccttccca
2941 atgtgagacc acgaagtggc tcttcagtgg acgaaagggc ctcgtgatac gcctattttt
3001 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
3061 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
3121 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgcgctcacg
3181 caactggtcc agaaccttga ccgaacgcag cggtggtaac ggcgcagtgg cggttttcat
3241 ggcttgttat gactgttttt ttttggggta cagtctatgc ctcgggcatc caagcagcaa
3301 gcgcgttacg ccgtgggtcg atgtttgatg ttatggagca gcaacgatgt tacgcagcag
3361 ggcagtcgcc ctaaaacaaa gttaaacatc atgagggaag cggtgatcgc cgaagtatcg
3421 a
//
| {
"Vector backbone": "pICH41295",
"Backbone size w/o insert (bp)": "",
"Vector type": "Synthetic Biology",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Spectinomycin, 50 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Building a pipeline to identify and engineer constitutive and repressible promoters",
"Publication": "Quant Plant Biol. 2023 Oct 19;4:e12. doi: 10.1017/qpb.2023.10. eCollection 2023.",
"DOI": "10.1017/qpb.2023.10.",
"Authors": "Yang EJY, Nemhauser JL /qpb",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/37901686",
"PubMed ID": "PubMed 37901686"
} | {
"Gene/Insert name": "pAT5G42470",
"Species": "A. thaliana (mustard weed)",
"Insert Size (bp)": "1168",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "unknown (unknown if destroyed)",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pMBPcyto-TEV-BseRI | 157739 | (Empty Backbone)empty vector adds TEV-protease-removable N-term His10 + maltose binding protein (without signal sequence); avoids unwanted residues in the final protein sequence by using the BseRI restriction sites | Debra Hansen | Empty vectors with BseRI sites avoids cloning artifacts in final protein sequence (unpublished) | https://media.addgene.org/snapgene-media/v2.0.0/sequences/334003/34ed7ddd-8800-49ae-8add-73f0997db2b3/addgene-plasmid-157739-sequence-334003.gbk | full | LOCUS sequence-334003- 6369 bp DNA circular SYN 26-AUG-2024
DEFINITION empty vector adds TEV-protease-removable N-term His10 + maltose
binding protein (without signal sequence); avoids unwanted residues
in the final protein sequence by using the BseRI restriction sites.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6369)
TITLE Empty vectors with BseRI sites avoids cloning artifacts in final
protein sequence
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 6369)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
FEATURES Location/Qualifiers
source 1..6369
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(18..36)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 104..208
/gene="bla"
/label=AmpR promoter
CDS 209..1069
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(427..446)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 1240..1828
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 1729..1748
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 1982..1999
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 2014..2153
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(2079..2098)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 2239..2261
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(2255..2446)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
primer_bind 3904..3923
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind 4007..4026
/label=T7
/note="T7 promoter, forward primer"
promoter 4007..4025
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
RBS 4114..4136
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 4156..4185
/codon_start=1
/product="10xHis affinity tag"
/label=10xHis
/translation="HHHHHHHHHH"
CDS 4195..5295
/codon_start=1
/gene="malE (mutated)"
/product="maltose binding protein from E. coli"
/label=MBP
/note="This version of the gene does not encode a signal
sequence, so MBP will remain in the cytosol."
/translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
WYAVRTAVINAASGRQTVDEALKDAQT"
primer_bind 5269..5292
/label=MBP-F
/note="Maltose binding protein, forward primer"
primer_bind complement(5905..5923)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 5919..5966
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
promoter complement(6332..6360)
/label=tet promoter
/note="E. coli promoter for tetracycline efflux protein
gene"
ORIGIN
1 ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
61 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
121 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
181 gcttcaataa cattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
241 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
301 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
361 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
421 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg
481 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
541 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
601 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
661 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
721 accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact
781 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
841 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
901 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
961 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
1021 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
1081 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
1141 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
1201 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
1261 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
1321 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
1381 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
1441 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
1501 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
1561 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
1621 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
1681 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
1741 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
1801 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
1861 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
1921 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
1981 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
2041 tctgtgcggt atttcacacc gcaatggtgc actctcagta caatctgctc tgatgccgca
2101 tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac
2161 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
2221 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
2281 aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt
2341 ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc
2401 tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt
2461 aagggggatt tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga
2521 tacgggttac tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg
2581 cggtatggat gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta
2641 atacagatgt aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca
2701 taatggtgca gggcgctgac ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga
2761 ccattcatgt tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct
2821 cgcgtatcgg tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc
2881 tcaacgacag gagcacgatc atgcgcaccc gtggccagga cccaacgctg cccgagatgc
2941 gccgcgtgcg gctgctggag atggcggacg cgatggatat gttctgccaa gggttggttt
3001 gcgcattcac agttctccgc aagaattgat tggctccaat tcttggagtg gtgaatccgt
3061 tagcgaggtg ccgccggctt ccattcaggt cgaggtggcc cggctccatg caccgcgacg
3121 caacgcgggg aggcagacaa ggtatagggc ggcgcctaca atccatgcca acccgttcca
3181 tgtgctcgcc gaggcggcat aaatcgccgt gacgatcagc ggtccaatga tcgaagttag
3241 gctggtaaga gccgcgagcg atccttgaag ctgtccctga tggtcgtcat ctacctgcct
3301 ggacagcatg gcctgcaacg cgggcatccc gatgccgccg gaagcgagaa gaatcataat
3361 ggggaaggcc atccagcctc gcgtcgcgaa cgccagcaag acgtagccca gcgcgtcggc
3421 cgccatgccg gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg gaccagtgac
3481 gaaggcttga gcgagggcgt gcaagattcc gaataccgca agcgacaggc cgatcatcgt
3541 cgcgctccag cgaaagcggt cctcgccgaa aatgacccag agcgctgccg gcacctgtcc
3601 tacgagttgc atgataaaga agacagtcat aagtgcggcg acgatagtca tgccccgcgc
3661 ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgac gctctccctt
3721 atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc
3781 cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc
3841 caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc
3901 atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc
3961 cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg cgaaattaat acgactcact
4021 atagggagac cacaacggtt tccctctagt gccggctccg gagagctctt taattaagcg
4081 gccgccctgc aggactcgag ttctagaaat aattttgttt aactttaaga aggagatata
4141 catatgaaat cttctcatca tcaccatcac catcaccacc atcacggttc ttctatgaaa
4201 atcgaagaag gtaaactggt aatctggatt aacggcgata aaggctataa cggtctcgct
4261 gaagtcggta agaaattcga gaaagatacc ggaattaaag tcaccgttga gcatccggat
4321 aaactggaag agaaattccc acaggttgcg gcaactggcg atggccctga cattatcttc
4381 tgggcacacg accgctttgg tggctacgct caatctggcc tgttggctga aatcaccccg
4441 gacaaagcgt tccaggacaa gctgtatccg tttacctggg atgccgtacg ttacaacggc
4501 aagctgattg cttacccgat cgctgttgaa gcgttatcgc tgatttataa caaagatctg
4561 ctgccgaacc cgccaaaaac ctgggaagag atcccggcgc tggataaaga actgaaagcg
4621 aaaggtaaga gcgcgctgat gttcaacctg caagaaccgt acttcacctg gccgctgatt
4681 gctgctgacg ggggttatgc gttcaagtat gaaaacggca agtacgacat taaagacgtg
4741 ggcgtggata acgctggcgc gaaagcgggt ctgaccttcc tggttgacct gattaaaaac
4801 aaacacatga atgcagacac cgattactcc atcgcagaag ctgcctttaa taaaggcgaa
4861 acagcgatga ccatcaacgg cccgtgggca tggtccaaca tcgacaccag caaagtgaat
4921 tatggtgtaa cggtactgcc gaccttcaag ggtcaaccat ccaaaccgtt cgttggcgtg
4981 ctgagcgcag gtattaacgc cgccagtccg aacaaagagc tggcaaaaga gttcctcgaa
5041 aactatctgc tgactgatga aggtctggaa gcggttaata aagacaaacc gctgggtgcc
5101 gtagcgctga agtcttacga ggaagagttg gcgaaagatc cacgtattgc cgccactatg
5161 gaaaacgccc agaaaggtga aatcatgccg aacatcccgc agatgtccgc tttctggtat
5221 gccgtgcgta ctgcggtgat caacgccgcc agcggtcgtc agactgtcga tgaagccctg
5281 aaagacgcgc agactaattc gagctcgaac aacaacaaca ataacaataa caacaacctc
5341 gggatcgagg aaaacctgta cttccaaaga tctgtcgctc ctcatgtaga caccgacact
5401 gtgaacaccg ccatcctggg cgtttcaagg tccggcaaag gcgagacaat cgtagtgcca
5461 atgatcgaca atctgagccg cgcaaagaac cagtctagca tggtcgtgaa cgaccccaag
5521 ggcgagctct actctgccag caaggaaaca cttgagaaac gcggatacga cgtgcaggtg
5581 ctgaacatcc tggatcccct gcagggcatg agttacaacc cactccagct cgtgatcgac
5641 gcctgggtga acggcgacga ccaggaggct gcaaagaggg caaacacact gactttctca
5701 ctgtacaaca acccaaatgc tggcgacaac gctttcttta acaccagcgc ccagaatgct
5761 atcaacggaa tcatcctcgc tatgaggagg cagaagcttg gatccgcgat cgcggcgcgc
5821 cacctggtgg ccggccggta ccacgcgtgc gcgctgatcc ggctgctaac aaagcccgaa
5881 aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct
5941 ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga tatccacagg
6001 acgggtgtgg tcgccatgat cgcgtagtcg atagtggctc caagtagcga agcgagcagg
6061 actgggcggc ggccaaagcg gtcggacagt gctccgagaa cgggtgcgca tagaaattgc
6121 atcaacgcat atagcgctag cagcacgcca tagtgactgg cgatgctgtc ggaatggacg
6181 atatcccgca agaggcccgg cagtaccggc ataaccaagc ctatgcctac agcatccagg
6241 gtgacggtgc cgaggatgac gatgagcgca ttgttagatt tcatacacgg tgcctgactg
6301 cgttagcaat ttaactgtga taaactaccg cattaaagct tatcgatgat aagctgtcaa
6361 acatgagaa
//
| {
"Vector backbone": "pET PPL His6 MBP LIC cloning vector (2K-T) = Addgene #37183",
"Backbone size w/o insert (bp)": "",
"Vector type": "Bacterial Expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "Low Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "",
"Publication": "",
"DOI": "",
"Authors": "",
"PubMed Link": "",
"PubMed ID": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Ligation Independent Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "GAAAGGTGAAATCATGCCGAACATC",
"3′ sequencing primer": "CTTCCTTTCGGGCTTTGTTAGC"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pET28a-PSAT | 74115 | Expression of PSAT | Lewis Cantley | Mullarky et al Proc Natl Acad Sci U S A. 2016 Feb 16;113(7):1778-83. doi: 10.1073/pnas.1521548113. Epub 2016 Feb 1. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/141328/6a4ed6d3-523b-45ef-b8ae-68694c97ad1d/addgene-plasmid-74115-sequence-141328.gbk | full | LOCUS sequence-141328- 6491 bp DNA circular SYN 26-AUG-2024
DEFINITION Expression of PSAT.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6491)
AUTHORS Mullarky E, Lucki NC, Beheshti Zavareh R, Anglin JL, Gomes AP,
Nicolay BN, Wong JC, Christen S, Takahashi H, Singh PK, Blenis J,
Warren JD, Fendt SM, Asara JM, DeNicola GM, Lyssiotis CA, Lairson
LL, Cantley LC
TITLE Identification of a small molecule inhibitor of 3-phosphoglycerate
dehydrogenase to target serine biosynthesis in cancers.
JOURNAL Proc Natl Acad Sci U S A. 2016 Feb 16;113(7):1778-83. doi:
10.1073/pnas.1521548113. Epub 2016 Feb 1.
PUBMED 26831078
REFERENCE 2 (bases 1 to 6491)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1073/pnas.1521548113"; journalName: "Proc Natl Acad Sci U S A";
date: "2016-02-16- 16"; volume: "113"; issue: "7"; pages: "1778-83"
FEATURES Location/Qualifiers
source 1..6491
/mol_type="other DNA"
/organism="synthetic DNA construct"
terminator complement(26..73)
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
primer_bind 69..87
/label=T7 Term
/note="T7 terminator, reverse primer"
CDS complement(140..157)
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS complement(1365..1382)
/codon_start=1
/product="thrombin recognition and cleavage site"
/label=thrombin site
/translation="LVPRGS"
CDS complement(1392..1409)
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
RBS 1428..1450
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
protein_bind 1465..1489
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
primer_bind complement(1489..1508)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(1490..1508)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(1592..1611)
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
promoter 1817..1894
/gene="lacI"
/label=lacI promoter
CDS 1895..2977
/codon_start=1
/gene="lacI"
/product="lac repressor"
/label=lacI
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
primer_bind complement(1914..1933)
/label=LacI-R
/note="LacI, reverse primer"
CDS 3786..3977
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
primer_bind complement(3971..3993)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
misc_feature 4079..4221
/label=bom
/note="basis of mobility region from pBR322"
primer_bind 4134..4153
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(4236..4253)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4407..4995)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4487..4506)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind complement(5106..5125)
/label=pENTR-R
/note="pENTR vectors, reverse primer"
CDS 5117..5932
/codon_start=1
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
CDS 5117..5932
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind complement(5190..5209)
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
rep_origin complement(6025..6480)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(6162..6183)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 6374..6393
/label=F1ori-R
/note="F1 origin, reverse primer"
ORIGIN
1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagcttgt
181 cgacggagct cgaattcgga tccactagta acggccgcca gtgtgctgga attcgccctt
241 ttggatcctc atagctgatg catctccaaa aattttttca tgaaggcggc cagcttctga
301 acgtcttcaa ttgtgacagc attatacaga gaggcccgga tgcctcccac agacctatgc
361 cctttcaagg acaacatatt gagttcaaga gctttatcaa gaaatctttt ttctaaagca
421 tcatctcctt tggcattgcc aatgcggaat ggaatattca tcttgcttct attttggggc
481 tccactggac aaacgtagaa tccttgagaa ttatcaataa tctcataaat tgtttgagat
541 ttgatggagc taagcttctc catggccgcg gcacctccat tgtttttaat ccactccaga
601 accaagccca tgacgtagat gctgaagcat ggaggcgtgt tgtacaagga gctgtttcca
661 gcctgcacct tgtattccag gaccgagggg cactctcgga gggcaaaccc cagcaggtca
721 tcacggacaa tcaccacggt gaccccagca gagccaacat tcttctgggc accagcaaaa
781 atcacaccaa acttggaaac atccactggc ttggacagga agtttgagga catgtcacaa
841 accagtactg ctcccttgac atcgggtata aagtcaaact ccacaccatg caccgtctca
901 tttgcgcaat aatacacgta ggaggcatct gggttgaggt tccaggtgct tggatctgga
961 atttttgtat aactcccaag tttagggtga acgatattta tagtcccaaa cttcttggct
1021 tcttctgcgg ccttagctga ccaagctcct gtcaccacat agtccgcaca ccttcctgct
1081 ttcaagccaa tgaggtttaa ggggacagca ctgaactggc cgcacccacc tccttgcaga
1141 aaaatcacct tatagttgtc tggaacagct agcaattccc gcacaagatt ctctgtattg
1201 ttaataatct tggcaaaatc tgatgacctg tgactcattt caagaacact aatgccaact
1261 cctttgtagt ctaataattc cttttgtatc tctaacaaca ctgagtgcgg cagcttggcg
1321 ggaccaggcc caaagttgac cacctgcctg ggggcgtcca tatggctgcc gcgcggcacc
1381 aggccgctgc tgtgatgatg atgatgatgg ctgctgccca tggtatatct ccttcttaaa
1441 gttaaacaaa attatttcta gaggggaatt gttatccgct cacaattccc ctatagtgag
1501 tcgtattaat ttcgcgggat cgagatctcg atcctctacg ccggacgcat cgtggccggc
1561 atcaccggcg ccacaggtgc ggttgctggc gcctatatcg ccgacatcac cgatggggaa
1621 gatcgggctc gccacttcgg gctcatgagc gcttgtttcg gcgtgggtat ggtggcaggc
1681 cccgtggccg ggggactgtt gggcgccatc tccttgcatg caccattcct tgcggcggcg
1741 gtgctcaacg gcctcaacct actactgggc tgcttcctaa tgcaggagtc gcataaggga
1801 gagcgtcgag atcccggaca ccatcgaatg gcgcaaaacc tttcgcggta tggcatgata
1861 gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt
1921 cgcagagtat gccggtgtct cttatcagac cgtttcccgc gtggtgaacc aggccagcca
1981 cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg gcggagctga attacattcc
2041 caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg ctgattggcg ttgccacctc
2101 cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca
2161 actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc
2221 ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg atcattaact atccgctgga
2281 tgaccaggat gccattgctg tggaagctgc ctgcactaat gttccggcgt tatttcttga
2341 tgtctctgac cagacaccca tcaacagtat tattttctcc catgaagacg gtacgcgact
2401 gggcgtggag catctggtcg cattgggtca ccagcaaatc gcgctgttag cgggcccatt
2461 aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat aaatatctca ctcgcaatca
2521 aattcagccg atagcggaac gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac
2581 catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca acgatcagat
2641 ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg cggatatctc
2701 ggtagtggga tacgacgata ccgaagacag ctcatgttat atcccgccgt taaccaccat
2761 caaacaggat tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc aactctctca
2821 gggccaggcg gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac
2881 cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
2941 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtaagtt
3001 agctcactca ttaggcaccg ggatctcgac cgatgccctt gagagccttc aacccagtca
3061 gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact gtcttcttta
3121 tcatgcaact cgtaggacag gtgccggcag cgctctgggt cattttcggc gaggaccgct
3181 ttcgctggag cgcgacgatg atcggcctgt cgcttgcggt attcggaatc ttgcacgccc
3241 tcgctcaagc cttcgtcact ggtcccgcca ccaaacgttt cggcgagaag caggccatta
3301 tcgccggcat ggcggcccca cgggtgcgca tgatcgtgct cctgtcgttg aggacccggc
3361 taggctggcg gggttgcctt actggttagc agaatgaatc accgatacgc gagcgaacgt
3421 gaagcgactg ctgctgcaaa acgtctgcga cctgagcaac aacatgaatg gtcttcggtt
3481 tccgtgtttc gtaaagtctg gaaacgcgga agtcagcgcc ctgcaccatt atgttccgga
3541 tctgcatcgc aggatgctgc tggctaccct gtggaacacc tacatctgta ttaacgaagc
3601 gctggcattg accctgagtg atttttctct ggtcccgccg catccatacc gccagttgtt
3661 taccctcaca acgttccagt aaccgggcat gttcatcatc agtaacccgt atcgtgagca
3721 tcctctctcg tttcatcggt atcattaccc ccatgaacag aaatccccct tacacggagg
3781 catcagtgac caaacaggaa aaaaccgccc ttaacatggc ccgctttatc agaagccaga
3841 cattaacgct tctggagaaa ctcaacgagc tggacgcgga tgaacaggca gacatctgtg
3901 aatcgcttca cgaccacgct gatgagcttt accgcagctg cctcgcgcgt ttcggtgatg
3961 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
4021 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg
4081 cagccatgac ccagtcacgt agcgatagcg gagtgtatac tggcttaact atgcggcatc
4141 agagcagatt gtactgagag tgcaccatat atgcggtgtg aaataccgca cagatgcgta
4201 aggagaaaat accgcatcag gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
4261 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
4321 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
4381 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
4441 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
4501 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
4561 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
4621 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
4681 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
4741 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
4801 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
4861 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
4921 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
4981 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
5041 gaaaactcac gttaagggat tttggtcatg aacaataaaa ctgtctgctt acataaacag
5101 taatacaagg ggtgttatga gccatattca acgggaaacg tcttgctcta ggccgcgatt
5161 aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca
5221 atcaggtgcg acaatctatc gattgtatgg gaagcccgat gcgccagagt tgtttctgaa
5281 acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct
5341 gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg
5401 gttactcacc actgcgatcc ccgggaaaac agcattccag gtattagaag aatatcctga
5461 ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc
5521 tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg
5581 aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt
5641 tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac
5701 tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat
5761 tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg
5821 cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa
5881 tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct aagaattaat
5941 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
6001 catttccccg aaaagtgcca cctgaaattg taaacgttaa tattttgtta aaattcgcgt
6061 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
6121 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
6181 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
6241 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
6301 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
6361 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
6421 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
6481 cccattcgcc a
//
| {
"Vector backbone": "pET28a",
"Backbone size w/o insert (bp)": "5369",
"Vector type": "Mammalian Expression, Bacterial Expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Kanamycin, 50 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "Unknown"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Identification of a small molecule inhibitor of 3-phosphoglycerate dehydrogenase to target serine biosynthesis in cancers",
"Publication": "Proc Natl Acad Sci U S A. 2016 Feb 16;113(7):1778-83. doi: 10.1073/pnas.1521548113. Epub 2016 Feb 1.",
"DOI": "10.1073/pnas.1521548113.",
"Authors": "Mullarky E, Lucki NC, Beheshti Zavareh R, Anglin JL, Gomes AP, Nicolay BN, Wong JC, Christen S, Takahashi H, Singh PK, Blenis J, Warren JD, Fendt SM, Asara JM, DeNicola GM, Lyssiotis CA, Lairson LL, Cantley LC /pnas",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/26831078",
"PubMed ID": "PubMed 26831078"
} | {
"Gene/Insert name": "pET28a-PSAT",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "1110",
"Mutation": "",
"Entrez Gene": "PSAT1",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pENTR-SYNZIP7 | 80663 | Encodes the synthetic coiled coil peptide SYNZIP7 | Amy Keating | Thompson et al ACS Synth Biol. 2012 Apr 20;1(4):118-29. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/171183/08eb38df-ee5b-40f3-83ba-f95da0ba5292/addgene-plasmid-80663-sequence-171183.gbk | full | LOCUS sequence-171183- 2738 bp DNA circular SYN 26-AUG-2024
DEFINITION Encodes the synthetic coiled coil peptide SYNZIP7.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2738)
AUTHORS Thompson KE, Bashor CJ, Lim WA, Keating AE
TITLE SYNZIP protein interaction toolbox: in vitro and in vivo
specifications of heterospecific coiled-coil interaction domains.
JOURNAL ACS Synth Biol. 2012 Apr 20;1(4):118-29.
PUBMED 22558529
REFERENCE 2 (bases 1 to 2738)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth
Biol."; date: "2012-04-20"; volume: "1(4)"; pages: "118-29"
FEATURES Location/Qualifiers
source 1..2738
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 92..109
/label=L4440
/note="L4440 vector, forward primer"
terminator complement(268..295)
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator complement(387..473)
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
primer_bind 522..544
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 536..553
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 537..553
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 569..668
/gene="mutant version of attL"
/label=attL1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
protein_bind complement(863..962)
/gene="mutant version of attL"
/label=attL2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
primer_bind complement(979..998)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(980..998)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(1003..1019)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1003..1019)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(1121..1140)
/label=pENTR-R
/note="pENTR vectors, reverse primer"
CDS 1132..1941
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind complement(1199..1218)
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
rep_origin 2088..2676
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2577..2596
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
661 agcaggctcc gcggccgccc ccttcaccat gaaagagatc gaatacctgg aaaaagaaat
721 tgaacgtctg aaagacctgc gtgaacacct gaaacaggac aacgcggctc accgtcagga
781 actgaacgcg ctgcgtctgg aagaagcgaa actggaattc atcctggcgc acctgctgtc
841 tacctgaagg gtgggcgcgc cgacccagct ttcttgtaca aagttggcat tataagaaag
901 cattgcttat caatttgttg caacgaacag gtcactatca gtcaaaataa aatcattatt
961 tgccatccag ctgatatccc ctatagtgag tcgtattaca tggtcatagc tgtttcctgg
1021 cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca
1081 tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt tatgagccat
1141 attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga tttatatggg
1201 tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg
1261 aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt
1321 acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag
1381 cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca
1441 gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca
1501 gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc
1561 gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat
1621 tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataaactt
1681 ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt
1741 tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga
1801 taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa
1861 cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg
1921 atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacactg gcagagcatt
1981 acgctgactt gacgggacgg cgcaagctca tgaccaaaat cccttaacgt gagttacgcg
2041 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
2101 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
2161 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
2221 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
2281 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
2341 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
2401 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
2461 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
2521 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
2581 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
2641 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
2701 cggttcctgg ccttttgctg gccttttgct cacatgtt
//
| {
"Vector backbone": "pENTR_D-TOPO",
"Backbone size w/o insert (bp)": "2586",
"Vector type": "Entry vector",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Kanamycin, 50 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "Unknown"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "SYNZIP protein interaction toolbox: in vitro and in vivo specifications of heterospecific coiled-coil interaction domains",
"Publication": "ACS Synth Biol. 2012 Apr 20;1(4):118-29.",
"DOI": "10.1021/sb200015u",
"Authors": "Thompson KE, Bashor CJ, Lim WA, Keating AE",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/22558529",
"PubMed ID": "PubMed 22558529"
} | {
"Gene/Insert name": "SYNZIP7",
"Species": "Synthetic",
"Insert Size (bp)": "156",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "TOPO Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "M13 (-21) Forward, TGTAAAACGACGGCCAGT",
"3′ sequencing primer": "M13 Reverse, CAGGAAACAGCTATGAC"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pAW212 | 172196 | P1 integration cassette for AT110 Polymerase for integration onto Orthorep for Nanobody evolution with different restriction sites | Chang Liu | Wellner et al Nat Chem Biol. 2021 Jun 24. pii: 10.1038/s41589-021-00832-4. doi: 10.1038/s41589-021-00832-4. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/338565/b91b0d20-cb8e-4d19-ab44-fda9ed5df96c/addgene-plasmid-172196-sequence-338565.gbk | full | LOCUS sequence-338565- 4614 bp DNA circular SYN 25-AUG-2024
DEFINITION P1 integration cassette for AT110 Polymerase for integration onto
Orthorep for Nanobody evolution with different restriction sites.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4614)
AUTHORS Wellner A, McMahon C, Gilman MSA, Clements JR, Clark S, Nguyen KM,
Ho MH, Hu VJ, Shin JE, Feldman J, Hauser BM, Caradonna TM, Wingler
LM, Schmidt AG, Marks DS, Abraham J, Kruse AC, Liu CC
TITLE Rapid generation of potent antibodies by autonomous hypermutation in
yeast.
JOURNAL Nat Chem Biol. 2021 Jun 24. pii: 10.1038/s41589-021-00832-4. doi:
10.1038/s41589-021-00832-4.
PUBMED 34168368
REFERENCE 2 (bases 1 to 4614)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat Chem
Biol. 2021 Jun 24. pii: 10.1038/s41589-021-00832-4. doi:
10.1038/s41589-021-00832-4."
FEATURES Location/Qualifiers
source 1..4614
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 118..137
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 371..388
/label=L4440
/note="L4440 vector, forward primer"
CDS 1311..1322
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
CDS 1323..1349
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
misc_RNA 1720..1771
/label=sTRSV HHRz
/note="hammerhead ribozyme from the tobacco ringspot virus
satellite RNA (Khvorova et al., 2003)"
terminator 1778..1943
/gene="S. cerevisiae ADH1"
/label=ADH1 terminator
/note="transcription terminator for the S. cerevisiae
alcohol dehydrogenase 1 (ADH1) gene"
primer_bind complement(1833..1857)
/label=pHybLex-R
/note="ADH terminator, reverse primer"
CDS complement(2015..2686)
/codon_start=1
/gene="S. cerevisiae TRP1"
/product="phosphoribosylanthranilate isomerase, required
for tryptophan biosynthesis"
/label=TRP1
/note="yeast auxotrophic marker"
/translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
KK"
primer_bind 2975..2997
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
promoter complement(3937..4028)
/gene="bla"
/label=AmpR promoter
terminator complement(4095..4142)
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
primer_bind 4138..4156
/label=T7 Term
/note="T7 terminator, reverse primer"
rep_origin join(4243..4614,1..217)
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
ORIGIN
1 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
61 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
121 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
181 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
241 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
301 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
361 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
421 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
481 agcgggcagt gagcgcaacg caatagtact ataatatatg aattacatta ttaatttaaa
541 aacgcgtgat gacctataca taggaagatc tatagaaaca aaaagattaa taactttcaa
601 atatcagaaa aatatagaaa catgtgataa gctcatagac atataaagga tccatgagat
661 tcccatctat cttcaccgct gttttgttcg ctgcttcttc tgctttggct gctccagcta
721 acaccaccac cgaagacgaa accgctcaaa tcccagctga agctgttatc gactactctg
781 acttggaagg tgacttcgac gctgctgctt tgccattgtc taactctacc aacaacggtt
841 tgtcttctac caacaccacc atcgcttcta tcgctgctaa ggaagaaggt gttcaattgg
901 acaagagaga agctgacgca ggtacccagg tgcagctgca ggaaagcggc ggcggcctgg
961 tgcaggcggg cggcagcctg cgcctgagct gcgcggcgag cggctatatt tctgacgctt
1021 actacatggg ctggtatcgc caggcgccgg gcaaagaacg cgaatttgtt gccactatta
1081 ctcatggtac taatacctat tatgcggata gcgtgaaagg ccgctttacc attagccgcg
1141 ataacgcgaa aaacaccgtg tatctgcaga tgaacagcct gaaaccggaa gataccgcgg
1201 tgtattattg cgcggttctg gaaactcgtt cttactcttt tcgttattgg ggccagggca
1261 cccaggtgac cgtgagcagc gtcgacggtt ccaaggacaa tagctcgacg attgaaggta
1321 gatacccata cgacgttcca gactacgctc tgcaggctag tggtggtggt ggttctggtg
1381 gtggtggttc tggtggtggt ggttctgcta gcgacgtcca ggaactgaca actatatgcg
1441 agcaaatccc ctcaccaact ttagaatcga cgccgtactc tttgtcaacg actactattt
1501 tggccaacgg gaaggcaatg caaggagttt ttgaatatta caaatcagta acgtttgtca
1561 gtaattgcgg ttctcacccc tcaacaacta gcaaaggcag ccccataaac acacagtatg
1621 ttttttgaag aagacttgtc atgaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
1681 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaac ctgtcaccgg atgtgttttc
1741 cggtctgatg agtccgtgag gacgaaacag ggaattcgcg aatttcttat gatttatgat
1801 ttttattatt aaataagtta taaaaaaaat aagtgtatac aaattttaaa gtgactctta
1861 ggttttaaaa cgaaaattct tattcttgag taactctttc ctgtaggtca ggttgctttc
1921 tcaggtatag catgaggtcg ctcgagctcc gtttctatta tgaatttcat ttataaagtt
1981 tatgtacaaa tatcataaaa aaagagaatc tttttttctt agcatttttg acgaaatttg
2041 ctattttgtt agagtctttt acaccatttg tctccacacc tccgcttaca tcaacaccaa
2101 taacgccatt taatctaagc gcatcaccaa cattttctgg cgtcagtcca ccagctaaca
2161 taaaatgtaa gctttcgggg ctctcttgcc ttccaaccca gtcagaaatc gagttccaat
2221 ccaaaagttc acctgtccca cctgcttctg aatcaaacaa gggaataaac gaatgaggtt
2281 tctgtgaagc tgcactgagt agtatgttgc agtcttttgg aaatacgagt cttttaataa
2341 ctggcaaacc gaggaactct tggtattctt gccacgactc atctccatgc agttggacga
2401 tatcaatgcc gtaatcattg accagagcca aaacatcctc cttaggttga ttacgaaaca
2461 cgccaaccaa gtatttcgga gtgcctgaac tatttttata tgcttttaca agacttgaaa
2521 ttttccttgc aataaccggg tcaattgttc tctttctatt gggcacacat ataataccca
2581 gcaagtcagc atcggaatct agagcacatt ctgcggcctc tgtgctctgc aagccgcaaa
2641 ctttcaccaa tggaccagaa ctacctgtga aattaataac agacatttta taattattat
2701 aaaaactttc atataagaga tataaaattt aatatggaag aaataagaca agaaagatac
2761 aaccaaatga aagaagctct aaatagtgtt gaaggttata aaggaaaaat tgtagcctca
2821 gactcagatt ggtgtttcaa agatcctcaa ggcaatagaa taacagattt tgatagtact
2881 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
2941 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
3001 tcaccgtcat caccgaaacg cgcgagacga aaggcaaaaa aaaaggctcc aaaaggagcc
3061 tttaattgta tcggtttacc aatgcttaat cagtgaagcc cctatttcag caatctgtct
3121 atttcgttcg tccatagttg cctgactacc cgtcgtgtag ataactacga tacgggaagg
3181 cttaccatct ggtccaagtg cggcaatgat accccgtgaa ccacgctctc ctgccccaga
3241 tttatcagca ataaaccagc cagcaggaag tgccgaacgc agtagtggtc ctgcaacttt
3301 atccgcttca agccagtcta ttaattgttg ccgtgaagcc agagtaagta gttcgccagt
3361 taatagtttg cgtaatgttg ttgccattgc cgcaggcatt gtggtgtctc gctcgtcgtt
3421 tggtatggct tcattcagtt ctggttccca acggtcaagg cgagttacat ggtctcccat
3481 attgtgcaaa aaagcggtta attccttcgg tccaccgatg gttgtcagaa gtaagttcgc
3541 cgcagtgtta tcactcatcg ttatggcggc actacataat tctcttaccg tcattccgtc
3601 cgtaaggtgc ttttctgtca ctggggagta ttcaaccaag tcattctgcg aatagtgtat
3661 tcgtcgtccg agttgctctt gccccgcatc tacacgggat aatactgctc cacatagcag
3721 aactttgaaa gtgctcatca ttggaaaacg ctcttcaggg cgaaaactct caaggatttt
3781 accgctattc aagtccagtt ctatgtatcc cactcgtgcc cccaattggt cctctgcgtc
3841 ttttactttc acgagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
3901 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
3961 aagcatttat cagggttatt gtctgatgag cggatacata tttgaatgta tttagaaaaa
4021 taaacaaata ggggaattaa aaaaaagccc gctcattagg cgggctacta gtcggatata
4081 gttcctcctt tcagcaaaaa acccctcaag acccgtttag aggccccaag gggttatgct
4141 agttattgct cagcggtggc agcagccaac tcagcttcct ttcgggcttt gttagcagcc
4201 ggatcttcta gaagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
4261 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
4321 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
4381 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
4441 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
4501 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
4561 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aact
//
| {
"Vector backbone": "ColE1",
"Backbone size w/o insert (bp)": "2244",
"Vector type": "Yeast Expression",
"Selectable markers": "TRP1"
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "Unknown"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Rapid generation of potent antibodies by autonomous hypermutation in yeast",
"Publication": "Nat Chem Biol. 2021 Jun 24. pii: 10.1038/s41589-021-00832-4. doi: 10.1038/s41589-021-00832-4.",
"DOI": "10.1038/s41589-021-00832-4.",
"Authors": "Wellner A, McMahon C, Gilman MSA, Clements JR, Clark S, Nguyen KM, Ho MH, Hu VJ, Shin JE, Feldman J, Hauser BM, Caradonna TM, Wingler LM, Schmidt AG, Marks DS, Abraham J, Kruse AC, Liu CC /s---",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/34168368",
"PubMed ID": "PubMed 34168368"
} | {
"Gene/Insert name": "p10B2-AT110",
"Species": "Synthetic",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "10B2 (P1 specific promoter)",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Gibson Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "gttggccgattcattaatgcag",
"3′ sequencing primer": "AACCTCTGACACATGCAGCTCC"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pCAGGS-S-B.1.617.1-FLAG | 177098 | B.1.617.1 variant S-protein with c-terminal FLAG tag | Daniel Conway | Spike variants with FLAG tags (unpublished) | https://media.addgene.org/snapgene-media/v2.0.0/sequences/346795/a0353188-0407-4784-a175-13fdf972bf95/addgene-plasmid-177098-sequence-346795.gbk | full | LOCUS sequence-346795- 8598 bp DNA circular SYN 26-AUG-2024
DEFINITION B.1.617.1 variant S-protein with c-terminal FLAG tag.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8598)
TITLE Spike variants with FLAG tags
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 8598)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
FEATURES Location/Qualifiers
source 1..8598
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 4..383
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
intron 662..1682
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
primer_bind 1602..1624
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 1690..1709
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
CDS complement(3055..3072)
/codon_start=1
/product="tetracysteine peptide that binds biarsenical
labeling reagents"
/label=tetracysteine tag
/translation="CCPGCC"
CDS 5568..5591
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
primer_bind complement(5619..5638)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_signal 5684..5739
/label=beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(5738..5757)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
primer_bind complement(6100..6116)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6100..6116)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(6113..6135)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 6124..6140
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(6148..6178)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6193..6214
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 6273..6468
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 6319..6454
/label=SV40 ori
/note="SV40 origin of replication"
polyA_signal 6474..6608
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(6511..6530)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 6565..6584
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(6676..6693)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(6847..7435)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6927..6946)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(7606..8466)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 8229..8248
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(8467..8571)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
361 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
421 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
481 cgatgggggc gggggggggg ggggnsgcgc gccrggsggg gsggggsggg gsgrggggsg
541 gggsggggsg rggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gkgggggtgc cgggcggggc
1261 ggggccgcct cgggccgggg rgggctcggg ggrggggcgc ggcggccccc ggagcgccgg
1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
1621 gaccggcggc tctagctaga gcctctgcta accatgttca tgccttcttc tttttcctac
1681 agctcctggg caacgtgctg gttattgtgc tgtctcatca ttttggcaaa gaattcggat
1741 ccatgtttgt gttcctggtg ctgctgccac tggtgtccag ccagtgtgtg aacctgacca
1801 ccaggaccca acttcctcct gcctacacca actccttcac caggggagtc tactaccctg
1861 acaaggtgtt caggtcctct gtgctgcaca gcacccagga cctgttcctg ccattcttca
1921 gcaatgtgac ctggttccat gccatccatg tgtctggcac caatggcacc aagaggtttg
1981 acaaccctgt gctgccattc aatgatggag tctactttgc cagcacagag aagagcaaca
2041 tcatcagggg ctggattttt ggcaccaccc tggacagcaa gacccagtcc ctgctgattg
2101 tgaacaatgc caccaatgtg gtgattaagg tgtgtgagtt ccagttctgt aatgacccat
2161 tcctggacgt ctactaccac aagaacaaca agtcctggat gaagtctgag ttcagggtct
2221 actcctctgc caacaactgt acctttgaat atgtgagcca accattcctg atggacttgg
2281 agggcaagca gggcaacttc aagaacctga gggagtttgt gttcaagaac attgatggct
2341 acttcaagat ttacagcaaa cacacaccaa tcaacctggt gagggacctg ccacagggct
2401 tctctgcctt ggaaccactg gtggacctgc caattggcat caacatcacc aggttccaga
2461 ccctgctggc tctgcacagg tcctacctga cacctggaga ctcctcctct ggctggacag
2521 caggagcagc agcctactat gtgggctacc tccaaccaag gaccttcctg ctgaaataca
2581 atgagaatgg caccatcaca gatgctgtgg actgtgccct ggacccactg tctgagacca
2641 agtgtaccct gaaatccttc acagtggaga agggcatcta ccagaccagc aacttcaggg
2701 tccaaccaac agagagcatt gtgaggtttc caaacatcac caacctgtgt ccatttggag
2761 aggtgttcaa tgccaccagg tttgcctctg tctatgcctg gaacaggaag aggattagca
2821 actgtgtggc tgactactct gtgctctaca actctgcctc cttcagcacc ttcaagtgtt
2881 atggagtgag cccaaccaaa ctgaatgacc tgtgtttcac caatgtctat gctgactcct
2941 ttgtgattag gggagatgag gtgagacaga ttgcccctgg acaaacaggc aagattgctg
3001 actacaacta caaactgcct gatgacttca caggctgtgt gattgcctgg aacagcaaca
3061 acctggacag caaggtggga ggcaactaca actacaggta cagactgttc aggaagagca
3121 acctgaaacc atttgagagg gacatcagca cagagattta ccaggctggc agcacaccat
3181 gtaatggagt gcagggcttc aactgttact ttccactcca atcctatggc ttccaaccaa
3241 ccaatggagt gggctaccaa ccatacaggg tggtggtgct gtcctttgaa ctgctccatg
3301 cccctgccac agtgtgtgga ccaaagaaga gcaccaacct ggtgaagaac aagtgtgtga
3361 acttcaactt caatggactg acaggcacag gagtgctgac agagagcaac aagaagttcc
3421 tgccattcca acagtttggc agggacattg ctgacaccac agatgctgtg agggacccac
3481 agaccttgga gattctggac atcacaccat gttcctttgg aggagtgtct gtgattacac
3541 ctggcaccaa caccagcaac caggtggctg tgctctacca gggtgtgaac tgtactgagg
3601 tgcctgtggc tatccatgct gaccaactta caccaacctg gagggtctac agcacaggca
3661 gcaatgtgtt ccagaccagg gctggctgtc tgattggagc agagcatgtg aacaactcct
3721 atgagtgtga catcccaatt ggagcaggca tctgtgcctc ctaccagacc cagaccaaca
3781 gcagaaggag ggcaaggtct gtggcaagcc agagcatcat tgcctacaca atgagtctgg
3841 gagcagagaa ctctgtggct tacagcaaca acagcattgc catcccaacc aacttcacca
3901 tctctgtgac cacagagatt ctgcctgtga gtatgaccaa gacctctgtg gactgtacaa
3961 tgtatatctg tggagacagc acagagtgta gcaacctgct gctccaatat ggctccttct
4021 gtacccaact taacagggct ctgacaggca ttgctgtgga acaggacaag aacacccagg
4081 aggtgtttgc ccaggtgaag cagatttaca agacacctcc aatcaaggac tttggaggct
4141 tcaacttcag ccagattctg cctgacccaa gcaagccaag caagaggtcc ttcattgagg
4201 acctgctgtt caacaaggtg accctggctg atgctggctt catcaagcaa tatggagact
4261 gtctgggaga cattgctgcc agggacctga tttgtgccca gaagttcaat ggactgacag
4321 tgctgcctcc actgctgaca gatgagatga ttgcccaata cacctctgcc ctgctggctg
4381 gcaccatcac ctctggctgg acctttggag caggagcagc cctccaaatc ccatttgcta
4441 tgcagatggc ttacaggttc aatggcattg gagtgaccca gaatgtgctc tatgagaacc
4501 agaaactgat tgccaaccag ttcaactctg ccattggcaa gattcaggac tccctgtcca
4561 gcacagcctc tgccctgggc aaactccaag atgtggtgaa ccagaatgcc caggctctga
4621 acaccctggt gaagcaactt tccagcaact ttggagccat ctcctctgtg ctgaatgaca
4681 tcctgagcag actggacaag gtggaggctg aggtccagat tgacagactg attacaggca
4741 gactccaatc cctccaaacc tatgtgaccc aacaacttat cagggctgct gagattaggg
4801 catctgccaa cctggctgcc accaagatga gtgagtgtgt gctgggacaa agcaagaggg
4861 tggacttctg tggcaagggc taccacctga tgagttttcc acagtctgcc cctcatggag
4921 tggtgttcct gcatgtgacc tatgtgcctg cccatgagaa gaacttcacc acagcccctg
4981 ccatctgcca tgatggcaag gctcactttc caagggaggg agtgtttgtg agcaatggca
5041 ccgactggtt tgtgacccag aggaacttct atgaaccaca gattatcacc acagacaaca
5101 cctttgtgtc tggcaactgt gatgtggtga ttggcattgt gaacaacaca gtctatgacc
5161 cactccaacc tgaactggac tccttcaagg aggaactgga caaatacttc aagaaccaca
5221 ccagccctga tgtggacctg ggagacatct ctggcatcaa tgcctctgtg gtgaacatcc
5281 agaaggagat tgacagactg aatgaggtgg ctaagaacct gaatgagtcc ctgattgacc
5341 tccaagaact gggcaaatat gaacaataca tcaagtggcc atggtacatc tggctgggct
5401 tcattgctgg actgattgcc attgtgatgg tgaccatcat gctgtgctgc atgacctcat
5461 gttgctcctg tcttaagggg tgctgctcct gcggttcctg ttgcaaattt gacgaggacg
5521 acagcgaacc ggtgcttaag ggtgtgaagc tgcattacac cggaggcgat tacaaggatg
5581 acgatgacaa gtgacggccg cctgcaggtt aattaagatc tttttccctc tgccaaaaat
5641 tatggggaca tcatgaagcc ccttgagcat ctgacttctg gctaataaag gaaatttatt
5701 ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggaaggac atatgggagg
5761 gcaaatcatt taaaacatca gaatgagtat ttggtttaga gtttggcaac atatgcccat
5821 atgctggctg ccatgaacaa aggttggcta taaagaggtc atcagtatat gaaacagccc
5881 cctgctgtcc attccttatt ccatagaaaa gccttgactt gaggttagat tttttttata
5941 ttttgttttg tgttattttt ttctttaaca tccctaaaat tttccttaca tgttttacta
6001 gccagatttt tcctcctctc ctgactactc ccagtcatag ctgtccctct tctcttatgg
6061 agatccctcg acctgcagcc caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
6121 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
6181 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
6241 ccagtcggga aacctgtcgt gccagcggat ccgcatctca attagtcagc aaccatagtc
6301 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
6361 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
6421 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctaacttgt
6481 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
6541 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
6601 tctggatccg ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
6661 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
6721 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
6781 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
6841 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
6901 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
6961 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
7021 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
7081 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
7141 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
7201 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
7261 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
7321 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
7381 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
7441 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
7501 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
7561 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
7621 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
7681 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
7741 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
7801 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
7861 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
7921 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
7981 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
8041 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
8101 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
8161 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
8221 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
8281 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
8341 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
8401 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
8461 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
8521 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
8581 ccgaaaagtg ccacctgg
//
| {
"Vector backbone": "pCAGGS",
"Backbone size w/o insert (bp)": "",
"Vector type": "Mammalian Expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Industry MTA"
} | {
"Title": "",
"Publication": "",
"DOI": "",
"Authors": "",
"PubMed Link": "",
"PubMed ID": ""
} | {
"Gene/Insert name": "S-protein B.1.617.1",
"Species": "SARS-CoV-2",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "S",
"Promoter": "",
"Tag / Fusion Protein": "FLAG (C terminal on insert)"
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "EcoRI (unknown if destroyed)",
"3′ cloning site": "BstEII (unknown if destroyed)",
"5′ sequencing primer": "pCAG-F",
"3′ sequencing primer": "Bglob-PA-R"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pNCX3.1 | 67147 | Expresses mRNA of NCX3.1 (Rat brain) | Nicoll et al J Biol Chem. 1996 Oct 4;271(40):24914-21. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/331642/95f1211e-183a-44bb-85bb-210a923fb415/addgene-plasmid-67147-sequence-331642.gbk | full | LOCUS sequence-331642- 6093 bp DNA circular SYN 26-AUG-2024
DEFINITION Expresses mRNA of NCX3.1 (Rat brain).
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6093)
AUTHORS Nicoll DA, Quednau BD, Qui Z, Xia YR, Lusis AJ, Philipson KD
TITLE Cloning of a third mammalian Na+-Ca2+ exchanger, NCX3.
JOURNAL J Biol Chem. 1996 Oct 4;271(40):24914-21.
PUBMED 8798769
REFERENCE 2 (bases 1 to 6093)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J Biol
Chem."; date: "1996-10-4"; volume: "271(40)"; pages: "24914-21"
FEATURES Location/Qualifiers
source 1..6093
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 36..58
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 50..67
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 51..67
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 74..93
/label=T7
/note="T7 promoter, forward primer"
promoter 74..92
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(3625..3642)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3796..4384)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3876..3895)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4555..5415)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 5178..5197
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5416..5520)
/gene="bla"
/label=AmpR promoter
rep_origin 5547..6002
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5634..5653)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 5844..5865
/label=F1ori-F
/note="F1 origin, forward primer"
ORIGIN
1 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
61 ggccagtgaa ttgtaatacg actcactata gggcgaattg ggtaccgggc cccccctcga
121 ggtcctggaa gctaagggga atcggtctca ggcctgtcgg gaagaccatt ggtgctgtgg
181 tggggccaga gcctaagtct tgtgtatggc gtggttacgg ctgcagcctc tcacctctgc
241 cttcctccat ttcgggctgg ttacttttgt gctcttcctg aatggtcttc gagcagaggc
301 tggtgacttg agggatgtgc ccagtgcagg acagaacaat gagtcctgtt cagggtcatc
361 agactgcaag gagggtgtca tcttgccaat ctggtatcca gagaaccctt cccttgggga
421 caagattgca agggtcattg tctattttgt ggccctgata tacatgtttc ttggagtgtc
481 tatcattgct gaccgattca tggcatctat tgaagttatt acctcccaag agagggaggt
541 gacaatcaag aagcccaatg gagagaccag cacaactaca attcgggtat ggaatgaaac
601 tgtctccaac ctgaccctga tggccctagg ctcttctgct ccggagattc tcctgtcttt
661 aattgaggtg tgtggtcacg ggttcattgc tggtgatttg ggaccgtcta ccattgtcgg
721 cagtgcagcc ttcaacatgt tcatcatcat tggcatctgt gtctatgtga tcccagatgg
781 ggagactcgc aagatcaagc accttcgagt cttctttgtc acggctgctt ggagcgtctt
841 tgcctatatt tggctctaca tgatcctggc agtcttctct cctggtgtgg tccaggtttg
901 ggaaggcctc cttactctct tcttctttcc agtgtgtgtc ctgctggctt gggtggcaga
961 taagcgactg ctcttctaca aatacatgca caaaagatac cgcacagata aacaccgagg
1021 aattatcatt gagacagagg gtgaacaccc taagggcatt gagatggatg ggaaaatgat
1081 gaattctcac ttcctggatg ggaaccttat acccttggaa ggaaaggagg tagatgaatc
1141 tcgcagggaa atgatccgaa ttcttaagga tctgaagcaa aaacacccag aaaaagactt
1201 agatcagctg gtggagatgg ccaattacta tgctctttcc caccaacaga agagccgtgc
1261 tttctaccgc atccaagcca cccggatgat gactggtgca ggcaatatac ttaagaaaca
1321 tgcagcagag caagccaaga agaccgccag catgagtgag gtgcatactg atgagcctga
1381 ggactttgcc tcgaaggtct tttttgaccc atgctcttac cagtgcctgg agaactgtgg
1441 agctgtcctc ctgactgtgg tgaggaaagg gggagatata tccaagacta tgtacgtgga
1501 ctacaaaaca gaggacggct ctgccaatgc aggggctgac tatgagttca cagaaggcac
1561 tgtggttctg aagccaggag agacccagaa ggagttctcc gtgggcatca tcgatgatga
1621 catttttgaa gaggatgaac acttcttcgt gagactgagc aatgtccgtg tagaagagga
1681 gcagctggaa gaggggatga cccccgccat cctcaatagt cttcctttgc cacgggctgt
1741 cctggcttcc ccttgtgtgg ccacagtaac catcttggat gatgaccatg caggaatttt
1801 cacttttgaa tgtgatacca ttcatgtcag tgaaagtatt ggtgttatgg aagtcaaggt
1861 tttgaggaca tcgggtgccc ggggcacagt catcgtccct tttaggacag tagaaggaac
1921 agccaagggt ggtggtgagg actttgaaga tacttatggg gagctggagt ttaagaatga
1981 tgaaacggtg aaaaccataa gggttaaaat agtagatgag gaggagtacg aaaggcaaga
2041 gaatttcttc attgcccttg gtgaaccgaa atggatggaa cgtggaatat cagcgctcct
2101 gttatctcca gaggtgacag acaggaagct gactatggag gaagaggagg ccaagagaat
2161 agcagagatg ggaaagccag tattgggtga acaccctaaa ctagaggtca tcattgaaga
2221 gtcctatgag ttcaagagta cagtggataa actgatcaag aagacaaacc tggcattggt
2281 tgtggggacc cattcctgga gggaccaatt catggaagcc atcactgtta gtgcagcagg
2341 agatgaggag gaagatgaat ctggagagga gaggctgcca tcatgctttg actatgtcat
2401 gcacttcctg acggtctttt ggaaggtgct ctttgcctgt gtgcccccca cagagtactg
2461 ccatggctgg gcctgcttcg tggtctccat cctcatcatc ggcatgctca ccgccatcat
2521 cggggacctg gcctctcact tcggctgcac catcgggctc aaggattcgg tcacagctgt
2581 tgtttttgtg gcattcggca cctctgtgcc agatacattt gccagcaaag ctgctgccct
2641 gcaggatgtg tatgcagatg cttctattgg caatgtcacc ggcagtaatg ctgtcaatgt
2701 cttcctgggt attggtttgg cctggtccgt cgccgccatc tactgggcca tgcagggaca
2761 ggagttccat gtgtccgctg gcactctggc cttctcggtc actcttttca ccatctttgc
2821 atttgtctgc ctcagtgtgc tcttgtaccg tcggcggccc cacctgggcg gggagcttgg
2881 aggtcctcgt ggctgcaagc ttgccacaac gtggctcttt gtgagcctgt ggctcctcta
2941 cgtactcttt gccacgctgg aggcctactg ctacatcaag gggttctgaa ccaccagaac
3001 aagacctcca cagggcaggc ctaggacttc tcttgagaga aaggcgcttt ctcaccaacc
3061 ttcccaggtt gagcatccct ggagaagaag catcgagacc tgagccccag gaacttcacc
3121 tgacctaggc cctggcactg cacagaaagg ggcaaaactt aaccaatcca atggaatgga
3181 gacgataccc actgtccaaa gtatctaatt acaaacaaac caacaacagc aacaaatcca
3241 cctctcccca tcctgccact cacacccctg agcctcccct tctattctac cactcgacta
3301 ttcctttgac tctagttcct caaggggcaa gatccactag taacggccgc cagtgtgctg
3361 gaattcggct ttctagaaaa ggaacaatca gatatagtaa atttatatat atataccgcg
3421 tactttgcct gagtcccggc cgctgctgcc actcggcaag gctggcgtgt gtactctctg
3481 cctgcctttc atcccatgct gtggtggcga aggggaatca atcccttgca aagtttgccc
3541 catttggata aatgtctata tgtaaattac aggctggttg gaggaaaaaa aaaaaaaaaa
3601 aagtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
3661 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
3721 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
3781 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
3841 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
3901 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
3961 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
4021 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
4081 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
4141 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
4201 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
4261 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
4321 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
4381 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
4441 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
4501 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
4561 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
4621 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
4681 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
4741 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
4801 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
4861 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
4921 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
4981 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
5041 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
5101 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
5161 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
5221 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
5281 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
5341 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
5401 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
5461 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
5521 cacatttccc cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg
5581 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
5641 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
5701 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
5761 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
5821 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
5881 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
5941 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat
6001 ttgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
6061 gctattacgc cagctggcga aagggggatg tgc
//
| {
"Vector backbone": "pBS?",
"Backbone size w/o insert (bp)": "",
"Vector type": "",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Cloning of a third mammalian Na+-Ca2+ exchanger, NCX3",
"Publication": "J Biol Chem. 1996 Oct 4;271(40):24914-21.",
"DOI": "",
"Authors": "Nicoll DA, Quednau BD, Qui Z, Xia YR, Lusis AJ, Philipson KD",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/8798769",
"PubMed ID": "PubMed 8798769"
} | {
"Gene/Insert name": "NCX3.1",
"Species": "R. norvegicus (rat)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
||
pT7-V5-SBP-C1-HseIF4EBP1-S65ET70E_AA | 148247 | Mammalian Expression of HseIF4EBP1-S65ET70E | Elisa Izaurralde | Izaurralde lab plasmids (unpublished) | https://media.addgene.org/snapgene-media/v2.0.0/sequences/290695/4bb3b03c-9505-4469-9d7a-c5bbe5ff570d/addgene-plasmid-148247-sequence-290695.gbk | full | LOCUS sequence-290695- 4555 bp DNA circular SYN 26-AUG-2024
DEFINITION Mammalian Expression of HseIF4EBP1-S65ET70E.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4555)
TITLE Izaurralde lab plasmids
REFERENCE 2 (bases 1 to 4555)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..4555
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 61..364
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 365..568
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 519..539
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 613..632
/label=T7
/note="T7 promoter, forward primer"
promoter 613..631
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory 659..668
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 668..709
/codon_start=1
/product="epitope tag from simian virus 5"
/label=V5 tag
/translation="GKPIPNPLLGLDST"
CDS 731..844
/codon_start=1
/product="streptavidin-binding peptide"
/label=SBP
/note="selected from a peptide library; binds streptavidin
with nanomolar affinity (Keefe et al., 2001)"
/translation="MDEKTTGWGGGHVVEGLAGELEQLRARLEHHPQGQREP"
polyA_signal 1343..1464
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1380..1399)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 1434..1453
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(1471..1926)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1608..1629)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 1820..1839
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 1953..2057
/gene="bla"
/label=AmpR promoter
promoter 2059..2416
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
enhancer 2059..2250
/label=SV40 enhancer
/note="enhancer for the SV40 early promoter (Herr, 1993)"
primer_bind complement(2082..2102)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 2267..2402
/label=SV40 ori
/note="SV40 origin of replication"
CDS 2451..3245
/codon_start=1
/product="aminoglycoside phosphotransferase"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
CDS 2451..3245
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(2505..2524)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3115..3134
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(3433..3452)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 3477..3524
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 3853..4441
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4342..4361
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
541 acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg
601 gcttatcgaa attaatacga ctcactatag ggagacccaa gctctagcgc taccggtcgc
661 caccatgggt aagcctatcc ctaaccctct cctcggtctc gattctacgc ccggaagcgg
721 tagcggtacg atggacgaga agaccaccgg ctgggggggc ggccacgtgg tggagggcct
781 ggccggcgag ctggagcagc tgcgggccag gctggagcac caccctcagg gccagcggga
841 gcccagatct cgagctatgt ccgggggcag cagctgcagc cagaccccaa gccgggccat
901 ccccgccact cgccgggtgg tgctcggcga cggcgtgcag ctcccgcccg gggactacag
961 cacgaccccc ggcggcacgc tcttcagcac caccccggga ggtaccagga tcatctatga
1021 ccggaaattc ctgatggagt gtcggaacga gcctgtgacc aaagagcccc caagggatct
1081 gcccaccatt ccgggggtca ccagcccttc cagtgatgag ccccccatgg aagccagcca
1141 gagccacctg cgcaatagcc cagaagataa gcgggcgggc ggtgaagagt cacagtttga
1201 gatggacatt taaggatcca ccggatctag ataactgatc ataatcagcc ataccacatt
1261 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
1321 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
1381 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
1441 gtccaaactc atcaatgtat cttaacgcgt aaattgtaag cgttaatatt ttgttaaaat
1501 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
1561 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
1621 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
1681 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
1741 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
1801 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
1861 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
1921 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
1981 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
2041 attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
2101 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
2161 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
2221 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
2281 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
2341 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
2401 cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac
2461 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
2521 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
2581 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg
2641 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
2701 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
2761 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
2821 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
2881 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
2941 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc
3001 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
3061 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
3121 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
3181 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
3241 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
3301 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
3361 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag
3421 ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc
3481 aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg
3541 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
3601 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
3661 caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga
3721 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
3781 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
3841 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
3901 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
3961 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
4021 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
4081 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
4141 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
4201 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
4261 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
4321 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
4381 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
4441 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
4501 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcat
//
| {
"Vector backbone": "Unknown",
"Backbone size w/o insert (bp)": "",
"Vector type": "Mammalian Expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Kanamycin, 50 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "",
"Publication": "",
"DOI": "",
"Authors": "",
"PubMed Link": "",
"PubMed ID": ""
} | {
"Gene/Insert name": "HseIF4EBP1-S65ET70E",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "EIF4EBP1",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Unknown",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pcDNA3.1-MS2-HA-HsNanos1_X | 147931 | Mammalian Expression of HsNanos1 | Elisa Izaurralde | Bhandari et al Genes Dev. 2014 Apr 15;28(8):888-901. doi: 10.1101/gad.237289.113. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/290378/c9db8a52-0a1b-4e2f-808f-6eed57ef3d0c/addgene-plasmid-147931-sequence-290378.gbk | full | LOCUS sequence-290378- 6760 bp DNA circular SYN 26-AUG-2024
DEFINITION Mammalian Expression of HsNanos1.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6760)
AUTHORS Bhandari D, Raisch T, Weichenrieder O, Jonas S, Izaurralde E
TITLE Structural basis for the Nanos-mediated recruitment of the CCR4-NOT
complex and translational repression.
JOURNAL Genes Dev. 2014 Apr 15;28(8):888-901. doi: 10.1101/gad.237289.113.
PUBMED 24736845
REFERENCE 2 (bases 1 to 6760)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1101/gad.237289"; journalName: "Genes Dev"; date: "2014-04-15-
15"; volume: "28"; issue: "8"; pages: "888-901"
FEATURES Location/Qualifiers
source 1..6760
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(44..63)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 235..614
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 615..818
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 769..789
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 863..882
/label=T7
/note="T7 promoter, forward primer"
promoter 863..881
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory 895..904
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 1333..1359
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
promoter complement(2316..2334)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(2317..2334)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(2354..2371)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2360..2584
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2630..3058
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2717..2736)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2927..2948
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(3067..3087)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 3072..3401
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3252..3387
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3314..3333
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3468..4262
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(3522..3541)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4132..4151
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 4436..4557
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4473..4492)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4527..4546
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4606..4622)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4606..4622)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4619..4641)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4630..4646
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(4654..4684)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4699..4720
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4837..4854)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5008..5593)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5088..5107)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5764..6624)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 6387..6406
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6625..6729)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttgccacc
901 atggcttcta actttactca gttcgttctc gtcgacaatg gcggaactgg cgacgtgact
961 gtcgccccaa gcaacttcgc taacggggtc gctgaatgga tcagctctaa ctcgcgatca
1021 caggcttaca aagtaacctg tagcgttcgt cagagctctg cgcagaatcg caaatacacc
1081 atcaaagtcg aggtgcctaa agtggcaacc cagactgttg gtggtgaaga gcttcctgta
1141 gccggatgga gatcttactt aaatatggaa ctaaccattc caattttcgc cacgaattcc
1201 gactgcgagc ttattgttaa ggcaatgcaa ggtctcctaa aagatggaaa cccgattccc
1261 tcggccatcg cggccaactc cggcatctac ggaggtggag gtggatctgg tggatctcct
1321 ccacatatgg cttacccata cgatgttcca gattacgctc ctccaaagct tggatccgaa
1381 ttcccgggtc gactcgagat ggaagccttc ccatgggccc ccagatcccc cagaagaggc
1441 agagcacctc ctcccatggc cctggtgcca agcgccagat atgtgtctgc ccctggacct
1501 gcccaccccc agcctttcag cagctggaac gattacctgg gcctggccac cctgatcacc
1561 aaggccgtgg atggcgagcc cagattcgga tgtgccagag gcggaaatgg cggcggaggc
1621 agccctccta gtagcagcag cagctcctgt tgcagccctc acacaggcgc tggacctggc
1681 gctctgggac ctgcactggg cccacctgac tacgacgagg acgacgacga tgacagcgac
1741 gagcctggca gcagaggcag atacctgggc tctgccctgg aactgcgggc tctggaactg
1801 tgtgccggac ctgctgaagc cggcctgctg gaagagagat tcgccgagct gagccccttc
1861 gccggcagag ctgcagcagt gctgctggga tgtgctcctg ctgccgcagc cgccgctaca
1921 acaacatctg aggccacccc tcgggaagag agggctcctg cttgggccgc tgaacctaga
1981 ctgcatgctg cctctggcgc cgctgctgca agactgctga aacccgaact gcaagtgtgc
2041 gtgttctgcc ggaacaacaa agaagctatg gccctgtaca ccacccacat cctgaagggc
2101 cccgacggca gagtgctgtg tcccgtgctg cggagataca catgccctct gtgtggcgcc
2161 agcggcgaca atgcccacac catcaagtac tgccccctga gcaaggtgcc ccctccacca
2221 gctagacccc ctccaagatc tgccagagat ggcccccctg gcaagaagct gagataatct
2281 agagtcgacc cgggcggccg cctagagggc cctattctat agtgtcacct aaatgctaga
2341 gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
2401 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
2461 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
2521 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
2581 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt
2641 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
2701 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
2761 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
2821 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
2881 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
2941 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
3001 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
3061 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
3121 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
3181 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
3241 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
3301 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
3361 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
3421 tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag
3481 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
3541 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
3601 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
3661 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
3721 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
3781 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
3841 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
3901 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
3961 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg
4021 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
4081 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
4141 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
4201 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
4261 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
4321 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
4381 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
4441 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
4501 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
4561 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
4621 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
4681 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
4741 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
4801 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
4861 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
4921 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
4981 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
5041 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
5101 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
5161 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
5221 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
5281 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
5341 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
5401 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
5461 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
5521 caaacaaacc accgctggta gcggtttttt tgtttgcaag cagcagatta cgcgcagaaa
5581 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
5641 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
5701 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
5761 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
5821 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
5881 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
5941 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
6001 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
6061 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
6121 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
6181 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
6241 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
6301 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
6361 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
6421 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
6481 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
6541 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
6601 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
6661 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
6721 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
| {
"Vector backbone": "Unknown",
"Backbone size w/o insert (bp)": "",
"Vector type": "Mammalian Expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Structural basis for the Nanos-mediated recruitment of the CCR4-NOT complex and translational repression",
"Publication": "Genes Dev. 2014 Apr 15;28(8):888-901. doi: 10.1101/gad.237289.113.",
"DOI": "10.1101/gad.237289.113.",
"Authors": "Bhandari D, Raisch T, Weichenrieder O, Jonas S, Izaurralde E /gad",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/24736845",
"PubMed ID": "PubMed 24736845"
} | {
"Gene/Insert name": "HsNanos1",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "NANOS1",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Unknown",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pDONR223_SERPINB5_p.A42F | 82738 | Gateway Donor vector containing SERPINB5, part of the Target Accelerator Plasmid Collection. | Berger et al Cancer Cell. 2016 Aug 8;30(2):214-28. doi: 10.1016/j.ccell.2016.06.022. Epub 2016 Jul 28. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/161720/748a53ba-890d-4689-be75-c24a0dfe0758/addgene-plasmid-82738-sequence-161720.gbk | full | LOCUS sequence-161720- 3486 bp DNA circular SYN 26-AUG-2024
DEFINITION Gateway Donor vector containing SERPINB5, part of the Target
Accelerator Plasmid Collection..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3486)
AUTHORS Berger AH, Brooks AN, Wu X, Shrestha Y, Chouinard C, Piccioni F,
Bagul M, Kamburov A, Imielinski M, Hogstrom L, Zhu C, Yang X, Pantel
S, Sakai R, Watson J, Kaplan N, Campbell JD, Singh S, Root DE,
Narayan R, Natoli T, Lahr DL, Tirosh I, Tamayo P, Getz G, Wong B,
Doench J, Subramanian A, Golub TR, Meyerson M, Boehm JS
TITLE High-throughput Phenotyping of Lung Cancer Somatic Mutations.
JOURNAL Cancer Cell. 2016 Aug 8;30(2):214-28. doi:
10.1016/j.ccell.2016.06.022. Epub 2016 Jul 28.
PUBMED 27478040
REFERENCE 2 (bases 1 to 3486)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1016/j.ccell.2016.06.022"; journalName: "Cancer Cell"; date:
"2016-08-8- 8"; volume: "30"; issue: "2"; pages: "214-28"
FEATURES Location/Qualifiers
source 1..3486
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 92..109
/label=L4440
/note="L4440 vector, forward primer"
terminator complement(268..295)
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator complement(387..473)
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
primer_bind 522..544
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 536..553
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 537..553
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 569..668
/gene="mutant version of attL"
/label=attL1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
protein_bind complement(1365..1464)
/gene="mutant version of attL"
/label=attL2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
primer_bind complement(1481..1500)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(1482..1500)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(1505..1521)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1505..1521)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
CDS 1952..2743
/codon_start=1
/gene="aadA"
/product="aminoglycoside adenylyltransferase (Murphy,
1985)"
/label=SmR
/note="confers resistance to spectinomycin and
streptomycin"
/translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
rep_origin 2836..3424
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3325..3344
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
661 agttggcatg gatgccctgc aactagcaaa ttcggctttt gccgttgatc tgttcaaaca
721 actatgtgaa aaggagccac tgggcaatgt cctcttctct ccaatctgtc tctccacctc
781 tctgtcactt tttcaagtgg gtgctaaagg tgacactgca aatgaaattg gacaggttct
841 tcattttgaa aatgtcaaag atgtaccctt tggatttcaa acagtaacat cggatgtaaa
901 caaacttagt tccttttact cactgaaact aatcaagcgg ctctacgtag acaaatctct
961 gaatctttct acagagttca tcagctctac gaagagaccc tatgcaaagg aattggaaac
1021 tgttgacttc aaagataaat tggaagaaac gaaaggtcag atcaacaact caattaagga
1081 tctcacagat ggccactttg agaacatttt agctgacaac agtgtgaacg accagaccaa
1141 aatccttgtg gttaatgctg cctactttgt tggcaagtgg atgaagaaat ttcctgaatc
1201 agaaacaaaa gaatgtcctt tcagagtcaa caaggtatgt ggggcagcat gtagcagtaa
1261 aaggagccca attatagatg tgaaaaatga cagggacaga gtggggcata aatccattcc
1321 aatgaggaac ctgagggcac ggccggcaaa gtgcctttcc tgattgccaa ctttcttgta
1381 caaagttggc attataagaa agcattgctt atcaatttgt tgcaacgaac aggtcactat
1441 cagtcaaaat aaaatcatta tttgccatcc agctgatatc ccctatagtg agtcgtatta
1501 catggtcata gctgtttcct ggcagctctg gcccgtgtct caaaatctct gatgttacat
1561 tgcacaagat aaaaatatat catcatgcct cctctagacc agccaggaca gaaatgcctc
1621 gacttcgctg ctacccaagg ttgccgggtg acgcacaccg tggaaacgga tgaaggcacg
1681 aacccagtgg acataagcct gttcggttcg taagctgtaa tgcaagtagc gtatgcgctc
1741 acgcaactgg tccagaacct tgaccgaacg cagcggtggt aacggcgcag tggcggtttt
1801 catggcttgt tatgactgtt tttttggggt acagtctatg cctcgggcat ccaagcagca
1861 agcgcgttac gccgtgggtc gatgtttgat gttatggagc agcaacgatg ttacgcagca
1921 gggcagtcgc cctaaaacaa agttaaacat tatgagggaa gcggtgatcg ccgaagtatc
1981 gactcaacta tcagaggtag ttggcgtcat cgagcgccat ctcgaaccga cgttgctggc
2041 cgtacatttg tacggctccg cagtggatgg cggcctgaag ccacacagtg atattgattt
2101 gctggttacg gtgaccgtaa ggcttgatga aacaacgcgg cgagctttga tcaacgacct
2161 tttggaaact tcggcttccc ctggagagag cgagattctc cgcgctgtag aagtcaccat
2221 tgttgtgcac gacgacatca ttccgtggcg ttatccagct aagcgcgaac tgcaatttgg
2281 agaatggcag cgcaatgaca ttcttgcagg tatcttcgag ccagccacga tcgacattga
2341 tctggctatc ttgctgacaa aagcaagaga acatagcgtt gccttggtag gtccagcggc
2401 ggaggaactc tttgatccgg ttcctgaaca ggatctattt gaggcgctaa atgaaacctt
2461 aacgctatgg aactcgccgc ccgactgggc tggcgatgag cgaaatgtag tgcttacgtt
2521 gtcccgcatt tggtacagcg cagtaaccgg caaaatcgcg ccgaaggatg tcgctgccga
2581 ctgggcaatg gagcgcctgc cggcccagta tcagcccgtc atacttgaag ctagacaggc
2641 ttatcttgga caagaagaag atcgcttggc ctcgcgcgca gatcagttgg aagaatttgt
2701 ccactacgtg aaaggcgaga tcaccaaggt agtcggcaaa taaccctcga gccacccatg
2761 accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa
2821 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
2881 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
2941 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
3001 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
3061 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
3121 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
3181 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
3241 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
3301 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
3361 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
3421 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
3481 catgtt
//
| {
"Vector backbone": "pDONR223",
"Backbone size w/o insert (bp)": "",
"Vector type": "Gateway Entry vector",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Spectinomycin, 50 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "High-throughput Phenotyping of Lung Cancer Somatic Mutations",
"Publication": "Cancer Cell. 2016 Aug 8;30(2):214-28. doi: 10.1016/j.ccell.2016.06.022. Epub 2016 Jul 28.",
"DOI": "10.1016/j.ccell.2016.06.022.",
"Authors": "Berger AH, Brooks AN, Wu X, Shrestha Y, Chouinard C, Piccioni F, Bagul M, Kamburov A, Imielinski M, Hogstrom L, Zhu C, Yang X, Pantel S, Sakai R, Watson J, Kaplan N, Campbell JD, Singh S, Root DE, Narayan R, Natoli T, Lahr DL, Tirosh I, Tamayo P, Getz G, Wong B, Doench J, Subramanian A, Golub TR, Meyerson M, Boehm JS /jccell",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/27478040",
"PubMed ID": "PubMed 27478040"
} | {
"Gene/Insert name": "SERPINB5",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "p.A42F; p.176S>P; p.190_192delTDTins25; p.194_196delPVQinsSIP; p.198M>R; p.200_201delMEinsLR; p.203_204delTFinsRPAK; p.206_375delinsLS",
"Entrez Gene": "SERPINB5",
"Promoter": "None",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Gateway Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "M13pUC-fwd",
"3′ sequencing primer": "M13-Rev"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
||
BS-Trp1 | 69196 | complementation of yeast auxotrophic marker TRP1. Although this is the plasmid described in the paper, the depositor notes that it has a low integration efficiency and Plasmid 69503 should be used. | Zhiping Xie | Li et al Autophagy. 2015;11(6):954-60. doi: 10.1080/15548627.2015.1040971. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/121984/d8b884d2-1c6a-4914-8570-0dbbe9909b8b/addgene-plasmid-69196-sequence-121984.gbk | full | LOCUS sequence-121984- 5330 bp DNA circular SYN 26-AUG-2024
DEFINITION complementation of yeast auxotrophic marker TRP1. Although this is
the plasmid described in the paper, the depositor notes that it has
a low integration efficiency and Plasmid 69503 should be used. .
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5330)
AUTHORS Li D, Song JZ, Shan MH, Li SP, Liu W, Li H, Zhu J, Wang Y, Lin J,
Xie Z
TITLE A fluorescent tool set for yeast Atg proteins.
JOURNAL Autophagy. 2015;11(6):954-60. doi: 10.1080/15548627.2015.1040971.
PUBMED 25998947
REFERENCE 2 (bases 1 to 5330)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Autophagy."; date: "2015"; volume: "11(6):954-60. doi"; pages: "
10.1080/15548627.2015.1040971"
FEATURES Location/Qualifiers
source 1..5330
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 24..128
/gene="bla"
/label=AmpR promoter
CDS 129..989
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(347..366)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 1160..1748
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 1649..1668
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 1902..1919
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 2036..2057
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2072..2102
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2110..2126
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
primer_bind 2115..2137
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 2134..2150
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 2134..2150
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 2169..2189
/label=T3
/note="T3 promoter, forward primer"
promoter 2171..2189
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 2260..2537
/gene="S. cerevisiae TRP1"
/label=TRP1 promoter
CDS 2538..3212
/codon_start=1
/gene="S. cerevisiae TRP1"
/product="phosphoribosylanthranilate isomerase, required
for tryptophan biosynthesis"
/label=TRP1
/note="yeast auxotrophic marker"
/translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
KK"
rep_origin 3051..3888
/label=ARS1
/note="S. cerevisiae autonomously replicating sequence
ARS1/ARS416"
primer_bind complement(4686..4705)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(4687..4705)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(4715..4732)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(4715..4731)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4724..4746)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 4873..5328
/direction=RIGHT
/label=M13 ori
/note="M13 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4960..4979)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 5170..5191
/label=F1ori-F
/note="F1 origin, forward primer"
ORIGIN
1 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
61 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
121 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
181 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
241 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
301 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
361 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
421 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
481 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
541 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
601 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
661 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
721 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
781 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
841 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
901 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
961 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
1021 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
1081 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
1141 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
1201 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
1261 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
1321 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
1381 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
1441 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
1501 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
1561 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
1621 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
1681 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
1741 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
1801 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
1861 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
1921 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
1981 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
2041 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
2101 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
2161 ccaagcgcgc aattaaccct cactaaaggg aacaaaagct ggagctccac cgcggtggcg
2221 gccgctctag aactagtcaa tcagtaaaaa tcaacggtta acgacattac tatatatata
2281 atataggaag catttaatag aacagcatcg taatatatgt gtactttgca gttatgacgc
2341 cagatggcag tagtggaaga tattctttat tgaaaaatag cttgtcacct tacgtacaat
2401 cttgatccgg agcttttctt tttttgccga ttaagaattc ggtcgaaaaa agaaaaggag
2461 agggccaaga gggagggcat tggtgactat tgagcacgtg agtatacgtg attaagcaca
2521 caaaggcagc ttggagtatg tctgttatta atttcacagg tagttctggt ccattggtga
2581 aagtttgcgg cttgcagagc acagaggccg cagaatgtgc tctagattcc gatgctgact
2641 tgctgggtat tatatgtgtg cccaatagaa agagaacaat tgacccggtt attgcaagga
2701 aaatttcaag tcttgtaaaa gcatataaaa atagttcagg cactccgaaa tacttggttg
2761 gcgtgtttcg taatcaacct aaggaggatg ttttggctct ggtcaatgat tacggcattg
2821 atatcgtcca actgcatgga gatgagtcgt ggcaagaata ccaagagttc ctcggtttgc
2881 cagttattaa aagactcgta tttccaaaag actgcaacat actactcagt gcagcttcac
2941 agaaacctca ttcgtttatt cccttgtttg attcagaagc aggtgggaca ggtgaacttt
3001 tggattggaa ctcgatttct gactgggttg gaaggcaaga gagccccgaa agcttacatt
3061 ttatgttagc tggtggactg acgccagaaa atgttggtga tgcgcttaga ttaaatggcg
3121 ttattggtgt tgatgtaagc ggaggtgtgg agacaaatgg tgtaaaagac tctaacaaaa
3181 tagcaaattt cgtcaaaaat gctaagaaat aggttattac tgagtagtat ttatttaagt
3241 attgtttgtg cacttgcctg caggcctttt gaaaagcaag cataaaagat ctaaacataa
3301 aatctgtaaa ataacaagat gtaaagataa tgctaaatca tttggctttt tgattgattg
3361 tacaggaaaa tatacatcgc agggggttga cttttaccat ttcaccgcaa tggaatcaaa
3421 cttgttgaag agaatgttca caggcgcata cgctacaatg acccgattct tgctagcctt
3481 ttctcggtct tgcaaacaac cgccggcagc ttagtatata aatacacatg tacatacctc
3541 tctccgtatc ctcgtaatca ttttcttgta tttatcgtct tttcgctgta aaaactttat
3601 cacacttatc tcaaatacac ttattaaccg cttttactat tatcttctac gctgacagta
3661 atatcaaaca gtgacacata ttaaacacag tggtttcttt gcataaacac catcagcctc
3721 aagtcgtcaa gtaaagattt cgtgttcatg cagatagata acaatctata tgttgataat
3781 tagcgttgcc tcatcaatgc gagatccgtt taaccggacc ctagtgcact taccccacgt
3841 tcggtccact gtgtgccgaa catgctcctt cactatttta acatgtggaa ttcttgaaag
3901 aatgaaatcg ccatgccaag ccatcacacg gtcttttatg caattgattg accgcctgca
3961 acacataggc agtaaaattt ttactgaaac gtatataatc atcataagcg acaagtgagg
4021 caacaccttt gttaccacat tgacaacccc aggtattcat acttcctatt agcggaatca
4081 ggagtgcaaa aagagaaaat aaaagtaaaa aggtagggca acacatagta tgaatacaaa
4141 cgttccaata ttcagttctc cggtcagaga tttaccaagg tctttcgaac aaaaacattt
4201 agcggttgta gatgcatttt tccaaacata ccatgtcaaa cctgatttta tcgctaggtc
4261 tcctggcaga gtaaatctga ttggtgagca tatagattat tgcgattttt cagttttgcc
4321 attagccatt gatgtggata tgctttgcgc agttaaaatt ttagacgaaa aaaatccatc
4381 cattacctta acaaatgcgg accctaaatt tgctcagcga aagtttgatc tgcctttaga
4441 tggttcctac atggccatag atccgtctgt gtcggaatgg tcgaattact ttaaatgcgg
4501 actacatgtg gcacattcat acttgaaaaa aattgctccg gaaagattta ataatacacc
4561 cttagtaggt gcgcagatct tttgccagag cgatattcct actggtggtg gactctcatc
4621 tgcatttact tgcgcggcag cactagccac aattagactc gagggggggc ccggtaccca
4681 attcgcccta tagtgagtcg tattacgcgc gctcactggc cgtcgtttta caacgtcgtg
4741 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
4801 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
4861 atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
4921 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
4981 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
5041 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
5101 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
5161 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
5221 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
5281 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag
//
| {
"Vector backbone": "Bluescript II SK+",
"Backbone size w/o insert (bp)": "",
"Vector type": "Yeast Expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "A fluorescent tool set for yeast Atg proteins",
"Publication": "Autophagy. 2015;11(6):954-60. doi: 10.1080/15548627.2015.1040971.",
"DOI": "10.1080/15548627.2015.1040971.",
"Authors": "Li D, Song JZ, Shan MH, Li SP, Liu W, Li H, Zhu J, Wang Y, Lin J, Xie Z /",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/25998947",
"PubMed ID": "PubMed 25998947"
} | {
"Gene/Insert name": "TRP1",
"Species": "S. cerevisiae (budding yeast)",
"Insert Size (bp)": "5300",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "SpeI (not destroyed)",
"3′ cloning site": "XhoI (not destroyed)",
"5′ sequencing primer": "T7",
"3′ sequencing primer": "SP6"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pLVTHM-mSox2-3xflag | 206376 | Expresses 3xflag fused mouse SOX2 in mammalian cells, for lentivirus generation. | Ralf Jauch | Hu et al Nucleic Acids Res. 2023 Aug 23:gkad597. doi: 10.1093/nar/gkad597. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/410024/d25a0fda-f929-47b9-8f08-fea08e7cd2ba/addgene-plasmid-206376-sequence-410024.gbk | full | LOCUS sequence-410024- 11322 bp DNA circular SYN 27-AUG-2024
DEFINITION Expresses 3xflag fused mouse SOX2 in mammalian cells, for lentivirus
generation..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11322)
AUTHORS Hu H, Ho DHH, Tan DS, MacCarthy CM, Yu CH, Weng M, Scholer HR, Jauch
R
TITLE Evaluation of the determinants for improved pluripotency induction
and maintenance by engineered SOX17.
JOURNAL Nucleic Acids Res. 2023 Aug 23:gkad597. doi: 10.1093/nar/gkad597.
PUBMED 37607832
REFERENCE 2 (bases 1 to 11322)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 27, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic
Acids Res. 2023 Aug 23:gkad597. doi: 10.1093/nar/gkad597."
FEATURES Location/Qualifiers
source 1..11322
/mol_type="other DNA"
/organism="synthetic DNA construct"
LTR 2..635
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 682..807
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1300..1533
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1718..1762
/codon_start=1
/product="antigenic peptide corresponding to amino acids
655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
et al., 2013)"
/label=gp41 peptide
/note="recognized by the 2H10 single-chain llama nanobody"
/translation="KNEQELLELDKWASL"
promoter 2147..3325
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 2378..3316
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 3273..3293
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
misc_feature 3373..3490
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
CDS 4495..4560
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
misc_feature 4578..5166
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(4631..4651)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(5049..5060)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
protein_bind 5334..5604
/label=tetracycline response element
/bound_moiety="tetracycline repressor TetR"
/note="contains seven copies of the tetracycline operator
tetO"
promoter 5633..5847
/label=H1 promoter
/note="human H1 RNA promoter"
primer_bind 5768..5787
/label=H1
/note="Human H1 promoter, forward primer"
protein_bind complement(5885..5918)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
LTR 5938..6118
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
promoter complement(6143..6161)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(6144..6161)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(6451..6470)
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind complement(6684..6703)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 6803..6825
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(6863..6881)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 6949..7053
/gene="bla"
/label=AmpR promoter
CDS 7054..7914
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(7272..7291)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 8085..8673
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 8574..8593
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 8827..8844
/label=L4440
/note="L4440 vector, forward primer"
primer_bind complement(8914..8934)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 8919..9248
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 9099..9234
/label=SV40 ori
/note="SV40 origin of replication"
intron 10382..10447
/label=small t intron
/note="SV40 (simian virus 40) small t antigen intron"
CDS 10577..10597
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
polyA_signal 11022..11156
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(11059..11078)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 11113..11132
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
ORIGIN
1 ttggaagggc taattcactc ccaaagaaga caagatatcc ttgatctgtg gatctaccac
61 acacaaggct acttccctga ttagcagaac tacacaccag ggccaggggt cagatatcca
121 ctgacctttg gatggtgcta caagctagta ccagttgagc cagataaggt agaagaggcc
181 aataaaggag agaacaccag cttgttacac cctgtgagcc tgcatgggat ggatgacccg
241 gagagagaag tgttagagtg gaggtttgac agccgcctag catttcatca cgtggcccga
301 gagctgcatc cggagtactt caagaactgc tgatatcgag cttgctacaa gggactttcc
361 gctggggact ttccagggag gcgtggcctg ggcgggactg gggagtggcg agccctcaga
421 tcctgcatat aagcagctgc tttttgcctg tactgggtct ctctggttag accagatctg
481 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
541 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
601 cagacccttt tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa
661 gcgaaaggga aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg
721 gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag
781 aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg
841 gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg
901 gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc
961 tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga
1021 tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac
1081 accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag
1141 caagcggccg ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga
1201 attatataaa tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa
1261 gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt
1321 cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag
1381 acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca
1441 acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc
1501 tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact
1561 catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat
1621 ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat
1681 acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga
1741 attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat
1801 aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact
1861 ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc
1921 aaccccgagg ggacccgaca ggcccgaagg aatagaagaa gaaggtggag agagagacag
1981 agacagatcc attcgattag tgaacggatc tcgacggtat cgatcacgag actagcctcg
2041 accatcgcga tgtcgacgat aagctttgca aagatggata aagttttaaa cagagaggaa
2101 tctttgcagc taatggacct tctaggtctt gaaaggagtg ggaattggct ccggtgcccg
2161 tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa
2221 ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg
2281 gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa
2341 cgttcttttt cgcaacgggt ttgccgccag aacacaggta agtgccgtgt gtggttcccg
2401 cgggcctggc ctctttacgg gttatggccc ttgcgtgcct tgaattactt ccacctggct
2461 gcagtacgtg attcttgatc ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc
2521 ttgcgcttaa ggagcccctt cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg
2581 ccgccgcgtg cgaatctggt ggcaccttcg cgcctgtctc gctgctttcg ataagtctct
2641 agccatttaa aatttttgat gacctgctgc gacgcttttt ttctggcaag atagtcttgt
2701 aaatgcgggc caagatctgc acactggtat ttcggttttt ggggccgcgg gcggcgacgg
2761 ggcccgtgcg tcccagcgca catgttcggc gaggcggggc ctgcgagcgc ggccaccgag
2821 aatcggacgg gggtagtctc aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc
2881 gtgtatcgcc ccgccctggg cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga
2941 aagatggccg cttcccggcc ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg
3001 agagcgggcg ggtgagtcac ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc
3061 ttcatgtgac tccacggagt accgggcgcc gtccaggcac ctcgattagt tctcgagctt
3121 ttggagtacg tcgtctttag gttgggggga ggggttttat gcgatggagt ttccccacac
3181 tgagtgggtg gagactgaag ttaggccagc ttggcacttg atgtaattct ccttggaatt
3241 tgcccttttt gagtttggat cttggttcat tctcaagcct cagacagtgg ttcaaagttt
3301 ttttcttcca tttcaggtgt cgtgaggaat ttcgacattt aaatttaatt aatctcgacg
3361 gtatcggtta acttttaaaa gaaaaggggg gattgggggg tacagtgcag gggaaagaat
3421 agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa ttacaaaaat
3481 tcaaaatttt atcgatcacg agactagcct cgaggtttaa acatgtataa catgatggag
3541 acggagctga agccgccggg cccgcagcaa gcttcggggg gcggcggcgg aggaggcaac
3601 gccacggcgg cggcgaccgg cggcaaccag aagaacagcc cggaccgcgt caagaggccc
3661 atgaacgcct tcatggtatg gtcccggggg cagcggcgta agatggccca ggagaacccc
3721 aagatgcaca actcggagat cagcaagcgc ctgggcgcgg agtggaaact tttgtccgag
3781 accgagaagc ggccgttcat cgacgaggcc aagcggctgc gcgctctgca catgaaggag
3841 cacccggatt ataaataccg gccgcggcgg aaaaccaaga cgctcatgaa gaaggataag
3901 tacacgcttc ccggaggctt gctggccccc ggcgggaaca gcatggcgag cggggttggg
3961 gtgggcgccg gcctgggtgc gggcgtgaac cagcgcatgg acagctacgc gcacatgaac
4021 ggctggagca acggcagcta cagcatgatg caggagcagc tgggctaccc gcagcacccg
4081 ggcctcaacg ctcacggcgc ggcacagatg caaccgatgc accgctacga cgtcagcgcc
4141 ctgcagtaca actccatgac cagctcgcag acctacatga acggctcgcc cacctacagc
4201 atgtcctact cgcagcaggg cacccccggt atggcgctgg gctccatggg ctctgtggtc
4261 aagtccgagg ccagctccag cccccccgtg gttacctctt cctcccactc cagggcgccc
4321 tgccaggccg gggacctccg ggacatgatc agcatgtacc tccccggcgc cgaggtgccg
4381 gagcccgctg cgcccagtag actgcacatg gcccagcact accagagcgg cccggtgccc
4441 ggcacggcca ttaacggcac actgcccctg tcgcacatga cgcgtggcag cggcgactac
4501 aaggaccacg acggcgacta caaggaccac gacatcgact acaaggacga cgacgacaag
4561 tagactagtc atatgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta
4621 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
4681 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
4741 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
4801 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
4861 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
4921 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt
4981 cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct
5041 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
5101 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
5161 ccccgcatcg gtacgtatgg ccaggtacct ttaagaccaa tgacttacaa ggcagctgta
5221 gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga
5281 agacaagatg ggatcaattc accatgctag tggatccccc tcgagtttac cactccctat
5341 cagtgataga gaaaagtgaa agtcgagttt accactccct atcagtgata gagaaaagtg
5401 aaagtcgagt ttaccactcc ctatcagtga tagagaaaag tgaaagtcga gtttaccact
5461 ccctatcagt gatagagaaa agtgaaagtc gagtttacca ctccctatca gtgatagaga
5521 aaagtgaaag tcgagtttac cactccctat cagtgataga gaaaagtgaa agtcgagttt
5581 accactccct atcagtgata gagaaaagtg aaagtcgggg ctgcaggaat tcgaacgctg
5641 acgtcatcaa cccgctccaa ggaatcgcgg gcccagtgtc actaggcggg aacacccagc
5701 gcgcgtgcgc cctggcagga agatggctgt gagggacagg ggagtggcgc cctgcaatat
5761 ttgcatgtcg ctatgtgttc tgggaaatca ccataaacgt gaaatgtctt tggatttggg
5821 aatcttataa gttctgtatg agaccacgcg gcgtctccaa gcttatcgat accgtcgcat
5881 gggaataact tcgtatagca tacattatac gaagttatgc tgctttttgc ttgtactggg
5941 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
6001 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
6061 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagc
6121 atctagaatt aattccgtgt attctatagt gtcacctaaa tcgtatgtgt atgatacata
6181 aggttatgta ttaattgtag ccgcgttcta acgacaatat gtacaagcct aattgtgtag
6241 catctggctt actgaagcag accctatcat ctctctcgta aactgccgtc agagtcggtt
6301 tggttggacg aaccttctga gtttctggta acgccgtccc gcacccggaa atggtcagcg
6361 aaccaatcag cagggtcatc gctagccaga tcctctacgc cggacgcatc gtggccggca
6421 tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc cgacatcacc gatggggaag
6481 atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg cgtgggtatg gtggcaggcc
6541 ccgtggccgg gggactgttg ggcgccatct ccttgcatgc accattcctt gcggcggcgg
6601 tgctcaacgg cctcaaccta ctactgggct gcttcctaat gcaggagtcg cataagggag
6661 agcgtcgaat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
6721 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
6781 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
6841 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
6901 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
6961 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
7021 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
7081 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
7141 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
7201 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
7261 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
7321 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
7381 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
7441 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
7501 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
7561 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
7621 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
7681 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
7741 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
7801 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
7861 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
7921 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
7981 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
8041 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
8101 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
8161 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
8221 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
8281 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
8341 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
8401 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
8461 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
8521 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
8581 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
8641 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
8701 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
8761 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
8821 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
8881 cccgcgcgtt ggccgattca ttaatgcagc tgtggaatgt gtgtcagtta gggtgtggaa
8941 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
9001 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
9061 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
9121 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
9181 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
9241 tttgcaaaaa gcttggacac aagacaggct tgcgagatat gtttgagaat accactttat
9301 cccgcgtcag ggagaggcag tgcgtaaaaa gacgcggact catgtgaaat actggttttt
9361 agtgcgccag atctctataa tctcgcgcaa cctattttcc cctcgaacac tttttaagcc
9421 gtagataaac aggctgggac acttcacatg agcgaaaaat acatcgtcac ctgggacatg
9481 ttgcagatcc atgcacgtaa actcgcaagc cgactgatgc cttctgaaca atggaaaggc
9541 attattgccg taagccgtgg cggtctgtac cgggtgcgtt actggcgcgt gaactgggta
9601 ttcgtcatgt cgataccgtt tgtatttcca gctacgatca cgacaaccag cgcgagctta
9661 aagtgctgaa acgcgcagaa ggcgatggcg aaggcttcat cgttattgat gacctggtgg
9721 ataccggtgg tactgcggtt gcgattcgtg aaatgtatcc aaaagcgcac tttgtcacca
9781 tcttcgcaaa accggctggt cgtccgctgg ttgatgacta tgttgttgat atcccgcaag
9841 atacctggat tgaacagccg tgggatatgg gcgtcgtatt cgtcccgcca atctccggtc
9901 gctaatcttt tcaacgcctg gcactgccgg gcgttgttct ttttaacttc aggcgggtta
9961 caatagtttc cagtaagtat tctggaggct gcatccatga cacaggcaaa cctgagcgaa
10021 accctgttca aaccccgctt taaacatcct gaaacctcga cgctagtccg ccgctttaat
10081 cacggcgcac aaccgcctgt gcagtcggcc cttgatggta aaaccatccc tcactggtat
10141 cgcatgatta accgtctgat gtggatctgg cgcggcattg acccacgcga aatcctcgac
10201 gtccaggcac gtattgtgat gagcgatgcc gaacgtaccg acgatgattt atacgatacg
10261 gtgattggct accgtggcgg caactggatt tatgagtggg ccccggatct ttgtgaagga
10321 accttacttc tgtggtgtga cataattgga caaactacct acagagattt aaagctctaa
10381 ggtaaatata aaatttttaa gtgtataatg tgttaaacta ctgattctaa ttgtttgtgt
10441 attttagatt ccaacctatg gaactgatga atgggagcag tggtggaatg cctttaatga
10501 ggaaaacctg ttttgctcag aagaaatgcc atctagtgat gatgaggcta ctgctgactc
10561 tcaacattct actcctccaa aaaagaagag aaaggtagaa gaccccaagg actttccttc
10621 agaattgcta agttttttga gtcatgctgt gtttagtaat agaactcttg cttgctttgc
10681 tatttacacc acaaaggaaa aagctgcact gctatacaag aaaattatgg aaaaatattc
10741 tgtaaccttt ataagtaggc ataacagtta taatcataac atactgtttt ttcttactcc
10801 acacaggcat agagtgtctg ctattaataa ctatgctcaa aaattgtgta cctttagctt
10861 tttaatttgt aaaggggtta ataaggaata tttgatgtat agtgccttga ctagagatca
10921 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
10981 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
11041 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
11101 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatcaact
11161 ggataactca agctaaccaa aatcatccca aacttcccac cccataccct attaccactg
11221 ccaattacct gtggtttcat ttactctaaa cctgtgattc ctctgaatta ttttcatttt
11281 aaagaaattg tatttgttaa atatgtacta caaacttagt ag
//
| {
"Vector backbone": "pLVTHM-3xflag",
"Backbone size w/o insert (bp)": "",
"Vector type": "Mammalian Expression, Lentiviral",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "NEB Stable",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Evaluation of the determinants for improved pluripotency induction and maintenance by engineered SOX17",
"Publication": "Nucleic Acids Res. 2023 Aug 23:gkad597. doi: 10.1093/nar/gkad597.",
"DOI": "10.1093/nar/gkad597.",
"Authors": "Hu H, Ho DHH, Tan DS, MacCarthy CM, Yu CH, Weng M, Scholer HR, Jauch R /nar/gkad",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/37607832",
"PubMed ID": "PubMed 37607832"
} | {
"Gene/Insert name": "Sox2",
"Species": "M. musculus (mouse)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "Sox2",
"Promoter": "",
"Tag / Fusion Protein": "3xFLAG (C terminal on insert)"
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "unknown (unknown if destroyed)",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
D23_Ld_170240 | 135839 | Expression of Leishmania donovani ectodomain in mammalian cells | Gavin Wright | Roberts et al PLoS Pathog. 2022 Feb 24;18(2):e1010364. doi: 10.1371/journal.ppat.1010364. eCollection 2022 Feb. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/274549/d59a6322-0db6-42a7-8818-350fd0a796e8/addgene-plasmid-135839-sequence-274549.gbk | full | LOCUS sequence-274549- 8643 bp DNA circular SYN 26-AUG-2024
DEFINITION Expression of Leishmania donovani ectodomain in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8643)
AUTHORS Roberts AJ, Ong HB, Clare S, Brandt C, Harcourt K, Franssen SU,
Cotton JA, Muller-Sienerth N, Wright GJ
TITLE Systematic identification of genes encoding cell surface and
secreted proteins that are essential for in vitro growth and
infection in Leishmania donovani.
JOURNAL PLoS Pathog. 2022 Feb 24;18(2):e1010364. doi:
10.1371/journal.ppat.1010364. eCollection 2022 Feb.
PUBMED 35202447
REFERENCE 2 (bases 1 to 8643)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "PLoS
Pathog."; date: "2022-02-24"; volume: "18(2):e1010364. doi"; pages:
" 10.1371/journal.ppat.1010364. eCollection 2022 Feb"
FEATURES Location/Qualifiers
source 1..8643
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 1..380
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 381..584
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 535..555
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 581..605
/label=LNCX
/note="Human CMV promoter, forward primer"
primer_bind 1086..1105
/label=pMT2-F
/note="Synthetic intron, forward primer"
regulatory 1128..1137
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 3801..3818
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
polyA_signal 3870..3925
/label=beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(3924..3943)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
primer_bind complement(6161..6179)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 6247..6351
/gene="bla"
/label=AmpR promoter
CDS 6352..7212
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(6570..6589)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 7383..7971
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 7872..7891
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 8125..8142
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 8259..8280
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 8295..8325
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 8333..8349
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
primer_bind 8338..8360
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 8357..8373
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 8357..8373
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
ORIGIN
1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
601 agatcctcac tctcttccgc atcgctgtct gcgagggcca gctgttgggc tcgcggttga
661 ggacaaactc ttcgcggtct ttccagtact cttggatcgg aaacccgtcg gcctccgaac
721 ggtactccgc caccgaggga cctgagcgag tccgcatcga ccggatcgga aaacctctcg
781 agaaaggcgt ctaaccagtc acagtcgcaa ggtaggctga gcaccgtggc gggcggcagc
841 gggtggcggt cggggttgtt tctggcggag gtgctgctga tgatgtaatt aaagtaggcg
901 gtcttgagac ggcggatggt cgaggtgagg tgtggcaggc ttgagatcca gctgttgggg
961 tgagtactcc ctctcaaaag cgggcattac ttctgcgcta agattgtcag tttccaaaaa
1021 cgaggaggat ttgatattca cctggcccga tctggccata cacttgagtg acaatgacat
1081 ccactttgcc tttctctcca caggtgtcca ctcccaggtc caagtttgcc accatggagt
1141 ttcagaccca ggtactcatg tccctgctgc tctgcatgtc tggtgcggcc gccacctttg
1201 atggcgacgc cgtgacagcc cctcctagcg ccagcaccgc cttcaccgag caccctttca
1261 ccctgcctac cgccgccagc ctgctggacg tgctggccag cgacttctgc accagaaccg
1321 gcgagggcga ggaaagcagc tgcaccccca gcgtgtccag actgcccacc caccccgtgc
1381 tgatcctgtt taccgagagc aacgccggcc acagcagcac cctggccgcc ctgagagcca
1441 aagtgcggca gatcgccacc gacgtgccca gcagcgtgct gcgggtgcac gagtacgtga
1501 tccccatgag cgccacagag cggcggctga tcgagctgct gctgatcggc gccagccacg
1561 tgccaacact ggccctgttt cacggccggg tggccaaggt gcaggtcgcc cctggatcca
1621 tcgtgtctgc cagccccttc cagacccctg tgctgtactc caccaccccc ctggcctccg
1681 ccagctacct ggaactgaga agctgggccc tgtccgagct gcccgccaga tacgtggacc
1741 ccgtgacctt ccacctgatc cctagcctgc agttcgtgtt cagccctgcc gaagtgcacg
1801 aaacactgca gctcgtgcgg gacgccctgg aagccagcga gagcagatcc gtgctgcctg
1861 ccttcgtgtc catggcctac atgcggctga cccggcacgg cagcgaagag gtgctggctg
1921 ccctgtctag cattgccacc caggccggca acgctgtgct gaccctggtc accgaaagcg
1981 ccgaggtggc cgctgcctgg ggactgagag ctgagcacac cttcaccatg gccccttggt
2041 ccagcacagt ggccgcctac ctgaacagca gcgcctccga tggcttcgcc gcctccgtgt
2101 ccgatctgag ccctccccag agcatcggca ccgtgaccga gctggccgaa gccacagccg
2161 gctctgccgc ttggagagcc gcttgtgccg cctctgtgcg ggccgagatc gcccagctgc
2221 agaactggcg gcaggccatg aacgccttca acaccaccag ccctctgcgg cggatcgaca
2281 gcgccgctca tctgctgcac gagctgaagg ccctgcagaa tgccctgaag atcgtgttcg
2341 tgctgagaga aagcgacgag atgtggtttc accaccacct ggaagtggct gccgctctgg
2401 ccagaaggct gcagagcacc accgtgttct acaacacaac cacactggct aagggcagca
2461 aggcccggtc ccgggtggaa agatcctgga cccccatcat gagatgcgac gtgttctggc
2521 tggacgccga gcagctccct gccgtggctg atggactgca cgtgtcccag gtgcccagtg
2581 tgctgattct ggtgcccctg cagtccagat tcggagaggg cggcgacggc gatagagccg
2641 atgctgctgc agccgctggc gacgacgatc aggcagctgc cggcgcttct gaggaagccg
2701 gcctgagaag gcgggacccc ttcatcggca tccacaccat caacagatac gacctgctga
2761 aagccgccta tatggacgac gcccagatga cccacgagcc tgtgtccggc aaagaggcac
2821 tgcctctgtt ccccagcgat tccgatgccc tgctgcggtt cctggctagc ggcagctttc
2881 tgggcgccat gcagtacacc atgtcctcca cccagctgtc taccctgcgg gctaaagtga
2941 cagagctgcc tgatgccgtg caggccgcta ccgccagcgg cggaaccagc aacaggcact
3001 acctgcagct ggaccggcgg tactaccctc tgagaggggc cgaggaaccc atggaaggcc
3061 ccgtgtacgt cagacagatc ctgaacggct ccagccccct gccagtgctg ggctccgacg
3121 acagcgacac cgaactgggc agacagcccg tgggcggcag aagcggcaca caccctagag
3181 gcaccacctc cgcctccccc gacgaggccg atgagagact gagacagcgg agagccagag
3241 aagaggccgc agccctgaaa cggaaggccg cctgggaggc caatctggcc aagcggagaa
3301 gggaaaaggc cgaccggatg cggagaaagg ctgccgagga tgccgccgag cgcgagaggc
3361 agcaggaagc cttccagcag gaagtgaacg ccgccctgaa ggccgaacag gaaggcgctg
3421 gcggcggagc tggcggaaca tgggcttctc agcctgaggg cctgctcgtc agaaggccca
3481 ctctggaaga tgccagcgaa ggcctggtgg aagagggcag acggaagggc aggccccaga
3541 ggcccttcgg ggaggaagtg gatagccagg acgctgccca gattctccgg cggaggcgga
3601 gacaccgcga gtacagagag tggctggctg acagacagct gatggtcaat agatgcgtga
3661 cagtgaccga aaagggcaag ctgagcctga tgaccagatg ggaaggcgcg cccggcagcc
3721 tgcaccacat cctggacgcc cagaagatgc tgtggaacca cagggacagg aacctgcctc
3781 ctctggcccc tctgggcccc caccaccatc accaccattg agtctagaca cgtgattaat
3841 taaggatccc ccgacctcga cctctggcta ataaaggaaa tttattttca ttgcaatagt
3901 gtgttggaat tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttggt
3961 cgagatccct cggagatctc tagctagagg atcgatcccc gccccggacg aactaaacct
4021 gactacgaca tctctgcccc ttcttcgcgg ggcagtgcat gtaatccctt cagttggttg
4081 gtacaacttg ccaactgggc cctgttccac atgtgacacg gggggggacc aaacacaaag
4141 gggttctctg actgtagttg acatccttat aaatggatgt gcacatttgc caacactgag
4201 tggctttcat cctggagcag actttgcagt ctgtggactg caacacaaca ttgcctttat
4261 gtgtaactct tggctgaagc tcttacacca atgctggggg acatgtacct cccaggggcc
4321 caggaagact acgggaggct acaccaacgt caatcagagg ggcctgtgta gctaccgata
4381 agcggaccct caagagggca ttagcaatag tgtttataag gcccccttgt taaccctaaa
4441 cgggtagcat atgcttcccg ggtagtagta tatactatcc agactaaccc taattcaata
4501 gcatatgtta cccaacggga agcatatgct atcgaattag ggttagtaaa agggtcctaa
4561 ggaacagcga tatctcccac cccatgagct gtcacggttt tatttacatg gggtcaggat
4621 tccacgaggg tagtgaacca ttttagtcac aagggcagtg gctgaagatc aaggagcggg
4681 cagtgaactc tcctgaatct tcgcctgctt cttcattctc cttcgtttag ctaatagaat
4741 aactgctgag ttgtgaacag taaggtgtat gtgaggtgct cgaaaacaag gtttcaggtg
4801 acgcccccag aataaaattt ggacgggggg ttcagtggtg gcattgtgct atgacaccaa
4861 tataaccctc acaaacccct tgggcaataa atactagtgt aggaatgaaa cattctgaat
4921 atctttaaca atagaaatcc atggggtggg gacaagccgt aaagactgga tgtccatctc
4981 acacgaattt atggctatgg gcaacacata atcctagtgc aatatgatac tggggttatt
5041 aagatgtgtc ccaggcaggg accaagacag gtgaaccatg ttgttacact ctatttgtaa
5101 caaggggaaa gagagtggac gccgacagca gcggactcca ctggttgtct ctaacacccc
5161 cgaaaattaa acggggctcc acgccaatgg ggcccataaa caaagacaag tggccactct
5221 tttttttgaa attgtggagt gggggcacgc gtcagccccc acacgccgcc ctgcggtttt
5281 ggactgtaaa ataagggtgt aataacttgg ctgattgtaa ccccgctaac cactgcggtc
5341 aaaccacttg cccacaaaac cactaatggc accccgggga atacctgcat aagtaggtgg
5401 gcgggccaag ataggggcgc gattgctgcg atctggagga caaattacac acacttgcgc
5461 ctgagcgcca agcacagggt tgttggtcct catattcacg aggtcgctga gagcacggtg
5521 ggctaatgtt gccatgggta gcatatacta cccaaatatc tggatagcat atgctatcct
5581 aatctatatc tgggtagcat aggctatcct aatctatatc tgggtagcat atgctatcct
5641 aatctatatc tgggtagtat atgctatcct aatttatatc tgggtagcat aggctatcct
5701 aatctatatc tgggtagcat atgctatcct aatctatatc tgggtagtat atgctatcct
5761 aatctgtatc cgggtagcat atgctatcct aatagagatt agggtagtat atgctatcct
5821 aatttatatc tgggtagcat atactaccca aatatctgga tagcatatgc tatcctaatc
5881 tatatctggg tagcatatgc tatcctaatc tatatctggg tagcataggc tatcctaatc
5941 tatatctggg tagcatatgc tatcctaatc tatatctggg tagtatatgc tatcctaatt
6001 tatatctggg tagcataggc tatcctaatc tatatctggg tagcatatgc tatcctaatc
6061 tatatctggg tagtatatgc tatcctaatc tgtatccggg tagcatatgc tatcctcatg
6121 ataagctgtc aaacatgaga attttcttga agacgaaagg gcctcgtgat acgcctattt
6181 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
6241 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
6301 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
6361 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
6421 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
6481 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
6541 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac
6601 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
6661 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
6721 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
6781 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
6841 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca
6901 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
6961 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
7021 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
7081 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
7141 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
7201 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
7261 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
7321 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
7381 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
7441 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
7501 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
7561 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
7621 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
7681 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
7741 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
7801 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
7861 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
7921 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
7981 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
8041 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
8101 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
8161 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
8221 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
8281 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
8341 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct ctagctagag
8401 gtcgaccaat tctcatgttt gacagcttat catcgcagat ccgggcaacg ttgttgccat
8461 tgctgcaggc gcagaactgg taggtatgga agatctatac attgaatcaa tattggcaat
8521 tagccatatt agtcattggt tatatagcat aaatcaatat tggctattgg ccattgcata
8581 cgttgtatct atatcataat atgtacattt atattggctc atgtccaata tgaccgccat
8641 gtt
//
| {
"Vector backbone": "pTT3",
"Backbone size w/o insert (bp)": "6132",
"Vector type": "Mammalian Expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Systematic identification of genes encoding cell surface and secreted proteins that are essential for in vitro growth and infection in Leishmania donovani",
"Publication": "PLoS Pathog. 2022 Feb 24;18(2):e1010364. doi: 10.1371/journal.ppat.1010364. eCollection 2022 Feb.",
"DOI": "10.1371/journal.ppat.1010364.",
"Authors": "Roberts AJ, Ong HB, Clare S, Brandt C, Harcourt K, Franssen SU, Cotton JA, Muller-Sienerth N, Wright GJ /journalppat",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/35202447",
"PubMed ID": "PubMed 35202447"
} | {
"Gene/Insert name": "hypothetical protein, unknown function",
"Species": "Leishmania donovani",
"Insert Size (bp)": "2511",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": "biotinylation sequence and His tag (C terminal on insert)"
} | {
"Cloning method": "Unknown",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
KIAA1804 gRNA (BRDN0001145266) | 77584 | 3rd generation lentiviral gRNA plasmid targeting human KIAA1804 | Doench et al Nat Biotechnol. 2016 Jan 18. doi: 10.1038/nbt.3437. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/188046/02184e88-b519-49e8-83a8-c82eb3f32d22/addgene-plasmid-77584-sequence-188046.gbk | full | LOCUS sequence-188046- 8323 bp DNA circular SYN 26-AUG-2024
DEFINITION 3rd generation lentiviral gRNA plasmid targeting human KIAA1804.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8323)
AUTHORS Doench JG, Fusi N, Sullender M, Hegde M, Vaimberg EW, Donovan KF,
Smith I, Tothova Z, Wilen C, Orchard R, Virgin HW, Listgarten J,
Root DE
TITLE Optimized sgRNA design to maximize activity and minimize off-target
effects of CRISPR-Cas9.
JOURNAL Nat Biotechnol. 2016 Jan 18. doi: 10.1038/nbt.3437.
PUBMED 26780180
REFERENCE 2 (bases 1 to 8323)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat
Biotechnol. 2016 Jan 18. doi: 10.1038/nbt.3437."
FEATURES Location/Qualifiers
source 1..8323
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 87..1265
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 318..1256
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 1213..1233
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
CDS 1278..1877
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind complement(1278..1297)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 1774..1794
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
misc_feature 1893..2481
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(1946..1966)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(2364..2375)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 2553..2786
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 2864..2985
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2901..2920)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2955..2974
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 3025..3160
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(3180..3199)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(3181..3199)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(3209..3226)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(3209..3225)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3218..3240)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 3367..3822
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3454..3473)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3664..3685
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 3848..3952
/gene="bla"
/label=AmpR promoter
CDS 3953..4813
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(4171..4190)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 4984..5572
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5473..5492
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 5726..5743
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 5860..5881
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 5896..5926
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5934..5950
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
primer_bind 5939..5961
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 5958..5974
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 5958..5974
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 5993..6013
/label=T3
/note="T3 promoter, forward primer"
promoter 5995..6013
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 6041..6267
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 6268..6448
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 6495..6620
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 7113..7346
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 7531..7575
/codon_start=1
/product="antigenic peptide corresponding to amino acids
655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
et al., 2013)"
/label=gp41 peptide
/note="recognized by the 2H10 single-chain llama nanobody"
/translation="KNEQELLELDKWASL"
promoter 7753..8001
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA
(Domitrovich & Kunkel, 2003)"
primer_bind 7753..7773
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 7924..7943
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
misc_RNA 8023..8098
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
misc_feature 8154..8271
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
ORIGIN
1 cccgggtgca aagatggata aagttttaaa cagagaggaa tctttgcagc taatggacct
61 tctaggtctt gaaaggagtg ggaattggct ccggtgcccg tcagtgggca gagcgcacat
121 cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgatccggt gcctagagaa
181 ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg
241 gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt
301 ttgccgccag aacacaggta agtgccgtgt gtggttcccg cgggcctggc ctctttacgg
361 gttatggccc ttgcgtgcct tgaattactt ccacctggct gcagtacgtg attcttgatc
421 ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa ggagcccctt
481 cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt
541 ggcaccttcg cgcctgtctc gctgctttcg ataagtctct agccatttaa aatttttgat
601 gacctgctgc gacgcttttt ttctggcaag atagtcttgt aaatgcgggc caagatctgc
661 acactggtat ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca
721 catgttcggc gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc
781 aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg
841 cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga aagatggccg cttcccggcc
901 ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac
961 ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac tccacggagt
1021 accgggcgcc gtccaggcac ctcgattagt tctcgagctt ttggagtacg tcgtctttag
1081 gttgggggga ggggttttat gcgatggagt ttccccacac tgagtgggtg gagactgaag
1141 ttaggccagc ttggcacttg atgtaattct ccttggaatt tgcccttttt gagtttggat
1201 cttggttcat tctcaagcct cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt
1261 cgtgacgtac ggccaccatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg
1321 acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc
1381 acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca
1441 cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gccgtggcgg
1501 tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca
1561 tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc
1621 cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggagtctcg cccgaccacc
1681 agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg
1741 gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg
1801 gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc
1861 gcaagcccgg tgcctgaacg cgttaagtcg acaatcaacc tctggattac aaaatttgtg
1921 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
1981 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
2041 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
2101 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
2161 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
2221 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
2281 cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg
2341 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
2401 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
2461 ccctttgggc cgcctccccg cgtcgacttt aagaccaatg acttacaagg cagctgtaga
2521 tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag
2581 acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga
2641 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
2701 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
2761 ttagtcagtg tggaaaatct ctagcagtac gtatagtagt tcatgtcatc ttattattca
2821 gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc
2881 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
2941 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta
3001 gctatcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
3061 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
3121 gagctattcc agaagtagtg aggaggcttt tttggaggcc tagggacgta cccaattcgc
3181 cctatagtga gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg
3241 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
3301 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
3361 aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
3421 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
3481 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
3541 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
3601 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
3661 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
3721 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
3781 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg
3841 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
3901 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
3961 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
4021 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
4081 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
4141 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
4201 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
4261 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
4321 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
4381 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
4441 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
4501 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
4561 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
4621 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
4681 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
4741 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
4801 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
4861 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
4921 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
4981 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
5041 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
5101 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
5161 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
5221 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
5281 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
5341 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
5401 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
5461 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
5521 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
5581 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
5641 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
5701 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
5761 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
5821 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
5881 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
5941 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa
6001 ccctcactaa agggaacaaa agctggagct gcaagcttaa tgtagtctta tgcaatactc
6061 ttgtagtctt gcaacatggt aacgatgagt tagcaacatg ccttacaagg agagaaaaag
6121 caccgtgcat gccgattggt ggaagtaagg tggtacgatc gtgccttatt aggaaggcaa
6181 cagacgggtc tgacatggat tggacgaacc actgaattgc cgcattgcag agatattgta
6241 tttaagtgcc tagctcgata cataaacggg tctctctggt tagaccagat ctgagcctgg
6301 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
6361 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
6421 ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag
6481 ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag
6541 gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga
6601 gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa
6661 ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca
6721 gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac
6781 aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat
6841 ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg
6901 aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg
6961 ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag tgaattatat
7021 aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc aaagagaaga
7081 gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg gttcttggga
7141 gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc cagacaatta
7201 ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc gcaacagcat
7261 ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct ggctgtggaa
7321 agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa actcatttgc
7381 accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca gatttggaat
7441 cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt aatacactcc
7501 ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt ggaattagat
7561 aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta tataaaatta
7621 ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt actttctata
7681 gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct cccaaccccg
7741 aggggaccca gagagggcct atttcccatg attccttcat atttgcatat acgatacaag
7801 gctgttagag agataattag aattaatttg actgtaaaca caaagatatt agtacaaaat
7861 acgtgacgta gaaagtaata atttcttggg tagtttgcag ttttaaaatt atgttttaaa
7921 atggactatc atatgcttac cgtaacttga aagtatttcg atttcttggc tttatatatc
7981 ttgtggaaag gacgaaacac cgaggataag acctctctcc gagttttaga gctagaaata
8041 gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt
8101 ttttaagctt ggcgtaacta gatcttgaga caaatggcag tattcatcca caattttaaa
8161 agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca
8221 gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat
8281 tacagggaca gcagagatcc actttggcgc cggctcgagg ggg
//
| {
"Vector backbone": "lentiGuide-Puro",
"Backbone size w/o insert (bp)": "",
"Vector type": "Mammalian Expression, Lentiviral, CRISPR",
"Selectable markers": "Puromycin"
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Optimized sgRNA design to maximize activity and minimize off-target effects of CRISPR-Cas9",
"Publication": "Nat Biotechnol. 2016 Jan 18. doi: 10.1038/nbt.3437.",
"DOI": "10.1038/nbt.3437.",
"Authors": "Doench JG, Fusi N, Sullender M, Hegde M, Vaimberg EW, Donovan KF, Smith I, Tothova Z, Wilen C, Orchard R, Virgin HW, Listgarten J, Root DE /nbt",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/26780180",
"PubMed ID": "PubMed 26780180"
} | {
"Gene/Insert name": "KIAA1804",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "hU6",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Unknown",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "U6-F",
"3′ sequencing primer": "pS2-R"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
||
pMSR:MSMEG_0372:Dendra | 159056 | T7 expression of C-term fluorescent fusion in E. coli | Judd et al mBio. 2021 Mar 2;12(2). pii: mBio.02401-20. doi: 10.1128/mBio.02401-20. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/316089/0642f3d1-bf60-40cb-bd5b-e30abe0083da/addgene-plasmid-159056-sequence-316089.gbk | full | LOCUS sequence-316089- 6230 bp DNA circular SYN 26-AUG-2024
DEFINITION T7 expression of C-term fluorescent fusion in E. coli.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6230)
AUTHORS Judd JA, Canestrari J, Clark R, Joseph A, Lapierre P,
Lasek-Nesselquist E, Mir M, Palumbo M, Smith C, Stone M, Upadhyay A,
Wirth SE, Dedrick RM, Meier CG, Russell DA, Dills A, Dove E, Kester
J, Wolf ID, Zhu J, Rubin ER, Fortune S, Hatfull GF, Gray TA, Wade
JT, Derbyshire KM
TITLE A Mycobacterial Systems Resource for the Research Community.
JOURNAL mBio. 2021 Mar 2;12(2). pii: mBio.02401-20. doi:
10.1128/mBio.02401-20.
PUBMED 33653882
REFERENCE 2 (bases 1 to 6230)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "mBio.";
date: "2021-03-2"; volume: "12(2). pii: mBio.02401-20. doi"; pages:
" 10.1128/mBio.02401-20"
FEATURES Location/Qualifiers
source 1..6230
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 46..65
/label=T7
/note="T7 promoter, forward primer"
promoter 46..64
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory 919..928
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 2174..2197
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS 2454..3254
/codon_start=1
/gene="aac(3)IV"
/product="aminoglycoside 3-N-acetyltransferase type IV
"
/label=ApmR
/note="confers resistance to apramycin (Paget and Davies,
1996)"
/translation="MSSAVECNVVQYEWRKAELIGQLLNLGVTPGGVLLVHSSFRSVRP
LEDGPLGLIEALRATLGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDTFWRLPNV
KRSAHPFAFAAAGPQAEQIISDPLPLPPYSPASPVARVHELDGQVLLLGVGHDANTTLH
LAELMAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGH
AFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG"
rep_origin 3379..3967
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3868..3887
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 tgatagagtt tgtcctccct atcagtgata gataggctta aggagtaata cgactcacta
61 tagggagaag gaggccatta tggcttcgga cctgttttcc caagtggtca attccgggcc
121 gggatcgttc ctggccaagc agctgggcgt gccgcagccc gagacgttgc gccgctaccg
181 ccccggtgac ccgccgctgg cgggctcgct gctgatcggc ggatccggcc gcgtggccga
241 acccctgcgc accgcattgg ccgacgacta caacctggtg tccaacaaca tcggtggccg
301 gtgggccgat tcgttcggcg gcgtggtgtt cgacgccacg ggcatcaccg aggccgaggg
361 cctcaaggag ctctacacgt tcttcacgcc gctgctgcgc aacctcgcgc cgtgtgcgcg
421 ggtggtcgtc gtcggcacca cgcccgcgga ggccggcagc gtccacgcac aggtggtgca
481 gcgggcgctg gaaggtttca cgcgctcgct gggcaaggag ctgcgccgtg gcgcgaccgt
541 ctccctggtc tatctctcgg ccgacgccaa gccgggcgcc acgggtctcg agtcgaccat
601 gcggttcatc ctgtcggcca agtcggccta cgtcgacggt caggtgttcc gcgtcggcgc
661 cgccgattcg actccgcccg ccgactggga caagccgctc gacggcaagg tcgccgtggt
721 caccggcgcg gcccgcggta tcggcgcgac gatcgccgag gtgttcgcgc gtgacggcgc
781 gaccgtggtg gcgatcgacg tcgacggtgc ggccgaggat ctcaagcgcg tcgccgacaa
841 ggtgggcggc accgcactca cgctcgacgt gaccgccgac gacgccgtcg acaagatcac
901 cgcgcacgtc accgaacacc atggcggcaa ggtcgacatc ctcgtcaaca acgcgggtat
961 cacgcgcgac aagctgctcg ccaacatgga cgagaagcgc tgggacgcgg tgatcgccgt
1021 caacctgctc gcgccgcagc gcctcaccga aggcctggtg ggcaacggaa ccatcggcga
1081 gggcggccgg gtgatcggcc tgtcgtcgat ggcgggcatc gcgggcaacc gcggccagac
1141 caattacgcc accaccaagg ccgggatgat cgggctggcc gaggcgctgg cgcccgtgct
1201 ggccgacaag ggcatcacga tcaacgccgt cgcacccggc ttcatcgaga ccaagatgac
1261 cgaggccatc ccgctcgcca cgcgtgaggt gggcaggcgg ctcaactcgc tgttccaggg
1321 cggccagccc gtcgacgtgg ccgagttgat cgcgtacttc gccagccccg cctcgaacgc
1381 ggtgacgggc aacaccatcc gcgtgtgcgg ccaggcgatg ctgggggcga agcttggtgc
1441 ggccgcgggc aacacgcctg gcatcaatct gatcaaggag gacatgcgtg tcaaggtgca
1501 tatggagggc aacgtgaacg gtcacgcctt cgtgatcgag ggggagggta agggtaagcc
1561 ttacgaaggg acccagaccg ccaacctcac cgtcaaggag ggcgcccctc tcccattctc
1621 gtacgatatc ctcactaccg cggtgcatta cgggaaccgc gtgttcacca agtaccccga
1681 ggatatccct gactacttca agcagagctt cccggaggga tactcctggg agcggacgat
1741 gaccttcgaa gacaagggta tctgtacgat ccgcagcgac atctcgctgg aaggggactg
1801 tttcttccag aacgtccgtt tcaaggggac caactttccg ccgaacggcc cggtcatgca
1861 gaagaagacc ctcaagtggg agccgagcac ggagaagctg cacgttcggg acggcctcct
1921 ggtcggcaac atcaacatgg cgctgctgct ggaaggcggc ggccactacc tatgcgactt
1981 caagaccacg tacaaagcga agaaggtcgt ccagctgccc gacgcgcact tcgtggacca
2041 ccgcatcgaa atcctgggga acgattcgga ctacaacaag gtcaagttgt acgagcacgc
2101 tgtggcacgc tatagcccgc tcccgtcgca ggtctggttc gaaggcggcg gcggggagaa
2161 cctgtatttc caggactaca aggacgacga cgacaagtga tttaaataac cccttggggc
2221 ctctaaacgg gtcttgaggg gttttttgtt aactagcgta cgatcgaatt cctgcagctc
2281 acggtaactg atgccgtatt tgcagtacca gcgtacggcc cacagaatga tgtcacgctg
2341 aaaatgccgg cctttgaatg ggttcatgtg cagctccatc agcaaaaggg gatgataagt
2401 ttatcaccac cgactatttg caacagtgcc gttgatcgtg ctatgatcga ctgatgtcat
2461 cagcggtgga gtgcaatgtc gtgcaatacg aatggcgaaa agccgagctc atcggtcagc
2521 ttctcaacct tggggttacc cccggcggtg tgctgctggt ccacagctcc ttccgtagcg
2581 tccggcccct cgaagatggg ccacttggac tgatcgaggc cctgcgtgct acgctgggtc
2641 cgggagggac gctcgtcatg ccctcgtggt caggtctgga cgacgagccg ttcgatcctg
2701 ccacgtcgcc cgttacaccg gaccttggag ttgtctctga cacattctgg cgcctgccaa
2761 atgtaaagcg cagcgcccat ccatttgcct ttgcggcagc ggggccacag gcagagcaga
2821 tcatctctga tccattgccc ctgccacctt actcgcctgc aagcccggtc gcccgtgtcc
2881 atgaactcga tgggcaggta cttctcctcg gcgtgggaca cgatgccaac acgacgctgc
2941 atcttgccga gttgatggca aaggttccct atggggtgcc gagacactgc accattcttc
3001 aggatggcaa gttggtacgc gtcgattatc tcgagaatga ccactgctgt gagcgctttg
3061 ccttggcgga caggtggctc aaggagaaga gccttcagaa ggaaggtcca gtcggtcatg
3121 cctttgctcg gttgatccgc tcccgcgaca ttgtggcgac agccctgggt caactgggcc
3181 gagatccgtt gatcttcctg catccgccag aggcgggatg cgaagaatgc gatgccgctc
3241 gccagtcgat tggctgagct catgagcgga gaacgagatg acgttggagg ggcaaggtcg
3301 cgctgattgc tggggcaaca cgggggatcc actagttcca tgagcgtcag accccgtaga
3361 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
3421 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
3481 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
3541 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
3601 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
3661 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
3721 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
3781 cgccacgctt cccgagggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
3841 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
3901 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
3961 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
4021 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
4081 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc aacgcgtgcg gccgcggtac
4141 ccggggatcc tctagagtcg accaccaagg gcaccatctc tgcttgggcc accccgttgg
4201 ccgcagccag ctcgctgaga gccgtgaacg acagggcgaa cgccagcccg ccgacggcga
4261 gggttccgac cgctgcaact cccggtgcaa ccttgtcccg gtctattctc ttcactgcac
4321 cagctccaat ctggtgtgaa tgcccctcgt ctgttcgcgc aggcgggggg ctctattcgt
4381 ttgtcagcat cgaaagtagc cagatcaggg atgcgttgca accgcgtatg cccaggtcag
4441 aagagtcgca caagagttgc agacccctgg aaagaaaaat ggccagaggg cgaaaacacc
4501 ctctgaccag cggagcgggc gacgggaatc gaacccgcgt agctagtttg gaagaatggg
4561 tgtctgccga ccacatatgg gccggtcaag ataggttttt accccctctc ggctgcatcc
4621 tctaagtgga aagaaattgc aggtcgtaga agcgcgttga agcctgagag ttgcacagga
4681 gttgcaaccc ggtagccttg ttcacgacga gaggagacct agttggcacg tcgcggatgg
4741 ggatcgctga agactcagcg cagcgggagg atccaagcct catacgtcaa cccgcaggac
4801 ggtgtgaggt actacgcgct gcagacctac gacaacaaga tggacgccga agcctggctc
4861 gcgggcgaga agcggctcat cgagatggag acctggaccc ctccacagga ccgggcgaag
4921 aaggcagccg ccagcgccat cacgctggag gagtacaccc ggaagtggct cgtggagcgc
4981 gacctcgcag acggcaccag ggatctgtac agcgggcacg cggagcgccg catctacccg
5041 gtgctaggtg aagtggcggt cacagagatg acgccagctc tggtgcgtgc gtggtgggcc
5101 gggatgggta ggaagcaccc gactgcccgc cggcatgcct acaacgtcct ccgggcggtg
5161 atgaacacag cggtcgagga caagctgatc gcagagaacc cgtgccggat cgagcagaag
5221 gcagccgatg agcgcgacgt agaggcgctg acgcctgagg agctggacat cgtcgccgct
5281 gagatcttcg agcactaccg gatcgcggca tacatcctgg cgtggacgag cctccggttc
5341 ggagagctga tcgagcttcg ccgcaaggac atcgtggacg acggcatgac gatgaagctc
5401 cgggtgcgcc gtggcgcttc ccgcgtgggg aacaagatcg tcgttggcaa cgccaagacc
5461 gtccggtcga agcgtcctgt gacggttccg cctcacgtcg cggagatgat ccgagcgcac
5521 atgaaggacc gtacgaagat gaacaagggc cccgaggcat tcctggtgac cacgacgcag
5581 ggcaaccggc tgtcgaagtc cgcgttcacc aagtcgctga agcgtggcta cgccaagatc
5641 ggtcggccgg aactccgcat ccacgacctc cgcgctgtcg gcgctacgtt cgccgctcag
5701 gcaggtgcga cgaccaagga gctgatggcc cgtctcggtc acacgactcc taggatggcg
5761 atgaagtacc agatggcgtc tgaggcccgc gacgaggcta tcgctgaggc gatgtccaag
5821 ctggccaaga cctcctgaaa cgcaaaaagc ccccctccca aggacactga gtcctaaaga
5881 ggggggtttc ttgtcagtac gcgaagaacc acgcctggcc gcgagcgcca gcaccgccgc
5941 tctgtgcgga gacctgggca ccagccccgc cgccgccagg agcattgccg ttcccgccag
6001 aaatctagac ggtgaccaca acgcgcccgc tttgatcggg gacgtcgaaa tattggatcg
6061 tcggcaccgt cacggccgtg ggaggcggca cgatccgcga cgtgatgatc ggccgcatcc
6121 ccacggtgct gcgcagtgag ctctacgcca tcccggcgtt gatctgtgcg ttcgcacgca
6181 caggcccggt gtgagaaggg tctctgacga gcgggagaac tccctatcag
//
| {
"Vector backbone": "pMSR:Dendra",
"Backbone size w/o insert (bp)": "",
"Vector type": "Bacterial Expression ; Integrating Mycobacterial Expression Vector attL5",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Apramycin, 25 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "A Mycobacterial Systems Resource for the Research Community",
"Publication": "mBio. 2021 Mar 2;12(2). pii: mBio.02401-20. doi: 10.1128/mBio.02401-20.",
"DOI": "10.1128/mBio.02401-20.",
"Authors": "Judd JA, Canestrari J, Clark R, Joseph A, Lapierre P, Lasek-Nesselquist E, Mir M, Palumbo M, Smith C, Stone M, Upadhyay A, Wirth SE, Dedrick RM, Meier CG, Russell DA, Dills A, Dove E, Kester J, Wolf ID, Zhu J, Rubin ER, Fortune S, Hatfull GF, Gray TA, Wade JT, Derbyshire KM /mBio-",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/33653882",
"PubMed ID": "PubMed 33653882"
} | {
"Gene/Insert name": "MSMEG_0372",
"Species": "M. smegmatis",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "pmycTetO",
"Tag / Fusion Protein": "Gly-Ala Linker/Dendra2/1x Flag Tag (C terminal on insert)"
} | {
"Cloning method": "Ligation Independent Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "TGATAGAGTTTGTCCTCCCTATCAG",
"3′ sequencing primer": "TCCATATGCACCTTGACACG"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
||
pSB8 | 82037 | Hybrid CitA[1-193]-Tar[204-553] expression plasmid | Victor Sourjik | Bi et al ACS Synth Biol. 2016 Jun 10. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/174335/4ae48c95-b3ac-47c9-bad1-373fa8da2157/addgene-plasmid-82037-sequence-174335.gbk | full | LOCUS sequence-174335- 5796 bp DNA circular SYN 26-AUG-2024
DEFINITION Hybrid CitA[1-193]-Tar[204-553] expression plasmid.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5796)
AUTHORS Bi S, Pollard AM, Yang Y, Jin F, Sourjik V
TITLE Engineering Hybrid Chemotaxis Receptors in Bacteria.
JOURNAL ACS Synth Biol. 2016 Jun 10.
PUBMED 27285081
REFERENCE 2 (bases 1 to 5796)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth
Biol. 2016 Jun 10."
FEATURES Location/Qualifiers
source 1..5796
/mol_type="other DNA"
/organism="synthetic DNA construct"
terminator 521..555
/label=lambda t0 terminator
/note="minimal transcription terminator from phage lambda
(Scholtissek and Grosse, 1987)"
CDS complement(974..1633)
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind 1548..1567
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
promoter complement(1634..1736)
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
primer_bind complement(2128..2145)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2262..2807)
/direction=LEFT
/label=p15A ori
/note="Plasmids containing the medium-copy-number p15A
origin of replication can be propagated in E. coli cells
that contain a second plasmid with the ColE1 origin.
"
primer_bind complement(2342..2361)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 aatcttgcca gtcgcagcgc ccaggcggca aaagagatca aagccctcat tgaagactcc
61 gtctcacgcg ttgataccgg ttcggtgctg gtcgaaagcg ccggggaaac aatgaacaat
121 atcgtcaatg ctgtcactcg cgtgactgac attatgggcg agattgcatc ggcatcggat
181 gaacagagcc gtggcatcga tcaagtcgca ttggcggttt cggaaatgga tcgcgtcacg
241 caacagaacg catcgctggt gcaggaatca gctgccgccg ccgctgcgct ggaagaacag
301 gcgagtcgtt taacgcaagc agtttccgcg ttccgtctgg cagccagccc actcaccaat
361 aaaccgcaaa caccatcccg tcctgccagt gagcaaccac cggctcagcc acgactgcga
421 attgctgaac aagatccaaa ctgggaaaca ttttgaggat ccgcggataa ataagtaacg
481 atccggtcca gtaatgacct cagaactcca tctggatttg ttcagaacgc tcggttgccg
541 ccgggcgttt tttattggtg agaatcgcag caacttgtcg cgccaatcga gccatgtcgt
601 cgtcaacgac cccccattca agaacagcaa gcagcattga gaactttgga atccagtccc
661 tcttccacct gctgaccgga tcagcagtcc ccggaacatc gtagctgacg ccttcgcgtt
721 gctcagttgt ccaaccccgg aaacgggaaa aagcaagttt tccccgctcc cggcgtttca
781 ataactgaaa accatactat ttcacagttt aaatcacatt aaacgacagt aatccccgtt
841 gatttgtgcg ccaacacaga tccgtcgacc gggtcgaatt tgctttcgaa tttctgccat
901 tcatccgctt attatcactt attcaggcgt agcaaccagg cgtttaaggg caccaataac
961 tgccttaaaa aaattacgcc ccgccctgcc actcatcgca gtactgttgt aattcattaa
1021 gcattctgcc gacatggaag ccatcacaaa cggcatgatg aacctgaatc gccagcggca
1081 tcagcacctt gtcgccttgc gtataatatt tgcccatggt gaaaacgggg gcgaagaagt
1141 tgtccatatt ggccacgttt aaatcaaaac tggtgaaact cacccaggga ttggctgaga
1201 cgaaaaacat attctcaata aaccctttag ggaaataggc caggttttca ccgtaacacg
1261 ccacatcttg cgaatatatg tgtagaaact gccggaaatc gtcgtggtat tcactccaga
1321 gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta acaagggtga acactatccc
1381 atatcaccag ctcaccgtct ttcattgcca tacggaattc cggatgagca ttcatcaggc
1441 gggcaagaat gtgaataaag gccggataaa acttgtgctt atttttcttt acggtcttta
1501 aaaaggccgt aatatccagc tgaacggtct ggttataggt acattgagca actgactgaa
1561 atgcctcaaa atgttcttta cgatgccatt gggatatatc aacggtggta tatccagtga
1621 tttttttctc cattttagct tccttagctc ctgaaaatct cgataactca aaaaatacgc
1681 ccggtagtga tcttatttca ttatggtgaa agttggaacc tcttacgtgc cgatcaacgt
1741 ctcattttcg ccaaaagttg gcccagggct tcccggtatc aacagggaca ccaggattta
1801 tttattctgc gaagtgatct tccgtcacag gtatttattc ggcgcaaagt gcgtcgggtg
1861 atgctgccaa cttactgatt tagtgtatga tggtgttttt gaggtgctcc agtggcttct
1921 gtttctatca gctgtccctc ctgttcagct actgacgggg tggtgcgtaa cggcaaaagc
1981 accgccggac atcagcgcta gcggagtgta tactggctta ctatgttggc actgatgagg
2041 gtgtcagtga agtgcttcat gtggcaggag aaaaaaggct gcaccggtgc gtcagcagaa
2101 tatgtgatac aggatatatt ccgcttcctc gctcactgac tcgctacgct cggtcgttcg
2161 actgcggcga gcggaaatgg cttacgaacg gggcggagat ttcctggaag atgccaggaa
2221 gatacttaac agggaagtga gagggccgcg gcaaagccgt ttttccatag gctccgcccc
2281 cctgacaagc atcacgaaat ctgacgctca aatcagtggt ggcgaaaccc gacaggacta
2341 taaagatacc aggcgtttcc ccctggcggc tccctcgtgc gctctcctgt tcctgccttt
2401 cggtttaccg gtgtcattcc gctgttatgg ccgcgtttgt ctcattccac gcctgacact
2461 cagttccggg taggcagttc gctccaagct ggactgtatg cacgaacccc ccgttcagtc
2521 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggaaa gacatgcaaa
2581 agcaccactg gcagcagcca ctggtaattg atttagagga gttagtcttg aagtcatgcg
2641 ccggttaagg ctaaactgaa aggacaagtt ttggtgactg cgctcctcca agccagttac
2701 ctcggttcaa agagttggta gctcagagaa ccttcgaaaa accgccctgc aaggcggttt
2761 tttcgttttc agagcaagag attacgcgca gaccaaaacg atctcaagaa gatcatctta
2821 ttaatcagat aaaatatttc tagatttcag tgcaatttat ctcttcaaat gtagcacctg
2881 aagtcagccc catacgatat aagttgtaat tctcatgttt gacagcttat catcgatccc
2941 gcaagaggcc cggcagtacc ggcataacca agcctatgcc tacagcatcc agggtgacgg
3001 tgccgaggat gacgatgagc gcattgttag atttcataca cggtgcctga ctgcgttagc
3061 aatttaactg tgataaacta ccgcattaaa gcttcttgcg aaacagctct cgaccaggtt
3121 tatactggtc gtgcagatga aagcagtttt tgccgaggtc gattccgacc agcgccatat
3181 cgctcatagt gatggtctcc gaattataaa caccctgcga aagcgtagcc tacgcagggt
3241 gtcgggtgct catcccatta cccgttttcc catctcttaa cttaaggggg gaacggtgtc
3301 ctgtctttca tggggctcgt tcactgctaa cttacctgct tccctgtctg tcaattctct
3361 ctatcctgcg atcccgcgaa gaaccaaaaa agctcgacag agggcgcggt cattttaggt
3421 cgggcggatc ggcgccgccg gctcggctgg tgtgccgcac agcaccgcct acgtgagctg
3481 ccagttgatg aacttccccc gttgccagct agggcgcaag cgggctgtat aagatcactg
3541 cccatcacat tgatcggctc ggattttttc tcaatccgta aacaggtcaa acatcagttg
3601 ccgcaaccaa atattggcta ggtccttgtg gtacttcgca tgccagaaca tgttgatggc
3661 tatttcaggc aagacgactg ggtgcggcaa ggcgcttagg ccgaagggct ccacacagca
3721 gtcggctaaa cgtatcggca cagtggcgag cagatcggtg cgctggagga tgtggccaac
3781 ggcggcgaag tgcggcactt ccagacggat gtcgcgccgg atgccgaccc gtgtcatgta
3841 cgtgtccacc tcgccgtggc cggtgccagc ggcgatgaca cgcacgtggc cgtaggaaca
3901 gaagcgctcc agagtcaggg gttcgcgggt gactggatgg tccttgcgac ataggcacac
3961 gtagtgattc tggagcagcc ggcgctgaaa gaagccagtt tgcagattgg gaagcaggcc
4021 cacggccaag tccacggttc cgttctgcaa ggcctgcatc aggctcatcg aactgtcgcg
4081 caccgtactg atcacgcaat tgggggcctg gtgagccagc acatccatca gccgcggcat
4141 gaagtagatc tcgccaatgt cggtcatggc cagggtgaag gtacgctcgc tggtcagcgg
4201 atcgaagctt tcatggtgct gtagggcgtt gcgcagtgcg tgcatggccg aagtgacggg
4261 ctcggccaga tgcgcggcat agggtgtggg ttccattccc tgatgtgtgc gcacgaagag
4321 tgggtcctgt agcgaggtgc gcaggcgttt cagcgcattg ctcacggcag gctgggtcag
4381 gcccaggttc tccgcagtga tagagacgcg tctgtcgacc agcaactggt tgaacaccac
4441 cagcaggttt aaatccaggt cacgcagttc catggggcct cgcttgggtt attgctggtg
4501 cccggccggg cgcaatattc atgttgatga tttattatat atcgagtggt gtatttatca
4561 atattgtttg ctccgttatc gttattaaca agtcatcaat aatgccataa ggagtaccat
4621 atgttgcagc ttaacgagaa taaacagttt gcatttttcc aaagactggc atttccgctg
4681 cgtatctttt tgctgattct ggtgttctca atatttgtca ttgcagccct ggcgcaatat
4741 tttacggcca gttttgagga ctatttaacg cttcatgtac gcgacatggc aatgaatcag
4801 gcgaaaatta ttgcctccaa tgacagtgtc atctctgcgg tgaaaacgcg tgactacaaa
4861 cggctggcga ccatcgctaa caaattacaa agagataccg attttgatta tgtggtgatt
4921 ggggaccggc actcgatccg cctttaccat cctaatccgg agaaaattgg ttatcctatg
4981 cagttcacca aacagggcgc gctggagaaa ggggagagct acttcattac cgggaaaggg
5041 tcaatgggga tggcgatgcg cgccaaaacg ccaatctttg atgacgatgg aaaagtcatc
5101 ggcgtggtgt cgattggcta cctggtgagt aaaatcgata gctggcgggc tgagttttta
5161 ttaccgatgg caggtgtgtt tgtcgtgctg ttagggatta ttctgctggt ggcgtggtac
5221 ggcattcgcc gtatgttgct tactccgctg gcaaaaatta ttgctcacat tcgcgaaatc
5281 gccggtggta acctggcgaa taccctgacc attgacgggc gcagtgaaat gggcgacctg
5341 gcgcagagcg tttcacacat gcaacgctct ttgactgaca ccgtcactca tgtccgcgaa
5401 ggttcagatg ccatctatgc cggtacccgt gaaattgcgg cgggcaacac cgatctttcc
5461 tcccgtactg aacagcaggc atccgcgctg gaagaaactg ccgccagcat ggagcagctc
5521 accgcgacag tgaagcaaaa cgccgataac gcccgccagg cctcgcaact ggcgcaaagt
5581 gcctccgaca ccgcccagca cggcggcaaa gtggtggatg gcgtagtgaa aacgatgcat
5641 gagatcgccg atagttcgaa gaaaattgcc gacattatca gcgttatcga cggtattgcc
5701 ttccagacta atatcctcgc gctgaatgcc gcggttgaag ccgcgcgtgc gggtgaacag
5761 ggccgtggtt ttgccgtggt ggcgggtgaa gtgcgt
//
| {
"Vector backbone": "pKG116",
"Backbone size w/o insert (bp)": "4211",
"Vector type": "",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Chloramphenicol, 25 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "Low Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Engineering Hybrid Chemotaxis Receptors in Bacteria",
"Publication": "ACS Synth Biol. 2016 Jun 10.",
"DOI": "10.1021/acssynbio.6b00053",
"Authors": "Bi S, Pollard AM, Yang Y, Jin F, Sourjik V",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/27285081",
"PubMed ID": "PubMed 27285081"
} | {
"Gene/Insert name": "Encodes the hybrid receptor of E. coli chemoreceptor Tar and histidine kinase CitA: CitA[1-193]-Tar[204-553]",
"Species": "E. coli",
"Insert Size (bp)": "1632",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "NdeI (not destroyed)",
"3′ cloning site": "BamHI (not destroyed)",
"5′ sequencing primer": "CGGGCGCAATATTCATGTTG",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pD2529-CAG-HA-mouse GARP | 180258 | Mouse full length GARP expression | Timothy Springer | Jiang et al Cancer Immunol Res. 2022 Feb 18. pii: 681626. doi: 10.1158/2326-6066.CIR-21-0593. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/357201/9b856a98-c655-4398-9b31-d06ba8aec817/addgene-plasmid-180258-sequence-357201.gbk | full | LOCUS sequence-357201- 9267 bp DNA circular SYN 25-AUG-2024
DEFINITION Mouse full length GARP expression.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9267)
AUTHORS Jiang A, Qin Y, Springer TA
TITLE Loss of LRRC33-dependent TGFbeta1 activation enhances anti-tumor
immunity and checkpoint blockade therapy.
JOURNAL Cancer Immunol Res. 2022 Feb 18. pii: 681626. doi:
10.1158/2326-6066.CIR-21-0593.
PUBMED 35181792
REFERENCE 2 (bases 1 to 9267)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Cancer
Immunol Res. 2022 Feb 18. pii: 681626. doi:
10.1158/2326-6066.CIR-21-0593."
FEATURES Location/Qualifiers
source 1..9267
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 3..306
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 308..583
/label=chicken beta-actin promoter
intron 584..1600
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
primer_bind 1524..1546
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 1608..1627
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
CDS 1785..1811
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
polyA_signal 5338..5393
/label=beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(5392..5411)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
rep_origin complement(6103..6691)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6183..6202)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6777..7586)
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind 7500..7519
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
promoter complement(7605..7652)
/label=EM7 promoter
/note="synthetic bacterial promoter "
CDS complement(8078..8677)
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind complement(8161..8181)
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
primer_bind 8658..8677
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
promoter complement(8687..9186)
/label=PGK promoter
/note="mouse phosphoglycerate kinase 1 promoter"
primer_bind complement(8735..8754)
/label=mPGK-F
/note="Mouse PGK promoter, forward primer"
primer_bind 9128..9151
/label=MSCV-rev
/note="Murine stem cell virus, reverse primer"
ORIGIN
1 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
61 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
121 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
181 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
241 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
301 accatggtcg aggtgagccc cacgttctgc ttcactctcc ccatctcccc cccctcccca
361 cccccaattt tgtatttatt tattttttaa ttattttgtg cagcgatggg ggcggggggg
421 gggggggcgc gcgccaggcg gggcggggsg gggsgrgggg sggggsgggg sgrggcggrg
481 aggtgcggcg gcagccaatc agagcggcgc gctccgaaag tttcctttta tggcgaggcg
541 gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgcgctg
601 ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac
661 cgcgttactc ccacaggtga gcgggcggga cggcccttct cctccgggct gtaattagcg
721 cttggtttaa tgacggcttg tttcttttct gtggctgcgt gaaagccttg aggggctccg
781 ggagggccct ttgtgcgggg ggagcggctc ggggggtgcg tgcgtgtgtg tgtgcgtggg
841 gagcgccgcg tgcggctccg cgctgcccgg cggctgtgag cgctgcgggc gcggcgcggg
901 gctttgtgcg ctccgcagtg tgcgcgaggg gagcgcggcc gggggcggtg ccccgcggtg
961 cggggggggc tgcgagggga acaaaggctg cgtgcggggt gtgtgcgtgg gggggtgagc
1021 agggggtgtg ggcgcgtcgg tcgggctgca accccccctg cacccccctc cccgagttgc
1081 tgagcacggc ccggcttcgg gtgcggggct ccgtacgggg cgtggcgcgg ggctcgccgt
1141 gccgggcggg gggtggcggc aggtgggggt gccgggcggg gcggggccgc ctcgggccgg
1201 ggagggctcg ggggaggggc gcggcggccc ccggagcgcc ggcggctgtc gaggcgcggc
1261 gagccgcagc cattgccttt tatggtaatc gtgcgagagg gcgcagggac ttcctttgtc
1321 ccaaatctgt gcggagccga aatctgggag gcgccgccgc accccctcta gcgggcgcgg
1381 ggcgaagcgg tgcggcgccg gcaggaagga aatgggcggg gagggccttc gtgcgtcgcc
1441 gcgccgccgt ccccttctcc ctctccagcc tcggggctgt ccgcgggggg acggctgcct
1501 tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg gctctagagc
1561 ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca acgtgctggt
1621 tattgtgctg tctcatcatt ttggcaaaga attcggcttg atcgaagccg tctcagggga
1681 gagaccacac ccaagctgtc tagagccgcc accttgtgaa gagcgccgcc accatgcctt
1741 tgctcctcct gttgcccttg ctctgggctg gcgccctcgc ggcctaccca tacgatgttc
1801 cagattacgc tctcgagatc tctcagcgtc gagagcaagt gccctgtagg acggtgaaca
1861 aggaggcctt gtgccacggc cttggcctgc tccaggtccc ctcggtgctc tcactggaca
1921 tccaagctct ctacttgtcc gggaaccaac tgcagagcat cctggtctcc ccactgggct
1981 tctatacagc gcttcgtcac ctggatttaa gtgacaacca gattagcttc ctccaggctg
2041 gggtcttcca ggccctgccc tacctggagc accttaacct ggcccacaat cggcttgcca
2101 caggcatggc tctcaacagt ggtggtctgg gccgcctgcc ccttttggtc tccctggacc
2161 tgtctgggaa cagcctgcat ggcaatctgg tggagcgact gttgggtgag accccgaggc
2221 tgcgtacgct ctcactggcg gagaacagcc tcactcgcct ggcacgccac accttctggg
2281 gtatgccagc ggtggagcaa ttagacctcc acagcaatgt cctcatggac atcgaggacg
2341 gtgcctttga ggccctgccc cacctgactc acctcaatct ctccagaaac tccctcacct
2401 gcatctcaga cttcagcctc cagcagctgc aggtactaga cttgagctgc aacagcattg
2461 aggccttcca gacggccccg gaaccacagg cccagttcca gttggcctgg ctcgacctcc
2521 gggagaacaa gctgctccac ttccctgacc tggccgtgtt cccgagactc atctacctga
2581 atgtgtctaa caacctcatc cagctccctg cggggctgcc ccggggcagt gaggacctcc
2641 acgcaccctc tgaaggctgg tcagcctctc cactgtccaa ccccagccgg aatgccagca
2701 cccaccctct ctcccagctc ctgaacctgg atctgagtta caatgagatc gaactggtcc
2761 ctgctagctt tcttgaacac ctgacctccc tgcgcttcct caacctcagc agaaactgcc
2821 tgcgatcctt tgaggctcga caagtggact ccctgccctg cctggtgctt ttggacttga
2881 gccacaatgt gctggaagca ttagaactgg gtaccaaagt cctagggtcc ctgcagacgt
2941 tgcttctgca ggacaatgcc ttgcaagaac ttccacccta tacctttgcc agcctggcca
3001 gcctgcagag gctaaaccta caggggaacc aggtcagtcc ctgtgggggg ccagcagaac
3061 caggtccccc gggctgtgtg gacttctctg ggatccccac ccttcatgtt cttaacatgg
3121 cagggaactc gatgggaatg ctcagggcgg gcagcttcct tcacacccca cttacagagc
3181 tggacctctc caccaatcct ggtctggatg tggccacggg agccctggtg ggcctggagg
3241 catccttaga ggtactggag cttcaaggca acgggctgac tgtcttgaga gtggacttgc
3301 cctgctttct ccgtctcaag cgccttaacc ttgctgagaa ccagttaagc cacctgcccg
3361 cttggacgcg ggctgtgtcc ctggaggtgc tagatctgcg gaacaacagc ttcagcctcc
3421 tgccaggcaa tgccatgggt ggcctggaga ccagtctccg gcggctgtac ttgcagggaa
3481 atccactcag ctgctgtggc aatggctggc tggcggccca gctgcaccag ggcagagtgg
3541 atgtggatgc tactcaggac ctaatctgcc gctttggctc ccaggaggag ttgtccctga
3601 gcctagtgcg tccagaggat tgcgagaagg gagggctgaa gaatgtcaac ctcatcctcc
3661 tcctcagctt cacactggtc tctgccatcg tcctcaccac gctggccacc atctgcttcc
3721 tccgccggca gaagctcagc caacaataca aagcctgata agctagcggt tgaggtctct
3781 aaaagcgtct tcctgttctc atcacatcat atcaaggtta tataccatca atattgccac
3841 agatgttact tagcctttta atatttctct aatttagtgt atatgcaatg atagttctct
3901 gatttctgag attgagtttc tcatgtgtaa tgattattta gagtttctct ttcatctgtt
3961 caaatttttg tctagtttta ttttttactg atttgtaaga cttcttttta taatctgcat
4021 attacaattc tctttactgg ggtgttgcaa atattttctg tcattctatg gcctgacttt
4081 tcttaatggt tttttaattt taaaaataag tcttaatatt catgcaatct aattaacaat
4141 cttttctttg tggttaggac tttgagtcat aagaaatttt tctctacact gaagtcatga
4201 tggcatgctt ctatattatt ttctaaaaga tttaaagttt tgccttctcc atttagactt
4261 ataattcact ggaatttttt tgtgtgtatg gtatgacata tgggttccct tttatttttt
4321 acatataaat atatttccct gtttttctaa aaaagaaaaa gatcatcatt ttcccattgt
4381 aaaatgccat atttttttca taggtcactt acatatatca atgggtctgt ttctgagctc
4441 tactctattt tatcagcctc actgtctatc cccacacatc tcatgctttg ctctaaatct
4501 tgatatttag tggaacattc tttcccattt tgttctacaa gaatattttt gttattgtct
4561 ttgggctttc tatatacatt ttgaaatgag gttgacaagt taataacagg cctattgatt
4621 ggaaagtttg tcaacgaatt gtgggtcttt tggggtttgc tgcccctttt acgcaatgtg
4681 gatatcctgc tttaatgcct ttatatgcat gtatacaagc aaaacaggct tttactttct
4741 cgccaactta caaggccttt ctcagtaaac agtatatgac cctttacccc gttgctcggc
4801 aacggcctgg tctgtgccaa gtgtttgctg acgcaacccc cactggttgg ggcttggcca
4861 taggccatca gcgcatgcgt ggaacctttg tgtctcctct gccgatccat actgcggaac
4921 tcctagccgc ttgttttgct cgcagcaggt ctggagcaaa cctcatcggg accgacaatt
4981 ctgtcgtact ctcccgcaag tatacatcgt ttccatggct gctaggctgt gctgccaact
5041 ggatcctgcg cgggacgtcc tttgtttacg tcccgtcggc gctgaatccc gcggacgacc
5101 cctcccgggg ccgcttgggg ctctaccgcc cgcttctccg tctgccgtac cgtccgacca
5161 cggggcgcac ctctctttac gcggactccc cgtctgtgcc ttctcatctg ccggaccgtg
5221 tgcacttcgc ttcacctctg cacgtcgcat ggaggccacc gtgaacgccc accggaacct
5281 gcccaaggtc ttgcataaga ggactcttgg actttcagca atgtcatctc agtggctaat
5341 aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa
5401 ggacatatgg gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg
5461 caacatatgc ccatatgctg gctgccatga acaaaggttg gctataaaga ggtcatcagt
5521 atatgaaaca gccccctgct gtccattcct tattccatag aaaagccttg acttgaggtt
5581 agattttttt tatattttgt tttgtgttat ttttttcttt aacatcccta aaattttcct
5641 tacatgtttt actagccaga tttttcctcc tctcctgact actcccagtc atagctgtcc
5701 ctcttctctt atggagatct ttgcattttt agacatttag aagcctatat cttgttacag
5761 aattggaatt acacaaaaat tctaccatat tttgaaagct taggttgttc tgaaaaaaac
5821 aatatattgt tttcctgggt aaactaaaag tcccctcgag gaaaggcccc taaagtgaaa
5881 cagtgcaaaa cgttcaaaaa ctgtctggca atacaagttc cactttgacc aaaacggctg
5941 gcagtaaaag ggttaagaag actgtcagcc ttgagcggta tcagctcact caaaggcggt
6001 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
6061 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
6121 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
6181 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
6241 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
6301 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
6361 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
6421 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
6481 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg gctaactacg gctacactag
6541 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
6601 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
6661 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
6721 tgacgctcag tggaacgacg cgcgcgtaac tcacgttaag ggattttggt catgagttag
6781 aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca
6841 tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg
6901 atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt
6961 aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa
7021 tccggtgaga atggcaaaag tttatgcatt tctttccaga cttgttcaac aggccagcca
7081 ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc
7141 tgagcgaggc gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgagtgc
7201 aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct
7261 tctaatacct ggaacgctgt ttttccgggg atcgcagtgg tgagtaacca tgcatcatca
7321 ggagtacgga taaaatgctt gatggtcgga agtggcataa attccgtcag ccagtttagt
7381 ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac
7441 tctggcgcat cgggcttccc atacaagcga tagattgtcg cacctgattg cccgacatta
7501 tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa tcgcggcctc
7561 gacgtttccc gttggatatg gctcattttt tacttcctca ccttgtcgta ttatactatg
7621 ccgatatact atgccgatga ttaattgtcg acactgcggg ggctctgtgt ggtaagcagg
7681 tcttaacctt tttactgcca atgacgcatg ggatacgtcg tggcagtaaa agggcttaaa
7741 tgccaacgac gcgtcccata cgttgttggc attttaattc ttctctctgc agcggcagca
7801 tgtgccgccg ctgcagagag tttctagcga tgacagcccc tctgggcaac gagccggggg
7861 ggctgtcttt ctttatgttt taaatgcact gacctcccac attccctttt tagtaaaata
7921 ttcagaaata atttaaatac atcattgcaa tgaaaataaa tgttttttat taggcagaat
7981 ccagatgctc aaggcccttc ataatatccc ccagtttagt agttggactt agggaacaaa
8041 ggaaccttta atagaaattg gacagcaaga aagcgagtca ggcaccgggc ttgcgggtca
8101 tgcaccaggt gcgcggtcct tcgggcacct cgacgtcggc ggtgacggtg aagccgagcc
8161 gctcgtagaa ggggaggttg cggggcgcgg atgtctccag gaaggcgggc accccggcgc
8221 gctcggccgc ctccactccg gggagcacga cggcgctgcc cagacccttg ccctggtggt
8281 cgggcgacac gccgacggtg gccaggaacc acgcgggctc cttgggccgg tgcggcgcca
8341 ggaggccttc catctgttgc tgcgcggcca gccgggaacc gctcaactcg gccatgcgcg
8401 ggccgatctc ggcgaacacc gcccccgctt cgacgctctc cggcgtggtc cagaccgcca
8461 ccgcggcgcc gtcgtccgcg acccacacct tgccgatgtc gagcccgacg cgcgtgagga
8521 agagttcttg cagctcggtg acccgctcga tgtggcggtc cggatcgacg gtgtggcgcg
8581 tggcggggta gtcggcgaac gcggcggcga gggtgcgtac ggccctgggg acgtcgtcgc
8641 gggtggcgag gcgcaccgtg ggcttgtact cggtcatggt ggcggacgaa aggcccggag
8701 atgaggaaga ggagaacagc gcggcagacg tgcgcttttg aagcgtgcag aatgccgggc
8761 ctccggagga ccttcgggcg cccgccccgc ccctgagccc gcccctgagc ccgcccccgg
8821 acccacccct tcccagcctc tgagcccaga aagcgaagga gcaaagctgc tattggccgc
8881 tgccccaaag gcctacccgc ttccattgct cagcggtgct gtccatctgc acgagactag
8941 tgagtcgtgc tacttccatt tgtcacgtcc tgcacgacgc gagctgcggg gcggggggga
9001 acttcctgac taggggagga gtagaaggtg gcgcgaaggg gccaccaaag aacggagccg
9061 gttggcgcct accggtggat gtggaatgtg tgcgaggcca gaggccactt gtgtagcgcc
9121 aagtgcccag cggggctgct aaagcgcatg ctccagactg ccttgggaaa agcgcctccc
9181 ctacccggta gagaaacttg atctgtcgcc gcaattcaaa cttcgtgagg ctccggtgcc
9241 cgtcagtgac ctgctatact ctggaga
//
| {
"Vector backbone": "pD2529 CAG",
"Backbone size w/o insert (bp)": "",
"Vector type": "Mammalian Expression",
"Selectable markers": "Puromycin"
} | {
"Bacterial Resistance(s)": "Kanamycin, 50 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Loss of LRRC33-dependent TGFbeta1 activation enhances anti-tumor immunity and checkpoint blockade therapy",
"Publication": "Cancer Immunol Res. 2022 Feb 18. pii: 681626. doi: 10.1158/2326-6066.CIR-21-0593.",
"DOI": "10.1158/2326-6066.CIR-21-0593.",
"Authors": "Jiang A, Qin Y, Springer TA /-CIR--",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/35181792",
"PubMed ID": "PubMed 35181792"
} | {
"Gene/Insert name": "GARP",
"Species": "M. musculus (mouse)",
"Insert Size (bp)": "1938",
"Mutation": "",
"Entrez Gene": "Lrrc32",
"Promoter": "",
"Tag / Fusion Protein": "HA (N terminal on insert)"
} | {
"Cloning method": "Gibson Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "TTTCCTACAGCTCCTGGGCAA",
"3′ sequencing primer": "TGCCACAGATGTTACTTAGCCTTT"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pcDNA-dCas9-p300 Core (Y1467F) | 61362 | encodes S. pyogenes dCas9 with c-terminal fusion of human p300 HAT core (aa 1048-1664; inavtivating mutation Y1467F) driven by CMV promoter | Charles Gersbach | Hilton et al Nat Biotechnol. 2015 May;33(5):510-7. doi: 10.1038/nbt.3199. Epub 2015 Apr 6. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/99796/504b1b82-98c3-4676-88d9-89a95994a872/addgene-plasmid-61362-sequence-99796.gbk | full | LOCUS sequence-99796-e 11475 bp DNA circular SYN 26-AUG-2024
DEFINITION encodes S. pyogenes dCas9 with c-terminal fusion of human p300 HAT
core (aa 1048-1664; inavtivating mutation Y1467F) driven by CMV
promoter.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11475)
AUTHORS Hilton IB, D'Ippolito AM, Vockley CM, Thakore PI, Crawford GE, Reddy
TE, Gersbach CA
TITLE Epigenome editing by a CRISPR-Cas9-based acetyltransferase activates
genes from promoters and enhancers.
JOURNAL Nat Biotechnol. 2015 May;33(5):510-7. doi: 10.1038/nbt.3199. Epub
2015 Apr 6.
PUBMED 25849900
REFERENCE 2 (bases 1 to 11475)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10.1038/nbt.3199";
journalName: "Nat Biotechnol"; date: "2015-05"; volume: "33"; issue:
"5"; pages: "510-7"
FEATURES Location/Qualifiers
source 1..11475
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(44..63)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 235..614
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 615..818
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 769..789
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 863..882
/label=T7
/note="T7 promoter, forward primer"
promoter 863..881
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 907..972
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
CDS 979..999
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 1009..5112
/codon_start=1
/product="catalytically dead mutant of the Cas9
endonuclease from the Streptococcus pyogenes Type II
CRISPR/Cas system"
/label=dCas9
/note="RNA-guided DNA-binding protein that lacks
endonuclease activity due to the D10A mutation in the RuvC
catalytic domain and the H840A mutation in the HNH
catalytic domain"
/translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
DATLIHQSITGLYETRIDLSQLGGD"
CDS 5137..5157
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 5167..7017
/codon_start=1
/gene="EP300"
/product="catalytic histone acetyltransferase core of the
human E1A-associated protein p300 (Delvecchio et al.,
2013)"
/label=p300 core
/translation="IFKPEELRQALMPTLEALYRQDPESLPFRQPVDPQLLGIPDYFDI
VKSPMDLSTIKRKLDTGQYQEPWQYVDDIWLMFNNAWLYNRKTSRVYKYCSKLSEVFEQ
EIDPVMQSLGYCCGRKLEFSPQTLCCYGKQLCTIPRDATYYSYQNRYHFCEKCFNEIQG
ESVSLGDDPSQPQTTINKEQFSKRKNDTLDPELFVECTECGRKMHQICVLHHEIIWPAG
FVCDGCLKKSARTRKENKFSAKRLPSTRLGTFLENRVNDFLRRQNHPESGEVTVRVVHA
SDKTVEVKPGMKARFVDSGEMAESFPYRTKALFAFEEIDGVDLCFFGMHVQEYGSDCPP
PNQRRVYISYLDSVHFFRPKCLRTAVYHEILIGYLEYVKKLGYTTGHIWACPPSEGDDY
IFHCHPPDQKIPKPKRLQEWFKKMLDKAVSERIVHDYKDIFKQATEDRLTSAKELPYFE
GDFWPNVLEESIKELEQEEEERKREENTSNESTDVTKGDSKNAKKKNNKKTSKNKSSLS
RGNKKKPGMPNVSNDLSQKLYATMEKHKEVFFVIRLIAGPAANSLPPIVDPDPLIPCDL
MDGRDAFLTLARDKHLEFSSLRRAQWSTMCMLVELHTQSQD"
regulatory 6781..6790
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 7018..7044
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
primer_bind complement(7069..7086)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 7075..7299
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 7345..7773
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(7432..7451)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 7642..7663
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(7782..7802)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 7787..8116
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 7967..8102
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 8029..8048
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 8183..8977
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(8237..8256)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 8847..8866
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 9151..9272
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(9188..9207)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 9242..9261
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(9321..9337)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(9321..9337)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(9334..9356)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 9345..9361
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(9369..9399)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 9414..9435
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(9552..9569)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(9723..10308)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(9803..9822)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(10479..11339)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 11102..11121
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(11340..11444)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
901 gccatggact acaaagacca tgacggtgat tataaagatc atgacatcga ttacaaggat
961 gacgatgaca agatggcccc caagaagaag aggaaggtgg gccgcggaat ggataagaaa
1021 tactcaatag gcttagctat cggcacaaat agcgtcggat gggcggtgat cactgatgaa
1081 tataaggttc cgtctaaaaa gttcaaggtt ctgggaaata cagaccgcca cagtatcaaa
1141 aaaaatctta taggggctct tttatttgac agtggagaga cagcggaagc gactcgtctc
1201 aaacggacag ctcgtagaag gtatacacgt cggaagaatc gtatttgtta tctacaggag
1261 attttttcaa atgagatggc gaaagtagat gatagtttct ttcatcgact tgaagagtct
1321 tttttggtgg aagaagacaa gaagcatgaa cgtcatccta tttttggaaa tatagtagat
1381 gaagttgctt atcatgagaa atatccaact atctatcatc tgcgaaaaaa attggtagat
1441 tctactgata aagcggattt gcgcttaatc tatttggcct tagcgcatat gattaagttt
1501 cgtggtcatt ttttgattga gggagattta aatcctgata atagtgatgt ggacaaacta
1561 tttatccagt tggtacaaac ctacaatcaa ttatttgaag aaaaccctat taacgcaagt
1621 ggagtagatg ctaaagcgat tctttctgca cgattgagta aatcaagacg attagaaaat
1681 ctcattgctc agctccccgg tgagaagaaa aatggcttat ttgggaatct cattgctttg
1741 tcattgggtt tgacccctaa ttttaaatca aattttgatt tggcagaaga tgctaaatta
1801 cagctttcaa aagatactta cgatgatgat ttagataatt tattggcgca aattggagat
1861 caatatgctg atttgttttt ggcagctaag aatttatcag atgctatttt actttcagat
1921 atcctaagag taaatactga aataactaag gctcccctat cagcttcaat gattaaacgc
1981 tacgatgaac atcatcaaga cttgactctt ttaaaagctt tagttcgaca acaacttcca
2041 gaaaagtata aagaaatctt ttttgatcaa tcaaaaaacg gatatgcagg ttatattgat
2101 gggggagcta gccaagaaga attttataaa tttatcaaac caattttaga aaaaatggat
2161 ggtactgagg aattattggt gaaactaaat cgtgaagatt tgctgcgcaa gcaacggacc
2221 tttgacaacg gctctattcc ccatcaaatt cacttgggtg agctgcatgc tattttgaga
2281 agacaagaag acttttatcc atttttaaaa gacaatcgtg agaagattga aaaaatcttg
2341 acttttcgaa ttccttatta tgttggtcca ttggcgcgtg gcaatagtcg ttttgcatgg
2401 atgactcgga agtctgaaga aacaattacc ccatggaatt ttgaagaagt tgtcgataaa
2461 ggtgcttcag ctcaatcatt tattgaacgc atgacaaact ttgataaaaa tcttccaaat
2521 gaaaaagtac taccaaaaca tagtttgctt tatgagtatt ttacggttta taacgaattg
2581 acaaaggtca aatatgttac tgaaggaatg cgaaaaccag catttctttc aggtgaacag
2641 aagaaagcca ttgttgattt actcttcaaa acaaatcgaa aagtaaccgt taagcaatta
2701 aaagaagatt atttcaaaaa aatagaatgt tttgatagtg ttgaaatttc aggagttgaa
2761 gatagattta atgcttcatt aggtacctac catgatttgc taaaaattat taaagataaa
2821 gattttttgg ataatgaaga aaatgaagat atcttagagg atattgtttt aacattgacc
2881 ttatttgaag atagggagat gattgaggaa agacttaaaa catatgctca cctctttgat
2941 gataaggtga tgaaacagct taaacgtcgc cgttatactg gttggggacg tttgtctcga
3001 aaattgatta atggtattag ggataagcaa tctggcaaaa caatattaga ttttttgaaa
3061 tcagatggtt ttgccaatcg caattttatg cagctgatcc atgatgatag tttgacattt
3121 aaagaagaca ttcaaaaagc acaagtgtct ggacaaggcg atagtttaca tgaacatatt
3181 gcaaatttag ctggtagccc tgctattaaa aaaggtattt tacagactgt aaaagttgtt
3241 gatgaattgg tcaaagtaat ggggcggcat aagccagaaa atatcgttat tgaaatggca
3301 cgtgaaaatc agacaactca aaagggccag aaaaattcgc gagagcgtat gaaacgaatc
3361 gaagaaggta tcaaagaatt aggaagtcag attcttaaag agcatcctgt tgaaaatact
3421 caattgcaaa atgaaaagct ctatctctat tatctccaaa atggaagaga catgtatgtg
3481 gaccaagaat tagatattaa tcgtttaagt gattatgatg tcgatgccat tgttccacaa
3541 agtttcctta aagacgattc aatagacaat aaggtcttaa cgcgttctga taaaaatcgt
3601 ggtaaatcgg ataacgttcc aagtgaagaa gtagtcaaaa agatgaaaaa ctattggaga
3661 caacttctaa acgccaagtt aatcactcaa cgtaagtttg ataatttaac gaaagctgaa
3721 cgtggaggtt tgagtgaact tgataaagct ggttttatca aacgccaatt ggttgaaact
3781 cgccaaatca ctaagcatgt ggcacaaatt ttggatagtc gcatgaatac taaatacgat
3841 gaaaatgata aacttattcg agaggttaaa gtgattacct taaaatctaa attagtttct
3901 gacttccgaa aagatttcca attctataaa gtacgtgaga ttaacaatta ccatcatgcc
3961 catgatgcgt atctaaatgc cgtcgttgga actgctttga ttaagaaata tccaaaactt
4021 gaatcggagt ttgtctatgg tgattataaa gtttatgatg ttcgtaaaat gattgctaag
4081 tctgagcaag aaataggcaa agcaaccgca aaatatttct tttactctaa tatcatgaac
4141 ttcttcaaaa cagaaattac acttgcaaat ggagagattc gcaaacgccc tctaatcgaa
4201 actaatgggg aaactggaga aattgtctgg gataaagggc gagattttgc cacagtgcgc
4261 aaagtattgt ccatgcccca agtcaatatt gtcaagaaaa cagaagtaca gacaggcgga
4321 ttctccaagg agtcaatttt accaaaaaga aattcggaca agcttattgc tcgtaaaaaa
4381 gactgggatc caaaaaaata tggtggtttt gatagtccaa cggtagctta ttcagtccta
4441 gtggttgcta aggtggaaaa agggaaatcg aagaagttaa aatccgttaa agagttacta
4501 gggatcacaa ttatggaaag aagttccttt gaaaaaaatc cgattgactt tttagaagct
4561 aaaggatata aggaagttaa aaaagactta atcattaaac tacctaaata tagtcttttt
4621 gagttagaaa acggtcgtaa acggatgctg gctagtgccg gagaattaca aaaaggaaat
4681 gagctggctc tgccaagcaa atatgtgaat tttttatatt tagctagtca ttatgaaaag
4741 ttgaagggta gtccagaaga taacgaacaa aaacaattgt ttgtggagca gcataagcat
4801 tatttagatg agattattga gcaaatcagt gaattttcta agcgtgttat tttagcagat
4861 gccaatttag ataaagttct tagtgcatat aacaaacata gagacaaacc aatacgtgaa
4921 caagcagaaa atattattca tttatttacg ttgacgaatc ttggagctcc cgctgctttt
4981 aaatattttg atacaacaat tgatcgtaaa cgatatacgt ctacaaaaga agttttagat
5041 gccactctta tccatcaatc catcactggt ctttatgaaa cacgcattga tttgagtcag
5101 ctaggaggtg acccaattgc cggatccaag gctagcccga aaaagaaacg caaagttggg
5161 cgcgccattt tcaaaccaga agaactacga caggcactga tgccaacttt ggaggcactt
5221 taccgtcagg atccagaatc ccttcccttt cgtcaacctg tggaccctca gcttttagga
5281 atccctgatt actttgatat tgtgaagagc cccatggatc tttctaccat taagaggaag
5341 ttagacactg gacagtatca ggagccctgg cagtatgtcg atgatatttg gcttatgttc
5401 aataatgcct ggttatataa ccggaaaaca tcacgggtat acaaatactg ctccaagctc
5461 tctgaggtct ttgaacaaga aattgaccca gtgatgcaaa gccttggata ctgttgtggc
5521 agaaagttgg agttctctcc acagacactg tgttgctacg gcaaacagtt gtgcacaata
5581 cctcgtgatg ccacttatta cagttaccag aacaggtatc atttctgtga gaagtgtttc
5641 aatgagatcc aaggggagag cgtttctttg ggggatgacc cttcccagcc tcaaactaca
5701 ataaataaag aacaattttc caagagaaaa aatgacacac tggatcctga actgtttgtt
5761 gaatgtacag agtgcggaag aaagatgcat cagatctgtg tccttcacca tgagatcatc
5821 tggcctgctg gattcgtctg tgatggctgt ttaaagaaaa gtgcacgaac taggaaagaa
5881 aataagtttt ctgctaaaag gttgccatct accagacttg gcacctttct agagaatcgt
5941 gtgaatgact ttctgaggcg acagaatcac cctgagtcag gagaggtcac tgttagagta
6001 gttcatgctt ctgacaaaac cgtggaagta aaaccaggca tgaaagcaag gtttgtggac
6061 agtggagaga tggcagaatc ctttccatac cgaaccaaag ccctctttgc ctttgaagaa
6121 attgatggtg ttgacctgtg cttctttggc atgcatgttc aagagtatgg ctctgactgc
6181 cctccaccca accagaggag agtatacata tcttacctcg atagtgttca tttcttccgt
6241 cctaaatgct tgaggactgc agtctatcat gaaatcctaa ttggatattt agaatatgtc
6301 aagaaattag gttacacaac agggcatatt tgggcatgtc caccaagtga gggagatgat
6361 tatatcttcc attgccatcc tcctgaccag aagataccca agcccaagcg actgcaggaa
6421 tggttcaaaa aaatgcttga caaggctgta tcagagcgta ttgtccatga ctacaaggat
6481 atttttaaac aagctactga agatagatta acaagtgcaa aggaattgcc ttatttcgag
6541 ggtgatttct ggcccaatgt tctggaagaa agcattaagg aactggaaca ggaggaagaa
6601 gagagaaaac gagaggaaaa caccagcaat gaaagcacag atgtgaccaa gggagacagc
6661 aaaaatgcta aaaagaagaa taataagaaa accagcaaaa ataagagcag cctgagtagg
6721 ggcaacaaga agaaacccgg gatgcccaat gtatctaacg acctctcaca gaaactatat
6781 gccaccatgg agaagcataa agaggtcttc tttgtgatcc gcctcattgc tggccctgct
6841 gccaactccc tgcctcccat tgttgatcct gatcctctca tcccctgcga tctgatggat
6901 ggtcgggatg cgtttctcac gctggcaagg gacaagcacc tggagttctc ttcactccga
6961 agagcccagt ggtccaccat gtgcatgctg gtggagctgc acacgcagag ccaggactac
7021 ccgtacgacg ttccggacta cgcttcttga gtttaaaccg ctgatcagcc tcgactgtgc
7081 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
7141 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
7201 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
7261 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
7321 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg
7381 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
7441 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
7501 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
7561 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
7621 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
7681 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
7741 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg
7801 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
7861 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
7921 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
7981 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
8041 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
8101 cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga
8161 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
8221 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
8281 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
8341 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
8401 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
8461 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
8521 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
8581 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
8641 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
8701 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
8761 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
8821 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
8881 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
8941 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
9001 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
9061 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
9121 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
9181 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
9241 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
9301 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
9361 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
9421 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
9481 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
9541 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
9601 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
9661 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
9721 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
9781 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
9841 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
9901 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
9961 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
10021 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
10081 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
10141 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
10201 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
10261 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
10321 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
10381 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
10441 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
10501 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
10561 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
10621 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
10681 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
10741 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
10801 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
10861 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
10921 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
10981 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
11041 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
11101 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
11161 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
11221 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
11281 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
11341 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
11401 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
11461 gtgccacctg acgtc
//
| {
"Vector backbone": "pcDNA3.1",
"Backbone size w/o insert (bp)": "",
"Vector type": "Mammalian Expression, CRISPR, Synthetic Biology",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "Stbl3",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Epigenome editing by a CRISPR-Cas9-based acetyltransferase activates genes from promoters and enhancers",
"Publication": "Nat Biotechnol. 2015 May;33(5):510-7. doi: 10.1038/nbt.3199. Epub 2015 Apr 6.",
"DOI": "10.1038/nbt.3199.",
"Authors": "Hilton IB, D'Ippolito AM, Vockley CM, Thakore PI, Crawford GE, Reddy TE, Gersbach CA /nbt",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/25849900",
"PubMed ID": "PubMed 25849900"
} | {
"Gene/Insert name": "S.pyogenes dCas9 with c-terminal human p300 Core effector fusion (aa 1048-1664 of human p300)",
"Species": "H. sapiens (human), Synthetic; S. pyogenes",
"Insert Size (bp)": "6138",
"Mutation": "D10A; H840A in S.pyogenes Cas9; Y1467F mutation in p300 Core",
"Entrez Gene": "EP300",
"Promoter": "CMV",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "AscI (not destroyed)",
"3′ cloning site": "PmeI (not destroyed)",
"5′ sequencing primer": "CMV F",
"3′ sequencing primer": "Bgh R"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pAcGFP1-N1-RvCAHS8-GS | 192461 | Expression of RvCAHS8-GS_linker-AcGFP1 in mammalian cells | Takekazu Kunieda | Tanaka et al PLoS Biol. 2022 Sep 6;20(9):e3001780. doi: 10.1371/journal.pbio.3001780. eCollection 2022 Sep. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/378554/bdba029f-112e-4316-b228-7bc9c786c995/addgene-plasmid-192461-sequence-378554.gbk | full | LOCUS sequence-378554- 5360 bp DNA circular SYN 26-AUG-2024
DEFINITION Expression of RvCAHS8-GS_linker-AcGFP1 in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5360)
AUTHORS Tanaka A, Nakano T, Watanabe K, Masuda K, Honda G, Kamata S, Yasui
R, Kozuka-Hata H, Watanabe C, Chinen T, Kitagawa D, Sawai S, Oyama
M, Yanagisawa M, Kunieda T
TITLE Stress-dependent cell stiffening by tardigrade tolerance proteins
that reversibly form a filamentous network and gel.
JOURNAL PLoS Biol. 2022 Sep 6;20(9):e3001780. doi:
10.1371/journal.pbio.3001780. eCollection 2022 Sep.
PUBMED 36067153
REFERENCE 2 (bases 1 to 5360)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "PLoS
Biol."; date: "2022-09-6"; volume: "20(9):e3001780. doi"; pages: "
10.1371/journal.pbio.3001780. eCollection 2022 Sep"
FEATURES Location/Qualifiers
source 1..5360
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 61..364
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 365..568
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 519..539
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
CDS 1308..2027
/codon_start=1
/product="Aequorea coerulescens GFP"
/label=AcGFP1
/note="mammalian codon-optimized"
/translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
GFVTAAAITHGMDELYK"
polyA_signal 2149..2270
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2186..2205)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2240..2259
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(2277..2732)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2414..2435)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2626..2645
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2759..2863
/gene="bla"
/label=AmpR promoter
promoter 2865..3222
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
enhancer 2865..3056
/label=SV40 enhancer
/note="enhancer for the SV40 early promoter (Herr, 1993)"
primer_bind complement(2888..2908)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 3073..3208
/label=SV40 ori
/note="SV40 origin of replication"
CDS 3257..4051
/codon_start=1
/product="aminoglycoside phosphotransferase"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
CDS 3257..4051
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(3311..3330)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3921..3940
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(4239..4258)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 4283..4330
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4659..5246
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5147..5166
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gccaccatgt
601 ctcagcgagc ttcggaacac tcatcatcca gccaccagtc gtctggccag tctggacagt
661 cgagccagca cgtccaagag cagaacacca gctacacaca caccgatgtc cgcaccaacg
721 ttcccaatgt ccctcctcca gtcatctcga ccagcgctgc tggtctcagt gagcagatgg
781 tcggtcaagg tttcaccgcc tcggctgcgc gagtctccgg ccagtccacc gaagcccatg
841 tccaaatgac ccctgagatg gaagccgagg cccgcaagga ccgcgaacga tacgagcgcg
901 aactggaagc catcaaccaa cgccaccaga aggatattga aggcaagacc gaggcttacc
961 gaaagcaagc cgaagctgag gcggaacgtc ttcgacggga actagagaag caacatcaac
1021 gagacgttga gttccgcaag tccctggttc aagggacaat cgacaaccag aagaaacagg
1081 ttgagctgga ggctatgatg gccaagcgcg agttggaccg agaagccaag ctggctcaag
1141 atgctctcga gaagagcaaa atggccaccg atgctcaagt cacgttcgag tcagcagtag
1201 gccacacgat ttctggtagc accaccgtgt cccagtccga gaaagtatcc cagtccaaga
1261 agggtggagg cggttcaggc ggaggtggct ctggcggtgg cggatcgatg gtgagcaagg
1321 gcgccgagct gttcaccggc atcgtgccca tcctgatcga gctgaatggc gatgtgaatg
1381 gccacaagtt cagcgtgagc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
1441 tgaagttcat ctgcaccacc ggcaagctgc ctgtgccctg gcccaccctg gtgaccaccc
1501 tgagctacgg cgtgcagtgc ttctcacgct accccgatca catgaagcag cacgacttct
1561 tcaagagcgc catgcctgag ggctacatcc aggagcgcac catcttcttc gaggatgacg
1621 gcaactacaa gtcgcgcgcc gaggtgaagt tcgagggcga taccctggtg aatcgcatcg
1681 agctgaccgg caccgatttc aaggaggatg gcaacatcct gggcaataag atggagtaca
1741 actacaacgc ccacaatgtg tacatcatga ccgacaaggc caagaatggc atcaaggtga
1801 acttcaagat ccgccacaac atcgaggatg gcagcgtgca gctggccgac cactaccagc
1861 agaatacccc catcggcgat ggccctgtgc tgctgcccga taaccactac ctgtccaccc
1921 agagcgccct gtccaaggac cccaacgaga agcgcgatca catgatctac ttcggcttcg
1981 tgaccgccgc cgccatcacc cacggcatgg atgagctgta caagtgagcg gccgcgactc
2041 tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
2101 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
2161 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
2221 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaat
2281 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
2341 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
2401 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
2461 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
2521 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
2581 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
2641 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
2701 cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg
2761 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
2821 ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac
2881 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
2941 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
3001 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
3061 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
3121 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
3181 aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg
3241 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
3301 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
3361 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
3421 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt
3481 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
3541 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
3601 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
3661 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
3721 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
3781 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
3841 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
3901 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
3961 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
4021 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
4081 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
4141 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
4201 atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata
4261 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg
4321 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
4381 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
4441 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca
4501 ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
4561 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
4621 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
4681 gtaatctgct gcttgcaaac aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
4741 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
4801 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
4861 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
4921 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
4981 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
5041 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
5101 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
5161 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
5221 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
5281 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
5341 accgtattac cgccatgcat
//
| {
"Vector backbone": "pAcGFP1-N1",
"Backbone size w/o insert (bp)": "4650",
"Vector type": "Mammalian Expression",
"Selectable markers": "Neomycin (select with G418)"
} | {
"Bacterial Resistance(s)": "Kanamycin, 50 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "Unknown"
} | {
"Academic/Nonprofit Terms": "UBMTA ; Takara Bio Limited Use Label License (formerly Clontech)",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Stress-dependent cell stiffening by tardigrade tolerance proteins that reversibly form a filamentous network and gel",
"Publication": "PLoS Biol. 2022 Sep 6;20(9):e3001780. doi: 10.1371/journal.pbio.3001780. eCollection 2022 Sep.",
"DOI": "10.1371/journal.pbio.3001780.",
"Authors": "Tanaka A, Nakano T, Watanabe K, Masuda K, Honda G, Kamata S, Yasui R, Kozuka-Hata H, Watanabe C, Chinen T, Kitagawa D, Sawai S, Oyama M, Yanagisawa M, Kunieda T /journalpbio",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/36067153",
"PubMed ID": "PubMed 36067153"
} | {
"Gene/Insert name": "CAHS8",
"Species": "Ramazzottius varieornatus",
"Insert Size (bp)": "711",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "CMV",
"Tag / Fusion Protein": "AcGFP1 (C terminal on backbone)"
} | {
"Cloning method": "Gibson Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "CMV",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pRPS5A-PTP-3Xflag-16S rRNA Left_G1397C-ABE | 189642 | 16S rRNA Left_G1397C-ABE | Jin-Soo Kim | Mok et al Nat Plants. 2022 Dec 1. doi: 10.1038/s41477-022-01279-8. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/370911/da96fb56-23c7-4f3a-9cfb-307a61ab52bd/addgene-plasmid-189642-sequence-370911.gbk | full | LOCUS sequence-370911- 13154 bp DNA circular SYN 27-AUG-2024
DEFINITION 16S rRNA Left_G1397C-ABE.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 13154)
AUTHORS Mok YG, Hong S, Bae SJ, Cho SI, Kim JS
TITLE Targeted A-to-G base editing of chloroplast DNA in plants.
JOURNAL Nat Plants. 2022 Dec 1. doi: 10.1038/s41477-022-01279-8.
PUBMED 36456803
REFERENCE 2 (bases 1 to 13154)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 27, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat Plants.
2022 Dec 1. doi: 10.1038/s41477-022-01279-8."
FEATURES Location/Qualifiers
source 1..13154
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind complement(5..25)
/gene="mutant version of attB"
/label=attB4
/bound_moiety="BP Clonase(TM)"
/note="core recombination site for the Gateway(R) BP
reaction"
CDS 1857..1922
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
misc_feature 5341..5365
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
CDS 6696..7325
/codon_start=1
/product="stability protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/label=pVS1 StaA
/translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
CDS 7754..8827
/codon_start=1
/product="replication protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/label=pVS1 RepA
/translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
LVMRYRNLIEGEASAGS"
rep_origin 8893..9087
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
primer_bind complement(9323..9345)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
misc_feature 9431..9571
/label=bom
/note="basis of mobility region from pBR322"
primer_bind 9486..9505
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(9586..9603)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(9757..10345)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(9837..9856)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(10591..11382)
/codon_start=1
/gene="aadA"
/product="aminoglycoside adenylyltransferase (Murphy,
1985)"
/label=SmR
/note="confers resistance to spectinomycin and
streptomycin"
/translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
misc_feature 11903..11927
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
terminator complement(12020..12272)
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
CDS complement(12292..12843)
/codon_start=1
/gene="Streptomyces hygroscopicus bar"
/product="phosphinothricin acetyltransferase"
/label=BlpR
/note="confers resistance to bialophos or phosphinothricin"
/translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
WQLDFSLPVPPRPVLPVTEI"
promoter complement(12885..13064)
/label=NOS promoter
/note="nopaline synthase promoter"
ORIGIN
1 cgttcaactt ttctatacaa agttgaagat ctgactcaac ttttgattcg ctatttgcag
61 tgcacctgtg gcgttcatca catcttttgt gacactgttt gcactggtca ttgctattac
121 aaaggacctt cctgatgttg aaggagatcg aaagtaagta actgcacgca taaccatttt
181 ctttccgctc tttggctcaa tccatttgac agtcaaagac aatgtttaac cagctccgtt
241 tgatatattg tctttatgtg tttgttcaag catgtttagt taatcatgcc tttgattgat
301 cttgaatagg ttccaaatat caaccctggc aacaaaactt ggagtgagaa acattgcatt
361 cctcggttct ggacttctgc tagtaaatta tgtttcagcc atatcactag ctttctacat
421 gcctcaggtg aattcatcta tttccgtctt aactatttcg gttaattaaa gcacgaacac
481 cattactgca tgtagaagct tgataaacta tcgccaccaa tttatttttg ttgcgatatt
541 gttactttcc tcagtatgca gctttgaaaa gaccaaccct cttatccttt aacaatgaac
601 aggtttttag aggtagcttg atgattcctg cacatgtgat cttggcttca ggcttaattt
661 tccaggtaaa gcattatgag atactcttat atctcttaca tacttttgag ataatgcaca
721 agaacttcat aactatatgc tttagtttct gcatttgaca ctgccaaatt cattaatctc
781 taatatcttt gttgttgatc tttggtagac atgggtacta gaaaaagcaa actacaccaa
841 ggtaaaatac ttttgtacaa acataaactc gttatcacgg aacatcaatg gagtgtatat
901 ctaacggagt gtagaaacat ttgattattg caggaagcta tctcaggata ttatcggttt
961 atatggaatc tcttctacgc agagtatctg ttattcccct tcctctagct ttcaatttca
1021 tggtgaggat atgcagtttt ctttgtatat cattcttctt cttctttgta gcttggagtc
1081 aaaatcggtt ccttcatgta catacatcaa ggatatgtcc ttctgaattt ttatatcttg
1141 caataaaaat gcttgtacca attgaaacac cagctttttg agttctatga tcactgactt
1201 ggttctaacc aaaaaaaaaa aaatgtttaa tttacatatc taaaagtagg tttagggaaa
1261 cctaaacagt aaaatatttg tatattattc gaatttcact catcataaaa acttaaattg
1321 caccataaaa ttttgtttta ctattaatga tgtaatttgt gtaacttaag ataaaaataa
1381 tattccgtaa gttaaccggc taaaaccacg tataaaccag ggaacctgtt aaaccggttc
1441 tttactggat aaagaaatga aagcccatgt agacagctcc attagagccc aaaccctaaa
1501 tttctcatct atataaaagg agtgacatta gggtttttgt tcgtcctctt aaagcttctc
1561 gttttctctg ccgtctctct cattcgcgcg acgcaaacga tcttcaggtg atcttctttc
1621 tccaaatcct ctctcataac tctgatttcg tacttgtgta tttgagctca cgctctgttt
1681 ctctcaccac agccatggat tcacagctag tcttgtctct gaagctgaat ccaagcttca
1741 ctcctctttc tcctctcttc cctttcactc catgttcttc tttttcgccg tcgctccggt
1801 tttcttcttg ctactcccgc cgcctctatt ctccggttac cgtctacgcc gcgaaggact
1861 acaaagacca tgacggtgat tataaagatc atgacatcga ttacaaggat gacgatgaca
1921 agcccgggga tctacgcacg ctcggctaca gccagcagca acaggagaag atcaaaccga
1981 aggttcgttc gacagtggcg cagcaccacg aggcactggt cggccatggg tttacacacg
2041 cgcacatcgt tgcgctcagc caacacccgg cagcgttagg gaccgtcgct gtcaagtatc
2101 aggacatgat cgcagcgttg ccagaggcga cacacgaagc gatcgttggc gtcggcaaac
2161 agtggtccgg cgcacgcgct ctggaggcct tgctcacggt ggcgggagag ttgagaggtc
2221 caccgttaca gttggacaca ggccaacttc tcaagattgc aaaacgtggc ggcgtgaccg
2281 cagtggaggc agtgcatgca tggcgcaatg cactgacggg tgcccccctg aacttgaccc
2341 ctgcacaagt ggtggctatc gccagcaaca tcggtggtaa gcaagcattg gaaactgtgc
2401 aacgcctttt gccagtcctt tgtcaggatc atggtcttac acccgaccag gtggtcgcaa
2461 tcgcctctaa tattgggggg aagcaggcct tggaaactgt tcaaagattg ctcccagtcc
2521 tctgccagga tcacggcctg acccccgatc aagtcgttgc cattgctagt aacaatggcg
2581 gaaaacaagc tctcgagaca gtgcagcggc tcctccctgt gctctgccaa gcacacgggt
2641 tgacccccgc tcaggttgtg gctatcgcaa gtaacaacgg tggaaaacag gcccttgaaa
2701 ctgtccaacg ccttcttccc gttctctgcc aggctcacgg cttgacaccc gaccaggtcg
2761 ttgctattgc ctcaaacaat ggaggaaaac aggcactcga aaccgtccag cggctgctcc
2821 ccgtcttgtg ccaggaccat ggactgacac cagagcaagt ggttgccatc gcctccaaca
2881 acggtggtaa gcaagcactt gagactgtgc agaggctttt gcctgtgctt tgtcaggccc
2941 atgggctcac accagcccag gtcgttgcca ttgcaagtca tgacggcggc aagcaagcat
3001 tggagacagt gcagagactc ttgcccgtcc tgtgtcaaga ccatgggttg actcctgatc
3061 aggtcgtcgc aattgcctcc aatatcgggg gcaaacaggc cttggagaca gtccaacgct
3121 tgctcccagt tctttgccaa gatcacggcc ttacacccgc acaggtggtc gccatcgcta
3181 gtaatggtgg gggcaaacag gctctcgaga ctgttcaacg gcttctccca gttttgtgtc
3241 aagcacatgg gcttacccca gcacaggttg tcgcaattgc ctccaataac ggtggtaaac
3301 aggctctgga aacagtccag agactcctcc ccgttttgtg tcaagaccac ggtctgaccc
3361 ccgcccaagt ggttgctatt gccagtaaca ttggtggaaa acaagccctc gaaactgttc
3421 agaggctttt gccagtcctg tgccaggccc atgggcttac acctgagcaa gtggttgcaa
3481 tcgcttctaa cggagggggc aagcaggctc ttgagacagt tcaacgcctg ttgccagttt
3541 tgtgccaggc tcacggtctg actcccgctc aagttgtcgc tatcgccagc aataatggag
3601 gcaaacaagc acttgaaaca gtgcagagat tgttgccagt gctgtgccaa gaccacggct
3661 tgacacccgc ccaagttgtt gctattgcca gtaatatagg gggtaagcag gctctcgaaa
3721 cagttcaacg cctccttcct gttttgtgtc aggctcatgg tcttacaccc gatcaggttg
3781 ttgcaatcgc atcacacgat ggcgggaaac aggccctcga aaccgtccag cggcttttgc
3841 ctgtgctgtg ccaagctcat ggcttgacac ccgaacaggt tgttgccatt gccagtaacg
3901 ggggcggtaa acaggcactt gaaacagtgc agagattgct cccagttttg tgccaagcac
3961 acgggttgac acctgcacag gtggtggcaa tcgcatctaa cggcggagga aagcaggctc
4021 tggagactgt tcaaaggctt cttcccgttt tgtgtcaggc tcatggactt actccagacc
4081 aagttgtggc cattgcaagc aacaatggcg ggaaacaggc cttggagact gttcaaagac
4141 tcctgcccgt cctgtgtcaa gatcacggtc tgactccgga acaggtggtg gcgattgcaa
4201 gcaacggcgg cggcaaacag gctctagaga gcattgttgc ccagctctcc agacctgatc
4261 cggcgctagc cgcgttgaca aatgatcatc ttgttgctct tgcttgtctt ggcggacgac
4321 ctgccctgga tgctgtgaaa aaggggttgg ggggatccgg cagtgccata cctgtgaagc
4381 ggggagcaac aggggagaca aaggtgttca caggcaactc taacagtcca aagagcccca
4441 ccaaaggcgg gtgttcagga agcgaaactc ctggtacctc agagtccgct actcccgaat
4501 cctctgaggt ggagttttcc cacgagtact ggatgagaca tgccctgacc ctggccaaga
4561 gggcacggga tgagagggag gtgcctgtgg gagccgtgct ggtgctgaac aatagagtga
4621 tcggcgaggg ctggaacaga gccatcggcc tgcacgaccc aacagcccat gccgaaatta
4681 tggccctgag acagggcggc ctggtcatgc agaactacag actgattgac gccaccctgt
4741 acgtgacatt cgagccttgc gtgatgtgcg ccggcgccat gatccactct aggatcggcc
4801 gcgtggtgtt tggcgtgagg aactcaaaaa gaggcgccgc aggctccctg atgaacgtgc
4861 tgaactaccc cggcatgaat caccgcgtcg aaattaccga gggaatcctg gcagatgaat
4921 gtgccgccct gctgtgcgat ttctatcgga tgcctagaca ggtgttcaat gctcagaaga
4981 aggcccagag ctccatcaac tagggtacct gataatcgat atcccgcggc catgctagag
5041 tccgcaaaaa tcaccagtct ctctctacaa atctatctct ctctattttt ctccagaata
5101 atgtgtgagt agttcccaga taagggaatt agggttctta tagggtttcg ctcatgtgtt
5161 gagcatataa gaaaccctta gtatgtattt gtatttgtaa aatacttcta tcaataaaat
5221 ttctaattcc taaaaccaaa atccagtgac ctgcaggcat gcgacgtccg tcgggcccaa
5281 gcttagcttg agcttggatc agattgtcgt ttcccgcctt cagtttaaac tatcagtgtt
5341 tgacaggata tattggcggg taaacctaag agaaaagagc gtttattaga ataatcggat
5401 atttaaaagg gcgtgaaaag gtttatccgt tcgtccattt gtatgtgcat gccaaccaca
5461 gggttcccct cgggatcaaa gtactttaaa gtactttaaa gtactttaaa gtactttgat
5521 ccaacccctc cgctgctata gtgcagtcgg cttctgacgt tcagtgcagc cgtcttctga
5581 aaacgacatg tcgcacaagt cctaagttac gcgacaggct gccgccctgc ccttttcctg
5641 gcgttttctt gtcgcgtgtt ttagtcgcat aaagtagaat acttgcgact agaaccggag
5701 acattacgcc atgaacaaga gcgccgccgc tggcctgctg ggctatgccc gcgtcagcac
5761 cgacgaccag gacttgacca accaacgggc cgaactgcac gcggccggct gcaccaagct
5821 gttttccgag aagatcaccg gcaccaggcg cgaccgcccg gagctggcca ggatgcttga
5881 ccacctacgc cctggcgacg ttgtgacagt gaccaggcta gaccgcctgg cccgcagcac
5941 ccgcgaccta ctggacattg ccgagcgcat ccaggaggcc ggcgcgggcc tgcgtagcct
6001 ggcagagccg tgggccgaca ccaccacgcc ggccggccgc atggtgttga ccgtgttcgc
6061 cggcattgcc gagttcgagc gttccctaat catcgaccgc acccggagcg ggcgcgaggc
6121 cgccaaggcc cgaggcgtga agtttggccc ccgccctacc ctcaccccgg cacagatcgc
6181 gcacgcccgc gagctgatcg accaggaagg ccgcaccgtg aaagaggcgg ctgcactgct
6241 tggcgtgcat cgctcgaccc tgtaccgcgc acttgagcgc agcgaggaag tgacgcccac
6301 cgaggccagg cggcgcggtg ccttccgtga ggacgcattg accgaggccg acgccctggc
6361 ggccgccgag aatgaacgcc aagaggaaca agcatgaaac cgcaccagga cggccaggac
6421 gaaccgtttt tcattaccga agagatcgag gcggagatga tcgcggccgg gtacgtgttc
6481 gagccgcccg cgcacgtctc aaccgtgcgg ctgcatgaaa tcctggccgg tttgtctgat
6541 gccaagctgg cggcctggcc ggccagcttg gccgctgaag aaaccgagcg ccgccgtcta
6601 aaaaggtgat gtgtatttga gtaaaacagc ttgcgtcatg cggtcgctgc gtatatgatg
6661 cgatgagtaa ataaacaaat acgcaagggg aacgcatgaa ggttatcgct gtacttaacc
6721 agaaaggcgg gtcaggcaag acgaccatcg caacccatct agcccgcgcc ctgcaactcg
6781 ccggggccga tgttctgtta gtcgattccg atccccaggg cagtgcccgc gattgggcgg
6841 ccgtgcggga agatcaaccg ctaaccgttg tcggcatcga ccgcccgacg attgaccgcg
6901 acgtgaaggc catcggccgg cgcgacttcg tagtgatcga cggagcgccc caggcggcgg
6961 acttggctgt gtccgcgatc aaggcagccg acttcgtgct gattccggtg cagccaagcc
7021 cttacgacat atgggccacc gccgacctgg tggagctggt taagcagcgc attgaggtca
7081 cggatggaag gctacaagcg gcctttgtcg tgtcgcgggc gatcaaaggc acgcgcatcg
7141 gcggtgaggt tgccgaggcg ctggccgggt acgagctgcc cattcttgag tcccgtatca
7201 cgcagcgcgt gagctaccca ggcactgccg ccgccggcac aaccgttctt gaatcagaac
7261 ccgagggcga cgctgcccgc gaggtccagg cgctggccgc tgaaattaaa tcaaaactca
7321 tttgagttaa tgaggtaaag agaaaatgag caaaagcaca aacacgctaa gtgccggccg
7381 tccgagcgca cgcagcagca aggctgcaac gttggccagc ctggcagaca cgccagccat
7441 gaagcgggtc aactttcagt tgccggcgga ggatcacacc aagctgaaga tgtacgcggt
7501 acgccaaggc aagaccatta ccgagctgct atctgaatac atcgcgcagc taccagagta
7561 aatgagcaaa tgaataaatg agtagatgaa ttttagcggc taaaggaggc ggcatggaaa
7621 atcaagaaca accaggcacc gacgccgtgg aatgccccat gtgtggagga acgggcggtt
7681 ggccaggcgt aagcggctgg gttgtctgcc ggccctgcaa tggcactgga acccccaagc
7741 ccgaggaatc ggcgtgagcg gtcgcaaacc atccggcccg gtacaaatcg gcgcggcgct
7801 gggtgatgac ctggtggaga agttgaaggc cgcgcaggcc gcccagcggc aacgcatcga
7861 ggcagaagca cgccccggtg aatcgtggca agcggccgct gatcgaatcc gcaaagaatc
7921 ccggcaaccg ccggcagccg gtgcgccgtc gattaggaag ccgcccaagg gcgacgagca
7981 accagatttt ttcgttccga tgctctatga cgtgggcacc cgcgatagtc gcagcatcat
8041 ggacgtggcc gttttccgtc tgtcgaagcg tgaccgacga gctggcgagg tgatccgcta
8101 cgagcttcca gacgggcacg tagaggtttc cgcagggccg gccggcatgg ccagtgtgtg
8161 ggattacgac ctggtactga tggcggtttc ccatctaacc gaatccatga accgataccg
8221 ggaagggaag ggagacaagc ccggccgcgt gttccgtcca cacgttgcgg acgtactcaa
8281 gttctgccgg cgagccgatg gcggaaagca gaaagacgac ctggtagaaa cctgcattcg
8341 gttaaacacc acgcacgttg ccatgcagcg tacgaagaag gccaagaacg gccgcctggt
8401 gacggtatcc gagggtgaag ccttgattag ccgctacaag atcgtaaaga gcgaaaccgg
8461 gcggccggag tacatcgaga tcgagctagc tgattggatg taccgcgaga tcacagaagg
8521 caagaacccg gacgtgctga cggttcaccc cgattacttt ttgatcgatc ccggcatcgg
8581 ccgttttctc taccgcctgg cacgccgcgc cgcaggcaag gcagaagcca gatggttgtt
8641 caagacgatc tacgaacgca gtggcagcgc cggagagttc aagaagttct gtttcaccgt
8701 gcgcaagctg atcgggtcaa atgacctgcc ggagtacgat ttgaaggagg aggcggggca
8761 ggctggcccg atcctagtca tgcgctaccg caacctgatc gagggcgaag catccgccgg
8821 ttcctaatgt acggagcaga tgctagggca aattgcccta gcaggggaaa aaggtcgaaa
8881 aggtctcttt cctgtggata gcacgtacat tgggaaccca aagccgtaca ttgggaaccg
8941 gaacccgtac attgggaacc caaagccgta cattgggaac cggtcacaca tgtaagtgac
9001 tgatataaaa gagaaaaaag gcgatttttc cgcctaaaac tctttaaaac ttattaaaac
9061 tcttaaaacc cgcctggcct gtgcataact gtctggccag cgcacagccg aagagctgca
9121 aaaagcgcct acccttcggt cgctgcgctc cctacgcccc gccgcttcgc gtcggcctat
9181 cgcggccgct ggccgctcaa aaatggctgg cctacggcca ggcaatctac cagggcgcgg
9241 acaagccgcg ccgtcgccac tcgaccgccg gcgcccacat caaggcaccc tgcctcgcgc
9301 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
9361 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
9421 ggtgtcgggg cgcagccatg acccagtcac gtagcgatag cggagtgtat actggcttaa
9481 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
9541 cagatgcgta aggagaaaat accgcatcag gcgctcttcc gcttcctcgc tcactgactc
9601 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
9661 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
9721 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
9781 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
9841 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
9901 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
9961 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
10021 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
10081 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
10141 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
10201 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
10261 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
10321 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
10381 tcagtggaac gaaaactcac gttaagggat tttggtcatg catgatatat ctcccaattt
10441 gtgtagggct tattatgcac gcttaaaaat aataaaagca gacttgacct gatagtttgg
10501 ctgtgagcaa ttatgtgctt agtgcatcta acgcttgagt taagccgcgc cgcgaagcgg
10561 cgtcggcttg aacgaatttc tagctagaca ttatttgccg actaccttgg tgatctcgcc
10621 tttcacgtag tggacaaatt cttccaactg atctgcgcgc gaggccaagc gatcttcttc
10681 ttgtccaaga taagcctgtc tagcttcaag tatgacgggc tgatactggg ccggcaggcg
10741 ctccattgcc cagtcggcag cgacatcctt cggcgcgatt ttgccggtta ctgcgctgta
10801 ccaaatgcgg gacaacgtaa gcactacatt tcgctcatcg ccagcccagt cgggcggcga
10861 gttccatagc gttaaggttt catttagcgc ctcaaataga tcctgttcag gaaccggatc
10921 aaagagttcc tccgccgctg gacctaccaa ggcaacgcta tgttctcttg cttttgtcag
10981 caagatagcc agatcaatgt cgatcgtggc tggctcgaag atacctgcaa gaatgtcatt
11041 gcgctgccat tctccaaatt gcagttcgcg cttagctgga taacgccacg gaatgatgtc
11101 gtcgtgcaca acaatggtga cttctacagc gcggagaatc tcgctctctc caggggaagc
11161 cgaagtttcc aaaaggtcgt tgatcaaagc tcgccgcgtt gtttcatcaa gccttacggt
11221 caccgtaacc agcaaatcaa tatcactgtg tggcttcagg ccgccatcca ctgcggagcc
11281 gtacaaatgt acggccagca acgtcggttc gagatggcgc tcgatgacgc caactacctc
11341 tgatagttga gtcgatactt cggcgatcac cgcttccccc atgatgttta actttgtttt
11401 agggcgactg ccctgctgcg taacatcgtt gctgctccat aacatcaaac atcgacccac
11461 ggcgtaacgc gcttgctgct tggatgcccg aggcatagac tgtaccccaa aaaaacagtc
11521 ataacaagcc atgaaaaccg ccactgcgcc gttaccaccg ctgcgttcgg tcaaggttct
11581 ggaccagttg cgtgagcgca tacgctactt gcattacagc ttacgaaccg aacaggctta
11641 tgtccactgg gttcgtgccc gaattgatca caggcagcaa cgctctgtca tcgttacaat
11701 caacatgcta ccctccgcga gatcatccgt gtttcaaacc cggcagctta gttgccgttc
11761 ttccgaatag catcggtaac atgagcaaag tctgccgcct tacaacggct ctcccgctga
11821 cgccgtcccg gactgatggg ctgcctgtat cgagtggtga ttttgtgccg agctgccggt
11881 cggggagctg ttggctggct ggtggcagga tatattgtgg tgtaaacaaa ttgacgctta
11941 gacaacttaa taacacattg cggacgtttt taatgtactg aattaacgcc gaattgaatt
12001 atcagcttgc atgccggtcg atctagtaac atagatgaca ccgcgcgcga taatttatcc
12061 tagtttgcgc gctatatttt gttttctatc gcgtattaaa tgtataattg cgggactcta
12121 atcataaaaa cccatctcat aaataacgtc atgcattaca tgttaattat tacatgctta
12181 acgtaattca acagaaatta tatgataatc atcgcaagac cggcaacagg attcaatctt
12241 aagaaacttt attgccaaat gtttgaacga tctgcttgac tctaggggtc atcagatttc
12301 ggtgacgggc aggaccggac ggggcggcac cggcaggctg aagtccagct gccagaaacc
12361 cacgtcatgc cagttcccgt gcttgaagcc ggccgcccgc agcatgccgc ggggggcata
12421 tccgagcgcc tcgtgcatgc gcacgctcgg gtcgttgggc agcccgatga cagcgaccac
12481 gctcttgaag ccctgtgcct ccagggactt cagaaggtgg gtgtagagcg tggagcccag
12541 tcccgtccgc tggtggcggg gggagacgta cacggtcgac tcggccgtcc agtcgtaggc
12601 gttgcgtgcc ttccagggac ccgcgtaggc gatgccggcg acctcgccgt ccacctcggc
12661 gacgagccag ggatagcgct cccgcagacg gacgaggtcg tccgtccact cctgcggttc
12721 ctgcggctcg gtacggaagt tgaccgtgct tgtctcgatg tagtggttga cgatggtgca
12781 gaccgccggc atgtccgcct cggtggcacg gcggatgtcg gccgggcgtc gttctgggct
12841 catggtagat cccctcgatc gagttgagag tgaatatgag actctaattg gataccgagg
12901 ggaatttatg gaacgtcagt ggagcatttt tgacaagaaa tatttgctag ctgatagtga
12961 ccttaggcga cttttgaacg cgcaataatg gtttctgacg tatgtgctta gctcattaaa
13021 ctccagaaac ccgcggctca gtggctcctt caacgttgcg gttctgtcag ttccaaacgt
13081 aaaacggctt gtcccgcgtc atcggcgggg gtcataacgt gactccctta attctccgct
13141 catgatcgat aatt
//
| {
"Vector backbone": "pBAtC",
"Backbone size w/o insert (bp)": "",
"Vector type": "Plant Expression, TALEN",
"Selectable markers": "Basta"
} | {
"Bacterial Resistance(s)": "Spectinomycin, 50 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "Unknown"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Targeted A-to-G base editing of chloroplast DNA in plants",
"Publication": "Nat Plants. 2022 Dec 1. doi: 10.1038/s41477-022-01279-8.",
"DOI": "10.1038/s41477-022-01279-8.",
"Authors": "Mok YG, Hong S, Bae SJ, Cho SI, Kim JS /s---",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/36456803",
"PubMed ID": "PubMed 36456803"
} | {
"Gene/Insert name": "TALE and DddAtox and TadA 8e",
"Species": "A. thaliana (mustard weed)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "RPS5A",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Gibson Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "CTCACCACAGCCATGGATTCACAGCTAGTCTTGTCTC",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pRS-puro shNEDD4L #2 | 27017 | Low | Joan Massague | Gao et al Mol Cell. 2009 Nov 13. 36(3):457-68. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/33249/640b1c3f-538b-4928-835b-1e3eb1b05cde/addgene-plasmid-27017-sequence-33249.gbk | partial | LOCUS sequence-33249-e 929 bp DNA linear UNA 26-AUG-2024
DEFINITION natural linear DNA
ACCESSION .
VERSION .
KEYWORDS .
SOURCE natural DNA sequence
ORGANISM unspecified
REFERENCE 1 (bases 1 to 929)
AUTHORS Gao S, Alarcon C, Sapkota G, Rahman S, Chen PY, Goerner N, Macias
MJ, Erdjument-Bromage H, Tempst P, Massague J
TITLE Ubiquitin ligase Nedd4L targets activated Smad2/3 to limit TGF-beta
signaling.
JOURNAL Mol Cell. 2009 Nov 13. 36(3):457-68.
PUBMED 19917253
REFERENCE 2 (bases 1 to 929)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Mol Cell.
2009 Nov 13. 36(3):457-68."
FEATURES Location/Qualifiers
source 1..929
/mol_type="genomic DNA"
/organism="unspecified"
primer_bind 40..56
/label=pBluescriptKS
/note="For pBluescript vector"
primer_bind 41..57
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
promoter complement(130..344)
/label=H1 promoter
/note="human H1 RNA promoter"
primer_bind complement(190..209)
/label=H1
/note="Human H1 promoter, forward primer"
promoter 355..854
/label=PGK promoter
/note="mouse phosphoglycerate kinase 1 promoter"
primer_bind complement(390..413)
/label=MSCV-rev
/note="Murine stem cell virus, reverse primer"
primer_bind 787..806
/label=mPGK-F
/note="Mouse PGK promoter, forward primer"
primer_bind complement(879..898)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
ORIGIN
1 cctcactcct tctctaggcg ccggaattag atcgatctct cgaggtcgac ggtatcgata
61 agcttttcca aaaatgagga tcatttgtcc tactctcttg aagtaggaca aatgatcctc
121 aggggatctg tggtctcata cagaacttat aagattccca aatccaaaga catttcacgt
181 ttatggtgat ttcccagaac acatagcgac atgcaaatat tgcagggcgc cactcccctg
241 tccctcacag ccatcttcct gccagggcgc acgcgcgctg ggtgttcccg cctagtgaca
301 ctgggcccgc gattccttgg agcgggttga tgacgtcagc gttcgaattc taccgggtag
361 gggaggcgct tttcccaagg cagtctggag catgcgcttt agcagccccg ctgggcactt
421 ggcgctacac aagtggcctc tggcctcgca cacattccac atccaccggt aggcgccaac
481 cggctccgtt ctttggtggc cccttcgcgc caccttctac tcctccccta gtcaggaagt
541 tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca aatggaagta gcacgtctca
601 ctagtctcgt gcagatggac agcaccgctg agcaatggaa gcgggtaggc ctttggggca
661 gcggccaata gcagctttgc tccttcgctt tctgggctca gaggctggga aggggtgggt
721 ccgggggcgg gctcaggggc gggctcaggg gcggggcggg cgcccgaagg tcctccggag
781 gcccggcatt ctgcacgctt caaaagcgca cgtctgccgc gctgttctcc tcttcctcat
841 ctccgggcct ttcgacctgc agcccaagct agcttaccat gaccgagtac aagcccacgg
901 tgcgcctcgc cacccgcgac gacgtcccc
//
| {
"Vector backbone": "pRetroSuper",
"Backbone size w/o insert (bp)": "6401",
"Vector type": "Mammalian Expression, Retroviral, RNAi",
"Selectable markers": "Puromycin"
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Ubiquitin ligase Nedd4L targets activated Smad2/3 to limit TGF-beta signaling",
"Publication": "Mol Cell. 2009 Nov 13. 36(3):457-68.",
"DOI": "10.1016/j.molcel.2009.09.043",
"Authors": "Gao S, Alarcon C, Sapkota G, Rahman S, Chen PY, Goerner N, Macias MJ, Erdjument-Bromage H, Tempst P, Massague J",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/19917253",
"PubMed ID": "PubMed 19917253"
} | {
"Gene/Insert name": "shRNA NEDD4L #2",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "NEDD4L",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "BglII (not destroyed)",
"3′ cloning site": "HindIII (not destroyed)",
"5′ sequencing primer": "pLXSN-5",
"3′ sequencing primer": "MSCV-rev"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pBetaActin-FKBP-mScarlet-Dync1h1motorK2599T | 191331 | Expresses the fluorescently labeled motor-deficient dynein motor domain fused to the dimerization domain FKBP to recruit it to the plasma membrane by using the chemical dimerization system FKBP-FRB | Frank Bradke | Schelski et al Sci Adv. 2022 Nov 4;8(44):eabo2336. doi: 10.1126/sciadv.abo2336. Epub 2022 Nov 4. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/381751/41513365-0c32-4ceb-8ac6-cf7d4f3d7602/addgene-plasmid-191331-sequence-381751.gbk | full | LOCUS sequence-381751- 16665 bp DNA circular SYN 27-AUG-2024
DEFINITION Expresses the fluorescently labeled motor-deficient dynein motor
domain fused to the dimerization domain FKBP to recruit it to the
plasma membrane by using the chemical dimerization system FKBP-FRB.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 16665)
AUTHORS Schelski M, Bradke F
TITLE Microtubule retrograde flow retains neuronal polarization in a
fluctuating state.
JOURNAL Sci Adv. 2022 Nov 4;8(44):eabo2336. doi: 10.1126/sciadv.abo2336.
Epub 2022 Nov 4.
PUBMED 36332023
REFERENCE 2 (bases 1 to 16665)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 27, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Sci Adv.";
date: "2022-11-4"; volume: "8(44):eabo2336. doi"; pages: "
10.1126/sciadv.abo2336. Epub 2022 Nov 4"
FEATURES Location/Qualifiers
source 1..16665
/mol_type="other DNA"
/organism="synthetic DNA construct"
polyA_signal 150..284
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(187..206)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 241..260
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(298..314)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(298..314)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(311..333)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 322..338
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(346..376)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 391..412
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(529..546)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(700..1288)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(780..799)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(1459..2319)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 2082..2101
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(2320..2424)
/gene="bla"
/label=AmpR promoter
primer_bind 2492..2510
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(2548..2570)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 2670..2689
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 2883..2905
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 2897..2914
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 2898..2914
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 4715..4990
/label=chicken beta-actin promoter
primer_bind 5933..5955
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
CDS 6007..6330
/codon_start=1
/product="human FK506-binding protein FKBP12"
/label=FKBP
/translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
LKLE"
CDS 6352..7044
/codon_start=1
/product="bright monomeric red fluorescent protein evolved
from a synthetic template (Bindels et al., 2016)"
/label=mScarlet
/translation="VSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAK
LKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGGA
VTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIKM
ALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRHS
TGGMDELYK"
primer_bind complement(6482..6500)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 6750..6769
/label=mCherry-F
/note="mCherry, forward primer"
regulatory 11014..11023
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
ORIGIN
1 acggtaccgc gggcccggga tccaccggat ctagataagt aatgatcata atcagccata
61 tcacatctgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
121 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
181 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
241 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatcaattcg taatcatgtc
301 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
361 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
421 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
481 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
541 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
601 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
661 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
721 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
781 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
841 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
901 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
961 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
1021 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
1081 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
1141 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
1201 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
1261 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
1321 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
1381 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
1441 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
1501 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
1561 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
1621 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
1681 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
1741 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
1801 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
1861 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
1921 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
1981 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
2041 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
2101 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
2161 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
2221 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
2281 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
2341 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
2401 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
2461 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
2521 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
2581 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
2641 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
2701 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca
2761 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
2821 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
2881 ttcccagtca cgacgttgta aaacgacggc cagtgccaag ttgggatctt tgcattggcc
2941 cacggctctc aggatgggga tgctcccctt cagcacccgg ttccccttgg aaactgatgg
3001 tcctggctct gtggcatggc agtggcactg tgaggagccc cctaccagca gcacacagtg
3061 gggttggcac tgccacgctc cggatgccgc gctctgatcc aaccccataa tcaagggaac
3121 ccgaattgcc ccatcattgc ccccaccacc cccatcctgc cgggccctca caccccacgc
3181 tgccttctgg tgacattccc cagcccaaac ccacggcttc atggctaccg cggggcattt
3241 cccattgccg ccccattatc agctctgcac acctcccgct gtacccatgc ctcgtggctg
3301 cccttctttg acgtataatc ttctaattaa tacccggcct tgtcaaagtg gagcacaaac
3361 gttaattaat tccccagcag gcaggtaatt aacagtgtga ctcccttttt gctgcgagtg
3421 gggctgatac agagagatgt ggcactatgg agcccacggg gtcctggcac tgggtgccca
3481 cggaggtccc catgtgctgc agtgtcaccg cctccgagga gacagtattg tccctgcggt
3541 gtccctgcag ctcagctctg tccacagggc cacctccagt ttggagggga cacaatgcag
3601 ccccgatgca acccatcctc gcagcatccc agggacaaag accccactgc aagaccgcac
3661 acagggctgg gtcccgctcc cctaatatct acagtgcttc ttgcatggcc ccttaatcaa
3721 tgcagttaat cagcatgcgc tcatgcaccg ctctggagct gcaaagcccc tcgcagcgct
3781 gctcaccaac accgcgcacc gccccggccc agcctgcagc acgcgctgca aacaggaaag
3841 aaacaaaata ttgcccaaat gtaggcaaag gcattcggct gccttgacct ccgccgggcc
3901 gggccctgcc tgactcagct ccttactcag cgctcgcttc ctccctccgg ctgccaccgc
3961 cgcagcgcac accctgacaa agagtggccc ttaacgggct ctgaggtgca cccagcagtg
4021 cactcagcag tccaagggcc ggcctggagg tttgcaccgc tacgtgctga cattagcatt
4081 gaacttggcc ctgggtagtg ctgcaggccg ggcggggtgg gtgtagagag tgcagcgcgc
4141 gttgcacccg gtgccccttc ccctcccttg catcccagca ggctgcaccc cagcaccagg
4201 cccgtgcatg catgctcctg gtgttattgc agcctggtgc atgcatgcgt cttgatggtg
4261 cagcgctgtg catgcatcct ccttggtgtg tagcagtcta gtgcatgcat acccctcggt
4321 gttattgctg ctctgtgcac gcacgctcat tgtatcactt catcccagtg catgcactca
4381 cactggagcg attgctgctc ggtgcacgca cactcattgt atcactgcag ctcagtggct
4441 gcacgcacac cggtgttatt gctgctcggt gcgtgcatgc acatcagtgt cgctgcagct
4501 cagtgcatgc acgctcattg cccatcgcta tccctgcctc tcctgctggc gctccccggg
4561 aggtgacttc aaggggaccg caggaccacc tcgggggtgg ggggagggct gcacacgcgg
4621 accccgctcc ccctccccaa caaagcactg tggaatcaaa aaggggggag gggggatgga
4681 ggggcgcgtc acacccccgc cccacaccct cacctcgagg tgagccccac gttctgcttc
4741 actctcccca tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta
4801 ttttgtgcag cgatgggggc gggggggggg ggggcgcgcg ccaggcgggg cggggcgggg
4861 cgrggggcgg ggcggggcga ggcggagagg tgcggcggca gccaatcaga gcggcgcgct
4921 ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc
4981 gcggcgggcg ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc
5041 gcgccgcccg ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg
5101 cccttctcct ccgggctgta attagcgctt ggtttaatga cggctcgttt cttttctgtg
5161 gctgcgtgaa agccttaaag ggctccggga gggccctttg tgcggggggg agcggctcgg
5221 ggggtgcgtg cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg
5281 gctgtgagcg ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga
5341 gcgcggccgg gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg
5401 tgcggggtgt gtgcgtgggg gggtgagcag ggggtgtggg cgcggcggtc gggctgtaac
5461 ccccccctgc acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc
5521 cgtgcggggc gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggkgggggtg
5581 ccgggcgggg cggggccgcc tcgggccggg grgggctcgg gggaggggcg cggcggcccc
5641 cggagcgccg gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg
5701 tgcgagaggg cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg
5761 cgccgccgca ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa
5821 atgggcgggg agggccttcg tgcgtcgccg cgccgccgtc cccttctcca tctccagcct
5881 cggggctgtc cgcaggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc
5941 ttctggcgtg tgaccggcgg ggtttatatc ttcccttctc tgttcctccg cagcccccaa
6001 gctagcatgg gagtgcaggt ggaaaccatc tccccaggag acgggcgcac cttccccaag
6061 cgcggccaga cctgcgtggt gcactacacc gggatgcttg aagatggaaa gaaatttgat
6121 tcctcccggg acagaaacaa gccctttaag tttatgctag gcaagcagga ggtgatccga
6181 ggctgggaag aaggggttgc ccagatgagt gtgggtcaga gagccaaact gactatatct
6241 ccagattatg cctatggtgc cactgggcac ccaggcatca tcccaccaca tgccactctc
6301 gtcttcgatg tggagcttct aaaactggaa ggaagttcag gaggttctag tgtgagcaag
6361 ggcgaggcag tgatcaagga gttcatgcgg ttcaaggtgc acatggaggg ctccatgaac
6421 ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc
6481 gccaagctga aggtgaccaa gggtggcccc ctgcccttct cctgggacat cctgtcccct
6541 cagttcatgt acggctccag ggccttcacc aagcaccccg ccgacatccc cgactactat
6601 aagcagtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc
6661 gccgtgaccg tgacccagga cacctccctg gaggacggca ccctgatcta caaggtgaag
6721 ctccgcggca ccaacttccc tcctgacggc cccgtaatgc agaagaagac aatgggctgg
6781 gaagcgtcca ccgagcggtt gtaccccgag gacggcgtgc tgaagggcga cattaagatg
6841 gccctgcgcc tgaaggacgg cggccgctac ctggcggact tcaagaccac ctacaaggcc
6901 aagaagcccg tgcagatgcc cggcgcctac aacgtcgacc gcaagttgga catcacctcc
6961 cacaacgagg actacaccgt ggtggaacag tacgaacgct ccgagggccg ccactccacc
7021 ggcggcatgg acgagctgta caagtctggt ggatcaagtg gtggaggcga gatggccttg
7081 gaagaatttt tgaagcagat acgagaagtg tggaacacat acgaactaga cttggtcaac
7141 tatcagaaca aatgccgcct catccgaggc tgggatgacc tcttcaacaa ggtcaaggag
7201 catatcaaca gcgtctctgc catgaaactc tctccctatt acaaggtttt tgaagaagat
7261 gccctgagct gggaggacaa gctgaaccga atcatggctc tctttgacgt gtggattgac
7321 gtgcagaggc gctgggtcta cttggaaggc atcttcacag gcagtgcgga catcaaacat
7381 ctgctgcctg tggaaaccca gcgcttccag agtatcagca cggagttttt ggctctgatg
7441 aaaaaagtct ccaagtctcc acttgtgatg gatgttctga acatacaggg agtccagagg
7501 tctctggaaa ggttggcaga cctgctgggg aagatccaga aagcattggg ggagtacctg
7561 gaacgggagc gctcgtcttt ccccaggttc tactttgtgg gtgatgagga tttgcttgag
7621 atcattggga atagcaagaa cgtggctaag ttgcagaagc acttcaagaa aatgttcgcc
7681 ggcgtttcca gcatcatctt aaatgaggat aactctgtgg tcctcggcat ctcatcccga
7741 gaaggggagg aggttatgtt taaaactcca gtgtccatca ctgaacaccc caaaatcaat
7801 gagtggctca ctctggtgga aaaggagatg agagtcacct tggctaaact tctggcagag
7861 tctgttacag aagtggagat ttttggaaaa gcaacttcta ttgacccaaa tacctacatt
7921 acttggattg ataaatacca ggcccagcta gtggtattgt cagcccagat agcctggtcc
7981 gagaacgtgg agaatgcgct gagcaacgtt gggggaggtg gtgatgtggg gcccttgcag
8041 tctgtgctga gcaatgtgga ggtcacactc aacgtcttag cggactctgt cctcatggag
8101 cagcctcctc tccggagacg gaagctggag cacttgatca cagagctggt tcaccaaaga
8161 gacgttacca ggtccttgat caaaagcaag atcgacaacg ccaaatcttt cgaatggctc
8221 agccagatgc gattttactt tgaccctaag caaacggacg tgttacagca gctgtccatt
8281 cagatggcca atgccaagtt taactacggc tttgagtacc tgggtgttca agacaagctg
8341 gttcagacgc ccctcactga ccgttgctat ttgacaatga cacaagcttt ggaggccagg
8401 cttgggggct ccccatttgg acctgctgga accggaaaga cagagtccgt caaagccctc
8461 ggccatcagc tggggcggtt cgtcttagtt ttcaactgtg acgaaacctt tgatttccag
8521 gcgatgggac gtatctttgt ggggctttgc caagtgggcg cgtggggctg ctttgatgag
8581 tttaaccgcc tggaggagcg gatgctctca gctgtatccc agcaggtgca gtgcatacaa
8641 gaggcactgc gggagcattc caaccccaac tacgacaaga cctcagcccc cattacttgt
8701 gagctgctga acaaacaagt caaggttagc ccagacatgg ctatcttcat caccatgaac
8761 ccaggctatg caggtcgctc aaacctcccc gacaacctca agaagctctt ccggagcttg
8821 gcgatgacca agcctgaccg gcagctgatc gcccaggtca tgctgtactc gcagggcttc
8881 cggactgctg aggtccttgc caacaagatc gtcccgttct tcaaactgtg tgatgagcaa
8941 ctgtcttctc aaagccatta tgactttggg cttcgggctt tgaagagtgt gcttgtaagt
9001 gcaggcaatg tgaagaggga gaggatccag aagataaaga gggagaaaga agaacggggg
9061 gaagcagttg atgaaggaga gattgctgag aacctacctg agcaggagat tctgatccag
9121 agtgtctgcg agacaatggt gcccaagctg gtggcagaag atatcccgct gctcttcagc
9181 ctcctgtctg acgtgtttcc tggagtccag taccaccgtg gggagatgac agcccttcga
9241 gaagagctga agaaagtgtg tcaggagatg tacttgacat atggcgacgg agaagaagtt
9301 ggcgggatgt gggttgaaaa ggttctccag ctctatcaga tcacgcagat taaccacggc
9361 ctgatgatgg ttgggccttc aggaagtggg aagagcatgg cctggcgagt gctgctgaag
9421 gctctagaga ggcttgaggg tgtggagggc gtggcccaca taattgaccc caaggccatc
9481 agcaaagacc acctctacgg aaccttggat cccaacacca gggagtggac agacggactc
9541 ttcacacacg tgctgagaaa gatcatagac aatgtgagag gtgagctgca gaaacgccag
9601 tggattgtct ttgatggcga tgtggacccc gagtgggtgg agaacctgaa ctccgtgctg
9661 gatgacaaca agctcctgac gctgcctaat ggagagcgcc ttagcctgcc tcccaatgtg
9721 cgaataatgt ttgaagtgca ggacttgaaa tacgcgacgc tggccacggt gtctcgctgt
9781 ggcatggtgt ggttcagtga ggacgtcctg agcacagaca tgatcttcaa taacttcctg
9841 gccaggctgc gcagcattcc cctggatgaa ggggaggacg aggcacagcg taggcgaaaa
9901 ggcaaagaag acgagggcga agaggccgct tctccaatgc tgcagattca aagagatgca
9961 gcaacgatca tgcagccgta cttcacgtcc aacggcctgg tcaccaaggc gctggagcac
10021 gccttcaagc tggagcacat catggacctg acgcgcctgc gctgcctggg ctcgctcttc
10081 tccatgctgc accaggcctg ccgcaacgtg gcgcagtaca atgccaacca tcctgatttc
10141 cccatgcaga ttgagcagct ggagcgctac attcagcgct atctggttta cgccatcctc
10201 tggtccttat ctggagacag ccggctgaag atgagagcag agctgggcga gtacatccgg
10261 aggatcacca ctgtgccgct gcccactgcg cccaacgtcc ccatcattga ctatgaggtg
10321 tccatcagtg gagagtggtc gccgtggcag gccaaggtgc ctcagattga agtggagaca
10381 cacaaggtgg cagccccaga tgtcgtcgtg ccaaccttag atacagtccg ccatgaagct
10441 ctcctgtaca cttggctggc tgaacacaaa cccctggtct tatgtggccc tcctgggtct
10501 ggcacgacca tgacactctt cagtgctctc cgggccttgc ctgacatgga ggttgtgggc
10561 ctaaacttct caagtgccac tacaccagag ctgttgctga agacgtttga ccactactgc
10621 gaatacaggc gtacacctaa cggtgttgtc ttggctcctg ttcagcttgg aaagtggctg
10681 gtcttgttct gtgatgaaat caacttgccc gatatggata aatacggcac acagagagtc
10741 atctccttca tcagacagat ggtggagcat ggaggcttct atcgtacctc agaccaaacg
10801 tgggtaaagc tagagagaat ccagtttgtt ggggcttgca atccacccac tgaccctggg
10861 agaaagcccc tctcacacag gttcttgcgc cacgtgcctg ttgtatatgt ggattaccca
10921 ggaccagcct ccctgacgca aatctatggc acttttaacc gcgccatgct gaggctcatt
10981 ccctcgctcc gtacctatgc agaacccctc actgccgcca tggtggagtt ctataccatg
11041 tcacaggagc ggtttactca ggacacacag ccacactata tctactcgcc ccgtgagatg
11101 actcggtggg taagaggaat ctttgaagca ctgagacccc tggagacatt gcctgttgaa
11161 ggtctcattc gtatttgggc acatgaagcc ctccgtctct tccaagacag gcttgtggag
11221 gatgaggagc ggcgctggac tgacgaaaac attgacatgg ttgccttgaa gcacttccct
11281 aacatagaca aagagaaagc catgagccgg cctatcctgt acagcaactg gctgtcaaag
11341 gattatatcc cagttgacca agaagagcta agggattatg tcaaggctag gctgaaggtc
11401 ttctatgaag aagaacttga tgttcccttg gtcttattca atgaagttct ggaccatgtg
11461 ctgaggatcg acagaatatt ccgacaacct caaggccatc tgcttctgat tggtgttagt
11521 ggagctggaa aaaccaccct gtctcgcttt gtcgcctgga tgaatggctt gagtgtatac
11581 cagattaagg tccacaggaa gtacactggg gaagacttcg atgaagatct taggacagtg
11641 ctgagacggt ctggctgtaa aaatgaaaag atagcattca taatggatga gtccaacgtc
11701 ttagattccg gattcctgga acgaatgaat acccttctgg ctaatggcga ggttcctggg
11761 ctctttgaag gggatgagta tgccacgctg atgacacagt gcaaagaggg ggcccagaag
11821 gaggggctca tgctggactc ccacgaggag ctctacaagt ggttcaccag ccaggtgatc
11881 cgcaacctgc acgtggtctt caccatgaac ccgtcctcag aggggctcaa ggacagggcg
11941 gccacctcac cagccctttt caacaggtgt gtgctgaact ggtttggtga ctggtccact
12001 gaagcactgt atcaagtcgg caaagaattc accagtaaga tggacctgga gaagccgaac
12061 tatatagtgc ctgactacat gccagttgtg tatgataaac tgccgcagcc cccaacacac
12121 cgggaagcca ttgtgaacag ctgtgtgttt gtccatcaga ctcttcacca ggctaatgcg
12181 cgcctggcga agcgaggtgg cagaacgatg gccatcacac ctcggcacta cctggacttc
12241 atcaaccact acgccaacct cttccatgag aagcgcagcg agttggagga gcagcagatg
12301 cacctgaatg ttgggctccg aaaaatcaaa gagaccgtgg accaggtgga agaactgcgc
12361 cgtgacctga ggatcaagag ccaagagctg gaggtcaaga acgcagcggc caatgacaag
12421 ctgaagaaaa tggtgaaaga ccagcaagag gctgagaaga agaaggtcat gagccaagaa
12481 atccaggagc agctacacaa acagcaggaa gtgatcgcag acaagcagat gagtgtcaag
12541 gaggacctgg ataaagtgga gcctgctgtc atcgaggccc agaacgctgt gaagtccatc
12601 aagaagcagc acctggtgga ggtgaggtcc atggccaacc ctcctgcagc tgtgaagctg
12661 gctctggagt ccatctgcct gctgcttggt gagagcacca cggactggaa gcagatccgc
12721 tccatcatca tgagagagaa cttcatcccc accatcgtca acttctcggc tgaggagatc
12781 agtgatgcca tacgagagaa gatgaagaaa aactacatgt ccaaccccag ttacaactat
12841 gagatcgtca accgggcttc cctggcttgt ggtcctatgg tgaagtgggc gattgcacag
12901 ctcaattatg cagacatgtt aaagcgagtg gagcccctga ggaatgagct gcagaagctg
12961 gaagatgacg ccaaggacaa ccagcagaaa gccaatgagg tggagcagat gatcagggac
13021 ctggaagcca gcattgcccg ctacaaggag gagtacgctg tcctcatctc tgaggcccag
13081 gccatcaagg cagacctggc agctgtggaa gcaaaggtaa accggagcac tgcacttcta
13141 aagagtttat ctgccgaaag ggaacgttgg gaaaagacga gtgaaacttt caaaaaccag
13201 atgtctacca ttgctgggga ctgcctcctg tcggcggcct tcattgctta cgctggctac
13261 tttgaccagc aaatgcgcca gaacttgttc accacctggt ctcatcactt acagcaagcc
13321 aacatccagt tccgcacaga cattgcaagg acggagtacc tctccaatgc tgacgaacgc
13381 ctccgctggc aggccagctc tctgcctgcc gacgacctgt gcacagagaa tgccatcatg
13441 ctaaaacgat tcaataggta cccgctgatc atcgacccct ccggacaggc cacagaattc
13501 atcatgaatg aatataagga ccggaagatc acaaggacca gcttcttaga tgatgccttc
13561 cggaagaatt tggagagtgc tctgagattt ggcaatcccc ttcttgtcca ggatgtggaa
13621 agctacgatc cagttttgaa tccagtactg aaccgtgaag ttcggcgaac aggagggagg
13681 gtgctgatta ccctcggtga ccaagacata gacctgtcac catcctttgt catctttctg
13741 tctacccgag acccaactgt ggagttccca ccagacctct gctcccgggt gacctttgtg
13801 aacttcaccg tcacccgcag cagtttacag agccagtgtc tcaatgaagt gcttaaagca
13861 gaaaggccag acgtggacga gaagcgctcc gacctcctga aactgcaagg ggagttccaa
13921 ctccgtttac ggcagcttga aaagtctctg ctccaagctc tgaatgaggt aaaaggccgc
13981 attctggatg acgacaccat cattacaacc ctggagaatc tgaagcggga agctgccgag
14041 gttaccagaa aggtggagga gacagatatt gtcatgcagg aggtggagac cgtgtcccag
14101 cagtacctgc cgctgtccac tgcctgcagc agcatctact tcactatgga gtccttgaag
14161 caggttcact tcctgtacca gtactctctg cagttcttcc tggacatcta ccacaacgtg
14221 ctgtatgaga accccaacct gaagggtgcc accgatcaca cacagcgcct ctccatcata
14281 accaaggacc tcttccaggt ggcctttaac cgagtggctc gaggcatgct gcaccaggac
14341 cacatcacct tcgccatgct gctagcgagg atcaagctga aaggcaccgt gggggagccc
14401 acctatgatg cagagttcca gcacttcctc agagggaagg agattgtgct gagtgctggc
14461 tccaccccca aaatccaggg cctgaccgta gagcaggcag aagctgtcgt gaggttgagc
14521 tgtctgcctg catttaaaga tctgattgcc aaggtgcagg cagatgagca gtttggcatt
14581 tggctggata gcagttcacc agagcagacc gtgccatacc tctggagcga ggaaacgcct
14641 acaactccca ttgggcaggc catccaccgc ctgctcctga ttcaagcctt ccgacccgac
14701 cgcctgttgg ccatggccca catgtttgtt tccacaaacc tcggagagtc cttcatgtcc
14761 atcatggagc agcctctcga cctcacccac attgtgggca cagaggtgaa gccaaacaca
14821 cctgtcctga tgtgctctgt gcctgggtat gatgccagcg gacatgttga ggacctcgca
14881 gcagagcaga acacgcagat cacgtctatt gccatcggct cagcagaagg ctttaaccaa
14941 gcagataagg ccatcaacac cgccgtgaag tcaggcaggt gggtgatgtt gaagaacgtg
15001 cacctggccc cgggctggct gatgcagctg gagaagaagc tgcactccct gcagccccac
15061 gcctgcttcc gacttttcct caccatggag atcaacccca aggtgcccgt gaacctgctc
15121 cgtgctggtc ggatctttgt gtttgagcca ccacccggag tgaaagccaa catgctgagg
15181 acatttagca gcatccctgt ctctaggata tgcaagtctc caaatgagcg cgcccgcttg
15241 tatttcctgc tggcttggtt ccatgccatc atccaagagc gcctgcgcta tgctccactg
15301 gggtggtcaa agaagtacga atttggagaa tctgaccttc gatcagcttg tgacacggtg
15361 gacacgtggc tggacgacac agccaagggc cggcagaaca tctcaccgga taagatccca
15421 tggtctgccc tgaagaccct gatggcgcag tccatctatg gtgggcgggt ggacaatgag
15481 ttcgaccagc gtctgctcaa caccttcctg gagcggctgt tcaccacccg cagcttcgac
15541 agtgaattca agctggcgtg caaggttgac gggcacaagg acatccagat gcctgatggt
15601 atcaggcgag aggagtttgt gcagtgggtg gagctgctgc ctgacgccca gacaccctca
15661 tggctgggcc tccccaacaa tgctgaaagg gtcctgctta ccacccaagg cgtggacatg
15721 atcagcaaga tgctgaagat gcagatgctg gaggacgagg atgacctggc ctacgcagag
15781 acggagaaga aggcaaggac ggactccacc tccgatggcc gtccagcctg gatgaggaca
15841 ctgcacacca cggcgtccaa ctggctgcac ctcatcccac agacactgag ccccctgaag
15901 cgcaccgtgg agaacatcaa ggatcctttg tttaggttct ttgagagaga agtaaagatg
15961 ggggccaagt tactccaaga tgtgcgccaa gatcttgctg acgtcgttca agtgtgcgag
16021 gggaaaaaga agcagacaaa ctacctgcgc acgctcatca atgagctggt gaaagggatc
16081 ttgcctcgga gctggtccca ttacacagtg cctgcaggca tgactgtgat ccagtgggtg
16141 tcagatttca gcgagaggat caaacagctg cagaacatct cacaagcagc tgcatctggt
16201 ggtgccaagg aactgaagaa catccacgtg tgcctgggtg gcctgtttgt acctgaggcc
16261 tacatcactg ccaccagaca gtatgtggca caagctaaca gctggtccct ggaggaactc
16321 tgcctggaag tcaatgtcac cgcctcacag agtgccaccc ttgatgcctg cagctttgga
16381 gtcacaggtc tgaagctcca aggggccacg tgcagcaaca acaagctatc actctccaat
16441 gccatctcca cagtcctccc cctcacacag ctgcgctggg tgaaacaaac aagcgccgag
16501 aagaaggcca gcgtggtgac cttacctgtc tacctgaact tcacccgggc agacctcatc
16561 ttcaccgtgg actttgaaat tgctacaaag gaagatcctc gcagcttcta tgagcgtgga
16621 gttgccgttc tgtgcactga gtaaaagctt cgaattctgc agtcg
//
| {
"Vector backbone": "P_actin",
"Backbone size w/o insert (bp)": "",
"Vector type": "Mammalian Expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Microtubule retrograde flow retains neuronal polarization in a fluctuating state",
"Publication": "Sci Adv. 2022 Nov 4;8(44):eabo2336. doi: 10.1126/sciadv.abo2336. Epub 2022 Nov 4.",
"DOI": "10.1126/sciadv.abo2336.",
"Authors": "Schelski M, Bradke F /sciadvabo",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/36332023",
"PubMed ID": "PubMed 36332023"
} | {
"Gene/Insert name": "FKBP-mScarlet-Dync1h1motorK2599T",
"Species": "M. musculus (mouse)",
"Insert Size (bp)": "",
"Mutation": "DYNC1H1 motor domain aa 1453-4644",
"Entrez Gene": "Dync1h1",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Ligation Independent Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
mito-PDCD10-mScarlet-I | 208887 | Express mScarlet-I-PDCD10 targeted to the mitochondria in mammalian cells. | Guillaume Jacquemet | Neil J. et al BioRxiv | https://media.addgene.org/snapgene-media/v2.0.0/sequences/417776/e8dd79a1-51c7-4af6-b849-fd73b760a8ad/addgene-plasmid-208887-sequence-417776.gbk | full | LOCUS sequence-417776- 5487 bp DNA circular SYN 27-AUG-2024
DEFINITION Express mScarlet-I-PDCD10 targeted to the mitochondria in mammalian
cells..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5487)
AUTHORS Neil J. Ball, Sujan Ghimire, Gautier Follain, Ada O. Pajari, Monika
Vaitkeviciute, Alana R. Cowell, Bence Berki, Johanna Ivaska,
ProfileIlkka Paatero, Benjamin T. Goult, Guillaume Jacquemet
TITLE TLNRD1 is a CCM complex component and regulates endothelial barrier
integrity
JOURNAL BioRxiv
REFERENCE 2 (bases 1 to 5487)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 27, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "BioRxiv"
FEATURES Location/Qualifiers
source 1..5487
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 526..545
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
promoter complement(634..991)
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 648..783
/label=SV40 ori
/note="SV40 origin of replication"
enhancer 800..991
/label=SV40 enhancer
/note="enhancer for the SV40 early promoter (Herr, 1993)"
primer_bind 948..968
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter complement(993..1097)
/gene="bla"
/label=AmpR promoter
rep_origin 1124..1579
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1211..1230)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 1421..1442
/label=F1ori-F
/note="F1 origin, forward primer"
polyA_signal 1586..1707
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1597..1616)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 1651..1670
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
CDS complement(2528..3223)
/codon_start=1
/product="bright monomeric red fluorescent protein evolved
from a synthetic template (Bindels et al., 2016)"
/label=mScarlet-I
/note="matures faster than mScarlet"
/translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
STGGMDELYK"
primer_bind complement(2803..2822)
/label=mCherry-F
/note="mCherry, forward primer"
primer_bind 3072..3090
/label=mCherry-R
/note="mCherry, reverse primer"
regulatory 3220..3229
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS complement(3257..3331)
/codon_start=1
/product="mitochondrial presequence of human cytochrome c
oxidase subunit VIII (Rizutto et al., 1989)"
/label=COX8 presequence
/translation="MSVLTPLLLRGLTGSARRLPVPRAK"
primer_bind complement(3350..3369)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(3351..3369)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter complement(3414..3617)
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind complement(3443..3463)
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
enhancer 3618..3921
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
rep_origin complement(4096..4684)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4176..4195)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
polyA_signal 5013..5060
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
primer_bind 5085..5104
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
CDS complement(join(5292..5487,1..599))
/codon_start=1
/product="aminoglycoside phosphotransferase"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
CDS complement(join(5292..5487,1..599))
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(5403..5422)
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
ORIGIN
1 ttttccacca tgatattcgg caagcaggca tcgccatggg tcacgacgag atcctcgccg
61 tcgggcatgc tcgccttgag cctggcgaac agttcggctg gcgcgagccc ctgatgctct
121 tcgtccagat catcctgatc gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg
181 cgatgtttcg cttggtggtc gaatgggcag gtagccggat caagcgtatg cagccgccgc
241 attgcatcag ccatgatgga tactttctcg gcaggagcaa ggtgagatga caggagatcc
301 tgccccggca cttcgcccaa tagcagccag tcccttcccg cttcagtgac aacgtcgagc
361 acagctgcgc aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc ctcgtcttgc
421 agttcattca gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct
481 gacagccgga acacggcggc atcagagcag ccgattgtct gttgtgccca gtcatagccg
541 aatagcctct ccacccaagc ggccggagaa cctgcgtgca atccatcttg ttcaatcatg
601 cgaaacgatc ctcatcctgt ctcttgatcg atctttgcaa aagcctaggc ctccaaaaaa
661 gcctcctcac tacttctgga atagctcaga ggccgaggcg gcctcggcct ctgcataaat
721 aaaaaaaatt agtcagccat ggggcggaga atgggcggaa ctgggcggag ttaggggcgg
781 gatgggcgga gttaggggcg ggactatggt tgctgactaa ttgagatgca tgctttgcat
841 acttctgcct gctggggagc ctggggactt tccacacctg gttgctgact aattgagatg
901 catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc ctaactgaca
961 cacattccac agctggttct ttccgcctca ggactcttcc tttttcaata ttattgaagc
1021 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
1081 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgcgcc ctgtagcggc
1141 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
1201 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
1261 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
1321 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
1381 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
1441 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
1501 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
1561 atattaacgc ttacaattta cgccttaaga tacattgatg agtttggaca aaccacaact
1621 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
1681 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
1741 gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatg
1801 gctgattatg atcagttatc tagatccggt ggatcccggg cccgcggtac cgtcgactgc
1861 agaattccta tgccactgtc ttaaacgttt gcaagatgag gttggtttgg tgaataaggc
1921 gattcgcgct tacgaagacg ttaattgctt ttccgtcttt gaagtaagtc ttcaaagtat
1981 cactgaaaga ctttgagtac ttcacgaact ccttcttttg gtgttcgagt gcccttctat
2041 tttgatactg gtacttcttg aatacgttgt tcacggtatc caacagttcc tttatggcgg
2101 aggcaatgtc tttgatagtt tgcagaaacc gtaccctatc gttgatctcg tccgggattt
2161 ttgagagaat ttgcttcagc gcccgagctt tttcgttgag atcttggaac tcaggctcag
2221 gtctctcgat catatactcc tcaacgtcat cggcggccat ccgcagcaga gactcggtga
2281 aatttacctc cacagatttt ttttcaagaa ttttcattat gatgtcttga gtcaggccgg
2341 ggttttcctt ttctgccttg atgaacgccg cccgcaatgt ctgtgctgct gaaagattca
2401 cccgttcgag ctcgttaaaa actggataca ttacggcgta caaaggcatg gataccatag
2461 acgtggtttc ggcttcattc ttcatttctt ccattgtcat cctcatagct cgagatctga
2521 gtccggactt gtacagctcg tccatgccgc cggtggagtg gcggccctcg gagcgttcgt
2581 actgttccac cacggtgtag tcctcgttgt gggaggtgat gtccaacttg cggtcgacgt
2641 tgtaggcgcc gggcatctgc acgggcttct tggccttgta ggtggtcttg aagtccgcca
2701 ggtagcggcc gccgtccttc aggcgcaggg ccatcttaat gtcgcccttc agcacgccgt
2761 cctcggggta caaccgctcg gtggacgctt cccagcccat tgtcttcttc tgcattacgg
2821 ggccgtcagg agggaagttg gtgccgcgga gcttcacctt gtagatcagg gtgccgtcct
2881 ccagggaggt gtcctgggtc acggtcacgg cgccgccgtc ctcgaagttc atcacgcgct
2941 cccacttgaa gccctcgggg aaggactgct tatagtagtc ggggatgtcg gcggggtgct
3001 tgatgaaggc cctggagccg tacatgaact gaggggacag gatgtcccag gagaagggca
3061 gggggccacc cttggtcacc ttcagcttgg cggtctgggt gccctcgtag gggcggccct
3121 cgccctcgcc ctcgatctcg aactcgtggc cgttcatgga gccctccatg tgcaccttga
3181 accgcatgaa ctccttgatc actgcctcgc ccttgctcac catggtggcg accggtggat
3241 cccccaacga atggatcttg gcgcgcggca ctgggagccg ccgggccgag cctgtcaagc
3301 cccgcagcag cagcggcgtc aggacggaca tgctagccag cttgggtctc cctatagtga
3361 gtcgtattaa tttcgataag ccagtaagca gtgggttctc tagttagcca gagagctctg
3421 cttatataga cctcccaccg tacacgccta ccgcccattt gcgtcaatgg ggcggagttg
3481 ttacgacatt ttggaaagtc ccgttgattt tggtgccaaa acaaactccc attgacgtca
3541 atggggtgga gacttggaaa tccccgtgag tcaaaccgct atccacgccc attgatgtac
3601 tgccaaaacc gcatcaccat ggtaatagcg atgactaata cgtagatgta ctgccaagta
3661 ggaaagtccc ataaggtcat gtactgggca taatgccagg cgggccattt accgtcattg
3721 acgtcaatag ggggcgtact tggcatatga tacacttgat gtactgccaa gtgggcagtt
3781 taccgtaaat actccaccca ttgacgtcaa tggaaagtcc ctattggcgt tactatggga
3841 acatacgtca ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca
3901 tttaccgtaa gttatgtaac gcggaactcc atatatgggc tatgaactaa tgaccccgta
3961 attgattact attaataact aatgcatggc ggtaatacgg ttatccacag aatcagggga
4021 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
4081 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
4141 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
4201 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
4261 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
4321 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
4381 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
4441 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
4501 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
4561 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
4621 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
4681 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
4741 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
4801 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa cctgaggcta tggcagggcc
4861 tgccgccccg acgttggctg cgagccctgg gccttcaccc gaacttgggg ggtggggtgg
4921 ggaaaaggaa gaaacgcggg cgtattggcc ccaatggggt ctcggtgggg tatcgacaga
4981 gtgccagccc tgggaccgaa ccccgcgttt atgaacaaac gacccaacac cgtgcgtttt
5041 attctgtctt tttattgccg tcatagcgcg ggttccttcc ggtattgtct ccttccgtgt
5101 ttcagttagc ctccccctag ggtgggcgaa gaactccagc atgagatccc cgcgctggag
5161 gatcatccag ccggcgtccc ggaaaacgat tccgaagccc aacctttcat agaaggcggc
5221 ggtggaatcg aaatctcgtg atggcaggtt gggcgtcgct tggtcggtca tttcgaaccc
5281 cagagtcccg ctcagaagaa ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg
5341 ggagcggcga taccgtaaag cacgaggaag cggtcagccc attcgccgcc aagctcttca
5401 gcaatatcac gggtagccaa cgctatgtcc tgatagcggt ccgccacacc cagccggcca
5461 cagtcgatga atccagaaaa gcggcca
//
| {
"Vector backbone": "pMTS_mScarlet-i_N1",
"Backbone size w/o insert (bp)": "",
"Vector type": "Mammalian Expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Kanamycin, 50 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "TLNRD1 is a CCM complex component and regulates endothelial barrier integrity",
"Publication": "BioRxiv",
"DOI": "10.1101/2023.09.29.559344",
"Authors": "Neil J Ball, Sujan Ghimire, Gautier Follain, Ada O Pajari, Monika Vaitkevičiūtė, Alana R Cowell, Bence Berki, Johanna Ivaska, ProfileIlkka Paatero, Benjamin T Goult, Guillaume Jacquemet",
"PubMed Link": "",
"PubMed ID": ""
} | {
"Gene/Insert name": "CCM3",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "PDCD10 targeted to the mitonchondria",
"Entrez Gene": "PDCD10",
"Promoter": "",
"Tag / Fusion Protein": "mScarlet-I"
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "XhoI (unknown if destroyed)",
"3′ cloning site": "EcoRI (unknown if destroyed)",
"5′ sequencing primer": "CMV",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pSLQ2853-4 pHR: U6-Sasgv2CXCR4-2 CMV-EGFP | 84255 | Expresses optimized Sa sgCXCR4-2 gRNA with an EGFP fluorescent marker | Stanley Qi | Gao et al Nat Methods. 2016 Oct 24. doi: 10.1038/nmeth.4042. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/161110/f206241f-5ab0-4365-ac2a-c21f093f9a9d/addgene-plasmid-84255-sequence-161110.gbk | full | LOCUS sequence-161110- 7646 bp DNA circular SYN 26-AUG-2024
DEFINITION Expresses optimized Sa sgCXCR4-2 gRNA with an EGFP fluorescent
marker.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7646)
AUTHORS Gao Y, Xiong X, Wong S, Charles EJ, Lim WA, Qi LS
TITLE Complex transcriptional modulation with orthogonal and inducible
dCas9 regulators.
JOURNAL Nat Methods. 2016 Oct 24. doi: 10.1038/nmeth.4042.
PUBMED 27776111
REFERENCE 2 (bases 1 to 7646)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat
Methods. 2016 Oct 24. doi: 10.1038/nmeth.4042."
FEATURES Location/Qualifiers
source 1..7646
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(56..75)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 247..626
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 628..826
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
LTR 844..1024
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 1071..1196
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1695..1928
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 2113..2157
/codon_start=1
/product="antigenic peptide corresponding to amino acids
655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
et al., 2013)"
/label=gp41 peptide
/note="recognized by the 2H10 single-chain llama nanobody"
/translation="KNEQELLELDKWASL"
misc_feature 2455..2572
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
protein_bind complement(2825..2858)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
primer_bind 2862..2882
/label=mU6-F
/note="Mouse U6 promoter, forward primer"
misc_RNA 2966..3041
/label=Sa gRNA scaffold
/note="guide RNA scaffold for the Staphylococcus aureus
CRISPR/Cas9 system"
enhancer 3166..3469
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 3470..3673
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
CDS 3707..4426
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(3752..3773)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(4013..4032)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 4360..4381
/label=EGFP-C
/note="EGFP, forward primer"
protein_bind complement(4446..4479)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
misc_feature 4535..5123
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(4588..4608)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(5006..5017)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(5126..5142)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5127..5143)
/label=pBluescriptKS
/note="For pBluescript vector"
LTR 5652..5832
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
rep_origin complement(5894..6482)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5974..5993)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6653..7513)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 7276..7295
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7514..7618)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gcttaagcgg tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa
61 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
121 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
181 tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata
241 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
301 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
361 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
421 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
481 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
541 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
601 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
661 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
721 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
781 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagcgcg ttttgcctgt
841 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
901 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
961 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
1021 agcagtggcg cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac
1081 gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta
1141 cgccaaaaat tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta
1201 ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga
1261 aaaaatataa attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta
1321 atcctggcct gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat
1381 cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca accctctatt
1441 gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag
1501 agcaaaacaa aagtaagacc accgcacagc aagcggccgg ccgcgctgat cttcagacct
1561 ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa
1621 attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa
1681 agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg
1741 ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag
1801 cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc
1861 tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa
1921 cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg
1981 aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag
2041 tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa
2101 aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg
2161 aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga
2221 ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag
2281 ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc
2341 gaaggaatag aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac
2401 ggatcggcac tgcgtgcgcc aattctgcag acaaatggca gtattcatcc acaattttaa
2461 aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac
2521 agacatacaa actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttcgggttta
2581 ttacagggac agcagagatc cagtttggtt agtaccgggc ccgctctaga gatccgacgc
2641 gccatctcta ggcccgcgcc ggccccctcg cacggacttg tgggagaagc tcggctactc
2701 ccctgccccg gttaatttgc atataatatt tcctagtaac tatagaggct taatgtgcga
2761 taaaagacag ataatctgtt ctttttaata ctagctacat tttacatgat aggcttggat
2821 ttctataact tcgtatagca tacattatac gaagttataa acagcacaaa aggaaactca
2881 ccctaactgt aaagtaattg tgtgttttga gactataagt atcccttgga gaaccacctt
2941 gttggcccag gggaccctgc tgtttgttat agtactctgg aaacagaatc tactataaca
3001 aggcaaaatg ccgtgtttat ctcgtcaact tgttggcgag atttttttct cgagtactag
3061 gatccattag gcggccgcgt ggataaccgt attaccgcca tgcattagtt attaatagta
3121 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
3181 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
3241 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
3301 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
3361 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
3421 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
3481 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
3541 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
3601 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
3661 tataagcaga gctggtttag tgaaccgtca gatccgctag cccaccatgg tgagcaaggg
3721 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
3781 ccacaagttc agcgtgtctg gcgagggcga gggcgatgcc acctacggca agctgaccct
3841 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
3901 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
3961 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
4021 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
4081 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
4141 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggcgaa
4201 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
4261 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
4321 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
4381 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaagaat tcgtcgaggg
4441 acctaataac ttcgtatagc atacattata cgaagttata catgtttaag ggttccggtt
4501 ccactaggta caattcgata tcaagcttat cgataatcaa cctctggatt acaaaatttg
4561 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
4621 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
4681 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
4741 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
4801 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
4861 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
4921 gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg
4981 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
5041 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
5101 ctccctttgg gccgcctccc cgcatcgata ccgtcgacct cgatcgagac ctagaaaaac
5161 atggagcaat cacaagtagc aatacagcag ctaccaatgc tgattgtgcc tggctagaag
5221 cacaagagga ggaggaggtg ggttttccag tcacacctca ggtaccttta agaccaatga
5281 cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc
5341 taattcactc ccaacgaaga caagatatcc ttgatctgtg gatctaccac acacaaggct
5401 acttccctga ttggcagaac tacacaccag ggccagggat cagatatcca ctgacctttg
5461 gatggtgcta caagctagta ccagttgagc aagagaaggt agaagaagcc aatgaaggag
5521 agaacacccg cttgttacac cctgtgagcc tgcatgggat ggatgacccg gagagagaag
5581 tattagagtg gaggtttgac agccgcctag catttcatca catggcccga gagctgcatc
5641 cggactgtac tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
5701 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
5761 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
5821 aaatctctag cagcatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
5881 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
5941 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
6001 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
6061 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
6121 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
6181 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
6241 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
6301 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
6361 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
6421 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
6481 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
6541 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
6601 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
6661 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
6721 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
6781 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
6841 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
6901 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
6961 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
7021 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
7081 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
7141 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
7201 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
7261 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
7321 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
7381 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
7441 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
7501 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
7561 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
7621 catttccccg aaaagtgcca cctgac
//
| {
"Vector backbone": "pHR",
"Backbone size w/o insert (bp)": "",
"Vector type": "Mammalian Expression, Lentiviral, CRISPR",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "NEB Stable",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Complex transcriptional modulation with orthogonal and inducible dCas9 regulators",
"Publication": "Nat Methods. 2016 Oct 24. doi: 10.1038/nmeth.4042.",
"DOI": "10.1038/nmeth.4042.",
"Authors": "Gao Y, Xiong X, Wong S, Charles EJ, Lim WA, Qi LS /nmeth",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/27776111",
"PubMed ID": "PubMed 27776111"
} | {
"Gene/Insert name": "Sa sgCXCR4-2",
"Species": "H. sapiens (human), Synthetic; S. aureus",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "mouse U6",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "BstXI (not destroyed)",
"3′ cloning site": "XhoI (not destroyed)",
"5′ sequencing primer": "GAGATCCAGTTTGGTTAGTACCGGG",
"3′ sequencing primer": "ATGCATGGCGGTAATACGGTTAT"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pGEMT-Pax7bait-T2a-Cherry-pA | 111154 | knock in bait for Axolotl Pax7 locus | Elly Tanaka | Fei et al Proc Natl Acad Sci U S A. 2017 Nov 21;114(47):12501-12506. doi: 10.1073/pnas.1706855114. Epub 2017 Oct 31. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/211552/b8f03d6e-52b8-4e9d-9d54-369f0b6cc778/addgene-plasmid-111154-sequence-211552.gbk | full | LOCUS sequence-211552- 5575 bp DNA circular SYN 26-AUG-2024
DEFINITION knock in bait for Axolotl Pax7 locus.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5575)
AUTHORS Fei JF, Schuez M, Knapp D, Taniguchi Y, Drechsel DN, Tanaka EM
TITLE Efficient gene knockin in axolotl and its use to test the role of
satellite cells in limb regeneration.
JOURNAL Proc Natl Acad Sci U S A. 2017 Nov 21;114(47):12501-12506. doi:
10.1073/pnas.1706855114. Epub 2017 Oct 31.
PUBMED 29087939
REFERENCE 2 (bases 1 to 5575)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1073/pnas.1706855114"; journalName: "Proc Natl Acad Sci U S A";
date: "2017-11-21- 21"; volume: "114"; issue: "47"; pages:
"12501-12506"
FEATURES Location/Qualifiers
source 1..5575
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 17..39
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 31..48
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 32..48
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
CDS 1447..1500
/codon_start=1
/product="2A peptide from Thosea asigna virus capsid
protein"
/label=T2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="EGRGSLLTCGDVEENPGP"
CDS 1501..2211
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
primer_bind complement(1646..1664)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 1914..1933
/label=mCherry-F
/note="mCherry, forward primer"
primer_bind complement(2321..2340)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_signal 2386..2441
/label=beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(2440..2459)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
promoter complement(2771..2789)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(2772..2789)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(2807..2823)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2807..2823)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(2820..2842)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 2831..2847
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(2855..2885)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2900..2921
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(3038..3055)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3209..3797)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3289..3308)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3968..4828)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 4591..4610
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(4829..4933)
/gene="bla"
/label=AmpR promoter
rep_origin complement(5011..5466)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5148..5169)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 5360..5379
/label=F1ori-R
/note="F1 origin, reverse primer"
ORIGIN
1 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga attgtaatac
61 gactcactat agacgcgtat ggcttgtctc cccggagctg tccccagaat gatgcggcca
121 ggaccagggc agaactaccc acggaccggt ttcccactag aagtgtccac tcctctgggc
181 cagggcagag tcaaccaact ggggggagtg ttcattaacg ggcgcccgtt gcccaaccac
241 atccgccaca agatcgtgga gatggcacac cacggcattc ggccctgcgt gatttcacgg
301 cagctacgtg tgtcccacgg ctgcgtgtcc aagatcctgt gccggtacca ggagacgggg
361 tccatccggc caggagccat cggaggcagc aagcccaggc aggtggccac tcctgacgtg
421 gagaagaaga ttgaggaata taagagagag aaccccggga tgttcagctg ggagatcagg
481 gacaggctcc taaaggatgg acactgcgac aggagcacgg tgccctcagg taaccggctg
541 gacgaaggtt cagatgttga gtcagagcca gacttgccct tgaaacgcaa gcagcgacgc
601 agccgcacca ccttcactgc tgagcagctg gaggagttgg agaaggcctt tgagagaacg
661 cactacccag atatctacac ccgtgaagag ttggcgcaga gaaccaagct caccgaggcg
721 agagtacagg tgtggttcag caatcgtaga gcaaggtggc gtaaacaggc aggagccaat
781 caactggcag ctttcaatca tctcctgccc ggcggatttc cacccacagg aatgccaagt
841 ttgccaccct atcagcttcc agactccggc tatgcaagtg ccctagcaca agacggggga
901 agcacagtgc acagacctca gcccctgcca ccctcctcca tgcaccaggg tggattgtca
961 gctgctgatg ccagttcagc ctacggaccc cgacacagct tcaccagcta ttcggacagc
1021 ttcatgaacc caggcgggcc aactaaccac atgaatccgg tcagcaatgg gctgtctcca
1081 caggtgatga gcatcctcag caaccctagt ggagtgcagt cccagtccca ggatttctcc
1141 atctcccctc tgcatggtgg cttggacact tcgaactcca tctcggcaag ctgcagccag
1201 cggagcgact ccatcaagtc agtcgatagc ctcgccacct cgcagtccta ctgtcctccg
1261 acctacagca ccaccggcta cagcgtggat cctgtggcca gctaccaata tgggcaatac
1321 gggcaaactg ctgtagacta cttggctaag aacgtgagct tgtcgacaca gaggaggatg
1381 aagctcgggg accactcagc cgtgttgggt ttgcttcctg tggagacggg acaagcatac
1441 gcatgcgagg gcagaggaag tcttctaaca tgcggtgacg tggaggagaa tcccggccct
1501 atggtgagca agggcgagga ggataacatg gccatcatca aggagttcat gcgcttcaag
1561 gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc
1621 cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc
1681 ttcgcctggg acatcctgtc ccctcagttc atgtacggct ccaaggccta cgtgaagcac
1741 cccgccgaca tccccgacta cttgaagctg tccttccccg agggcttcaa gtgggagcgc
1801 gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac
1861 ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgta
1921 atgcagaaga agaccatggg ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc
1981 gccctgaagg gcgagatcaa gcagaggctg aagctgaagg acggcggcca ctacgacgct
2041 gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc
2101 aacatcaagt tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgaa
2161 cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaagta agaattctgc
2221 agtcgagtca tcgcgattaa ttcactcctc aggtgcaggc tgcctatcag aaggtggtgg
2281 ctggtgtggc caatgccctg gctcacaaat accactgaga tctttttccc tctgccaaaa
2341 attatgggga catcatgaag ccccttgagc atctgacttc tggctaataa aggaaattta
2401 ttttcattgc aatagtgtgt tggaattttt tgtgtctctc actcggaagg acatatggga
2461 gggcaaatca tttaaaacat cagaatgagt atttggttta gagtttggca acatatgccc
2521 atatgctggc tgccatgaac aaaggttggc tataaagagg tcatcagtat atgaaacagc
2581 cccctgctgt ccattcctta ttccatagaa aagccttgac ttgaggttag atttttttta
2641 tattttgttt tgtgttattt ttttctttaa catccctaaa attttcctta catgttttac
2701 tagccagatt tttcctcctc tcctgactac tcccagtcat agctgtccct cttctcttat
2761 gaagatctat tctatagtgt cacctaaata gcttggcgta atcatggtca tagctgtttc
2821 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
2881 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
2941 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
3001 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
3061 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
3121 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
3181 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
3241 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
3301 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
3361 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
3421 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
3481 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
3541 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
3601 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
3661 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
3721 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
3781 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
3841 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
3901 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
3961 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
4021 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
4081 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
4141 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
4201 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
4261 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
4321 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
4381 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
4441 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
4501 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
4561 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
4621 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
4681 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
4741 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
4801 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
4861 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
4921 taggggttcc gcgcacattt ccccgaaaag tgccacctga tgcggtgtga aataccgcac
4981 agatgcgtaa ggagaaaata ccgcatcagg aaattgtaag cgttaatatt ttgttaaaat
5041 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
5101 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
5161 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
5221 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
5281 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
5341 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
5401 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
5461 gcgcgtccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
5521 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgg
//
| {
"Vector backbone": "pGEMT",
"Backbone size w/o insert (bp)": "",
"Vector type": "Mammalian Expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA ; Takara Bio Limited Use Label License (formerly Clontech)",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Efficient gene knockin in axolotl and its use to test the role of satellite cells in limb regeneration",
"Publication": "Proc Natl Acad Sci U S A. 2017 Nov 21;114(47):12501-12506. doi: 10.1073/pnas.1706855114. Epub 2017 Oct 31.",
"DOI": "10.1073/pnas.1706855114.",
"Authors": "Fei JF, Schuez M, Knapp D, Taniguchi Y, Drechsel DN, Tanaka EM /pnas",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/29087939",
"PubMed ID": "PubMed 29087939"
} | {
"Gene/Insert name": "Pax7bait-T2a-Cherry-pA",
"Species": "Ambystoma mexicanum",
"Insert Size (bp)": "1344",
"Mutation": "wt",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "Unknown (unknown if destroyed)",
"3′ cloning site": "Unknown (unknown if destroyed)",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
LCK gRNA (BRDN0001145503) | 76412 | 3rd generation lentiviral gRNA plasmid targeting human LCK | Doench et al Nat Biotechnol. 2016 Jan 18. doi: 10.1038/nbt.3437. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/185788/dbd07d90-fcb2-4174-8f3a-2a4cf9c39121/addgene-plasmid-76412-sequence-185788.gbk | full | LOCUS sequence-185788- 8323 bp DNA circular SYN 26-AUG-2024
DEFINITION 3rd generation lentiviral gRNA plasmid targeting human LCK.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8323)
AUTHORS Doench JG, Fusi N, Sullender M, Hegde M, Vaimberg EW, Donovan KF,
Smith I, Tothova Z, Wilen C, Orchard R, Virgin HW, Listgarten J,
Root DE
TITLE Optimized sgRNA design to maximize activity and minimize off-target
effects of CRISPR-Cas9.
JOURNAL Nat Biotechnol. 2016 Jan 18. doi: 10.1038/nbt.3437.
PUBMED 26780180
REFERENCE 2 (bases 1 to 8323)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat
Biotechnol. 2016 Jan 18. doi: 10.1038/nbt.3437."
FEATURES Location/Qualifiers
source 1..8323
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 87..1265
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 318..1256
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 1213..1233
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
CDS 1278..1877
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind complement(1278..1297)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 1774..1794
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
misc_feature 1893..2481
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(1946..1966)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(2364..2375)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 2553..2786
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 2864..2985
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2901..2920)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2955..2974
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 3025..3160
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(3180..3199)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(3181..3199)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(3209..3226)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(3209..3225)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3218..3240)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 3367..3822
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3454..3473)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3664..3685
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 3848..3952
/gene="bla"
/label=AmpR promoter
CDS 3953..4813
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(4171..4190)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 4984..5572
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5473..5492
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 5726..5743
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 5860..5881
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 5896..5926
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5934..5950
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
primer_bind 5939..5961
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 5958..5974
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 5958..5974
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 5993..6013
/label=T3
/note="T3 promoter, forward primer"
promoter 5995..6013
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 6041..6267
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 6268..6448
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 6495..6620
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 7113..7346
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 7531..7575
/codon_start=1
/product="antigenic peptide corresponding to amino acids
655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
et al., 2013)"
/label=gp41 peptide
/note="recognized by the 2H10 single-chain llama nanobody"
/translation="KNEQELLELDKWASL"
promoter 7753..8001
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA
(Domitrovich & Kunkel, 2003)"
primer_bind 7753..7773
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 7924..7943
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
misc_RNA 8023..8098
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
misc_feature 8154..8271
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
ORIGIN
1 cccgggtgca aagatggata aagttttaaa cagagaggaa tctttgcagc taatggacct
61 tctaggtctt gaaaggagtg ggaattggct ccggtgcccg tcagtgggca gagcgcacat
121 cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgatccggt gcctagagaa
181 ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg
241 gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt
301 ttgccgccag aacacaggta agtgccgtgt gtggttcccg cgggcctggc ctctttacgg
361 gttatggccc ttgcgtgcct tgaattactt ccacctggct gcagtacgtg attcttgatc
421 ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa ggagcccctt
481 cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt
541 ggcaccttcg cgcctgtctc gctgctttcg ataagtctct agccatttaa aatttttgat
601 gacctgctgc gacgcttttt ttctggcaag atagtcttgt aaatgcgggc caagatctgc
661 acactggtat ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca
721 catgttcggc gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc
781 aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg
841 cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga aagatggccg cttcccggcc
901 ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac
961 ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac tccacggagt
1021 accgggcgcc gtccaggcac ctcgattagt tctcgagctt ttggagtacg tcgtctttag
1081 gttgggggga ggggttttat gcgatggagt ttccccacac tgagtgggtg gagactgaag
1141 ttaggccagc ttggcacttg atgtaattct ccttggaatt tgcccttttt gagtttggat
1201 cttggttcat tctcaagcct cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt
1261 cgtgacgtac ggccaccatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg
1321 acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc
1381 acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca
1441 cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gccgtggcgg
1501 tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca
1561 tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc
1621 cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggagtctcg cccgaccacc
1681 agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg
1741 gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg
1801 gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc
1861 gcaagcccgg tgcctgaacg cgttaagtcg acaatcaacc tctggattac aaaatttgtg
1921 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
1981 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
2041 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
2101 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
2161 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
2221 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
2281 cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg
2341 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
2401 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
2461 ccctttgggc cgcctccccg cgtcgacttt aagaccaatg acttacaagg cagctgtaga
2521 tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag
2581 acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga
2641 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
2701 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
2761 ttagtcagtg tggaaaatct ctagcagtac gtatagtagt tcatgtcatc ttattattca
2821 gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc
2881 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
2941 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta
3001 gctatcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
3061 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
3121 gagctattcc agaagtagtg aggaggcttt tttggaggcc tagggacgta cccaattcgc
3181 cctatagtga gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg
3241 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
3301 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
3361 aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
3421 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
3481 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
3541 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
3601 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
3661 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
3721 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
3781 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg
3841 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
3901 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
3961 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
4021 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
4081 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
4141 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
4201 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
4261 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
4321 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
4381 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
4441 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
4501 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
4561 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
4621 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
4681 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
4741 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
4801 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
4861 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
4921 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
4981 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
5041 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
5101 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
5161 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
5221 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
5281 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
5341 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
5401 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
5461 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
5521 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
5581 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
5641 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
5701 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
5761 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
5821 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
5881 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
5941 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa
6001 ccctcactaa agggaacaaa agctggagct gcaagcttaa tgtagtctta tgcaatactc
6061 ttgtagtctt gcaacatggt aacgatgagt tagcaacatg ccttacaagg agagaaaaag
6121 caccgtgcat gccgattggt ggaagtaagg tggtacgatc gtgccttatt aggaaggcaa
6181 cagacgggtc tgacatggat tggacgaacc actgaattgc cgcattgcag agatattgta
6241 tttaagtgcc tagctcgata cataaacggg tctctctggt tagaccagat ctgagcctgg
6301 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
6361 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
6421 ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag
6481 ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag
6541 gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga
6601 gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa
6661 ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca
6721 gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac
6781 aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat
6841 ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg
6901 aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg
6961 ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag tgaattatat
7021 aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc aaagagaaga
7081 gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg gttcttggga
7141 gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc cagacaatta
7201 ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc gcaacagcat
7261 ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct ggctgtggaa
7321 agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa actcatttgc
7381 accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca gatttggaat
7441 cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt aatacactcc
7501 ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt ggaattagat
7561 aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta tataaaatta
7621 ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt actttctata
7681 gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct cccaaccccg
7741 aggggaccca gagagggcct atttcccatg attccttcat atttgcatat acgatacaag
7801 gctgttagag agataattag aattaatttg actgtaaaca caaagatatt agtacaaaat
7861 acgtgacgta gaaagtaata atttcttggg tagtttgcag ttttaaaatt atgttttaaa
7921 atggactatc atatgcttac cgtaacttga aagtatttcg atttcttggc tttatatatc
7981 ttgtggaaag gacgaaacac cggctggttc ggctctacgc tggttttaga gctagaaata
8041 gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt
8101 ttttaagctt ggcgtaacta gatcttgaga caaatggcag tattcatcca caattttaaa
8161 agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca
8221 gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat
8281 tacagggaca gcagagatcc actttggcgc cggctcgagg ggg
//
| {
"Vector backbone": "lentiGuide-Puro",
"Backbone size w/o insert (bp)": "",
"Vector type": "Mammalian Expression, Lentiviral, CRISPR",
"Selectable markers": "Puromycin"
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Optimized sgRNA design to maximize activity and minimize off-target effects of CRISPR-Cas9",
"Publication": "Nat Biotechnol. 2016 Jan 18. doi: 10.1038/nbt.3437.",
"DOI": "10.1038/nbt.3437.",
"Authors": "Doench JG, Fusi N, Sullender M, Hegde M, Vaimberg EW, Donovan KF, Smith I, Tothova Z, Wilen C, Orchard R, Virgin HW, Listgarten J, Root DE /nbt",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/26780180",
"PubMed ID": "PubMed 26780180"
} | {
"Gene/Insert name": "LCK",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "LCK",
"Promoter": "hU6",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Unknown",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "U6-F",
"3′ sequencing primer": "pS2-R"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
||
TFAP4_BHLH_3-2 | 185055 | Deletion of BHLH domain of mouse TFAP4 in combination with TFAP4_BHLH_5-1 or TFAP4_BHLH_5-2 | Constance Ciaudo | Schaefer et al EMBO Rep. 2022 Jul 28:e54762. doi: 10.15252/embr.202254762. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/363843/1f4b0de1-c67d-4315-a6fc-396c4fea43a3/addgene-plasmid-185055-sequence-363843.gbk | full | LOCUS sequence-363843- 11688 bp DNA circular SYN 27-AUG-2024
DEFINITION Deletion of BHLH domain of mouse TFAP4 in combination with
TFAP4_BHLH_5-1 or TFAP4_BHLH_5-2.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11688)
AUTHORS Schaefer M, Nabih A, Spies D, Hermes V, Bodak M, Wischnewski H,
Stalder P, Ngondo RP, Liechti LA, Sajic T, Aebersold R, Gatfield D,
Ciaudo C
TITLE Global and precise identification of functional miRNA targets in
mESCs by integrative analysis.
JOURNAL EMBO Rep. 2022 Jul 28:e54762. doi: 10.15252/embr.202254762.
PUBMED 35899551
REFERENCE 2 (bases 1 to 11688)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 27, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "EMBO Rep.
2022 Jul 28:e54762. doi: 10.15252/embr.202254762."
FEATURES Location/Qualifiers
source 1..11688
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 118..137
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 371..388
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 505..526
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 541..571
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 579..595
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
primer_bind 584..606
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 603..619
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 603..619
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 638..658
/label=T3
/note="T3 promoter, forward primer"
promoter 640..658
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 686..912
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 913..1093
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 1140..1265
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1758..1991
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 2176..2220
/codon_start=1
/product="antigenic peptide corresponding to amino acids
655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
et al., 2013)"
/label=gp41 peptide
/note="recognized by the 2H10 single-chain llama nanobody"
/translation="KNEQELLELDKWASL"
promoter 2398..2646
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA
(Domitrovich & Kunkel, 2003)"
primer_bind 2398..2418
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 2569..2588
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
misc_RNA 2668..2743
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
misc_feature 2791..2908
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2977..3188
/label=EF-1-alpha core promoter
/note="core promoter for human elongation factor
EF-1-alpha"
regulatory 3208..3217
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 3217..3240
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS 3247..3267
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 3292..7392
/codon_start=1
/product="Cas9 (Csn1) endonuclease from the Streptococcus
pyogenes Type II CRISPR/Cas system"
/label=Cas9
/note="generates RNA-guided double strand breaks in DNA"
/translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
ATLIHQSITGLYETRIDLSQLGGD"
CDS 3292..7392
/codon_start=1
/product="Cas9 (Csn1) endonuclease from the Streptococcus
pyogenes Type II CRISPR/Cas system"
/label=Cas9
/note="generates RNA-guided double strand breaks in DNA"
/translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
ATLIHQSITGLYETRIDLSQLGGD"
CDS 7393..7440
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
CDS 7447..7503
/codon_start=1
/product="2A peptide from porcine teschovirus-1
polyprotein"
/label=P2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="ATNFSLLKQAGDVEENPGP"
CDS 7504..8211
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
GRHSTGGMDELYK"
primer_bind complement(7646..7664)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 7914..7933
/label=mCherry-F
/note="mCherry, forward primer"
misc_feature 8226..8814
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(8279..8299)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(8697..8708)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 8886..9119
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 9197..9318
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(9234..9253)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 9288..9307
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 9358..9493
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(9513..9532)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(9514..9532)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(9542..9559)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(9542..9558)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(9551..9573)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 9700..10155
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(9787..9806)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 9997..10018
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 10181..10285
/gene="bla"
/label=AmpR promoter
CDS 10286..11146
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(10504..10523)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin join(11317..11688,1..217)
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
ORIGIN
1 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
61 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
121 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
181 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
241 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
301 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
361 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
421 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
481 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
541 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
601 cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc actaaaggga
661 acaaaagctg gagctgcaag cttaatgtag tcttatgcaa tactcttgta gtcttgcaac
721 atggtaacga tgagttagca acatgcctta caaggagaga aaaagcaccg tgcatgccga
781 ttggtggaag taaggtggta cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca
841 tggattggac gaaccactga attgccgcat tgcagagata ttgtatttaa gtgcctagct
901 cgatacataa acgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
961 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
1021 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
1081 aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc
1141 tctctcgacg caggactcgg cttgctgaag cgcgcacggc aagaggcgag gggcggcgac
1201 tggtgagtac gccaaaaatt ttgactagcg gaggctagaa ggagagagat gggtgcgaga
1261 gcgtcagtat taagcggggg agaattagat cgcgatggga aaaaattcgg ttaaggccag
1321 ggggaaagaa aaaatataaa ttaaaacata tagtatgggc aagcagggag ctagaacgat
1381 tcgcagttaa tcctggcctg ttagaaacat cagaaggctg tagacaaata ctgggacagc
1441 tacaaccatc ccttcagaca ggatcagaag aacttagatc attatataat acagtagcaa
1501 ccctctattg tgtgcatcaa aggatagaga taaaagacac caaggaagct ttagacaaga
1561 tagaggaaga gcaaaacaaa agtaagacca ccgcacagca agcggccgct gatcttcaga
1621 cctggaggag gagatatgag ggacaattgg agaagtgaat tatataaata taaagtagta
1681 aaaattgaac cattaggagt agcacccacc aaggcaaaga gaagagtggt gcagagagaa
1741 aaaagagcag tgggaatagg agctttgttc cttgggttct tgggagcagc aggaagcact
1801 atgggcgcag cgtcaatgac gctgacggta caggccagac aattattgtc tggtatagtg
1861 cagcagcaga acaatttgct gagggctatt gaggcgcaac agcatctgtt gcaactcaca
1921 gtctggggca tcaagcagct ccaggcaaga atcctggctg tggaaagata cctaaaggat
1981 caacagctcc tggggatttg gggttgctct ggaaaactca tttgcaccac tgctgtgcct
2041 tggaatgcta gttggagtaa taaatctctg gaacagattt ggaatcacac gacctggatg
2101 gagtgggaca gagaaattaa caattacaca agcttaatac actccttaat tgaagaatcg
2161 caaaaccagc aagaaaagaa tgaacaagaa ttattggaat tagataaatg ggcaagtttg
2221 tggaattggt ttaacataac aaattggctg tggtatataa aattattcat aatgatagta
2281 ggaggcttgg taggtttaag aatagttttt gctgtacttt ctatagtgaa tagagttagg
2341 cagggatatt caccattatc gtttcagacc cacctcccaa ccccgagggg acccagagag
2401 ggcctatttc ccatgattcc ttcatatttg catatacgat acaaggctgt tagagagata
2461 attagaatta atttgactgt aaacacaaag atattagtac aaaatacgtg acgtagaaag
2521 taataatttc ttgggtagtt tgcagtttta aaattatgtt ttaaaatgga ctatcatatg
2581 cttaccgtaa cttgaaagta tttcgatttc ttggctttat atatcttgtg gaaaggacga
2641 aacaccgaac caaacgccct gcaggcggtt ttagagctag aaatagcaag ttaaaataag
2701 gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttg aattctagat
2761 cttgagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga ttggggggta
2821 cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca
2881 aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca gagatccact
2941 ttggcgccgg ctcgagtggc tccggtgccc gtcagtgggc agagcgcaca tcgcccacag
3001 tccccgagaa gttgggggga ggggtcggca attgaaccgg tgcctagaga aggtggcgcg
3061 gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag
3121 aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca
3181 gaacacaggt gtcgtgacgc gggatccgcc accatggatt acaaagacga tgacgataag
3241 atggccccaa agaagaagcg gaaggtcggt atccacggag tcccagcagc cgacaagaag
3301 tacagcatcg gcctggacat cggcaccaac tctgtgggct gggccgtgat caccgacgag
3361 tacaaggtgc ccagcaagaa attcaaggtg ctgggcaaca ccgaccggca cagcatcaag
3421 aagaacctga tcggagccct gctgttcgac agcggcgaaa cagccgaggc cacccggctg
3481 aagagaaccg ccagaagaag atacaccaga cggaagaacc ggatctgcta tctgcaagag
3541 atcttcagca acgagatggc caaggtggac gacagcttct tccacagact ggaagagtcc
3601 ttcctggtgg aagaggataa gaagcacgag cggcacccca tcttcggcaa catcgtggac
3661 gaggtggcct accacgagaa gtaccccacc atctaccacc tgagaaagaa actggtggac
3721 agcaccgaca aggccgacct gcggctgatc tatctggccc tggcccacat gatcaagttc
3781 cggggccact tcctgatcga gggcgacctg aaccccgaca acagcgacgt ggacaagctg
3841 ttcatccagc tggtgcagac ctacaaccag ctgttcgagg aaaaccccat caacgccagc
3901 ggcgtggacg ccaaggccat cctgtctgcc agactgagca agagcagacg gctggaaaat
3961 ctgatcgccc agctgcccgg cgagaagaag aatggcctgt tcggaaacct gattgccctg
4021 agcctgggcc tgacccccaa cttcaagagc aacttcgacc tggccgagga tgccaaactg
4081 cagctgagca aggacaccta cgacgacgac ctggacaacc tgctggccca gatcggcgac
4141 cagtacgccg acctgtttct ggccgccaag aacctgtccg acgccatcct gctgagcgac
4201 atcctgagag tgaacaccga gatcaccaag gcccccctga gcgcctctat gatcaagaga
4261 tacgacgagc accaccagga cctgaccctg ctgaaagctc tcgtgcggca gcagctgcct
4321 gagaagtaca aagagatttt cttcgaccag agcaagaacg gctacgccgg ctacattgac
4381 ggcggagcca gccaggaaga gttctacaag ttcatcaagc ccatcctgga aaagatggac
4441 ggcaccgagg aactgctcgt gaagctgaac agagaggacc tgctgcggaa gcagcggacc
4501 ttcgacaacg gcagcatccc ccaccagatc cacctgggag agctgcacgc cattctgcgg
4561 cggcaggaag atttttaccc attcctgaag gacaaccggg aaaagatcga gaagatcctg
4621 accttccgca tcccctacta cgtgggccct ctggccaggg gaaacagcag attcgcctgg
4681 atgaccagaa agagcgagga aaccatcacc ccctggaact tcgaggaagt ggtggacaag
4741 ggcgcttccg cccagagctt catcgagcgg atgaccaact tcgataagaa cctgcccaac
4801 gagaaggtgc tgcccaagca cagcctgctg tacgagtact tcaccgtgta taacgagctg
4861 accaaagtga aatacgtgac cgagggaatg agaaagcccg ccttcctgag cggcgagcag
4921 aaaaaggcca tcgtggacct gctgttcaag accaaccgga aagtgaccgt gaagcagctg
4981 aaagaggact acttcaagaa aatcgagtgc ttcgactccg tggaaatctc cggcgtggaa
5041 gatcggttca acgcctccct gggcacatac cacgatctgc tgaaaattat caaggacaag
5101 gacttcctgg acaatgagga aaacgaggac attctggaag atatcgtgct gaccctgaca
5161 ctgtttgagg acagagagat gatcgaggaa cggctgaaaa cctatgccca cctgttcgac
5221 gacaaagtga tgaagcagct gaagcggcgg agatacaccg gctggggcag gctgagccgg
5281 aagctgatca acggcatccg ggacaagcag tccggcaaga caatcctgga tttcctgaag
5341 tccgacggct tcgccaacag aaacttcatg cagctgatcc acgacgacag cctgaccttt
5401 aaagaggaca tccagaaagc ccaggtgtcc ggccagggcg atagcctgca cgagcacatt
5461 gccaatctgg ccggcagccc cgccattaag aagggcatcc tgcagacagt gaaggtggtg
5521 gacgagctcg tgaaagtgat gggccggcac aagcccgaga acatcgtgat cgaaatggcc
5581 agagagaacc agaccaccca gaagggacag aagaacagcc gcgagagaat gaagcggatc
5641 gaagagggca tcaaagagct gggcagccag atcctgaaag aacaccccgt ggaaaacacc
5701 cagctgcaga acgagaagct gtacctgtac tacctgcaga atgggcggga tatgtacgtg
5761 gaccaggaac tggacatcaa ccggctgtcc gactacgatg tggaccatat cgtgcctcag
5821 agctttctga aggacgactc catcgacaac aaggtgctga ccagaagcga caagaaccgg
5881 ggcaagagcg acaacgtgcc ctccgaagag gtcgtgaaga agatgaagaa ctactggcgg
5941 cagctgctga acgccaagct gattacccag agaaagttcg acaatctgac caaggccgag
6001 agaggcggcc tgagcgaact ggataaggcc ggcttcatca agagacagct ggtggaaacc
6061 cggcagatca caaagcacgt ggcacagatc ctggactccc ggatgaacac taagtacgac
6121 gagaatgaca agctgatccg ggaagtgaaa gtgatcaccc tgaagtccaa gctggtgtcc
6181 gatttccgga aggatttcca gttttacaaa gtgcgcgaga tcaacaacta ccaccacgcc
6241 cacgacgcct acctgaacgc cgtcgtggga accgccctga tcaaaaagta ccctaagctg
6301 gaaagcgagt tcgtgtacgg cgactacaag gtgtacgacg tgcggaagat gatcgccaag
6361 agcgagcagg aaatcggcaa ggctaccgcc aagtacttct tctacagcaa catcatgaac
6421 tttttcaaga ccgagattac cctggccaac ggcgagatcc ggaagcggcc tctgatcgag
6481 acaaacggcg aaaccgggga gatcgtgtgg gataagggcc gggattttgc caccgtgcgg
6541 aaagtgctga gcatgcccca agtgaatatc gtgaaaaaga ccgaggtgca gacaggcggc
6601 ttcagcaaag agtctatcct gcccaagagg aacagcgata agctgatcgc cagaaagaag
6661 gactgggacc ctaagaagta cggcggcttc gacagcccca ccgtggccta ttctgtgctg
6721 gtggtggcca aagtggaaaa gggcaagtcc aagaaactga agagtgtgaa agagctgctg
6781 gggatcacca tcatggaaag aagcagcttc gagaagaatc ccatcgactt tctggaagcc
6841 aagggctaca aagaagtgaa aaaggacctg atcatcaagc tgcctaagta ctccctgttc
6901 gagctggaaa acggccggaa gagaatgctg gcctctgccg gcgaactgca gaagggaaac
6961 gaactggccc tgccctccaa atatgtgaac ttcctgtacc tggccagcca ctatgagaag
7021 ctgaagggct cccccgagga taatgagcag aaacagctgt ttgtggaaca gcacaagcac
7081 tacctggacg agatcatcga gcagatcagc gagttctcca agagagtgat cctggccgac
7141 gctaatctgg acaaagtgct gtccgcctac aacaagcacc gggataagcc catcagagag
7201 caggccgaga atatcatcca cctgtttacc ctgaccaatc tgggagcccc tgccgccttc
7261 aagtactttg acaccaccat cgaccggaag aggtacacca gcaccaaaga ggtgctggac
7321 gccaccctga tccaccagag catcaccggc ctgtacgaga cacggatcga cctgtctcag
7381 ctgggaggcg acaagcgtcc tgctgctact aagaaagctg gtcaagctaa gaaaaagaaa
7441 gctagcgcca cgaacttctc tctgttaaag caagcaggcg acgtggaaga aaaccccggt
7501 cccgtgagca agggcgagga ggataacatg gccatcatca aggagttcat gcgcttcaag
7561 gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc
7621 cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc
7681 ttcgcctggg acatcctgtc ccctcagttc atgtacggct ccaaggccta cgtgaagcac
7741 cccgccgaca tccccgacta cttgaagctg tccttccccg agggcttcaa gtgggagcgc
7801 gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac
7861 ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgta
7921 atgcagaaga agaccatggg ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc
7981 gccctgaagg gcgagatcaa gcagaggctg aagctgaagg acggcggcca ctacgacgct
8041 gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc
8101 aacatcaagt tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgaa
8161 cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaagtg acgcgttaag
8221 tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
8281 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
8341 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
8401 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
8461 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
8521 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
8581 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
8641 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
8701 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
8761 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcgtcgac
8821 tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg
8881 gggactggaa gggctaattc actcccaacg aagacaagat ctgctttttg cttgtactgg
8941 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
9001 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
9061 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
9121 tacgtatagt agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat
9181 atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag
9241 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
9301 actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccatc
9361 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
9421 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
9481 ttttttggag gcctagggac gtacccaatt cgccctatag tgagtcgtat tacgcgcgct
9541 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
9601 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
9661 gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat
9721 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
9781 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
9841 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
9901 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
9961 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
10021 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
10081 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
10141 taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
10201 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
10261 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
10321 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
10381 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
10441 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
10501 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
10561 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
10621 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
10681 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
10741 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
10801 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
10861 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
10921 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
10981 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
11041 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
11101 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
11161 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
11221 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
11281 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
11341 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
11401 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
11461 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
11521 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
11581 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
11641 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaact
//
| {
"Vector backbone": "pLentiCRISPR-mCherry",
"Backbone size w/o insert (bp)": "",
"Vector type": "Lentiviral",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "NEB Stable",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA ; Takara Bio Limited Use Label License (formerly Clontech)",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Global and precise identification of functional miRNA targets in mESCs by integrative analysis",
"Publication": "EMBO Rep. 2022 Jul 28:e54762. doi: 10.15252/embr.202254762.",
"DOI": "10.15252/embr.202254762.",
"Authors": "Schaefer M, Nabih A, Spies D, Hermes V, Bodak M, Wischnewski H, Stalder P, Ngondo RP, Liechti LA, Sajic T, Aebersold R, Gatfield D, Ciaudo C /embr",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/35899551",
"PubMed ID": "PubMed 35899551"
} | {
"Gene/Insert name": "TFAP4_3_2_gRNA",
"Species": "M. musculus (mouse)",
"Insert Size (bp)": "20",
"Mutation": "",
"Entrez Gene": "Tcfap4",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Ligation Independent Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "GACTATCATATGCTTACCGT",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
mPlum-EB3-7 | 55964 | Localization: MT End Binding Protein, Excitation: 590, Emission: 649 | Michael Davidson | Davidson Red fluorescent proteins (unpublished) | https://media.addgene.org/snapgene-media/v2.0.0/sequences/131799/476cb93b-4bb1-4c24-a285-46559ae3eadd/addgene-plasmid-55964-sequence-131799.gbk | partial | LOCUS sequence-131799- 759 bp DNA linear UNA 26-AUG-2024
DEFINITION Localization: MT End Binding Protein, Excitation: 590, Emission:
649.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE natural DNA sequence
ORGANISM unspecified
REFERENCE 1 (bases 1 to 759)
TITLE Davidson Red fluorescent proteins
REFERENCE 2 (bases 1 to 759)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..759
/mol_type="genomic DNA"
/organism="unspecified"
regulatory 68..77
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
ORIGIN
1 accgtcagat ccgctagcgc taccggactc agatctcgag ctcaagcttc gaattctgca
61 gtcgactgcc accatggccg tcaatgtgta ctccacatct gtgaccagtg aaaatctgag
121 tcgccatgat atgcttgcat gggtcaacga ctccctgcac ctcaactata ccaagataga
181 acagctttgt tcaggggcag cctactgcca gttcatggac atgctcttcc ccggctgtgt
241 gcacttgagg aaagtgaagt tccaggccaa actagagcat gaatacatcc acaacttcaa
301 ggtgctgcaa gcagctttca agaagatggg tgttgacaaa atcattcctg tagagaaatt
361 agtgaaagga aaattccaag ataattttga gtttattcag tggtttaaga aattctttga
421 cgcaaactat gatggaaagg attacaaccc tctgctggcg cggcagggcc aggacgtagc
481 gccacctcct aacccaggtg atcagatctt caacaaatcc aagaaactca ttggcacagc
541 agttccacag aggacgtccc ccacaggccc aaaaaacatg cagacctctg gccggctgag
601 caatgtggcc cccccctgca ttctccggaa gaatcctcca tcagcccgaa atggcggcca
661 tgagactgat gcccaaattc ttgaactcaa ccaacagctg gtggacttga agctgacagt
721 ggatgggctg gagaaggaac gtgacttcta cttcagcaa
//
| {
"Vector backbone": "mPlum",
"Backbone size w/o insert (bp)": "4750",
"Vector type": "Mammalian Expression",
"Selectable markers": "Neomycin (select with G418)"
} | {
"Bacterial Resistance(s)": "Kanamycin, 50 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA ; Takara Bio Limited Use Label License (formerly Clontech)",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "",
"Publication": "",
"DOI": "",
"Authors": "",
"PubMed Link": "",
"PubMed ID": ""
} | {
"Gene/Insert name": "EB3",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "MAPRE3",
"Promoter": "CMV",
"Tag / Fusion Protein": "mPlum"
} | {
"Cloning method": "Unknown",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "CMV-F",
"3′ sequencing primer": "SV40pA-R"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
PCDH-METTL14-R298P | 141113 | Overexpression of METTL14 mutant in mammalian cells. | Jianjun Chen | Weng et al Cell Stem Cell. 2018 Feb 1;22(2):191-205.e9. doi: 10.1016/j.stem.2017.11.016. Epub 2017 Dec 28. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/302062/f9fa12d0-1fc6-4750-8e4e-8a32c9f6f4b9/addgene-plasmid-141113-sequence-302062.gbk | full | LOCUS sequence-302062- 9590 bp DNA circular SYN 25-AUG-2024
DEFINITION Overexpression of METTL14 mutant in mammalian cells..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9590)
AUTHORS Weng H, Huang H, Wu H, Qin X, Zhao BS, Dong L, Shi H, Skibbe J, Shen
C, Hu C, Sheng Y, Wang Y, Wunderlich M, Zhang B, Dore LC, Su R, Deng
X, Ferchen K, Li C, Sun M, Lu Z, Jiang X, Marcucci G, Mulloy JC,
Yang J, Qian Z, Wei M, He C, Chen J
TITLE METTL14 Inhibits Hematopoietic Stem/Progenitor Differentiation and
Promotes Leukemogenesis via mRNA m(6)A Modification.
JOURNAL Cell Stem Cell. 2018 Feb 1;22(2):191-205.e9. doi:
10.1016/j.stem.2017.11.016. Epub 2017 Dec 28.
PUBMED 29290617
REFERENCE 2 (bases 1 to 9590)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1016/j.stem.2017.11.016"; journalName: "Cell Stem Cell"; date:
"2018-02-1- 1"; volume: "22"; issue: "2"; pages: "191-205.e9"
FEATURES Location/Qualifiers
source 1..9590
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 52..168
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 258..461
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 412..432
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 458..482
/label=LNCX
/note="Human CMV promoter, forward primer"
CDS 541..567
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
promoter 1977..2188
/label=EF-1-alpha core promoter
/note="core promoter for human elongation factor
EF-1-alpha"
LTR 2201..2469
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from human
T-cell leukemia virus (HTLV) type 1"
regulatory 2498..2507
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 2528..3190
/codon_start=1
/product="green fluorescent protein 2 from Pontellina
plumata, also known as ppluGFP2 (Shagin et al., 2004)"
/label=CopGFP
/translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT
FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY
EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG
YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA"
CDS 3260..3313
/codon_start=1
/product="2A peptide from Thosea asigna virus capsid
protein"
/label=T2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="EGRGSLLTCGDVEENPGP"
CDS 3314..3913
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind complement(3314..3333)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 3810..3830
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
misc_feature 3914..4502
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(3967..3987)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(4385..4396)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 4576..4809
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 4881..5002
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4918..4937)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4972..4991
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 5042..5177
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(5210..5226)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5210..5226)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(5223..5245)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 5234..5250
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(5258..5288)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5303..5324
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(5441..5458)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5612..6200)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6371..7231)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 6994..7013
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7232..7336)
/gene="bla"
/label=AmpR promoter
primer_bind 7404..7422
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(7460..7482)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 7582..7601
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 7795..7817
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 7809..7826
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 7810..7826
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 7841..8067
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 8068..8248
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 8295..8420
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 8913..9146
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 9331..9375
/codon_start=1
/product="antigenic peptide corresponding to amino acids
655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
et al., 2013)"
/label=gp41 peptide
/note="recognized by the 2H10 single-chain llama nanobody"
/translation="KNEQELLELDKWASL"
ORIGIN
1 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcggttaa cttttaaaag
61 aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga
121 catacaaact aaagaattac aaaaacaaat tacaaaattc aaaattttat cgatactagt
181 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
241 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
301 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
361 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
421 gcggtaggcg tgtacggtgg gaggtttata taagcagagc tcgtttagtg aaccgtcaga
481 tcgcctggag acgccatcca cgctgttttg acctccatag aagattctag aaccaccatg
541 tacccatacg atgttccaga ttacgctgat agccgcttgc aggagatccg ggagcggcag
601 aagttacggc gacagctcct cgcgcagcag ttgggagctg aaagtgccga cagcattggt
661 gccgtgttaa atagcaaaga tgagcagaga gaaattgctg aaacaagaga aacttgcagg
721 gcttcctatg atacctctgc tccaaatgca aaacgtaagt atctggatga aggagagaca
781 gatgaagaca aaatggaaga atataaggat gaactagaaa tgcaacagga tgaagaaaat
841 ttgccatatg aagaagagat ttacaaagat tctagtactt ttcttaaggg aacacagagc
901 ttaaatcccc ataatgatta ctgccaacat tttgtagaca ctggacatag acctcagaat
961 ttcatcaggg atgtaggttt agctgacaga tttgaagaat atcctaaact gagggagctc
1021 atcaggctaa aggatgagtt aatagctaaa tctaacactc ctcccatgta cttacaagcc
1081 gatatagaag cctttgacat cagagaacta acacccaaat ttgatgtgat tcttctggaa
1141 ccccctttag aagaatatta cagagaaact ggcatcactg ctaatgaaaa atgctggact
1201 tgggatgata ttatgaagtt agaaattgat gagattgcag cacctcgatc atttattttt
1261 ctctggtgtg gttctgggga ggggttggac cttggaagag tgtgtttacg aaaatggggt
1321 tacagaagat gtgaagatat ttgttggatt aaaaccaata aaaacaatcc tgggaagact
1381 aagactttag atccaaaggc tgtctttcag agaacaaagg aacactgcct catggggatc
1441 aaaggaactg tgaagcctag cacagacggg gacttcattc atgctaatgt tgacattgac
1501 ttaattatca cagaagaacc tgaaattggc aatatagaaa aacctgtaga aatttttcat
1561 ataattgagc atttttgtct tggtagaaga cgccttcatc tatttggaag agatagtaca
1621 attcgaccag gctggctcac agttggacca acgcttacaa atagcaacta caatgcagaa
1681 acatatgcat cctatttcag tgctcctaat tcctacttga ctggttgtac agaagaaatt
1741 gagagacttc gaccaaaatc gcctcctccc aaatctaaat ctgaccgagg aggtggagct
1801 cccagaggtg gaggaagagg tggaacttct gctggccgtg gacgagaaag aaatagatct
1861 aacttccgag gagaaagagg tggctttaga gggggccgtg gaggagcaca cagaggtggc
1921 tttccacctc gataagcggc cgcgaaggat ctgcgatcgc tccggtgccc gtcagtgggc
1981 agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaacggg
2041 tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct
2101 ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt
2161 tcgcaacggg tttgccgcca gaacacagct gaagcttcga ggggctcgca tctctccttc
2221 acgcgcccgc cgccctacct gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc
2281 tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc
2341 gagaccgggc ctttgtccgg cgctcccttg gagcctacct agactcagcc ggctctccac
2401 gctttgcctg accctgcttg ctcaactcta cgtctttgtt tcgttttctg ttctgcgccg
2461 ttacagatcc aagctgtgac cggcgcctac gctagacgcc accatggaga gcgacgagag
2521 cggcctgccc gccatggaga tcgagtgccg catcaccggc accctgaacg gcgtggagtt
2581 cgagctggtg ggcggcggag agggcacccc caagcagggc cgcatgacca acaagatgaa
2641 gagcaccaaa ggcgccctga ccttcagccc ctacctgctg agccacgtga tgggctacgg
2701 cttctaccac ttcggcacct accccagcgg ctacgagaac cccttcctgc acgccatcaa
2761 caacggcggc tacaccaaca cccgcatcga gaagtacgag gacggcggcg tgctgcacgt
2821 gagcttcagc taccgctacg aggccggccg cgtgatcggc gacttcaagg tggtgggcac
2881 cggcttcccc gaggacagcg tgatcttcac cgacaagatc atccgcagca acgccaccgt
2941 ggagcacctg caccccatgg gcgataacgt gctggtgggc agcttcgccc gcaccttcag
3001 cctgcgcgac ggcggctact acagcttcgt ggtggacagc cacatgcact tcaagagcgc
3061 catccacccc agcatcctgc agaacggggg ccccatgttc gccttccgcc gcgtggagga
3121 gctgcacagc aacaccgagc tgggcatcgt ggagtaccag cacgccttca agacccccat
3181 cgccttcgcc agatcccgcg ctcagtcgtc caattctgcc gtggacggca ccgccggacc
3241 cggctccacc ggatctcgcg agggcagagg aagtcttcta acatgcggtg acgtggagga
3301 gaatcccggc cctatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt
3361 ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac
3421 cgtcgatccg gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg
3481 cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg
3541 gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc
3601 cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca
3661 ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg
3721 caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt
3781 gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt
3841 caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa
3901 gcccggtgcc tgaaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
3961 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
4021 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
4081 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
4141 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
4201 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
4261 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt
4321 tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
4381 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
4441 tcttccgcgt ctccgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
4501 gcctggtacc tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa
4561 aagaaaaggg gggactggaa gggctaattc actcccaacg aagataagat ctgctttttg
4621 cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag
4681 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc
4741 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa
4801 tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat
4861 gaatatcaga gagtgagagg aacttgttta ttgcagctta taatggttac aaataaagca
4921 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
4981 ccaaactcat caatgtatct tatcatgtct ggctctagct atcccgcccc taactccgcc
5041 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
5101 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
5161 aggctttttt ggaggcctag acttttgcag agaccaaatt cgtaatcatg tcatagctgt
5221 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
5281 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
5341 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
5401 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
5461 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
5521 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
5581 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
5641 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
5701 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
5761 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
5821 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
5881 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
5941 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
6001 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
6061 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
6121 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
6181 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
6241 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
6301 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
6361 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
6421 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
6481 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
6541 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
6601 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
6661 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
6721 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
6781 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
6841 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
6901 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
6961 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
7021 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
7081 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
7141 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
7201 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
7261 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
7321 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
7381 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt
7441 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
7501 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
7561 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc
7621 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat
7681 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
7741 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
7801 cacgacgttg taaaacgacg gccagtgcca agctgacgcg tgtagtctta tgcaatactc
7861 ttgtagtctt gcaacatggt aacgatgagt tagcaacatg ccttacaagg agagaaaaag
7921 caccgtgcat gccgattggt ggaagtaagg tggtacgatc gtgccttatt aggaaggcaa
7981 cagacgggtc tgacatggat tggacgaacc actgaattgc cgcattgcag agatattgta
8041 tttaagtgcc tagctcgata cataaacggg tctctctggt tagaccagat ctgagcctgg
8101 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
8161 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
8221 ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag
8281 ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag
8341 gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga
8401 gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa
8461 ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca
8521 gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac
8581 aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat
8641 ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg
8701 aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg
8761 ccactgatct tcagacctgg aggaggagat atgagggaca attggagaag tgaattatat
8821 aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc aaagagaaga
8881 gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg gttcttggga
8941 gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc cagacaatta
9001 ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc gcaacagcat
9061 ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct ggctgtggaa
9121 agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa actcatttgc
9181 accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca gatttggaat
9241 cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt aatacactcc
9301 ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt ggaattagat
9361 aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta tataaaatta
9421 ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt actttctata
9481 gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct cccaaccccg
9541 aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
//
| {
"Vector backbone": "PCDH",
"Backbone size w/o insert (bp)": "",
"Vector type": "Mammalian Expression",
"Selectable markers": "Puromycin"
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "NEB Stable",
"Copy number": "Unknown"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "METTL14 Inhibits Hematopoietic Stem/Progenitor Differentiation and Promotes Leukemogenesis via mRNA m(6)A Modification",
"Publication": "Cell Stem Cell. 2018 Feb 1;22(2):191-205.e9. doi: 10.1016/j.stem.2017.11.016. Epub 2017 Dec 28.",
"DOI": "10.1016/j.stem.2017.11.016.",
"Authors": "Weng H, Huang H, Wu H, Qin X, Zhao BS, Dong L, Shi H, Skibbe J, Shen C, Hu C, Sheng Y, Wang Y, Wunderlich M, Zhang B, Dore LC, Su R, Deng X, Ferchen K, Li C, Sun M, Lu Z, Jiang X, Marcucci G, Mulloy JC, Yang J, Qian Z, Wei M, He C, Chen J /jstem",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/29290617",
"PubMed ID": "PubMed 29290617"
} | {
"Gene/Insert name": "HMETTL14",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "Substitution - Missense, position 298, R➞P",
"Entrez Gene": "METTL14",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pXP842 | 46068 | (Empty Backbone) | Nancy DaSilva | Shen et al Yeast. 2012 Dec;29(12):495-503. doi: 10.1002/yea.2930. Epub 2012 Nov 19. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/65296/f82d9b50-3547-4f9e-a4aa-e7bffa2e84de/addgene-plasmid-46068-sequence-65296.gbk | full | LOCUS sequence-65296-e 5866 bp DNA circular SYN 26-AUG-2024
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5866)
AUTHORS Shen MW, Fang F, Sandmeyer S, Da Silva NA
TITLE Development and characterization of a vector set with regulated
promoters for systematic metabolic engineering in Saccharomyces
cerevisiae.
JOURNAL Yeast. 2012 Dec;29(12):495-503. doi: 10.1002/yea.2930. Epub 2012 Nov
19.
PUBMED 23166051
REFERENCE 2 (bases 1 to 5866)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10.1002/yea.2930";
journalName: "Yeast"; date: "2012-12"; volume: "29"; issue: "12";
pages: "495-503"
FEATURES Location/Qualifiers
source 1..5866
/mol_type="other DNA"
/organism="synthetic DNA construct"
terminator 602..849
/gene="S. cerevisiae CYC1"
/label=CYC1 terminator
/note="transcription terminator for CYC1"
primer_bind complement(618..636)
/label=CYC1
/note="CYC1 transcription termination signal, reverse
primer"
protein_bind complement(895..928)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
promoter 946..1162
/gene="S. cerevisiae URA3"
/label=URA3 promoter
CDS 1163..1966
/codon_start=1
/gene="S. cerevisiae URA3"
/product="orotidine-5'-phosphate decarboxylase, required
for uracil biosynthesis"
/label=URA3
/note="yeast auxotrophic marker, counterselectable with
5-fluoroorotic acid (5-FOA)"
/translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLATGE
YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
protein_bind complement(2045..2078)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
rep_origin 2101..3443
/label=2u ori
/note="yeast 2u plasmid origin of replication"
protein_bind complement(3064..3111)
/label=FRT
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
primer_bind complement(3463..3479)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3463..3479)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3476..3498)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3487..3503
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(3511..3541)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3556..3577
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(3694..3711)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3865..4453)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3945..3964)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4624..5484)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 5247..5266
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5485..5589)
/gene="bla"
/label=AmpR promoter
primer_bind 5657..5675
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(5713..5735)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 5835..5854
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 catatgattt agcggccgca aaacgtaggg gcaaacaaac ggaaaaatcg tttctcaaat
61 tttctgatgc caagaactct aaccagtctt atctaaaaat tgccttatga tccgtctctc
121 cggttacagc ctgtgtaact gattaatcct gcctttctaa tcaccattct aatgttttaa
181 ttaagggatt ttgtcttcat taacggcttt cgctcataaa aatgttatga cgttttgccc
241 gcaggcggga aaccatccac ttcacgagac tgatctcctc tgccggaaca ccgggcatct
301 ccaacttata agttggagaa ataagagaat ttcagattga gagaatgaaa aaaaaaaaaa
361 aaaaaaaggc agaggagagc atagaaatgg ggttcacttt ttggtaaagc tatagcatgc
421 ctatcacata taaatagagt gccagtagcg acttttttca cactcgaaat actcttacta
481 ctgctctctt gttgttttta tcacttcttg tttcttcttg gtaaatagaa tatcaagcta
541 caaaaagcat acaatcaact atcaactatt aactatatcg taatacacaa ctagtctcga
601 gtcatgtaat tagttatgtc acgcttacat tcacgccctc cccccacatc cgctctaacc
661 gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt tatagttatg
721 ttagtattaa gaacgttatt tatatttcaa atttttcttt tttttctgta cagacgcgtg
781 tacgcatgta acattatact gaaaaccttg cttgagaagg ttttgggacg ctcgaaggct
841 ttaatttgcg gccaagcttg catgcctgca ggtcgactct agaggatccc cgggataact
901 tcgtatagca tacattatac gaagttatca gggtccataa agcttttcaa ttcatctttt
961 ttttttttgt tctttttttt gattccggtt tctttgaaat ttttttgatt cggtaatctc
1021 cgagcagaag gaagaacgaa ggaaggagca cagacttaga ttggtatata tacgcatatg
1081 tggtgttgaa gaaacatgaa attgcccagt attcttaacc caactgcaca gaacaaaaac
1141 ctgcaggaaa cgaagataaa tcatgtcgaa agctacatat aaggaacgtg ctgctactca
1201 tcctagtcct gttgctgcca agctatttaa tatcatgcac gaaaagcaaa caaacttgtg
1261 tgcttcattg gatgttcgta ccaccaagga attactggag ttagttgaag cattaggtcc
1321 caaaatttgt ttactaaaaa cacatgtgga tatcttgact gatttttcca tggagggcac
1381 agttaagccg ctaaaggcat tatccgccaa gtacaatttt ttactcttcg aagacagaaa
1441 atttgctgac attggtaata cagtcaaatt gcagtactct gcgggtgtat acagaatagc
1501 agaatgggca gacattacga atgcacacgg tgtggtgggc ccaggtattg ttagcggttt
1561 gaagcaggcg gcggaagaag taacaaagga acctagaggc cttttgatgt tagcagaatt
1621 gtcatgcaag ggctccctag ctactggaga atatactaag ggtactgttg acattgcgaa
1681 gagcgacaaa gattttgtta tcggctttat tgctcaaaga gacatgggtg gaagagatga
1741 aggttacgat tggttgatta tgacacccgg tgtgggttta gatgacaagg gagacgcatt
1801 gggtcaacag tatagaaccg tggatgatgt ggtctctaca ggatctgaca ttattattgt
1861 tggaagagga ctatttgcaa agggaaggga tgctaaggta gagggtgaac gttacagaaa
1921 agcaggctgg gaagcatatt tgagaagatg cggccagcaa aactaaaaaa ctgtattata
1981 agtaaatgca tgtatactaa actcacaaat tagagcttca atttaattat atcagttatt
2041 acccataact tcgtatagca tacattatac gaagttatcc cgggtaccga gctcgaattc
2101 aacgaagcat ctgtgcttca ttttgtagaa caaaaatgca acgcgagagc gctaattttt
2161 caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgaa agcgctattt
2221 taccaacgaa gaatctgtgc ttcatttttg taaaacaaaa atgcaacgcg agagcgctaa
2281 tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgagagcgc
2341 tattttacca acaaagaatc tatacttctt ttttgttcta caaaaatgca tcccgagagc
2401 gctatttttc taacaaagca tcttagatta ctttttttct cctttgtgcg ctctataatg
2461 cagtctcttg ataacttttt gcactgtagg tccgttaagg ttagaagaag gctactttgg
2521 tgtctatttt ctcttccata aaaaaagcct gactccactt cccgcgttta ctgattacta
2581 gcgaagctgc gggtgcattt tttcaagata aaggcatccc cgattatatt ctataccgat
2641 gtggattgcg catactttgt gaacagaaag tgatagcgtt gatgattctt cattggtcag
2701 aaaattatga acggtttctt ctattttgtc tctatatact acgtatagga aatgtttaca
2761 ttttcgtatt gttttcgatt cactctatga atagttctta ctacaatttt tttgtctaaa
2821 gagtaatact agagataaac ataaaaaatg tagaggtcga gtttagatgc aagttcaagg
2881 agcgaaaggt ggatgggtag gttatatagg gatatagcac agagatatat agcaaagaga
2941 tacttttgag caatgtttgt ggaagcggta ttcgcaatat tttagtagct cgttacagtc
3001 cggtgcgttt ttggtttttt gaaagtgcgt cttcagagcg cttttggttt tcaaaagcgc
3061 tctgaagttc ctatactttc tagagaatag gaacttcgga ataggaactt caaagcgttt
3121 ccgaaaacga gcgcttccga aaatgcaacg cgagctgcgc acatacagct cactgttcac
3181 gtcgcaccta tatctgcgtg ttgcctgtat atatatatac atgagaagaa cggcatagtg
3241 cgtgtttatg cttaaatgcg tacttatatg cgtctattta tgtaggatga aaggtagtct
3301 agtacctcct gtgatattat cccattccat gcggggtatc gtatgcttcc ttcagcacta
3361 ccctttagct gttctatatg ctgccactcc tcaattggat tagtctcatc cttcaatgct
3421 atcatttcct ttgatattgg atcatacgaa ttcgtaatca tggtcatagc tgtttcctgt
3481 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
3541 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
3601 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
3661 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
3721 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
3781 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
3841 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
3901 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
3961 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
4021 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
4081 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
4141 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
4201 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
4261 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
4321 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
4381 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
4441 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
4501 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
4561 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
4621 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
4681 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
4741 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
4801 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
4861 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
4921 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
4981 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
5041 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
5101 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
5161 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
5221 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
5281 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
5341 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
5401 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
5461 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
5521 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
5581 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
5641 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
5701 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
5761 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
5821 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcac
//
| {
"Vector backbone": "pUC18",
"Backbone size w/o insert (bp)": "",
"Vector type": "Yeast Expression",
"Selectable markers": "URA3"
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "XL1 Blue",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Development and characterization of a vector set with regulated promoters for systematic metabolic engineering in Saccharomyces cerevisiae",
"Publication": "Yeast. 2012 Dec;29(12):495-503. doi: 10.1002/yea.2930. Epub 2012 Nov 19.",
"DOI": "10.1002/yea.2930.",
"Authors": "Shen MW, Fang F, Sandmeyer S, Da Silva NA /yea",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/23166051",
"PubMed ID": "PubMed 23166051"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
CMV500 A-C/EBP | 33352 | Low | Charles Vinson | Ahn et al Mol Cell Biol. 1998 Feb . 18(2):967-77. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/41495/fa6d4790-7e25-4fc3-a90e-0f6c65e4764a/addgene-plasmid-33352-sequence-41495.gbk | partial | LOCUS sequence-41495-e 735 bp DNA linear UNA 26-AUG-2024
DEFINITION natural linear DNA
ACCESSION .
VERSION .
KEYWORDS .
SOURCE natural DNA sequence
ORGANISM unspecified
REFERENCE 1 (bases 1 to 735)
AUTHORS Ahn S, Olive M, Aggarwal S, Krylov D, Ginty DD, Vinson C
TITLE A dominant-negative inhibitor of CREB reveals that it is a general
mediator of stimulus-dependent transcription of c-fos.
JOURNAL Mol Cell Biol. 1998 Feb . 18(2):967-77.
PUBMED 9447994
REFERENCE 2 (bases 1 to 735)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Mol Cell
Biol. 1998 Feb . 18(2):967-77."
FEATURES Location/Qualifiers
source 1..735
/mol_type="genomic DNA"
/organism="unspecified"
CDS complement(282..314)
/codon_start=1
/product="leader peptide from bacteriophage T7 gene 10"
/label=T7 tag (gene 10 leader)
/note="promotes efficient translation in E. coli"
/translation="MASMTGGQQMG"
CDS complement(318..341)
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
primer_bind complement(361..380)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(362..380)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter complement(425..628)
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind complement(454..474)
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
ORIGIN
1 tatagaatag ggccctctag aagcttgaat tcgcctcacg cgcagttgcc catggccttg
61 accaaggagc tctcaggcag ctggcggaag ataccccgca gcgtgtccag ttcacggctc
121 agctgttcca cccgcttgcg caggcggtca ttgtcactgg tcaactccaa caccttctgc
181 tcgagttcag cgttttcctg ctccagctct tcggcctctt tttccagctc ttcgttttca
241 cgggccagtt cctcagcacg ttgttccagg tcaggatccc gacccatttg ctgtccacca
301 gtcatgctag ccatatgctt gtcatcgtcg tccttgtagt ccatggtgga gcttgggtct
361 ccctatagtg agtcgtatta atttcgataa gccagtaagc agtgggttct ctagttagcc
421 agagagctct gcttatatag acctcccacc gtacacgcct accgcccatt tgcgtcaatg
481 gggcggagtt gttacgacat tttggaaagt cccgttgatt ttggtgccaa aacaaactcc
541 cattgacgtc aatggggtgg agacttggaa atccccgtga gtcaaaccgc tatccacgcc
601 cattgatgta ctgccaaaac cgcatcacca tggtaatagc gatgactaat acgtagatgt
661 actgccaagt aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt
721 taccgtcatt gacgt
//
| {
"Vector backbone": "CMV500",
"Backbone size w/o insert (bp)": "5500",
"Vector type": "Mammalian Expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "A dominant-negative inhibitor of CREB reveals that it is a general mediator of stimulus-dependent transcription of c-fos",
"Publication": "Mol Cell Biol. 1998 Feb . 18(2):967-77.",
"DOI": "",
"Authors": "Ahn S, Olive M, Aggarwal S, Krylov D, Ginty DD, Vinson C",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/9447994",
"PubMed ID": "PubMed 9447994"
} | {
"Gene/Insert name": "C/EBP",
"Species": "M. musculus (mouse)",
"Insert Size (bp)": "",
"Mutation": "Dominant Negative (see notes)",
"Entrez Gene": "Cebpa",
"Promoter": "CMV",
"Tag / Fusion Protein": "FLAG (N terminal on insert)"
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "NdeI (not destroyed)",
"3′ cloning site": "HindIII (not destroyed)",
"5′ sequencing primer": "CMV-Fwd",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
INCTbiosyn-pEF-GFP(rc)13 | 127509 | Plasmid has an EF1 alpha promoter in a forward sequence orientation and an inverted EGFP coding sequence (reverse complement) flanked by attB and attP integrase 13 attachment sites. | Martin Bonamino | Gomide et al Commun Biol. 2020 May 22;3(1):255. doi: 10.1038/s42003-020-0971-8. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/250751/5610af93-4ddd-4baa-ad89-7f55a6bb4790/addgene-plasmid-127509-sequence-250751.gbk | full | LOCUS sequence-250751- 5182 bp DNA circular SYN 26-AUG-2024
DEFINITION Plasmid has an EF1 alpha promoter in a forward sequence orientation
and an inverted EGFP coding sequence (reverse complement) flanked by
attB and attP integrase 13 attachment sites..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5182)
AUTHORS Gomide MS, Sales TT, Barros LRC, Limia CG, de Oliveira MA,
Florentino LH, Barros LMG, Robledo ML, Jose GPC, Almeida MSM, Lima
RN, Rehen SK, Lacorte C, Melo EO, Murad AM, Bonamino MH, Coelho CM,
Rech E
TITLE Genetic switches designed for eukaryotic cells and controlled by
serine integrases.
JOURNAL Commun Biol. 2020 May 22;3(1):255. doi: 10.1038/s42003-020-0971-8.
PUBMED 32444777
REFERENCE 2 (bases 1 to 5182)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Commun
Biol."; date: "2020-05-22"; volume: "3(1):255. doi"; pages: "
10.1038/s42003-020-0971-8"
FEATURES Location/Qualifiers
source 1..5182
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 43..1225
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 274..1216
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 1173..1193
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
CDS complement(1341..2060)
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS complement(1344..2060)
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(1386..1407)
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind 1735..1754
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1994..2015
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(2211..2230)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_signal 2276..2331
/label=beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(2330..2349)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
primer_bind complement(2692..2708)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2692..2708)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(2705..2727)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 2716..2732
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(2740..2770)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2785..2806
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2865..3060
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 2911..3046
/label=SV40 ori
/note="SV40 origin of replication"
polyA_signal 3066..3200
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3103..3122)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3157..3176
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(3268..3285)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3439..4027)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3519..3538)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4198..5058)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 4821..4840
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5059..5163)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 ccacctgggt cgacattgat tattgactag atcatcgcgt gaggctccgg tgcccgtcag
61 tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga
121 accggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc
181 cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt
241 ctttttcgca acgggtttgc cgccagaaca caggtaagtg ccgtgtgtgg ttcccgcggg
301 cctggcctct ttacgggtta tggcccttgc gtgccttgaa ttacttccac gcccctggct
361 gcagtacgtg attcttgatc ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc
421 ttgcgcttaa ggagcccctt cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg
481 ccgccgcgtg cgaatctggt ggcaccttcg cgcctgtctc gctgctttcg ataagtctct
541 agccatttaa aatttttgat gacctgctgc gacgcttttt ttctggcaag atagtcttgt
601 aaatgcgggc caagatctgc acactggtat ttcggttttt ggggccgcgg gcggcgacgg
661 ggcccgtgcg tcccagcgca catgttcggc gaggcggggc ctgcgagcgc ggccaccgag
721 aatcggacgg gggtagtctc aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc
781 gtgtatcgcc ccgccctggg cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga
841 aagatggccg cttcccggcc ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg
901 agagcgggcg ggtgagtcac ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc
961 ttcatgtgac tccacggagt accgggcgcc gtccaggcac ctcgattagt tctcgagctt
1021 ttggagtacg tcgtctttag gttgggggga ggggttttat gcgatggagt ttccccacac
1081 tgagtgggtg gagactgaag ttaggccagc ttggcacttg atgtaattct ccttggaatt
1141 tgcccttttt gagtttggat cttggttcat tctcaagcct cagacagtgg ttcaaagttt
1201 ttttcttcca tttcaggtgt cgtgaggaat tctgcagtcg acggtaccgc gggcccggga
1261 tccaccggtc gccaccgcat acattgttgt tgtttttcca gatccagttg gtcctgtaaa
1321 tataagcaat ccatgtgagt ttacttgtac agctcgtcca tgccgagagt gatcccggcg
1381 gcggtcacga actccagcag gaccatgtga tcgcgcttct cgttggggtc tttgctcagg
1441 gcggactggg tgctcaggta gtggttgtcg ggcagcagca cggggccgtc gccgatgggg
1501 gtgttctgct ggtagtggtc ggcgagctgc acgctgccgt cctcgatgtt gtggcggatc
1561 ttgaagttca ccttgatgcc gttcttctgc ttgtcggcca tgatatagac gttgtggctg
1621 ttgtagttgt actccagctt gtgccccagg atgttgccgt cctccttgaa gtcgatgccc
1681 ttcagctcga tgcggttcac cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg
1741 tagttgccgt cgtccttgaa gaagatggtg cgctcctgga cgtagccttc gggcatggcg
1801 gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc ggctgaagca ctgcacgccg
1861 taggtcaggg tggtcacgag ggtgggccag ggcacgggca gcttgccggt ggtgcagatg
1921 aacttcaggg tcagcttgcc gtaggtggca tcgccctcgc cctcgccgga cacgctgaac
1981 ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg gcaccacccc ggtgaacagc
2041 tcctcgccct tgctcaccat tattcggagt tgtatttatg ttactaaaac aactggtcaa
2101 gttctacaaa tacaaccgtt attgagcggc cgcactcctc aggtgcaggc tgcctatcag
2161 aaggtggtgg ctggtgtggc caatgccctg gctcacaaat accactgaga tctttttccc
2221 tctgccaaaa attatgggga catcatgaag ccccttgagc atctgacttc tggctaataa
2281 aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc actcggaagg
2341 acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta gagtttggca
2401 acatatgccc atatgctggc tgccatgaac aaaggttggc tataaagagg tcatcagtat
2461 atgaaacagc cccctgctgt ccattcctta ttccatagaa aagccttgac ttgaggttag
2521 atttttttta tattttgttt tgtgttattt ttttctttaa catccctaaa attttcctta
2581 catgttttac tagccagatt tttcctcctc tcctgactac tcccagtcat agctgtccct
2641 cttctcttat ggagatccct cgacctgcag cccaagcttg gcgtaatcat ggtcatagct
2701 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
2761 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
2821 actgcccgct ttccagtcgg gaaacctgtc gtgccagcgg atccgcatct caattagtca
2881 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
2941 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
3001 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
3061 aagctaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
3121 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
3181 tatcttatca tgtctggatc cgctgcatta atgaatcggc caacgcgcgg ggagaggcgg
3241 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
3301 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
3361 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
3421 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
3481 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
3541 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
3601 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
3661 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
3721 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
3781 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
3841 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
3901 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
3961 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
4021 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
4081 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
4141 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
4201 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
4261 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
4321 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
4381 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
4441 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
4501 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
4561 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
4621 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
4681 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
4741 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
4801 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
4861 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
4921 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
4981 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
5041 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
5101 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
5161 gcgcacattt ccccgaaaag tg
//
| {
"Vector backbone": "pEF-GFP",
"Backbone size w/o insert (bp)": "",
"Vector type": "Mammalian Expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Genetic switches designed for eukaryotic cells and controlled by serine integrases",
"Publication": "Commun Biol. 2020 May 22;3(1):255. doi: 10.1038/s42003-020-0971-8.",
"DOI": "10.1038/s42003-020-0971-8.",
"Authors": "Gomide MS, Sales TT, Barros LRC, Limia CG, de Oliveira MA, Florentino LH, Barros LMG, Robledo ML, Jose GPC, Almeida MSM, Lima RN, Rehen SK, Lacorte C, Melo EO, Murad AM, Bonamino MH, Coelho CM, Rech E /s---",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/32444777",
"PubMed ID": "PubMed 32444777"
} | {
"Gene/Insert name": "EGFP reverse complement coding sequence flanked by attB/attP Integrase 13 attachment sites.",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "848",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Unknown",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pGBW-m4133579 | 152181 | Mammalian expression plasmid for SARS-CoV-2 E (envelope) | SARS-CoV-2 protein Mammalian expression constructs, tagged, for protein expression and/or purification (unpublished) | https://media.addgene.org/snapgene-media/v2.0.0/sequences/300230/39f55573-df3c-4ae4-8103-1537f48160c6/addgene-plasmid-152181-sequence-300230.gbk | full | LOCUS sequence-300230- 6410 bp DNA circular SYN 26-AUG-2024
DEFINITION Mammalian expression plasmid for SARS-CoV-2 E (envelope).
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6410)
TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for
protein expression and/or purification
REFERENCE 2 (bases 1 to 6410)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6410
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 65..136
/label=5' beta-globin insulator
/note="insulator upstream of the human beta-globin locus
(Farrell et al., 2002)"
terminator complement(141..187)
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
polyA_signal 205..253
/note="synthetic polyadenylation signal"
enhancer 334..637
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 639..841
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 792..812
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
CDS 1141..1161
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQS"
CDS 1165..1230
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
polyA_signal 1283..1404
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1294..1313)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 1348..1367
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
terminator 1519..1565
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
terminator 1595..1731
/label=rrnG terminator
/note="transcription terminator from the E. coli ribosomal
RNA rrnG operon (Albrechtsen et al., 1991)"
misc_feature 1776..1847
/label=3' beta-globin insulator
/note="insulator downstream of the human beta-globin locus
(Farrell et al., 2002)"
rep_origin complement(2073..2661)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2153..2172)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
promoter 3161..4370
/label=UbC promoter
/note="human ubiquitin C promoter"
primer_bind 4207..4226
/label=hUBCpro-F
/note="Human Ubiquitin C (UbC) promoter, forward primer"
CDS 4374..4973
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind complement(4374..4393)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 4870..4890
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
polyA_signal 5000..5048
/note="synthetic polyadenylation signal"
CDS 5314..6174
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(5532..5551)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
terminator 6187..6323
/label=rrnG terminator
/note="transcription terminator from the E. coli ribosomal
RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
1 gtcgggcatc gaatctcgtc agctatcgtc agcttacctt tttggcagcg atcgcggctc
61 ccgacatctt ggaccattag ctccacaggt atcttcttcc ctctagtggt cataacagca
121 gcttcagcta cctctcaatt caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt
181 tatgctatca atcgttgcgt tacacacaca aaaaaccaac acacatccat cttcgatgga
241 tagcgatttt attatctaac tgctgatcga gtgtagccag atctagtaat caattacggg
301 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
361 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
421 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
481 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
541 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
601 gcagtacatc tacgtattag tcatcgctat taccatgctg atgcggtttt ggcagtacat
661 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
721 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
781 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
841 tggtttagtg aaccgtcaga tcagatcttt gtcgatccta ccatccactc gacacacccg
901 ccagcggccg ccaccatgta ctcattcgtt tcggaagaga caggtacgtt aatagttaat
961 agcgtacttc tttttcttgc tttcgtggta ttcttgctag ttacactagc catccttact
1021 gcgcttcgat tgtgtgcgta ctgctgcaat attgttaacg tgagtcttgt aaaaccttct
1081 ttttacgttt actctcgtgt taaaaatctg aattcttcta gagttcctga tcttctggtc
1141 gaaaaccttt actttcagtc gatggattat aaggaccacg atggagacta caaagatcat
1201 gacattgatt acaaggacga cgatgacaaa taagctagct tgactgactg agatacagcg
1261 taccttcagc tcacagacat gataagatac attgatgagt ttggacaaac cacaactaga
1321 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
1381 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
1441 cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtattggc
1501 ccatctctat cggtatcgta gcataacccc ttggggcctc taaacgggtc ttgaggggtt
1561 ttttgtgccc ctcgggccgg attgctatct accggcattg gcgcagaaaa aaatgcctga
1621 tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga tacggcttcc
1681 ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta aggtcgtcag
1741 ctatcctgca ggcgatctct cgatttcgat caagacattc ctttaatggt cttttctgga
1801 caccactagg ggtcagaagt agttcatcaa actttcttcc ctccctaatc tcattggtta
1861 ccttgggcta tcgaaactta attaaccagt caagtcagct acttggcgag atcgacttgt
1921 ctgggtttcg actacgctca gaattgcgtc agtcaagttc gatctggtcc ttgctattgc
1981 acccgttctc cgattacgag tttcatttaa atcatgtgag caaaaggcca gcaaaaggcc
2041 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
2101 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
2161 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
2221 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
2281 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
2341 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
2401 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
2461 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
2521 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
2581 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
2641 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
2701 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
2761 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
2821 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
2881 cgttcatcca tagttgcatt taaatttccg aactctccaa ggccctcgtc ggaaaatctt
2941 caaacctttc gtccgatcca tcttgcaggc tacctctcga acgaactatc gcaagtctct
3001 tggccggcct tgcgccttgg ctattgcttg gcagcgccta tcgccaggta ttactccaat
3061 cccgaatatc cgagatcggg atcacccgag agaagttcaa cctacatcct caatcccgat
3121 ctatccgaga tccgaggaat atcgaaatcg gggcgcgcct ggcctccgcg ccgggttttg
3181 gcgcctcccg cgggcgcccc cctcgtcacg gcgagcgctg ccacgtcaga cgaagggcgc
3241 aggagcgtcc tgatccttcc gcccggacgc tcaggacagc ggcccgctgc tcataagact
3301 cggccttaga accccagtat cagcagaagg acattttagg acgggacttg ggtgactcta
3361 gggcactggt tttctttcca gagagcggaa caggcgagga aaagtagtcc cttctcggcg
3421 attctgcgga gggatctccg tggggcggtg aacgccgatg attatataag gacgcgccgg
3481 gtgtggcaca gctagttccg tcgcagccgg gatttgggtc gcggttcttg tttgtggatc
3541 gctgtgatcg tcacttggtg agtagcgggc tgctgggctg gccggggctt tcgtggccgc
3601 cgggccgctc ggtgggacgg aagcgtgtgg agagaccgcc aagggctgta gtctgggtcc
3661 gcgagcaagg ttgccctgaa ctgggggttg gggggagcgc agcaaaatgg cggctgttcc
3721 cgagtcttga atggaagacg cttgtgaggc gggctgtgag gtcgttgaaa caaggtgggg
3781 ggcatggtgg gcggcaagaa cccaaggtct tgagcccttc gctaatgcgg gaaagctctt
3841 attcgggtga gatgggctgg gcaccatctg gggaccctga cgtgaagttt gtcactgact
3901 ggagaactcg gtttgtcgtc tgttgcgggg gcggcagtta tggcggtgcc gttgggcagt
3961 gcacccgtac ctttgggagc gcgcgccctc gtcgtgtcgt gacgtcaccc gttctgttgg
4021 cttataatgc agggtggggc cacctgccgg taggtgtgcg gtaggctttt ctccgtcgca
4081 ggacgcaggg ttcgggccta gggtaggctc tcctgaatcg acaggcgccg gacctctggt
4141 gaggggaggg ataagtgagg cgtcagtttc tttggtcggt tttatgtacc tatcttctta
4201 agtagctgaa gctccggttt tgaactatgc gctcggggtt ggcgagtgtg ttttgtgaag
4261 ttttttaggc accttttgaa atgtaatcat ttgggtcaat atgtaatttt cagtgttaga
4321 cttgtaaatt gtccgctaaa ttctggccgt ttttggcttt tttgttagac aacatgaccg
4381 agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc
4441 tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca
4501 tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca
4561 aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg
4621 aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc
4681 tggccgcgca gcaacagatg gagggcctcc tggcgccgca ccggcccaag gagcccgcgt
4741 ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg
4801 tcgtgctccc cggagtggag gccgccgagc gcgccggggt gcccgccttc ctggagacct
4861 ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg
4921 aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc taggctagta
4981 tgtaagccta gtcttagata ataaaatcgc tatccatcga agatggatgt gtgttggttt
5041 tttgtgtgtg taacgctagg cgcgcctggt gtaccgagaa cgatcctctc agtgcgagtc
5101 tcgacgatcc atatcgttgc ttggcagtca gccagtcgga atccagcttg ggacccagga
5161 agtccaatcg tcagatattg tactcaagcc tggtcacggc agcgtaccga tctgtttaaa
5221 cctagatatt gatagtctga tcggtcaacg tataatcgag tcctagcttt tgcaaacatc
5281 tatcaagaga caggatcagc aggaggcttt cgcatgagta ttcaacattt ccgtgtcgcc
5341 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
5401 aaagtaaaag atgctgaaga tcagttgggt gcgcgagtgg gttacatcga actggatctc
5461 aacagcggta agatccttga gagttttcgc cccgaagaac gctttccaat gatgagcact
5521 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
5581 ggtcgccgca tacactattc tcagaatgac ttggttgagt attcaccagt cacagaaaag
5641 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
5701 aacactgcgg ccaacttact tctgacaacg attggaggac cgaaggagct aaccgctttt
5761 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
5821 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aaccttgcgt
5881 aaactattaa ctggcgaact acttactcta gcttcccggc aacagttgat agactggatg
5941 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
6001 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
6061 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
6121 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaaccgatt
6181 ctaggtgcat tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac
6241 atatgccaga ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct
6301 ccttaccaga aatttatcct taagatcccg aatcgtttaa actcgactct ggctctatcg
6361 aatctccgtc gtttcgagct tacgcgaaca gccgtggcgc tcatttgctc
//
| {
"Vector backbone": "pTwist_CMV_Puro_6",
"Backbone size w/o insert (bp)": "6080",
"Vector type": "Mammalian Expression",
"Selectable markers": "Puromycin"
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "OpenMTA",
"Industry Terms": "OpenMTA"
} | {
"Title": "",
"Publication": "",
"DOI": "",
"Authors": "",
"PubMed Link": "",
"PubMed ID": ""
} | {
"Gene/Insert name": "SARS-CoV-2 E (envelope)",
"Species": "Severe acute respiratory syndrome coronavirus 2",
"Insert Size (bp)": "221",
"Mutation": "wt",
"Entrez Gene": "E",
"Promoter": "CMV",
"Tag / Fusion Protein": "Cleavable TEV;3xFlag (C terminal on backbone)"
} | {
"Cloning method": "Unknown",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
||
pUC57-Kan_FR13_00020841 | 121295 | Level 0 Golden-Gate compatible vector | Sophien Kamoun | Petit-Houdenot et al Mol Plant Microbe Interact. 2020 Aug;33(8):1032-1035. doi: 10.1094/MPMI-03-20-0052-A. Epub 2020 Jun 22. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/231364/234e1668-ba35-4670-8719-f4867f1dab38/addgene-plasmid-121295-sequence-231364.gbk | full | LOCUS sequence-231364- 3007 bp DNA circular SYN 26-AUG-2024
DEFINITION Level 0 Golden-Gate compatible vector.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3007)
AUTHORS Petit-Houdenot Y, Langner T, Harant A, Win J, Kamoun S
TITLE A Clone Resource of Magnaporthe oryzae Effectors That Share Sequence
and Structural Similarities Across Host-Specific Lineages.
JOURNAL Mol Plant Microbe Interact. 2020 Aug;33(8):1032-1035. doi:
10.1094/MPMI-03-20-0052-A. Epub 2020 Jun 22.
PUBMED 32460610
REFERENCE 2 (bases 1 to 3007)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1094/MPMI-03-20-0052-A"; journalName: "Mol Plant Microbe
Interact"; date: "2020-08"; volume: "33"; issue: "8"; pages:
"1032-1035"
FEATURES Location/Qualifiers
source 1..3007
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(29..51)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 151..170
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 364..386
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 378..395
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 379..395
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(831..847)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(831..847)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(844..866)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 855..871
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(879..909)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 924..945
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(1062..1079)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(1233..1821)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1313..1332)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(1992..2807)
/codon_start=1
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
CDS complement(1992..2807)
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind 2715..2734
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
promoter complement(2808..2912)
/gene="bla"
/label=AmpR promoter
primer_bind 2980..2998
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
ORIGIN
1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt gacgcgtatt gggatggaac
421 ttggtctcta atgattaatg aagattgtat tgtttctttg gttccagatg gtggttcttt
481 tgctagaaaa actttgagat cttctccagg tcaacaacat gatttgcaat ggggtaaata
541 tgcttttact attagagttg aaaatggttg tgctggtaaa aaaactgctg gtactattcc
601 accagctcat tttgtttctg ttggtagagc tgcttctttg aaaccaactt ctattttgag
661 agatggtgat ccaagaccac caactgctgt ttatccacca agacaaccac caagaactga
721 aggtagaaga actagatatg gttcttctgg tggtgctgtt actgttggtg ctagatgagc
781 ttagagacca agttccatcc caatggcgcg ccgagcttgg ctcgagcatg gtcatagctg
841 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
901 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
961 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
1021 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
1081 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
1141 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
1201 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
1261 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
1321 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
1381 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
1441 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
1501 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
1561 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
1621 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
1681 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
1741 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
1801 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
1861 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
1921 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
1981 tggtctgaca gttagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca
2041 ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg
2101 aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca
2161 tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca
2221 tgagtgacga ctgaatccgg tgagaatggc aaaagtttat gcatttcttt ccagacttgt
2281 tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc
2341 attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg acaattacaa
2401 acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct
2461 gaatcaggat attcttctaa tacctggaat gctgttttcc cagggatcgc agtggtgagt
2521 aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc
2581 gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca
2641 tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat tgtcgcacct
2701 gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa
2761 tttaatcgcg gcctagagca agacgtttcc cgttgaatat ggctcatact cttccttttt
2821 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
2881 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
2941 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
3001 tttcgtc
//
| {
"Vector backbone": "pUC57-Kan",
"Backbone size w/o insert (bp)": "2626",
"Vector type": "Golden Gate",
"Selectable markers": "lacZ"
} | {
"Bacterial Resistance(s)": "Kanamycin, 50 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "A Clone Resource of Magnaporthe oryzae Effectors That Share Sequence and Structural Similarities Across Host-Specific Lineages",
"Publication": "Mol Plant Microbe Interact. 2020 Aug;33(8):1032-1035. doi: 10.1094/MPMI-03-20-0052-A. Epub 2020 Jun 22.",
"DOI": "10.1094/MPMI-03-20-0052-A.",
"Authors": "Petit-Houdenot Y, Langner T, Harant A, Win J, Kamoun S /MPMI----A",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/32460610",
"PubMed ID": "PubMed 32460610"
} | {
"Gene/Insert name": "FR13_00020841",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "Magnaporthe oryzae",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "EcoRV (unknown if destroyed)",
"3′ cloning site": "EcoRV (unknown if destroyed)",
"5′ sequencing primer": "M13Fw",
"3′ sequencing primer": "M13Rv"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pCMV-PE7-P2A-GFP | 222996 | Mammalian expression of SpCas9 PE7 prime editor with P2A-EGFP marker | Brittany Adamson | Yan et al Nature. 2024 Apr 3. doi: 10.1038/s41586-024-07259-6. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/442553/7ec4f654-d53f-43f5-8e9c-6feb13f8e465/addgene-plasmid-222996-sequence-442553.gbk | full | LOCUS sequence-442553- 11256 bp DNA circular SYN 25-AUG-2024
DEFINITION Mammalian expression of SpCas9 PE7 prime editor with P2A-EGFP
marker.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11256)
AUTHORS Yan J, Oyler-Castrillo P, Ravisankar P, Ward CC, Levesque S, Jing Y,
Simpson D, Zhao A, Li H, Yan W, Goudy L, Schmidt R, Solley SC,
Gilbert LA, Chan MM, Bauer DE, Marson A, Parsons LR, Adamson B
TITLE Improving prime editing with an endogenous small RNA-binding
protein.
JOURNAL Nature. 2024 Apr 3. doi: 10.1038/s41586-024-07259-6.
PUBMED 38570691
REFERENCE 2 (bases 1 to 11256)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nature.
2024 Apr 3. doi: 10.1038/s41586-024-07259-6."
FEATURES Location/Qualifiers
source 1..11256
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 1..380
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 381..584
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 535..555
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 626..645
/label=T7
/note="T7 promoter, forward primer"
promoter 626..644
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 694..714
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 715..4815
/codon_start=1
/product="Cas9 (Csn1) endonuclease from the Streptococcus
pyogenes Type II CRISPR/Cas system"
/label=Cas9
/note="generates RNA-guided double strand breaks in DNA"
/translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRKLE
NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLKREDLLR
KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
ATLIHQSITGLYETRIDLSQLGGD"
CDS 4873..4893
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 7672..7692
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 7702..7728
/codon_start=1
/product="nuclear localization signal of human c-Myc
proto-oncogene (Dang and Lee, 1988)"
/label=c-myc NLS
/translation="PAAKRVKLD"
CDS 7738..7794
/codon_start=1
/product="2A peptide from porcine teschovirus-1
polyprotein"
/label=P2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="ATNFSLLKQAGDVEENPGP"
CDS 7795..8514
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 7795..8511
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(8101..8120)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 8448..8469
/label=EGFP-C
/note="EGFP, forward primer"
CDS 8520..8537
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(8560..8577)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 8566..8790
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(8861..8877)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(8861..8877)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(8874..8896)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 8885..8901
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(8909..8939)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 8954..8975
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(9092..9109)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(9263..9851)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(9343..9362)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(10022..10882)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 10645..10664
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(10883..10987)
/gene="bla"
/label=AmpR promoter
primer_bind complement(11066..11085)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc
601 agatccgcta gagatccgcg gccgctaata cgactcacta tagggagagc cgccaccatg
661 aaacggacag ccgacggaag cgagttcgag tcaccaaaga agaagcggaa agtcgacaag
721 aagtacagca tcggcctgga catcggcacc aactctgtgg gctgggccgt gatcaccgac
781 gagtacaagg tgcccagcaa gaaattcaag gtgctgggca acaccgaccg gcacagcatc
841 aagaagaacc tgatcggagc cctgctgttc gacagcggcg aaacagccga ggccacccgg
901 ctgaagagaa ccgccagaag aagatacacc agacggaaga accggatctg ctatctgcaa
961 gagatcttca gcaacgagat ggccaaggtg gacgacagct tcttccacag actggaagag
1021 tccttcctgg tggaagagga taagaagcac gagcggcacc ccatcttcgg caacatcgtg
1081 gacgaggtgg cctaccacga gaagtacccc accatctacc acctgagaaa gaaactggtg
1141 gacagcaccg acaaggccga cctgcggctg atctatctgg ccctggccca catgatcaag
1201 ttccggggcc acttcctgat cgagggcgac ctgaaccccg acaacagcga cgtggacaag
1261 ctgttcatcc agctggtgca gacctacaac cagctgttcg aggaaaaccc catcaacgcc
1321 agcggcgtgg acgccaaggc catcctgtct gccagactga gcaagagcag aaagctggaa
1381 aatctgatcg cccagctgcc cggcgagaag aagaatggcc tgttcggaaa cctgattgcc
1441 ctgagcctgg gcctgacccc caacttcaag agcaacttcg acctggccga ggatgccaaa
1501 ctgcagctga gcaaggacac ctacgacgac gacctggaca acctgctggc ccagatcggc
1561 gaccagtacg ccgacctgtt tctggccgcc aagaacctgt ccgacgccat cctgctgagc
1621 gacatcctga gagtgaacac cgagatcacc aaggcccccc tgagcgcctc tatgatcaag
1681 agatacgacg agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg
1741 cctgagaagt acaaagagat tttcttcgac cagagcaaga acggctacgc cggctacatt
1801 gacggcggag ccagccagga agagttctac aagttcatca agcccatcct ggaaaagatg
1861 gacggcaccg aggaactgct cgtgaagctg aagagagagg acctgctgcg gaagcagcgg
1921 accttcgaca acggcagcat cccccaccag atccacctgg gagagctgca cgccattctg
1981 cggcggcagg aagattttta cccattcctg aaggacaacc gggaaaagat cgagaagatc
2041 ctgaccttcc gcatccccta ctacgtgggc cctctggcca ggggaaacag cagattcgcc
2101 tggatgacca gaaagagcga ggaaaccatc accccctgga acttcgagga agtggtggac
2161 aagggcgctt ccgcccagag cttcatcgag cggatgacca acttcgataa gaacctgccc
2221 aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt gtataacgag
2281 ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc ccgccttcct gagcggcgag
2341 cagaaaaagg ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag
2401 ctgaaagagg actacttcaa gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg
2461 gaagatcggt tcaacgcctc cctgggcaca taccacgatc tgctgaaaat tatcaaggac
2521 aaggacttcc tggacaatga ggaaaacgag gacattctgg aagatatcgt gctgaccctg
2581 acactgtttg aggacagaga gatgatcgag gaacggctga aaacctatgc ccacctgttc
2641 gacgacaaag tgatgaagca gctgaagcgg cggagataca ccggctgggg caggctgagc
2701 cggaagctga tcaacggcat ccgggacaag cagtccggca agacaatcct ggatttcctg
2761 aagtccgacg gcttcgccaa cagaaacttc atgcagctga tccacgacga cagcctgacc
2821 tttaaagagg acatccagaa agcccaggtg tccggccagg gcgatagcct gcacgagcac
2881 attgccaatc tggccggcag ccccgccatt aagaagggca tcctgcagac agtgaaggtg
2941 gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt gatcgaaatg
3001 gccagagaga accagaccac ccagaaggga cagaagaaca gccgcgagag aatgaagcgg
3061 atcgaagagg gcatcaaaga gctgggcagc cagatcctga aagaacaccc cgtggaaaac
3121 acccagctgc agaacgagaa gctgtacctg tactacctgc agaatgggcg ggatatgtac
3181 gtggaccagg aactggacat caaccggctg tccgactacg atgtggacgc tatcgtgcct
3241 cagagctttc tgaaggacga ctccatcgac aacaaggtgc tgaccagaag cgacaagaac
3301 cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa gaactactgg
3361 cggcagctgc tgaacgccaa gctgattacc cagagaaagt tcgacaatct gaccaaggcc
3421 gagagaggcg gcctgagcga actggataag gccggcttca tcaagagaca gctggtggaa
3481 acccggcaga tcacaaagca cgtggcacag atcctggact cccggatgaa cactaagtac
3541 gacgagaatg acaagctgat ccgggaagtg aaagtgatca ccctgaagtc caagctggtg
3601 tccgatttcc ggaaggattt ccagttttac aaagtgcgcg agatcaacaa ctaccaccac
3661 gcccacgacg cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag
3721 ctggaaagcg agttcgtgta cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc
3781 aagagcgagc aggaaatcgg caaggctacc gccaagtact tcttctacag caacatcatg
3841 aactttttca agaccgagat taccctggcc aacggcgaga tccggaagcg gcctctgatc
3901 gagacaaacg gcgaaaccgg ggagatcgtg tgggataagg gccgggattt tgccaccgtg
3961 cggaaagtgc tgagcatgcc ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc
4021 ggcttcagca aagagtctat cctgcccaag aggaacagcg ataagctgat cgccagaaag
4081 aaggactggg accctaagaa gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg
4141 ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt gaaagagctg
4201 ctggggatca ccatcatgga aagaagcagc ttcgagaaga atcccatcga ctttctggaa
4261 gccaagggct acaaagaagt gaaaaaggac ctgatcatca agctgcctaa gtactccctg
4321 ttcgagctgg aaaacggccg gaagagaatg ctggcctctg ccggcgaact gcagaaggga
4381 aacgaactgg ccctgccctc caaatatgtg aacttcctgt acctggccag ccactatgag
4441 aagctgaagg gctcccccga ggataatgag cagaaacagc tgtttgtgga acagcacaag
4501 cactacctgg acgagatcat cgagcagatc agcgagttct ccaagagagt gatcctggcc
4561 gacgctaatc tggacaaagt gctgtccgcc tacaacaagc accgggataa gcccatcaga
4621 gagcaggccg agaatatcat ccacctgttt accctgacca atctgggagc ccctgccgcc
4681 ttcaagtact ttgacaccac catcgaccgg aagaggtaca ccagcaccaa agaggtgctg
4741 gacgccaccc tgatccacca gagcatcacc ggcctgtacg agacacggat cgacctgtct
4801 cagctgggag gtgactccgg cggaagctct ggtggcagca agcggaccgc cgacggctct
4861 gaattcgaga gccctaagaa gaaaagaaag gtgagcggag gctctagcgg cggaagcacc
4921 ctgaacattg aagacgagta tagactgcat gaaacaagca aggaacccga cgtgtccctg
4981 ggctccacct ggctgtccga ctttccccag gcctgggccg agacaggagg aatgggcctg
5041 gccgtgcggc aggcacccct gatcatccct ctgaaggcca cctctacacc cgtgagcatc
5101 aagcagtacc ctatgtctca ggaggccaga ctgggcatca agcctcacat ccagaggctg
5161 ctggaccagg gcatcctggt gccatgccag agcccctgga acacaccact gctgcccgtg
5221 aagaagccag gcaccaatga ctatagaccc gtgcaggatc tgagagaggt gaacaagagg
5281 gtggaggata tccaccccac cgtgcccaac ccttacaatc tgctgtccgg cctgccccct
5341 tctcaccagt ggtatacagt gctggacctg aaggatgcct tcttttgtct gagactgcac
5401 cctaccagcc agccactgtt cgcctttgag tggagggacc ctgagatggg catctctggc
5461 cagctgacct ggacacgcct gcctcagggc ttcaagaata gcccaacact gtttaacgag
5521 gccctgcacc gcgacctggc agatttccgg atccagcacc cagatctgat cctgctgcag
5581 tacgtggacg atctgctgct ggccgccacc agcgagctgg attgccagca gggaacacgc
5641 gccctgctgc agaccctggg aaacctggga tatagggcat ccgccaagaa ggcccagatc
5701 tgtcagaagc aggtgaagta cctgggctat ctgctgaagg agggccagag atggctgaca
5761 gaggccagga aggagacagt gatgggccag ccaacaccca agaccccaag acagctgagg
5821 gagttcctgg gcaaagcagg attttgcagg ctgttcatcc caggattcgc agagatggca
5881 gcacctctgt acccactgac caagccgggc accctgttta attggggccc tgaccagcag
5941 aaggcctatc aggagatcaa gcaggccctg ctgacagcac cagccctggg cctgccagac
6001 ctgaccaagc ctttcgagct gtttgtggat gagaagcagg gctacgccaa gggcgtgctg
6061 acccagaagc tgggaccatg gagacggccc gtggcctatc tgtccaagaa gctggaccca
6121 gtggcagcag gatggccacc atgcctgagg atggtggcag caatcgccgt gctgacaaag
6181 gatgccggca agctgaccat gggacagcca ctggtcatcc tggcaccaca cgcagtggag
6241 gccctggtga agcagcctcc agatcgctgg ctgtctaacg cccggatgac acactaccag
6301 gccctgctgc tggacaccga tcgcgtgcag tttggccctg tggtggccct gaatccagcc
6361 accctgctgc ctctgccaga ggagggcctg cagcacaact gtctggacat cctggcagag
6421 gcacacggaa caaggccaga cctgaccgat cagcccctgc ctgacgccga tcacacatgg
6481 tataccgatg gaagctccct gctgcaggag ggccagagga aggcaggagc agcagtgacc
6541 acagagacag aagtgatctg ggccaaggcc ctgccagcag gcacatccgc ccagcgggcc
6601 gagctgatcg ccctgaccca ggccctgaag atggccgagg gcaagaagct gaacgtgtac
6661 acagactcca gatatgcctt cgccaccgca cacatccacg gagagatcta caggcgccgg
6721 ggctggctga cctctgaggg caaggagatc aagaacaagg atgagatcct ggccctgctg
6781 aaggccctgt ttctgcccaa gcggctgagc atcatccact gtcctggaca ccagaaggga
6841 cactccgccg aggcaagggg caatcggatg gccgaccagg ccgccagaaa ggctgctatt
6901 actgaaactc ccgacacttc cactctgctg attgaaaact cctccccttc cggaggatct
6961 agcggaggct cctctggctc tgagacacct ggcacaagcg agagcgcaac acctgaaagc
7021 agcgggggca gcagcggggg gtcaatggct gaaaatggtg ataatgaaaa gatggctgcc
7081 ctggaggcca aaatctgtca tcaaattgag tattattttg gcgacttcaa tttgccacgg
7141 gacaagtttc taaaggaaca gataaaactg gatgaaggct gggtaccttt ggagataatg
7201 ataaaattca acaggttgaa ccgtctaaca acagacttta atgtaattgt ggaagcattg
7261 agcaaatcca aggcagaact catggaaatc agtgaagata aaactaaaat cagaaggtct
7321 ccaagcaaac ccctacctga agtgactgat gagtataaaa atgatgtaaa aaacagatct
7381 gtttatatta aaggcttccc aactgatgca actcttgatg acataaaaga atggttagaa
7441 gataaaggtc aagtactaaa tattcagatg agaagaacat tgcataaagc atttaaggga
7501 tcaatttttg ttgtgtttga tagcattgaa tctgctaaga aatttgtaga gacccctggc
7561 cagaagtaca aagaaacaga cctgctaata cttttcaagg acgattactt tgccaaaaaa
7621 aatgaatctg gcggctcaaa aagaaccgcc gacggcagcg aattcgagtc tcccaagaag
7681 aagaggaaag tcggctctgg ccctgccgct aagagagtga agctggacgg atccggcgca
7741 acaaacttct ctctgctgaa acaagccgga gatgtcgaag agaatcctgg accgatggtg
7801 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
7861 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
7921 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
7981 accaccctga cctatggagt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
8041 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
8101 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
8161 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
8221 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
8281 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
8341 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
8401 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
8461 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaaccggtc
8521 atcatcacca tcaccattga gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt
8581 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
8641 ccactgtcct ttcctaataa aatgagaaaa ttgcatcgca ttgtctgagt aggtgtcatt
8701 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
8761 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
8821 cgataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
8881 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
8941 cctagggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
9001 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
9061 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
9121 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
9181 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
9241 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
9301 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
9361 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
9421 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
9481 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
9541 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
9601 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
9661 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
9721 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
9781 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
9841 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacactcag tggaacgaaa
9901 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
9961 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
10021 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
10081 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
10141 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
10201 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
10261 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
10321 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
10381 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
10441 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
10501 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
10561 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
10621 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
10681 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
10741 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
10801 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
10861 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
10921 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
10981 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatcgatctc
11041 ccgatcccct agggtcgact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
11101 atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta
11161 caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg
11221 cgctgcttcg cgatgtacgg gccagatata cgcgtt
//
| {
"Vector backbone": "pCMV",
"Backbone size w/o insert (bp)": "3402",
"Vector type": "Mammalian Expression",
"Selectable markers": "EGFP"
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "NEB Stable",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Improving prime editing with an endogenous small RNA-binding protein",
"Publication": "Nature. 2024 Apr 3. doi: 10.1038/s41586-024-07259-6.",
"DOI": "10.1038/s41586-024-07259-6.",
"Authors": "Yan J, Oyler-Castrillo P, Ravisankar P, Ward CC, Levesque S, Jing Y, Simpson D, Zhao A, Li H, Yan W, Goudy L, Schmidt R, Solley SC, Gilbert LA, Chan MM, Bauer DE, Marson A, Parsons LR, Adamson B /s---",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/38570691",
"PubMed ID": "PubMed 38570691"
} | {
"Gene/Insert name": "PE7-P2A-EGFP",
"Species": "Synthetic; SpCas9 is from Streptococcus pyogenes; MMLV_RT is from the Moloney murine leukemia virus; La(SSB)1-194 is from Homo sapiens",
"Insert Size (bp)": "7854",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "CMV",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Gibson Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "CMV Forward, T7",
"3′ sequencing primer": "BGHR"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pDONR221 Ub-NLRP1 L1215 | 166828 | pDONR221 encoding ubiquitin-tagged NLRP1-CT to generate a L1215 N-terminus (no stop) | Daniel Bachovchin | Hollingsworth et al Nature. 2021 Mar 17. pii: 10.1038/s41586-021-03350-4. doi: 10.1038/s41586-021-03350-4. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/328303/4fa92d13-8ec2-40f9-a0a7-f5273c7bc2f4/addgene-plasmid-166828-sequence-328303.gbk | full | LOCUS sequence-328303- 3567 bp DNA circular SYN 26-AUG-2024
DEFINITION pDONR221 encoding ubiquitin-tagged NLRP1-CT to generate a L1215
N-terminus (no stop).
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3567)
AUTHORS Hollingsworth LR, Sharif H, Griswold AR, Fontana P, Mintseris J,
Dagbay KB, Paulo JA, Gygi SP, Bachovchin DA, Wu H
TITLE DPP9 sequesters the C terminus of NLRP1 to repress inflammasome
activation.
JOURNAL Nature. 2021 Mar 17. pii: 10.1038/s41586-021-03350-4. doi:
10.1038/s41586-021-03350-4.
PUBMED 33731932
REFERENCE 2 (bases 1 to 3567)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nature.
2021 Mar 17. pii: 10.1038/s41586-021-03350-4. doi:
10.1038/s41586-021-03350-4."
FEATURES Location/Qualifiers
source 1..3567
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 92..109
/label=L4440
/note="L4440 vector, forward primer"
terminator complement(268..295)
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator complement(387..473)
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
primer_bind 522..544
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 536..553
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 537..553
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 569..668
/gene="mutant version of attL"
/label=attL1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
CDS 686..913
/codon_start=1
/product="human ubiquitin"
/label=ubiquitin
/translation="MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIF
AGKQLEDGRTLSDYNIQKESTLHLVLRLRGG"
protein_bind complement(1692..1791)
/gene="mutant version of attL"
/label=attL2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
primer_bind complement(1808..1827)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(1809..1827)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(1832..1848)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1832..1848)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(1950..1969)
/label=pENTR-R
/note="pENTR vectors, reverse primer"
CDS 1961..2770
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind complement(2028..2047)
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
rep_origin 2917..3505
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3406..3425
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
661 agcaggcttc gaaggagata gaaccatgca gatcttcgtc aagacgttaa ccggtaaaac
721 cataactctg gaagttgaac catccgatac catcgaaaac gttaaggcta aaattcaaga
781 caaggaagga attccacctg atcaacaaag attgatcttt gccggtaagc agctcgagga
841 cggtagaacg ctgtctgatt acaacattca gaaggagtcg accttacatc ttgtcttaag
901 actaagaggt ggtttgggag tcctcctgaa aatgatccat aatgccctgc gcttcattcc
961 cgtcacctct gtggtgttgc tttaccaccg cgtccatcct gaggaagtca ccttccacct
1021 ctacctgatc ccaagtgact gctccattcg gaaggccata gatgatctag aaatgaaatt
1081 ccagtttgtg cgaatccaca agccaccccc gctgacccca ctttatatgg gctgtcgtta
1141 cactgtgtct gggtctggtt cagggatgct ggaaatactc cccaaggaac tggagctctg
1201 ctatcgaagc cctggagaag accagctgtt ctcggagttc tacgttggcc acttgggatc
1261 agggatcagg ctgcaagtga aagacaagaa agatgagact ctggtgtggg aggccttggt
1321 gaaaccagga gatctcatgc ctgcaactac tctgatccct ccagcccgca tagccgtacc
1381 ttcacctctg gatgccccgc agttgctgca ctttgtggac cagtatcgag agcagctgat
1441 agcccgagtg acatcggtgg aggttgtctt ggacaaactg catggacagg tgctgagcca
1501 ggagcagtac gagagggtgc tggctgagaa cacgaggccc agccagatgc ggaagctgtt
1561 cagcttgagc cagtcctggg accggaagtg caaagatgga ctctaccaag ccctgaagga
1621 gacccatcct cacctcatta tggaactctg ggagaagggc agcaaaaagg gactcctgcc
1681 actcagcagc gacccagctt tcttgtacaa agttggcatt ataagaaagc attgcttatc
1741 aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc
1801 tgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc
1861 cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat
1921 aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga
1981 aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
2041 tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc
2101 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
2161 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
2221 tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt
2281 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
2341 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
2401 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
2461 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc
2521 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
2581 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
2641 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
2701 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
2761 gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg
2821 acgggacggc gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg
2881 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
2941 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
3001 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
3061 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
3121 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
3181 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
3241 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
3301 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
3361 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
3421 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
3481 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
3541 cttttgctgg ccttttgctc acatgtt
//
| {
"Vector backbone": "pDONR221",
"Backbone size w/o insert (bp)": "",
"Vector type": "Gateway shuttling vector",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Kanamycin, 50 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "DPP9 sequesters the C terminus of NLRP1 to repress inflammasome activation",
"Publication": "Nature. 2021 Mar 17. pii: 10.1038/s41586-021-03350-4. doi: 10.1038/s41586-021-03350-4.",
"DOI": "10.1038/s41586-021-03350-4.",
"Authors": "Hollingsworth LR, Sharif H, Griswold AR, Fontana P, Mintseris J, Dagbay KB, Paulo JA, Gygi SP, Bachovchin DA, Wu H /s---",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/33731932",
"PubMed ID": "PubMed 33731932"
} | {
"Gene/Insert name": "NLRP1",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "NLRP1",
"Promoter": "",
"Tag / Fusion Protein": "Ubiquitin (N terminal on insert)"
} | {
"Cloning method": "Gateway Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "M13 F",
"3′ sequencing primer": "M13 R"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pcDNA1 Flag ATXN1[30Q] | 33236 | Low | Huda Zoghbi | Skinner et al Nature. 1997 Oct 30;389(6654):971-4. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/82872/22359c4f-1105-44c7-b3b7-eb7ea88a0133/addgene-plasmid-33236-sequence-82872.gbk | partial | LOCUS sequence-82872-e 909 bp DNA linear UNA 25-AUG-2024
DEFINITION natural linear DNA
ACCESSION .
VERSION .
KEYWORDS .
SOURCE natural DNA sequence
ORGANISM unspecified
REFERENCE 1 (bases 1 to 909)
AUTHORS Skinner PJ, Koshy BT, Cummings CJ, Klement IA, Helin K, Servadio A,
Zoghbi HY, Orr HT
TITLE Ataxin-1 with an expanded glutamine tract alters nuclear
matrix-associated structures.
JOURNAL Nature. 1997 Oct 30;389(6654):971-4.
PUBMED 9353120
REFERENCE 2 (bases 1 to 909)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nature.";
date: "1997-10-30"; volume: "389(6654)"; pages: "971-4"
FEATURES Location/Qualifiers
source 1..909
/mol_type="genomic DNA"
/organism="unspecified"
primer_bind 42..61
/label=T7
/note="T7 promoter, forward primer"
promoter 42..60
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 205..228
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
ORIGIN
1 tggctactag agaacccact gtcttactgg cttatcgaaa ttaatacgac tcactatagg
61 gagacccaag cttggtaccg agctcggatc cactagtaac ggccgccagt gtgctggaat
121 tccgtgactt tccgtttatc agactaaaat cagagccatc cagacagtga aacagtcacc
181 gtggaggggg gacggcgaaa aatggactac aaagacgacg acgacaaaaa atccaaccaa
241 gagcggagca acgaatgcct gcctcccaag aagcgcgaga tccccgccac cagccggtcc
301 tccgaggaga aggcccctac cctgcccagc gacaaccacc gggtggaggg cacagcatgg
361 ctcccgggca accctggtgg ccggggccac gggggcggga ggcatgggcc ggcagggacc
421 tcggtggagc ttggtttaca acagggaata ggtttacaca aagcattgtc cacagggctg
481 gactactccc cgcccagcgc tcccaggtct gtccccgtgg ccaccacgct gcctgccgcg
541 tacgccaccc cgcagccagg gaccccggtg tcccccgtac agtacgctca cctgccgcac
601 accttccagt tcattgggtc ctcccaatac agtggaacct atgccagctt catcccatca
661 cagctgatcc ccccaaccgc caaccccgtc accagtgcag tggcctcggc cgcaggggcc
721 accactccat cccagcgctc ccagctggag gcctattcca ctctgctggc caacatgggc
781 agtctgagcc agacgccggg acacaaggct gagcagcagc agcagcagca gcagcagcag
841 cagcagcatc agcatcagca gcagcagcag cagcagcagc agcagcagca gcagcagcag
901 cacctcagc
//
| {
"Vector backbone": "pcDNA1.1",
"Backbone size w/o insert (bp)": "",
"Vector type": "Mammalian Expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH10B",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Ataxin-1 with an expanded glutamine tract alters nuclear matrix-associated structures",
"Publication": "Nature. 1997 Oct 30;389(6654):971-4.",
"DOI": "10.1038/40153",
"Authors": "Skinner PJ, Koshy BT, Cummings CJ, Klement IA, Helin K, Servadio A, Zoghbi HY, Orr HT",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/9353120",
"PubMed ID": "PubMed 9353120"
} | {
"Gene/Insert name": "Ataxin-1",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "2448",
"Mutation": "insert contains a stretch of 29 amino acids (27 Q interrupted by 2 H) and no stop codon",
"Entrez Gene": "ATXN1",
"Promoter": "CMV",
"Tag / Fusion Protein": "Flag (N terminal on backbone)"
} | {
"Cloning method": "Unknown",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "CMV-F",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pXF87 | 139027 | (Empty Backbone)MosSCI backbone for MosSCI-SapTrap assembly | Erik Griffin | Fan et al G3 (Bethesda). 2019 Dec 17. pii: g3.119.400822. doi: 10.1534/g3.119.400822. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/273781/04939a46-1468-4a0e-a5a0-f32edc43679d/addgene-plasmid-139027-sequence-273781.gbk | full | LOCUS sequence-273781- 7534 bp DNA circular SYN 26-AUG-2024
DEFINITION MosSCI backbone for MosSCI-SapTrap assembly.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7534)
AUTHORS Fan X, Henau S, Feinstein J, Miller SI, Han B, Frokjaer-Jensen C,
Griffin EE
TITLE SapTrap Assembly of Caenorhabditis elegans MosSCI Transgene Vectors.
JOURNAL G3 (Bethesda). 2019 Dec 17. pii: g3.119.400822. doi:
10.1534/g3.119.400822.
PUBMED 31848219
REFERENCE 2 (bases 1 to 7534)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "G3
(Bethesda). 2019 Dec 17. pii: g3.119.400822. doi:
10.1534/g3.119.400822."
FEATURES Location/Qualifiers
source 1..7534
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter complement(199..303)
/gene="bla"
/label=AmpR promoter
primer_bind 371..389
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(427..449)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
protein_bind 772..792
/gene="mutant version of attB"
/label=attB4
/bound_moiety="BP Clonase(TM)"
/note="core recombination site for the Gateway(R) BP
reaction"
protein_bind 2130..2154
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
CDS complement(2300..2311)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind 4251..4268
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 4252..4268
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4481..4497)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4481..4497)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
protein_bind complement(4504..4528)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
protein_bind complement(5956..5976)
/gene="mutant version of attB"
/label=attB3
/bound_moiety="BP Clonase(TM)"
/note="core recombination site for the Gateway(R) BP
reaction"
rep_origin complement(6113..6701)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6193..6212)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(join(6872..7534,1..198))
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGSDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 7495..7514
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
ORIGIN
1 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
61 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
121 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
181 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
241 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
301 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
361 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
421 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
481 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
541 taactatgcg gcatcagagc agattgcact gagagtgcac catatgcggt gtgaaatacc
601 gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa
661 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
721 atgtgctgca aggcgattaa gttgggtaac gccagggatc cgccaagcta tcaactttgt
781 atagaaaagt tgcttgtcgg attatgggca cttcttttat cctctacttt ctttctagaa
841 aatgcaaatg ttgagtcaga aattgctgaa tggagtacaa ctgcaatggg tttagcttat
901 gtttctggaa caattactgg tggaattgtg attgaaaagt aaggttatat ttcatttcaa
961 taaatgttac cacagtataa caaaaacaag ttacattgtt cagagtatgt ctgaatgtat
1021 tattattatt gtatttgtgc atcagtactg tagttgtact gaagaagggt ttaaaataat
1081 ctcaacgttt ttaagacgag ttagtgagaa tacggtcgca gttaggatgt ttggacatag
1141 ttttattaga taatcgtgaa ctaactcact ataaaaaccc gctaactcat tgaataagat
1201 agttttaaac gttcaaaatt tgaagacaaa attgtatgaa taagtttaca gagttggtcg
1261 acgcaagctt ctccttctct tcacatttct caataattta gctctaattg cattcgtatt
1321 cttcgcaaaa atacgtattc tgattgaccc gatgaaatac ggatgcctag ttgcattgat
1381 catctacggt tatacttatg ggtgaggttt ttcaaatata ttttgattta atcgaataat
1441 gaaatttcag aactggagtt ggtccaattt cgtggtttat atcctctgaa ctggtacctc
1501 aaaaacatcg tagtattgct caatctgtgg catattcaat gaatactgta atggtggtta
1561 tcagtacatt tacagtatta cctctatatt ccgttattgg tagttatgct ttccttattc
1621 tatattcaat tccttcattt ataagtatgc ttatattatt tcgctactta ccggaaacca
1681 aaggacgaga gattcatgat attgtgaatg aacttaaaaa aaaatgaaaa cctttgaaaa
1741 cgttcatacg gccgaaattt tacttgatcg agccaattga acttttgtta agacttacag
1801 aaaagtgaga gattttgtta caaaaaagtg agaaaaaagc gaaaatgaac aggaaaaaga
1861 agaaaaatac acccgaagta tcatataaaa attaaaaaga ttgtttgacc tggcggaact
1921 tttttgcaaa aactctgaat atccatggca catgcttcgt gcaaaacagg atccaaatga
1981 atatcaacaa gagattcttg aagacgacga gccacttgct cagagcattg cttatctcga
2041 atgagaccct tgttaaaatc atttttcaaa cattatatga cattgtcgaa atgtcctcct
2101 gattccatga tggtagcaaa ctcacttcgt caagtttgta caaaaaagca ggcttgagcc
2161 aatttatcca agtccttgta aaaagtattc gaaaattgtt aacggataaa atgtttatta
2221 taatatcaaa aacaatttgt cagttgacca ctttttgata tatttgacag aaaccgggat
2281 gaattggctc aaagttaggg cgcccttcta ttacagggtt tctgataaca aaccggttat
2341 taactcccaa caagggatga tttcaattca tcatgctcaa attgacccaa attaagttac
2401 atgacaaatt catcgccctt ttcaactctt cttggctcat catctgttat tctgttccta
2461 ttctgtctgc accctatacc cctttgcata ctctctcgtc attcctttct tggatagtgt
2521 cttcttctcc ccagctctgc tacttctatg acttgcgccc gctgcttttc cgctcgttcc
2581 tctcctctct gacgtcttct tctgcttctt ttctccttcc tttttccctt ctcaatcttc
2641 ctctttttcc catttctgtc aatcattcga agaagaagaa gaagaagacc ctcattcatt
2701 tcattttttt ttctgtcggt gtgtgctgct cggttaagag tgagctctct attccacgtc
2761 ttcttctttt tcttcttcga ttcgaatcaa tcactccacc aaaacgcatt cgtttttggg
2821 attcaccccg cggttcgcaa taggtttctt tttcaaatta ttagcgttat aaatagaaaa
2881 atggtggagt ttcaataaaa atgataattt acaaagtgta ttttgattac atgtactcaa
2941 aaaggttgaa atttttcata ccagttttcc ggaaatccat ctgatatcat tatcgtattt
3001 ttctttttaa aaatgttttc aaaaaaaaac aaaatatagc tggttatttg gcaccctcta
3061 attaccattt tcctgtcaca ccacactctt ttccttcttc cacttctttc cgttttcagc
3121 cgcttccaac caaaccgata tgaaagccga gcaacaacga tcgattccac ccggctcggc
3181 gaccttcccg tcgcaggtga gactcagaaa actagagaaa ccgctcaact aactcttgat
3241 atccgatttc attcttttct cttttctttt tgttgaactt ttccatattt ccagatgcca
3301 cggccaccac caagcaccga acaaggaatc acaacggaat cggagcttgc gaagaaagct
3361 caaatcactc cgaacgacgt tttagcactt ccgggaatca ctcaaggtat cttgctccct
3421 ttggatttca gtttaaaaca taaatttagg attcttatgc tccccatctg cgaacatcta
3481 taacatcgag ttcaccaagt tccaaatccg tgatctggac actgagcaag tgcttttcga
3541 gatcgccaaa ccggagaacg atcaggagaa tgatgagtcg ccacaggagt cggcaagata
3601 cgtgcgttat agatttgctc caaacttttt gaaactcaaa acggtcggag caactgtgga
3661 attcaaagta ggagacatcc caatccatca tttccgaatg atcgaacgtc acttcttcaa
3721 agatcgcctt ctgaagtgtt ttgactttga attcggattc tgtattccga attcacgaaa
3781 caactgtgaa catatctatg agttccctca actctctcaa caactcagtg agtcattatt
3841 ctaaaaagta caattcaaaa gactaatctc tttttcagtg gacgacatga tcaacaatcc
3901 aaacgagact cgttctgaca gcttctattt cgtcgataac aaactcgtca tgcacaacaa
3961 agccgactac tcatatgatg cataaatatt taatacaaaa atgttctgga taattattct
4021 gtcgaataga aaaaaaactc caaatgtgat taaattccaa taattcctgt ctagtttgtt
4081 ccttcccctt ccccttctca tgttcaatgc attcctaagc ttttcagttc cccccttgtt
4141 tctatatttt ttcgctgtcc tgtcacactc gctaaaaaca ctaatcacac ggaaatctgt
4201 tttcaataaa aactccaact ttaactcatt ttcaatttca actgcacggg tgtaaaacga
4261 cggccagtcc gtaatacgac tcacttaagg ccttgactag agggtaccag agctcaccta
4321 ggtatctaga accggtgacg tcaccatggg aagcttcgtg gatccagata tcctgcagga
4381 attcctcgag tggcgaagag cccatggatc ccatatggaa ttctgcaggc ctgctcttcg
4441 gtaactagtg agtcgtatta cgtagcttgg cgtaatcatg gtcatagctg tttcctgccc
4501 gtgacccagc tttcttgtac aaagtggtaa gtgcaagtaa gatcagtgtt tgtttcgata
4561 tcagtctgtt tcgtaacggt cttctgtata actacaaaaa agaataaaaa accgtatcac
4621 aactgagaaa aaagacaaaa cttattgaac agtgttcctt tcaaaaaatt attttgtaaa
4681 aaatcgagtt tttctccaaa ataacgcacg caatcaaaaa caagaaaact aacgtgttct
4741 cctgggcgtg gaactcctta tcaagattac cattattgtc tgtcagaatt gatatttcaa
4801 ttatttcata tctagacaaa cccgggtgtc ttgcatttat tagtactgtg agtttacata
4861 caaacgaggc ttactcagat aacatagagt cacctctaat tagcaaaaac ccatttctaa
4921 ttagctattt cgaccagttc taaagtcacc gtaattagaa atggtttttg tagcttttct
4981 aattagaaat gaccccgaga taagtgaaat atggcatttt ctgggttttc tagtaagaaa
5041 tgatcccgca cgtagtgatt ccgcgttaca agccaaattc aaattaaaaa agtatattac
5101 acaattcaag ttacattatt ttcaggaaac aatggcgaag tgcaaagcag agattcaaaa
5161 ggaggaaaga gacaacagat atgaacgaag acttcttgaa aagagaaaga gacatgaggt
5221 gagtaaccat gataataaat aatcaagtaa gggctataga tttataaatt ctggagttaa
5281 cagtttcgaa aaaaagtagt tttcatatgt gcgaaggaat atcaaaaata ggggaatagc
5341 ctaaacgagt gagaaaatta aatgtttgtt attttttcaa aaatgtctgc agggccgtta
5401 tcctttcaaa tgcactctat caaacataga tcatatccat catatcttat ccagaattta
5461 tatcatatcc aagattgaga gcgtttcaag gaaaatctca aacaaccata caattttgta
5521 gatcgacaac aaggcctcca cttcttcatc gatggcctct gcagcacctg caccaccaag
5581 tttgtatttt tcatattaat ataaagtgag ccacgcatag tcgtttttag gatttttgcc
5641 agatcgatgt cgcgcaaaca gactgttttt ttttctttga aagccgtgcg tgacatgggc
5701 cacggattgt aacgccgcgc tggatggata aaaatttatt cagtttttta gcctttacaa
5761 aatatggttc gactactttc tttattttta catttttata ggtgctcatt gagctgagat
5821 attttgggta taatcggaaa atcgctgatt ttggtcattt ttctgcacta aagattacac
5881 aaaaagttca gtatttagat tcattgttga aaattgggga tgatctacag atacatatcg
5941 ccgtgttcaa aaacacaact ttattataca tagttgataa ttcgatggta tcagctcact
6001 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
6061 caaaaggcca gcaaaagacc aggaaccgta aaaaggccgc gttgctggcg tttttccata
6121 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
6181 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
6241 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
6301 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
6361 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
6421 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
6481 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
6541 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
6601 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
6661 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
6721 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
6781 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
6841 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
6901 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
6961 ctacgatacg ggagggctta ccatctgacc ccagtgctgc aatgataccg cgagacccac
7021 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
7081 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
7141 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
7201 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
7261 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
7321 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
7381 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
7441 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
7501 ccgcgccaca tagcagaact ttaaaagtgc tcat
//
| {
"Vector backbone": "original backbone pCFJ350; 2 unwanted SapI sites mutated, 2 SapI sites introduced for SapTrap assembly",
"Backbone size w/o insert (bp)": "",
"Vector type": "Worm Expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "SapTrap Assembly of Caenorhabditis elegans MosSCI Transgene Vectors",
"Publication": "G3 (Bethesda). 2019 Dec 17. pii: g3.119.400822. doi: 10.1534/g3.119.400822.",
"DOI": "10.1534/g3.119.400822.",
"Authors": "Fan X, Henau S, Feinstein J, Miller SI, Han B, Frokjaer-Jensen C, Griffin EE /g",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/31848219",
"PubMed ID": "PubMed 31848219"
} | {
"Gene/Insert name": "None",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
UbcH10 | 8504 | Joan Ruderman | Townsley et al Proc Natl Acad Sci U S A 1997 Mar 18;94(6):2362-7. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/17080/5b6e7a68-bd99-40be-abb1-7748a181a1c4/addgene-plasmid-8504-sequence-17080.gbk | partial | LOCUS sequence-17080-e 908 bp DNA linear UNA 26-AUG-2024
DEFINITION natural linear DNA
ACCESSION .
VERSION .
KEYWORDS .
SOURCE natural DNA sequence
ORGANISM unspecified
REFERENCE 1 (bases 1 to 908)
AUTHORS Townsley FM, Aristarkhov A, Beck S, Hershko A, Ruderman JV
TITLE Dominant-negative cyclin-selective ubiquitin carrier protein
E2-C/UbcH10 blocks cells in metaphase.
JOURNAL Proc Natl Acad Sci U S A 1997 Mar 18;94(6):2362-7.
PUBMED 9122200
REFERENCE 2 (bases 1 to 908)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Proc Natl
Acad Sci U S A"; date: "1997-03-18"; volume: "94(6)"; pages:
"2362-7"
FEATURES Location/Qualifiers
source 1..908
/mol_type="genomic DNA"
/organism="unspecified"
ORIGIN
1 ttaactttaa gaaggagata tacatatggc ttcccaaaac cgcgacccag ccgccactag
61 cgtcgccgcc gcccgtaaag gagctgagcc gagcgggggc gccgcccggg gtccggtggg
121 caaaaggcta cagcaggagc tgatgaccct catgatgtct ggcgataaag ggatttctgc
181 cttccctgaa tcagacaacc ttttcaaatg ggtagggacc atccatggag cagctggaac
241 agtatatgaa gacctgaggt ataagctctc gctagagttc cccagtggct acccttacaa
301 tgcgcccaca gtgaagttcc tcacgccctg ctatcacccc aacgtggaca cccagggtaa
361 catatgcctg gacatcctga aggaaaagtg gtctgccctg tatgatgtca ggaccattct
421 gctctccatc cagagccttc taggagaacc caacattgat agtcccttga acacacatgc
481 tgccgagctc tggaaaaacc ccacagcttt taagaagtac ctgcaagaaa cctactcaaa
541 gcaggtcacc agccaggagc cctgataagc ttatcgatga taagctgtca aacatgagaa
601 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
661 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
721 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
781 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
841 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
901 gggaagct
//
| {
"Vector backbone": "pT7-7",
"Backbone size w/o insert (bp)": "2488",
"Vector type": "Bacterial Expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Dominant-negative cyclin-selective ubiquitin carrier protein E2-C/UbcH10 blocks cells in metaphase",
"Publication": "Proc Natl Acad Sci U S A 1997 Mar 18;94(6):2362-7.",
"DOI": "10.1073/pnas.94.6.2362",
"Authors": "Townsley FM, Aristarkhov A, Beck S, Hershko A, Ruderman JV",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/9122200",
"PubMed ID": "PubMed 9122200"
} | {
"Gene/Insert name": "UbcH10",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "543",
"Mutation": "",
"Entrez Gene": "UBE2C",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "NdeI (not destroyed)",
"3′ cloning site": "HindIII (not destroyed)",
"5′ sequencing primer": "T7",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
||
pMXs-3XMyc-EGFP-OMP25 | 83355 | High | For tagging mitochondria with Myc epitopes (Control-MITO construct) | David Sabatini | Chen et al Cell. 2016 Aug 25;166(5):1324-1337.e11. doi: 10.1016/j.cell.2016.07.040. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/418753/edc85173-eb81-4e07-b33a-417ba11ac4a8/addgene-plasmid-83355-sequence-418753.gbk | full | LOCUS sequence-418753- 6605 bp DNA circular SYN 25-AUG-2024
DEFINITION For tagging mitochondria with Myc epitopes (Control-MITO construct).
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6605)
AUTHORS Chen WW, Freinkman E, Wang T, Birsoy K, Sabatini DM
TITLE Absolute Quantification of Matrix Metabolites Reveals the Dynamics
of Mitochondrial Metabolism.
JOURNAL Cell. 2016 Aug 25;166(5):1324-1337.e11. doi:
10.1016/j.cell.2016.07.040.
PUBMED 27565352
REFERENCE 2 (bases 1 to 6605)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1016/j.cell.2016.07"; journalName: "Cell"; date: "2016-08-25-
25"; volume: "166"; issue: "5"; pages: "1324-1337.e11"
FEATURES Location/Qualifiers
source 1..6605
/mol_type="other DNA"
/organism="synthetic DNA construct"
LTR 22..615
/note="long terminal repeat from Moloney murine leukemia
virus"
misc_feature 678..1035
/label=MMLV Psi
/note="packaging signal of Moloney murine leukemia virus
(MMLV)"
CDS 1094..1510
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
primer_bind 1440..1462
/label=pLXSN 5'
/note="Murine stem cell virus, forward primer. Also called
MSCV"
primer_bind 1478..1494
/label=pBABE 5'
/note="Psi packaging signal, forward primer for pBABE
vectors"
misc_feature 1520..1894
/label=pol region
/note="Moloney murine leukemia virus (MMLV) pol region
containing the splice acceptor site"
primer_bind 1832..1848
/label=pBMN 5'
/note="MMLV sequence, for inserts in pBMN retroviral
vector"
regulatory 1959..1968
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 1968..1997
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 2010..2039
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 2052..2081
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 2094..2807
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
FVTAAGITLGMDELYK"
CDS 2094..2807
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
FVTAAGITLGMDELYK"
primer_bind complement(2136..2157)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(2397..2416)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 2744..2765
/label=EGFP-C
/note="EGFP, forward primer"
misc_feature 2957..3543
/label=IRES2
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(3124..3141)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 3351..3370
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 3544..3939
/codon_start=1
/gene="Aspergillus terreus BSD"
/product="blasticidin S deaminase"
/label=BSD
/note="confers resistance to blasticidin"
/translation="MVPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
AIVKDSDGQPTAVGIRELLPSGYVWEG"
LTR 4110..4579
/note="long terminal repeat from Moloney murine leukemia
virus"
primer_bind complement(4709..4726)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4880..5468)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4960..4979)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5632..6492)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 6255..6274
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6493..6597)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 cccgaaaagt gccacctgca taatgaaaga ccccacctgt aggtttggca agctagctta
61 agtaacgcca ttttgcaagg catggaaaaa tacataactg agaatagaaa agttcagatc
121 aaggtcagga acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag
181 ttcctgcccc ggctcagggc caagaacaga tggaacagct gaatatgggc caaacaggat
241 atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg
301 gtccagccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct
361 gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg
421 cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggc gcgccagtcc
481 tccgattgac tgagtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc
541 cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag
601 cgggggtctt tcatttgggg gctcgtccgg gatcgggaga cccctgccca gggaccaccg
661 acccaccacc gggaggtaag ctggccagca acttatctgt gtctgtccga ttgtctagtg
721 tctatgactg attttatgcg cctgcgtcgg tactagttag ctaactagct ctgtatctgg
781 cggacccgtg gtggaactga cgagttcgga acacccggcc gcaaccctgg gagacgtccc
841 agggacttcg ggggccgttt ttgtggcccg acctgagtcc aaaaatcccg atcgttttgg
901 actctttggt gcacccccct aataggaggg atatgtggtt ctggtaggag acgagaacct
961 aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tgggaccgaa gccgcgccgc
1021 gcgtcttgtc tgctgcagca tcgttctgtg ttgtctctgt ctgactgtgt ttctgtattt
1081 gtctgaaaat tagggccaga ctgttaccac tcccttaagt ttgaccttag gtcactggaa
1141 agatgtcgag cggatcgctc acaaccagtc ggtagatgtc aagaagagac gttgggttac
1201 cttctgctct gcagaatggc caacctttaa cgtcggatgg ccgcgagacg gcacctttaa
1261 ccgagacctc atcacccagg ttaagatcaa ggtcttttca cctggcccgc atggacaccc
1321 agaccaggtc ccctacatcg tgacctggga agccttggct tttgaccccc ctccctgggt
1381 caagcccttt gtacacccta agcctccgcc tcctcttcct ccatccgccc cgtctctccc
1441 ccttgaacct cctcgttcga ccccgcctcg atcctccctt tatccagccc tcactccttc
1501 tctaggcgcc cccatatggc catatgagat cttatatggg gcacccccgc cccttgtaaa
1561 cttccctgac cctgacatga caagagttac taacagcccc tctctccaag ctcacttaca
1621 ggctctctac ttagtccagc acgaagtctg gagacctctg gcggcagcct accaagaaca
1681 actggaccga ccggtggtac ctcaccctta ccgagtcggc gacacagtgt gggtccgccg
1741 acaccagact aagaacctag aacctcgctg gaaaggacct tacacagtcc tgctgaccac
1801 ccccaccgcc ctcaaagtag acggcatcgc agcttggata cacgccgccc acgtgaaggc
1861 tgccgacccc gggggtggac catcctctag actgccggat ctagctagtt aattaaggat
1921 cccagtgtgg tggtacggga attcctgcag gcctcgaggc caccatggag cagaagctga
1981 tttctgagga agatctgggc acaggatccg aacagaaact gatttctgag gaagatctgg
2041 gcagcgccgg agagcagaag ctgatttctg aagaggatct gggagggagc ggcgtgagca
2101 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
2161 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
2221 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
2281 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
2341 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
2401 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
2461 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
2521 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
2581 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
2641 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
2701 cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
2761 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagccg cggcatcgag
2821 gcgacggaga gcctagtgga gttcctgtag ctgtggtgct gctgccagtg tttgccctta
2881 ccctggtagc agtttgggcc ttcgtgagat accgaaagca gctctgagcg gccgctacgt
2941 aaattccgcc cccccccccc tctccctccc ccccccctaa cgttactggc cgaagccgct
3001 tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg ccgtcttttg
3061 gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct aggggtcttt
3121 cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg
3181 aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg aaccccccac
3241 ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct gcaaaggcgg
3301 cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa tggctctcct
3361 caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt atgggatctg
3421 atctggggcc tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa aaacgtctag
3481 gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa tatggccaca
3541 accatggttc ctttgtctca agaagaatcc accctcattg aaagagcaac ggctacaatc
3601 aacagcatcc ccatctctga agactacagc gtcgccagcg cagctctctc tagcgacggc
3661 cgcatcttca ctggtgtcaa tgtatatcat tttactgggg gaccttgtgc agaactcgtg
3721 gtgctgggca ctgctgctgc tgcggcagct ggcaacctga cttgtatcgt cgcgatcgga
3781 aatgagaaca ggggcatctt gagcccctgc ggacggtgcc gacaggtgct tctcgatctg
3841 catcctggga tcaaagccat agtgaaggac agtgatggac agccgacggc agttgggatt
3901 cgtgaattgc tgccctctgg ttatgtgtgg gagggctaag tcgacgataa aataaaagat
3961 tttatttagt ctccagaaaa aggggggaat gaaagacccc acctgtaggt ttggcaagct
4021 agcttaagta acgccatttt gcaaggcatg gaaaaataca taactgagaa tagagaagtt
4081 cagatcaagg tcaggaacag atggaacagc tgaatatggg ccaaacagga tatctgtggt
4141 aagcagttcc tgccccggct cagggccaag aacagatgga acagctgaat atgggccaaa
4201 caggatatct gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggtcccca
4261 gatgcggtcc agccctcagc agtttctaga gaaccatcag atgtttccag ggtgccccaa
4321 ggacctgaaa tgaccctgtg ccttatttga actaaccaat cagttcgctt ctcgcttctg
4381 ttcgcgcgct tctgctcccc gagctcaata aaagagccca caacccctca ctcggggcgc
4441 cagtcctccg attgactgag tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt
4501 tgcatccgac ttgtggtctc gctgttcctt gggagggtct cctctgagtg attgactacc
4561 cgtcagcggg ggtctttcac atgcagcatg tatcaaaatt aatttggttt tttttcttaa
4621 gtatttacat taaatggcca tagttgcatt aatgaatcgg ccaacgcgcg gggagaggcg
4681 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
4741 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
4801 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
4861 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
4921 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
4981 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
5041 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
5101 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
5161 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
5221 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
5281 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
5341 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
5401 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
5461 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
5521 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttgc
5581 ggccggccgc aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
5641 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
5701 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
5761 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
5821 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
5881 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
5941 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
6001 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
6061 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
6121 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
6181 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
6241 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
6301 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
6361 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
6421 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
6481 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
6541 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
6601 atttc
//
| {
"Vector backbone": "pMXs rtv-016",
"Backbone size w/o insert (bp)": "5600",
"Vector type": "Mammalian Expression, Retroviral",
"Selectable markers": "Blasticidin"
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "30°C",
"Growth Strain(s)": "NEB Stable",
"Copy number": "Unknown"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Absolute Quantification of Matrix Metabolites Reveals the Dynamics of Mitochondrial Metabolism",
"Publication": "Cell. 2016 Aug 25;166(5):1324-1337.e11. doi: 10.1016/j.cell.2016.07.040.",
"DOI": "10.1016/j.cell.2016.07.040.",
"Authors": "Chen WW, Freinkman E, Wang T, Birsoy K, Sabatini DM /jcell",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/27565352",
"PubMed ID": "PubMed 27565352"
} | {
"Gene/Insert name": "3XMyc-EGFP-OMP25",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
TFORF0361 | 142255 | Lentiviral vector for overexpressing transcription factor ORFs with unique 24-bp barcodes. Barcodes facilitate identification of transcription factors in pooled screens. | Feng Zhang | Joung et al Cell. 2023 Jan 5;186(1):209-229.e26. doi: 10.1016/j.cell.2022.11.026. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/395748/38327c22-f513-4b40-8482-dc6f41140ca2/addgene-plasmid-142255-sequence-395748.gbk | full | LOCUS sequence-395748- 10762 bp DNA circular SYN 25-AUG-2024
DEFINITION Lentiviral vector for overexpressing transcription factor ORFs with
unique 24-bp barcodes. Barcodes facilitate identification of
transcription factors in pooled screens..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10762)
AUTHORS Joung J, Ma S, Tay T, Geiger-Schuller KR, Kirchgatterer PC, Verdine
VK, Guo B, Arias-Garcia MA, Allen WE, Singh A, Kuksenko O, Abudayyeh
OO, Gootenberg JS, Fu Z, Macrae RK, Buenrostro JD, Regev A, Zhang F
TITLE A transcription factor atlas of directed differentiation.
JOURNAL Cell. 2023 Jan 5;186(1):209-229.e26. doi:
10.1016/j.cell.2022.11.026.
PUBMED 36608654
REFERENCE 2 (bases 1 to 10762)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1016/j.cell.2022.11"; journalName: "Cell"; date: "2023-01-5- 5";
volume: "186"; issue: "1"; pages: "209-229.e26"
FEATURES Location/Qualifiers
source 1..10762
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 5..231
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 232..412
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 459..584
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1077..1310
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1495..1539
/codon_start=1
/product="antigenic peptide corresponding to amino acids
655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
et al., 2013)"
/label=gp41 peptide
/note="recognized by the 2H10 single-chain llama nanobody"
/translation="KNEQELLELDKWASL"
promoter 1770..1980
/label=SV40 promoter
/note="SV40 early promoter"
promoter 1792..1988
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 1839..1974
/label=SV40 ori
/note="SV40 origin of replication"
CDS 2020..2622
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGAS"
primer_bind complement(2020..2039)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 2516..2536
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
misc_feature 2665..2782
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2920..4098
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 3151..4089
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 4046..4066
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
misc_feature 6619..7207
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(6672..6692)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(7090..7101)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 7279..7512
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 7590..7711
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(7627..7646)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 7681..7700
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 7751..7886
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(7906..7925)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(7907..7925)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(7935..7952)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(7935..7951)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(7944..7966)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 8093..8548
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(8180..8199)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 8390..8411
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 8574..8678
/gene="bla"
/label=AmpR promoter
CDS 8679..9539
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(8897..8916)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 9710..10298
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 10199..10218
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 10452..10469
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 10586..10607
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 10622..10652
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 10660..10676
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
primer_bind 10665..10687
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 10684..10700
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 10684..10700
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 10719..10739
/label=T3
/note="T3 promoter, forward primer"
promoter 10721..10739
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
1 ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat gagttagcaa
61 catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt aaggtggtac
121 gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg aaccactgaa
181 ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa cgggtctctc
241 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
301 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
361 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
421 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
481 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
541 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
601 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
661 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
721 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
781 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
841 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
901 gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg
961 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta
1021 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga
1081 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg
1141 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg
1201 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc
1261 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg
1321 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat
1381 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac
1441 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat
1501 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca
1561 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga
1621 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg
1681 tttcagaccc acctcccaac cccgagggga cccatgcatt gcatctcaat tagtcagcaa
1741 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcgtctca
1801 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
1861 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
1921 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
1981 tttgcaaaaa gctttctaga ggtaccaccg gagcttacca tgaccgagta caagcccacg
2041 gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc
2101 gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc
2161 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg
2221 gacgacggcg ccgccgtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg
2281 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa
2341 cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc
2401 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga
2461 gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac
2521 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga
2581 ccgcgcacct ggtgcatgac ccgcaagccc ggtgccagtt aacgaattct agatcttgag
2641 acaaatggca gtattcatcc acaattttaa aagaaaaggg gggattgggg ggtacagtgc
2701 aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat tacaaaaaca
2761 aattacaaaa attcaaaatt ttcgggttta ttacagggac agcagagatc cactttggcg
2821 ccggctcgag gcctgcaggt gcaaagatgg ataaagtttt aaacagagag gaatctttgc
2881 agctaatgga ccttctaggt cttgaaagga gtgggaattg gctccggtgc ccgtcagtgg
2941 gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacc
3001 ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc
3061 ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt
3121 tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc ccgcgggcct
3181 ggcctcttta cgggttatgg cccttgcgtg ccttgaatta cttccacctg gctgcagtac
3241 gtgattcttg atcccgagct tcgggttgga agtgggtggg agagttcgag gccttgcgct
3301 taaggagccc cttcgcctcg tgcttgagtt gaggcctggc ctgggcgctg gggccgccgc
3361 gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc tctagccatt
3421 taaaattttt gatgacctgc tgcgacgctt tttttctggc aagatagtct tgtaaatgcg
3481 ggccaagatc tgcacactgg tatttcggtt tttggggccg cgggcggcga cggggcccgt
3541 gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc gagaatcgga
3601 cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg gcctcgcgcc gccgtgtatc
3661 gccccgccct gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc ggaaagatgg
3721 ccgcttcccg gccctgctgc agggagctca aaatggagga cgcggcgctc gggagagcgg
3781 gcgggtgagt cacccacaca aaggaaaagg gcctttccgt cctcagccgt cgcttcatgt
3841 gactccacgg agtaccgggc gccgtccagg cacctcgatt agttctcgag cttttggagt
3901 acgtcgtctt taggttgggg ggaggggttt tatgcgatgg agtttcccca cactgagtgg
3961 gtggagactg aagttaggcc agcttggcac ttgatgtaat tctccttgga atttgccctt
4021 tttgagtttg gatcttggtt cattctcaag cctcagacag tggttcaaag tttttttctt
4081 ccatttcagg tgtcgtgagg ctagcaccgc caccatgcac ggagggcgga gctgcggccc
4141 gaggacgcga cgcgagccca gttccggcga ggaggccgcg ccagtgacag cgatggcggc
4201 ggagtcggcg ctccaagttg tggagaagct gcaggcgcgc ctggccgcga acccggaccc
4261 taagaagcta ttgaaatatt tgaagaaact ctccaccctg cctattacag tagacattct
4321 tgcggagact ggggttggga aaacagtaaa tagcttgcga aaacacgagc atgttggaag
4381 ctttgccagg gacctagtgg cccagtggaa gaagctggtt cctgtggaac gaaatgctga
4441 gcctgatgaa caggactttg agaagagcaa ttcccgaaag cgccctcggg atgccctgca
4501 gaaggaggag gagatggagg gggactacca agaaacctgg aaagccacgg ggagccgatc
4561 ctatagccct gaccacaggc agaagaaaca taggaaactc tcggagctcg agagacctca
4621 caaagtgtct cacggtcatg agaggagaga tgagagaaag aggtgtcaca gaatgtcacc
4681 aacttactct tcagaccctg agtcttctga ttatggccat gttcaatccc ctccatcttg
4741 taccagtcct catcagatgt acgtcgacca ctacagatcc ctggaggagg accaggagcc
4801 cattgtttca caccagaagc ctgggaaagg ccacagcaat gcctttcagg acagactcgg
4861 ggccagccaa gaacgacacc tgggtgaacc ccatgggaaa ggggttgtga gtcaaaacaa
4921 ggagcacaaa tcttcccaca aggacaaacg ccccgtggat gccaagagtg atgagaaggc
4981 ctctgtggtg agcagagaga aatcacacaa ggccctctcc aaagaggaga accgaaggcc
5041 accctcaggg gacaatgcaa gggagaaacc gccctctagt ggcgtaaaga aagagaagga
5101 cagagagggc agcagcctga agaagaagtg tttgcctccc tcagaggccg cttcagacaa
5161 ccacctgaaa aagccaaagc acagagaccc agagaaagcc aaattggaca aaagcaagca
5221 aggtctggac agctttgaca caggaaaagg agcaggagac ctgttgccca aggtaaaaga
5281 gaagggttct aacaacctaa agactccaga agggaaagtc aaaactaatt tggatagaaa
5341 gtcactgggc tccctcccta aagttgagga gacagatatg gaggatgaat tcgagcagcc
5401 aaccatgtct tttgaatcct acctcagcta tgaccagccc cggaagaaaa agaaaaagat
5461 tgtgaaaact tcagccacgg cacttggaga taaaggactt aaaaaaaatg actctaaaag
5521 cactggtaaa aacttggact cagttcagaa attacccaag gtgaacaaaa ccaagtcaga
5581 gaagccggct ggagctgatt tagccaagct gagaaaggtg cctgatgtgt tgccagtgtt
5641 gccagacctc ccgttacccg cgatacaggc caattaccgt ccactgcctt ccctcgagct
5701 gatatcctcc ttccagccaa agcgaaaagc gttctcttca ccccaggaag aagaagaagc
5761 tggatttact gggcgcagaa tgaattccaa gatgcaggtg tattctggtt ccaagtgtgc
5821 ctatctccct aaaatgatga ccttgcacca gcaatgcatc cgagtactta aaaacaacat
5881 cgattcaatc tttgaagtgg gaggagtccc atactctgtt cttgaacccg ttttggagag
5941 gtgtacacct gatcagctgt atcgcataga ggaatacaat catgtattaa ttgaagaaac
6001 agatcaatta tggaaagttc attgtcaccg agactttaag gaagaaagac ccgaagagta
6061 tgagtcgtgg cgagagatgt acctgcggct tcaggacgcc cgagagcagc ggctacgagt
6121 actaacaaag aatatccagt tcgcacatgc caataagccc aaaggccgac aagcaaagat
6181 ggcctttgtc aactctgtgg ccaagccacc tcgtgacgtc cggaggaggc aggaaaagtt
6241 tggaacggga ggagcagctg tccctgagaa aatcaagatc aagccagccc cgtaccccat
6301 gggaagcagc catgcttccg ccagtagcat cagctttaac cccagccctg aggagccggc
6361 ctatgatggc ccaagcacca gcagtgccca cttggcacca gtggtcagca gcactgtttc
6421 ctatgatcct aggaaaccca ctgtgaagaa aattgcccca atgatggcca agacaattaa
6481 agctttcaag aacagattct cccgacgata gtaatgaact agtccgtaac ttgaaagtat
6541 ttcgatttct tggctttata tatcttgtgg aaaggacgac cgtcttcagt catctggatc
6601 ctaacgcgtt aagtcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt
6661 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
6721 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
6781 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
6841 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
6901 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
6961 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
7021 tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
7081 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
7141 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
7201 tccccgcgtc gactttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt
7261 taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagacaa gatctgcttt
7321 ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
7381 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
7441 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
7501 aaatctctag cagtacgtat agtagttcat gtcatcttat tattcagtat ttataacttg
7561 caaagaaatg aatatcagag agtgagagga acttgtttat tgcagcttat aatggttaca
7621 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
7681 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gctctagcta tcccgcccct
7741 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
7801 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
7861 gtagtgagga ggcttttttg gaggcctagg gacgtaccca attcgcccta tagtgagtcg
7921 tattacgcgc gctcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
7981 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
8041 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg ggacgcgccc
8101 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
8161 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
8221 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
8281 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
8341 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
8401 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
8461 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
8521 tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa
8581 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
8641 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
8701 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
8761 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
8821 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
8881 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
8941 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
9001 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
9061 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
9121 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
9181 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
9241 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
9301 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
9361 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
9421 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
9481 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
9541 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
9601 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
9661 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
9721 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
9781 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
9841 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
9901 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
9961 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
10021 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
10081 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
10141 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
10201 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
10261 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
10321 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
10381 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
10441 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
10501 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
10561 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg
10621 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
10681 acacaggaaa cagctatgac catgattacg ccaagcgcgc aattaaccct cactaaaggg
10741 aacaaaagct ggagctgcaa gc
//
| {
"Vector backbone": "pLX_TRC317",
"Backbone size w/o insert (bp)": "8264",
"Vector type": "Mammalian Expression, Lentiviral",
"Selectable markers": "Puromycin"
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "NEB Stable",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "A transcription factor atlas of directed differentiation",
"Publication": "Cell. 2023 Jan 5;186(1):209-229.e26. doi: 10.1016/j.cell.2022.11.026.",
"DOI": "10.1016/j.cell.2022.11.026.",
"Authors": "Joung J, Ma S, Tay T, Geiger-Schuller KR, Kirchgatterer PC, Verdine VK, Guo B, Arias-Garcia MA, Allen WE, Singh A, Kuksenko O, Abudayyeh OO, Gootenberg JS, Fu Z, Macrae RK, Buenrostro JD, Regev A, Zhang F /jcell",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/36608654",
"PubMed ID": "PubMed 36608654"
} | {
"Gene/Insert name": "TCEB3",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "2498",
"Mutation": "",
"Entrez Gene": "ELOA",
"Promoter": "EF1a",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Unknown",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "GGGTGGAGACTGAAGTTAGGCCAG",
"3′ sequencing primer": "CACATAGCGTAAAAGGAGCAACATAG"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
TFORF1947 | 142294 | Lentiviral vector for overexpressing transcription factor ORFs with unique 24-bp barcodes. Barcodes facilitate identification of transcription factors in pooled screens. | Feng Zhang | Joung et al Cell. 2023 Jan 5;186(1):209-229.e26. doi: 10.1016/j.cell.2022.11.026. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/395772/f06c33bb-5bc4-4b9e-a3ba-8af6b7faf78f/addgene-plasmid-142294-sequence-395772.gbk | full | LOCUS sequence-395772- 8611 bp DNA circular SYN 26-AUG-2024
DEFINITION Lentiviral vector for overexpressing transcription factor ORFs with
unique 24-bp barcodes. Barcodes facilitate identification of
transcription factors in pooled screens..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8611)
AUTHORS Joung J, Ma S, Tay T, Geiger-Schuller KR, Kirchgatterer PC, Verdine
VK, Guo B, Arias-Garcia MA, Allen WE, Singh A, Kuksenko O, Abudayyeh
OO, Gootenberg JS, Fu Z, Macrae RK, Buenrostro JD, Regev A, Zhang F
TITLE A transcription factor atlas of directed differentiation.
JOURNAL Cell. 2023 Jan 5;186(1):209-229.e26. doi:
10.1016/j.cell.2022.11.026.
PUBMED 36608654
REFERENCE 2 (bases 1 to 8611)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1016/j.cell.2022.11"; journalName: "Cell"; date: "2023-01-5- 5";
volume: "186"; issue: "1"; pages: "209-229.e26"
FEATURES Location/Qualifiers
source 1..8611
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 5..231
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 232..412
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 459..584
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1077..1310
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1495..1539
/codon_start=1
/product="antigenic peptide corresponding to amino acids
655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
et al., 2013)"
/label=gp41 peptide
/note="recognized by the 2H10 single-chain llama nanobody"
/translation="KNEQELLELDKWASL"
promoter 1770..1980
/label=SV40 promoter
/note="SV40 early promoter"
promoter 1792..1988
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 1839..1974
/label=SV40 ori
/note="SV40 origin of replication"
CDS 2020..2622
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGAS"
primer_bind complement(2020..2039)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 2516..2536
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
misc_feature 2665..2782
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2920..4098
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 3151..4089
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 4046..4066
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
misc_feature 4468..5056
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(4521..4541)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(4939..4950)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 5128..5361
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 5439..5560
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5476..5495)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 5530..5549
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 5600..5735
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(5755..5774)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(5756..5774)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(5784..5801)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(5784..5800)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5793..5815)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 5942..6397
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(6029..6048)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 6239..6260
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 6423..6527
/gene="bla"
/label=AmpR promoter
CDS 6528..7388
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(6746..6765)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 7559..8147
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 8048..8067
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 8301..8318
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 8435..8456
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 8471..8501
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 8509..8525
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
primer_bind 8514..8536
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 8533..8549
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 8533..8549
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 8568..8588
/label=T3
/note="T3 promoter, forward primer"
promoter 8570..8588
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
1 ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat gagttagcaa
61 catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt aaggtggtac
121 gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg aaccactgaa
181 ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa cgggtctctc
241 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
301 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
361 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
421 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
481 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
541 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
601 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
661 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
721 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
781 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
841 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
901 gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg
961 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta
1021 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga
1081 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg
1141 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg
1201 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc
1261 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg
1321 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat
1381 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac
1441 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat
1501 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca
1561 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga
1621 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg
1681 tttcagaccc acctcccaac cccgagggga cccatgcatt gcatctcaat tagtcagcaa
1741 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcgtctca
1801 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
1861 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
1921 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
1981 tttgcaaaaa gctttctaga ggtaccaccg gagcttacca tgaccgagta caagcccacg
2041 gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc
2101 gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc
2161 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg
2221 gacgacggcg ccgccgtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg
2281 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa
2341 cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc
2401 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga
2461 gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac
2521 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga
2581 ccgcgcacct ggtgcatgac ccgcaagccc ggtgccagtt aacgaattct agatcttgag
2641 acaaatggca gtattcatcc acaattttaa aagaaaaggg gggattgggg ggtacagtgc
2701 aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat tacaaaaaca
2761 aattacaaaa attcaaaatt ttcgggttta ttacagggac agcagagatc cactttggcg
2821 ccggctcgag gcctgcaggt gcaaagatgg ataaagtttt aaacagagag gaatctttgc
2881 agctaatgga ccttctaggt cttgaaagga gtgggaattg gctccggtgc ccgtcagtgg
2941 gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacc
3001 ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc
3061 ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt
3121 tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc ccgcgggcct
3181 ggcctcttta cgggttatgg cccttgcgtg ccttgaatta cttccacctg gctgcagtac
3241 gtgattcttg atcccgagct tcgggttgga agtgggtggg agagttcgag gccttgcgct
3301 taaggagccc cttcgcctcg tgcttgagtt gaggcctggc ctgggcgctg gggccgccgc
3361 gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc tctagccatt
3421 taaaattttt gatgacctgc tgcgacgctt tttttctggc aagatagtct tgtaaatgcg
3481 ggccaagatc tgcacactgg tatttcggtt tttggggccg cgggcggcga cggggcccgt
3541 gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc gagaatcgga
3601 cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg gcctcgcgcc gccgtgtatc
3661 gccccgccct gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc ggaaagatgg
3721 ccgcttcccg gccctgctgc agggagctca aaatggagga cgcggcgctc gggagagcgg
3781 gcgggtgagt cacccacaca aaggaaaagg gcctttccgt cctcagccgt cgcttcatgt
3841 gactccacgg agtaccgggc gccgtccagg cacctcgatt agttctcgag cttttggagt
3901 acgtcgtctt taggttgggg ggaggggttt tatgcgatgg agtttcccca cactgagtgg
3961 gtggagactg aagttaggcc agcttggcac ttgatgtaat tctccttgga atttgccctt
4021 tttgagtttg gatcttggtt cattctcaag cctcagacag tggttcaaag tttttttctt
4081 ccatttcagg tgtcgtgagg ctagcaccgc caccatgttt atgtttaatt ggtttacaga
4141 ctgtctgtgg actcttttcc tgtcaaatta ccagccatct gttgaatctt caagtccagg
4201 aggttcagca acatcagatg accatgaatt tgatccatca gctgacatgc tggttcatga
4261 ttttgatgat gaacgaacat tagaagagga agaaatgatg gaaggagaaa caaacttcag
4321 ctctgaaata gaagatcttg caagggtaaa taacatgtag taatgaacta gtccgtaact
4381 tgaaagtatt tcgatttctt ggctttatat atcttgtgga aaggacgatg tcccttaggc
4441 tatgcattgg gaacgcgtta agtcgacaat caacctctgg attacaaaat ttgtgaaaga
4501 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
4561 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
4621 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
4681 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
4741 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
4801 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
4861 aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
4921 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
4981 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
5041 tgggccgcct ccccgcgtcg actttaagac caatgactta caaggcagct gtagatctta
5101 gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag
5161 atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct
5221 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
5281 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
5341 cagtgtggaa aatctctagc agtacgtata gtagttcatg tcatcttatt attcagtatt
5401 tataacttgc aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata
5461 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
5521 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat
5581 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
5641 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
5701 attccagaag tagtgaggag gcttttttgg aggcctaggg acgtacccaa ttcgccctat
5761 agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac
5821 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
5881 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
5941 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
6001 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
6061 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
6121 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
6181 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
6241 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
6301 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
6361 aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg
6421 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
6481 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
6541 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
6601 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
6661 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
6721 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
6781 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
6841 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
6901 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
6961 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
7021 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
7081 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
7141 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
7201 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
7261 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
7321 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
7381 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
7441 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
7501 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
7561 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
7621 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
7681 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
7741 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
7801 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
7861 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
7921 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
7981 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
8041 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
8101 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
8161 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
8221 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
8281 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
8341 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
8401 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
8461 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
8521 aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc
8581 actaaaggga acaaaagctg gagctgcaag c
//
| {
"Vector backbone": "pLX_TRC317",
"Backbone size w/o insert (bp)": "8264",
"Vector type": "Mammalian Expression, Lentiviral",
"Selectable markers": "Puromycin"
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "NEB Stable",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "A transcription factor atlas of directed differentiation",
"Publication": "Cell. 2023 Jan 5;186(1):209-229.e26. doi: 10.1016/j.cell.2022.11.026.",
"DOI": "10.1016/j.cell.2022.11.026.",
"Authors": "Joung J, Ma S, Tay T, Geiger-Schuller KR, Kirchgatterer PC, Verdine VK, Guo B, Arias-Garcia MA, Allen WE, Singh A, Kuksenko O, Abudayyeh OO, Gootenberg JS, Fu Z, Macrae RK, Buenrostro JD, Regev A, Zhang F /jcell",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/36608654",
"PubMed ID": "PubMed 36608654"
} | {
"Gene/Insert name": "MIER1",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "347",
"Mutation": "",
"Entrez Gene": "MIER1",
"Promoter": "EF1a",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Unknown",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "GGGTGGAGACTGAAGTTAGGCCAG",
"3′ sequencing primer": "CACATAGCGTAAAAGGAGCAACATAG"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pcDNA3.1-Peredox-mCherry-NLS | 32384 | High | Gary Yellen | Hung et al Cell Metab. 2011 Oct 5;14(4):545-54. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/37048/4acf70bd-8625-4215-a635-a20310c1dfd9/addgene-plasmid-32384-sequence-37048.gbk | full | LOCUS sequence-37048-e 8155 bp DNA circular SYN 25-AUG-2024
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8155)
AUTHORS Hung YP, Albeck JG, Tantama M, Yellen G
TITLE Imaging Cytosolic NADH-NAD(+) Redox State with a Genetically Encoded
Fluorescent Biosensor.
JOURNAL Cell Metab. 2011 Oct 5;14(4):545-54.
PUBMED 21982714
REFERENCE 2 (bases 1 to 8155)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Cell
Metab."; date: "2011-10-5"; volume: "14(4)"; pages: "545-54"
FEATURES Location/Qualifiers
source 1..8155
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 66..85
/label=T7
/note="T7 promoter, forward primer"
promoter 66..84
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 133..2178
/codon_start=1
/product="genetically encoded fluorescent biosensor of the
cytosolic NADH-NAD+ redox state (Hung et al., 2011)"
/label=Peredox
/note="consists of two pH-insensitive mutant copies of the
Thermus aquaticus redox-sensing repressor Rex (T-Rex)
separated by circularly permuted monomeric T-Sapphire
(cpmTS), a pH-resistant GFP variant"
/translation="KVPEAAISRLITYLRILEELEAQGVHRTASEQLGELAQVTAFQVD
KDLSYFGSYGTDGVGYTVPVLKRELRHILGLNRKWGLCIVGMGRLGSALADWPGFGESF
ELRGFFDVDPGMVGRPVRGGVIEHVDLLPQRVPGRIEIALLTVPREAAQKAADLLVAAG
IKGILNFAPVVLEVPKEVAVENVDILAGLTRLSFAILNPTWSAAGGHGFTAHNVYIMAD
KQKNGIKANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSIQSKLSKDPNEK
RDHMVLLEFVTAAGITHGMDELYKGGTGGSMVSKGEELFTGVVPILVELDGDVNGHKFS
VSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTFSYGVMVFARYPDHMKQHDFFKSA
MPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNTK
VPEAAISRLITYLRILEELEAQGVHRTASEQLGELAQVTAFQVDEDLSYFGSYGTDGVG
YTVPVLKRELRHILGLNRKWGLCIVGMGRLGSALADWPGFGESFELRGFFDVDPGMVGR
PVRGGVIEHVDLLPQRVPGRIEIALLTVPREAAQKAADLLVAAGIKGILNFAPVVLEVP
KEVAVENVDFLAGLTRLSFAILNPKWREEMMGSGTGGNASDGGGSGG"
CDS 2179..2886
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
primer_bind complement(2324..2342)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 2592..2611
/label=mCherry-F
/note="mCherry, forward primer"
CDS 2887..2907
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
primer_bind complement(2952..2969)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2958..3182
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 3228..3656
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3315..3334)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3525..3546
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(3665..3685)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 3670..3999
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3850..3985
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3912..3931
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 4066..4860
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(4120..4139)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4730..4749
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 5034..5155
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5071..5090)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 5125..5144
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(5204..5220)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5204..5220)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(5217..5239)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 5228..5244
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(5252..5282)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5297..5318
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(5435..5452)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5606..6191)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5686..5705)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6362..7222)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 6985..7004
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7223..7327)
/gene="bla"
/label=AmpR promoter
primer_bind complement(7402..7421)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 7593..7972
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter join(7973..8155,1..21)
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 8127..8147
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
ORIGIN
1 gaggtctata taagcagagc tctctggcta actagagaac ccactgctta ctggcttatc
61 gaaattaata cgactcacta tagggagacc caagctggct agctctagat tgggcccacc
121 atggttgcta gcaaagttcc tgaagcagcc atttccagac tgattactta cctgagaatt
181 ctggaagagc tggaagccca aggtgtacac cgcaccgcct ccgagcaact gggcgagctg
241 gcccaggtca ccgccttcca ggttgataag gacctgtcct acttcggctc ctacggcact
301 gacggcgttg gctacactgt accggtcctc aagagagaac tgcgccatat cctcggtctg
361 aaccgcaaat ggggcctgtg catcgtgggg atgggccgcc tgggatccgc tcttgctgat
421 tggcctggtt ttggcgagag cttcgagctg cgcggtttct tcgatgttga cccaggtatg
481 gttggtcggc cggttcgcgg tggtgttatc gaacacgttg atctgctgcc ccagcgcgta
541 cctggtcgca tcgaaatcgc tctgctgacg gttccacgcg aggcagcaca gaaggctgcc
601 gacctgctgg ttgccgctgg catcaagggc atcctgaact tcgctccagt tgtactggag
661 gttcccaagg aagtggctgt tgagaacgtt gacatcctgg ccggtctgac gcgtctgagc
721 tttgccattc tgaaccccac gtggagcgca gcaggtgggc atggtttcac tgctcacaac
781 gtgtatatca tggctgacaa acagaagaac ggtatcaagg ccaacttcaa gatccgccac
841 aacatcgaag acggcggcgt tcagctggct gatcactacc agcagaacac cccaatcggc
901 gacggccctg ttctgctgcc tgacaaccac tacttaagca tccagtctaa actgtccaaa
961 gacccaaacg agaagcgcga ccacatggtt ctgctggaat ttgttaccgc cgctggcatc
1021 acccacggca tggacgaact gtacaagggc ggcaccgggg gatctatggt ttccaaaggc
1081 gaggaactgt tcactggtgt tgttcctatc ctggttgaac tggacggcga cgttaacggc
1141 cacaaattca gcgttagcgg ggaaggcgaa ggagacgcta cctacggcaa gctgactctg
1201 aagttcatct gcaccactgg caagctgcct gttccttggc ctaccctggt tactaccttc
1261 tcctatggtg ttatggtttt cgccagatat cctgaccaca tgaagcagca cgacttcttc
1321 aagagcgcca tgcctgaggg ctacgttcag gaacgcacca tctttttcaa ggacgacggc
1381 aactataaga cacgcgctga ggttaagttc gagggcgaca ctctggttaa ccgcatcgag
1441 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg gccataagct tgaatataac
1501 accaaggtac ctgaagccgc catcagccgc ctcatcactt acctccgcat cctggaggag
1561 ctcgaagccc aaggcgttca ccgcactgca agcgagcagc tgggcgaact ggcacaggtt
1621 acagcattcc aggttgacga agatctgtcc tatttcggtt cctacggcac cgacggtgtt
1681 ggctacacag tccctgtcct gaagcgcgag ctgcgccaca tcctgggcct gaaccgcaag
1741 tggggcttat gcatcgttgg catgggccgc ctcggtagtg cactggcaga ctggccgggc
1801 tttggtgaat ctttcgaact gcgcggcttc ttcgacgttg atccaggcat ggttgggcgc
1861 cctgttcgcg gtggcgttat cgagcatgtt gacctcctgc cacagcgcgt cccgggtcgc
1921 atcgagatcg ccctgctgac cgttcctcgc gaagctgcac agaaggcagc cgacttacta
1981 gtcgccgctg gtatcaaggg aattctgaac ttcgcccctg ttgttctgga ggttcctaag
2041 gaagttgctg ttgaaaacgt tgatttcctg gccggcctga cccgcctgag cttcgcaatc
2101 ctgaatccta agtggcgcga agaaatgatg ggttccggaa ctgggggcaa cgcctctgac
2161 ggtggtgggt ctggtggtat ggtgagcaag ggcgaggagg ataacatggc catcatcaag
2221 gagttcatgc gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc
2281 gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc
2341 aagggtggcc ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc
2401 aaggcctacg tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag
2461 ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag
2521 gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc
2581 ccctccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc ctccgagcgg
2641 atgtaccccg aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac
2701 ggcggccact acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg
2761 cccggcgcct acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc
2821 atcgtggaac agtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgaactg
2881 tacaagccta aaaagaaacg taaagttgaa gatgcttgag tcgagaagct taagtttaaa
2941 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
3001 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
3061 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
3121 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
3181 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
3241 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
3301 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
3361 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
3421 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
3481 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
3541 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
3601 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
3661 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
3721 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
3781 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
3841 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
3901 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
3961 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
4021 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
4081 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
4141 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
4201 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
4261 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
4321 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
4381 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
4441 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
4501 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
4561 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
4621 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
4681 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
4741 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
4801 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
4861 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
4921 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
4981 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
5041 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
5101 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
5161 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
5221 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
5281 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
5341 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
5401 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
5461 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
5521 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
5581 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
5641 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
5701 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
5761 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
5821 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
5881 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
5941 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
6001 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
6061 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
6121 aacaaaccac cgctggtagc ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
6181 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
6241 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
6301 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
6361 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
6421 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
6481 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
6541 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
6601 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
6661 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
6721 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
6781 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
6841 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
6901 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
6961 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
7021 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
7081 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
7141 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
7201 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
7261 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
7321 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga
7381 tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct
7441 gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac
7501 aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct
7561 gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata
7621 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
7681 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
7741 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
7801 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
7861 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
7921 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
7981 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
8041 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
8101 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgg
//
| {
"Vector backbone": "pcDNA3.1",
"Backbone size w/o insert (bp)": "5400",
"Vector type": "Mammalian Expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA ; Takara Bio Limited Use Label License (formerly Clontech)",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Imaging Cytosolic NADH-NAD(+) Redox State with a Genetically Encoded Fluorescent Biosensor",
"Publication": "Cell Metab. 2011 Oct 5;14(4):545-54.",
"DOI": "10.1016/j.cmet.2011.08.012",
"Authors": "Hung YP, Albeck JG, Tantama M, Yellen G",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/21982714",
"PubMed ID": "PubMed 21982714"
} | {
"Gene/Insert name": "Peredox-mCherry-NLS",
"Species": "",
"Insert Size (bp)": "2800",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "CMV",
"Tag / Fusion Protein": "Nuclear Localization Seq (C terminal on insert)"
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "XbaI (not destroyed)",
"3′ cloning site": "XhoI (destroyed during cloning)",
"5′ sequencing primer": "T7",
"3′ sequencing primer": "mCherry-R"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pJM522 EF1α intC-ZF10-NLS-HA in TUPV2 | 161552 | Constitutive expression of intC-ZF10-NLS-HA under the EF1α promoter | Joshua Leonard | Muldoon et al Sci Adv. 2021 Feb 19;7(8). pii: 7/8/eabe9375. doi: 10.1126/sciadv.abe9375. Print 2021 Feb. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/317924/5ac4ee8c-3294-45cb-a35b-b2c5b5231917/addgene-plasmid-161552-sequence-317924.gbk | full | LOCUS sequence-317924- 6295 bp DNA circular SYN 26-AUG-2024
DEFINITION Constitutive expression of intC-ZF10-NLS-HA under the EF1-alpha
promoter.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6295)
AUTHORS Muldoon JJ, Kandula V, Hong M, Donahue PS, Boucher JD, Bagheri N,
Leonard JN
TITLE Model-guided design of mammalian genetic programs.
JOURNAL Sci Adv. 2021 Feb 19;7(8). pii: 7/8/eabe9375. doi:
10.1126/sciadv.abe9375. Print 2021 Feb.
PUBMED 33608279
REFERENCE 2 (bases 1 to 6295)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Sci Adv.";
date: "2021-02-19"; volume: "7(8). pii: 7/8/eabe9375. doi"; pages: "
10.1126/sciadv.abe9375. Print 2021 Feb"
FEATURES Location/Qualifiers
source 1..6295
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin 834..1262
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(921..940)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 1131..1152
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 1285..1481
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 1332..1467
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 1394..1413
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
polyA_signal 1601..1722
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1638..1657)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 1692..1711
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(1771..1787)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1771..1787)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(1784..1806)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 1795..1811
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(1819..1849)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 1864..1885
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(2002..2019)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2173..2761)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2253..2272)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(2932..3792)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 3555..3574
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(3793..3897)
/gene="bla"
/label=AmpR promoter
promoter 4278..5460
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 4509..5451
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 5408..5428
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
primer_bind 5462..5486
/label=LNCX
/note="Human CMV promoter, forward primer"
CDS 6023..6043
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 6056..6082
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
primer_bind complement(6130..6147)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal join(6136..6295,1..65)
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
ORIGIN
1 ggacagcaag ggggaggatt gggaagtcaa tagcaggcat gctggggatg cggtgggctc
61 tatggcttct gaggcaggtc gaagaactat ttcctggctg ttacgcgagc gcatcgcgat
121 aagctctaga gggacagccc ccccccaaag cccccaggga tgtaattacg tccctccccc
181 gctagggggc agcagcgagc cgcccggggc tccgctccgg tccggcgctc cccccgcatc
241 cccgagccgg cagcgtgcgg ggacagcccg ggcacgggga aggtggcacg ggatcgcttt
301 cctctgaacg cttctcgctg ctctttgagc ctgcagacac ctggggggat acggggaaaa
361 agctttaggc tgaaagagag atttagaatg acagaatcat agaacggcct gcgtacggat
421 ggggctctag ctctagagct tgatatcgaa ttcctgcagc cccgctagag ggacagcccc
481 cccccaaagc ccccagggat gtaattacgt ccctcccccg ctagggggca gcagcgagcc
541 gcccggggct ccgctccggt ccggcgctcc ccccgcatcc ccgagccggc agcgtgcggg
601 gacagcccgg gcacggggaa ggtggcacgg gatcgctttc ctctgaacgc ttctcgctgc
661 tctttgagcc tgcagacacc tggggggata cggggaaaaa gctttaggct gaaagagaga
721 tttagaatga cagaatcata gaacggcctg ggcgcgcctg gtggcgaacg atcaacagac
781 gggagaggac taaagtcttc gctgacagcg accggactct agggggtatc cccacgcgcc
841 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
901 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
961 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
1021 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
1081 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
1141 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
1201 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
1261 ttaattctgt ggaatgtgtg tcagttaggc tcaattagtc agcaaccata gtcccgcccc
1321 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
1381 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
1441 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
1501 tatccatttt cggaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
1561 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
1621 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
1681 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
1741 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
1801 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
1861 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
1921 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
1981 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
2041 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
2101 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
2161 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
2221 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
2281 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
2341 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
2401 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
2461 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
2521 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
2581 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
2641 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
2701 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
2761 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
2821 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
2881 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
2941 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
3001 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
3061 aatgataccg cgagatccac gctcaccggc tccagattta tcagcaataa accagccagc
3121 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
3181 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
3241 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
3301 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
3361 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
3421 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
3481 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
3541 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
3601 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
3661 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
3721 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
3781 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
3841 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
3901 atttccccga aaagtgccac ctgacgtcga cggatcggga gatcgaagac gggcaaaacc
3961 gatgttgacg gactaatcct gacaagatct ggggaggtcg ctgagtagtg cgcgagcaaa
4021 atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt
4081 aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgtgagta attcatacaa
4141 aaggactcgc ccctgccttg gggaatccca gggaccgtcg ttaaactccc actaacgtag
4201 aacccagaga tcgctgcgtt cccgccccct cacccgcccg ctctcgtcat cactgaggtg
4261 gagaagagca tgcgtgaggc tccggtgccc gtcagtgggc agagcgcaca tcgcccacag
4321 tccccgagaa gttgggggga ggggtcggca attgaaccgg tgcctagaga aggtggcgcg
4381 gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag
4441 aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca
4501 gaacacaggt aagtgccgtg tgtggttccc gcgggcctgg cctctttacg ggttatggcc
4561 cttgcgtgcc ttgaattact tccacgcccc tggctgcagt acgtgattct tgatcccgag
4621 cttcgggttg gaagtgggtg ggagagttcg aggccttgcg cttaaggagc cccttcgcct
4681 cgtgcttgag ttgaggcctg gcttgggcgc tggggccgcc gcgtgcgaat ctggtggcac
4741 cttcgcgcct gtctcgctgc tttcgataag tctctagcca tttaaaattt ttgatgacct
4801 gctgcgacgc tttttttctg gcaagatagt cttgtaaatg cgggccaaga tctgcacact
4861 ggtatttcgg tttttggggc cgcgggcggc gacggggccc gtgcgtccca gcgcacatgt
4921 tcggcgaggc ggggcctgcg agcgcggcca ccgagaatcg gacgggggta gtctcaagct
4981 ggccggcctg ctctggtgcc tggcctcgcg ccgccgtgta tcgccccgcc ctgggcggca
5041 aggctggccc ggtcggcacc agttgcgtga gcggaaagat ggccgcttcc cggccctgct
5101 gcagggagct caaaatggag gacgcggcgc tcgggagagc gggcgggtga gtcacccaca
5161 caaaggaaaa gggcctttcc gtcctcagcc gtcgcttcat gtgactccac ggagtaccgg
5221 gcgccgtcca ggcacctcga ttagttctcg agcttttgga gtacgtcgtc tttaggttgg
5281 ggggaggggt tttatgcgat ggagtttccc cacactgagt gggtggagac tgaagttagg
5341 ccagcttggc acttgatgta attctccttg gaatttgccc tttttgagtt tggatcttgg
5401 ttcattctca agcctcagac agtggttcaa agtttttttc ttccatttca ggtgtcgtga
5461 gagctcgttt agtgaaccgt cagatcgcct ggagacgcca tccacgctgt tttgacctcc
5521 atagaagaga acgggaccga tccaaattct cgaggctagc gccaccatga tgctgaagaa
5581 gatcctgaag atcgaggagc tggacgagag agagctgatc gacatcgagg tgagcggcaa
5641 ccacctgttc tacgccaacg acatcctgac ccacaacagc ggatccggtg gtggcgggtc
5701 tggaggagga ggtagtggtg gaggtggctc tggtaccggt ggagctagac ccggggagcg
5761 ccccttccag tgtcgcattt gcatgcggaa cttttcgact aatcaaaaat tggaagttca
5821 tacccgtact cataccggtg aaaaaccgtt tcagtgtcgg atctgtatgc gaaatttctc
5881 cgttagacat aatttgcaaa gacatctacg tacgcacacc ggcgagaagc cattccaatg
5941 ccgaatatgc atgcgcaact tcagtcaaca tccaaatttg actagacacc taaaaaccca
6001 cctgagagga tctggctctg gccctaagaa gaagcgcaaa gttggctctg gcatgtaccc
6061 atacgacgta ccagattacg cttaggcggc cgctcgagtc tagagggccc gtttaaaccc
6121 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
6181 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
6241 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggca
//
| {
"Vector backbone": "pJM451 EF1α EBFP2-P2A-BlastR (TUPV2)",
"Backbone size w/o insert (bp)": "",
"Vector type": "Mammalian Expression, Synthetic Biology",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Model-guided design of mammalian genetic programs",
"Publication": "Sci Adv. 2021 Feb 19;7(8). pii: 7/8/eabe9375. doi: 10.1126/sciadv.abe9375. Print 2021 Feb.",
"DOI": "10.1126/sciadv.abe9375.",
"Authors": "Muldoon JJ, Kandula V, Hong M, Donahue PS, Boucher JD, Bagheri N, Leonard JN /sciadvabe",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/33608279",
"PubMed ID": "PubMed 33608279"
} | {
"Gene/Insert name": "intC-ZF10-NLS-HA",
"Species": "Synthetic",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "EF1α",
"Tag / Fusion Protein": "HA (C terminal on insert)"
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "NheI (not destroyed)",
"3′ cloning site": "NotI (not destroyed)",
"5′ sequencing primer": "CCATTTCAGGTGTCGTGAGAGCTC",
"3′ sequencing primer": "CAGTGGGAGTGGCACCTTCC"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pLV_PGK_3xHA-LTB4R(T308A) | 208092 | Mammalian expression of the human LTB4 receptor LTB4R with the T308A mutation with an N-terminal 3x HA tag. | Sean R Collins | Lundgren et al Sci Signal. 2023 Oct 3;16(805):eadd1845. doi: 10.1126/scisignal.add1845. Epub 2023 Oct 3. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/410790/1065717c-ee33-4bc8-b078-f2414a768cfd/addgene-plasmid-208092-sequence-410790.gbk | full | LOCUS sequence-410790- 9590 bp DNA circular SYN 27-AUG-2024
DEFINITION Mammalian expression of the human LTB4 receptor LTB4R with the T308A
mutation with an N-terminal 3x HA tag..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9590)
AUTHORS Lundgren SM, Rocha-Gregg BL, Akdogan E, Mysore MN, Hayes S, Collins
SR
TITLE Signaling dynamics distinguish high- and low-priority neutrophil
chemoattractant receptors.
JOURNAL Sci Signal. 2023 Oct 3;16(805):eadd1845. doi:
10.1126/scisignal.add1845. Epub 2023 Oct 3.
PUBMED 37788324
REFERENCE 2 (bases 1 to 9590)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 27, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Sci
Signal."; date: "2023-10-3"; volume: "16(805):eadd1845. doi"; pages:
" 10.1126/scisignal.add1845. Epub 2023 Oct 3"
FEATURES Location/Qualifiers
source 1..9590
/mol_type="other DNA"
/organism="synthetic DNA construct"
LTR 1..634
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 681..806
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1303..1536
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1721..1765
/codon_start=1
/product="antigenic peptide corresponding to amino acids
655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
et al., 2013)"
/label=gp41 peptide
/note="recognized by the 2H10 single-chain llama nanobody"
/translation="KNEQELLELDKWASL"
misc_feature 2028..2143
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1 (lacking the first T)"
promoter 2189..2699
/label=hPGK promoter
/note="human phosphoglycerate kinase 1 promoter"
primer_bind 2618..2636
/label=hPGK-F
/note="Human PGK promoter, forward primer"
CDS 2721..2801
/codon_start=1
/product="three tandem HA epitope tags"
/label=3xHA
/translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
misc_feature 3958..4507
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(4102..4119)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 4329..4348
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 4510..5535
/codon_start=1
/product="hygromycin B phosphotransferase"
/label=HygR
/note="confers resistance to hygromycin"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
E"
CDS 4510..5535
/codon_start=1
/gene="aph(4)-Ia"
/product="aminoglycoside phosphotransferase from E. coli"
/label=HygR
/note="confers resistance to hygromycin"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
E"
misc_feature 5559..6147
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(5612..5632)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(6030..6041)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 6354..6987
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
primer_bind complement(7115..7131)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(7115..7131)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(7128..7150)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 7139..7155
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(7163..7193)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 7208..7229
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(7346..7363)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(7517..8105)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(7597..7616)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(8276..9136)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 8899..8918
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(9137..9241)
/gene="bla"
/label=AmpR promoter
polyA_signal 9289..9423
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(9326..9345)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 9380..9399
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
ORIGIN
1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag
2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
2161 acagcagaga tccagtttat cgatccacgg ggttggggtt gcgccttttc caaggcagcc
2221 ctgggtttgc gcagggacgc ggctgctctg ggcgtggttc cgggaaacgc agcggcgccg
2281 accctgggtc tcgcacattc ttcacgtccg ttcgcagcgt cacccggatc ttcgccgcta
2341 cccttgtggg ccccccggcg acgcttcctg ctccgcccct aagtcgggaa ggttccttgc
2401 ggttcgcggc gtgccggacg tgacaaacgg aagccgcacg tctcactagt accctcgcag
2461 acggacagcg ccagggagca atggcagcgc gccgaccgcg atgggctgtg gccaatagcg
2521 gctgctcagc agggcgcgcc gagagcagcg gccgggaagg ggcggtgcgg gaggcggggt
2581 gtggggcggt agtgtgggcc ctgttcctgc ccgcgcggtg ttccgcattc tgcaagcctc
2641 cggagcgcac gtcggcagtc ggctccctcg ttgaccgaat caccgacctc tctccccagg
2701 acgcgggatc cgccaccatg tacccatacg atgttccaga ttacgcttac ccatacgatg
2761 ttccagatta cgcttaccca tacgatgttc cagattacgc taacactaca tcttctgcag
2821 cacccccctc actaggtgta gagttcatct ctctgctggc tatcatcctg ctgtcagtgg
2881 cgctggctgt ggggcttccc ggcaacagct ttgtggtgtg gagtatcctg aaaaggatgc
2941 agaagcgctc tgtcactgcc ctgatggtgc tgaacctggc cctggccgac ctggccgtat
3001 tgctcactgc tccctttttc cttcacttcc tggcccaagg cacctggagt tttggactgg
3061 ctggttgccg cctgtgtcac tatgtctgcg gagtcagcat gtacgccagc gtcctgctta
3121 tcacggccat gagtctagac cgctcactgg cggtggcccg cccctttgtg tcccagaagc
3181 tacgcaccaa ggcgatggcc cggcgggtgc tggcaggcat ctgggtgttg tcctttctgc
3241 tggccacacc cgtcctcgcg taccgcacag tagtgccctg gaaaacgaac atgagcctgt
3301 gcttcccgcg gtaccccagc gaagggcacc gggccttcca tctaatcttc gaggctgtca
3361 cgggcttcct gctgcccttc ctggctgtgg tggccagcta ctcggacata gggcgtcggc
3421 tacaggcccg gcgcttccgc cgcagccgcc gcaccggccg cctggtggtg ctcatcatcc
3481 tgaccttcgc cgccttctgg ctgccctacc acgtggtgaa cctggctgag gcgggccgcg
3541 cgctggccgg ccaggccgcc gggttagggc tcgtggggaa gcggctgagc ctggcccgca
3601 acgtgctcat cgcactcgcc ttcctgagca gcagcgtgaa ccccgtgctg tacgcgtgcg
3661 ccggcggcgg cctgctgcgc tcggcgggcg tgggcttcgt cgccaagctg ctggagggcg
3721 cgggctccga ggcgtccagc acgcgccgcg ggggcagcct gggccagacc gctaggagcg
3781 gccccgccgc tctggagccc ggcccttccg agagcctcac tgcctccagc cctctcaagt
3841 taaacgaact gaactaagcg gccgctctag agtcgacggg ccgcggtaac aattgttaac
3901 taacttaagc tagcaacggt ttccctctag cgggatcaat tccgcccccc ccccctaacg
3961 ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca
4021 ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga
4081 gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga
4141 aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca
4201 ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag
4261 atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa
4321 gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac
4381 cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga
4441 ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac
4501 gataatacca tgaaaaagcc tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa
4561 aagttcgaca gcgtctccga cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc
4621 agcttcgatg taggagggcg tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc
4681 tacaaagatc gttatgttta tcggcacttt gcatcggccg cgctcccgat tccggaagtg
4741 cttgacattg gggaattcag cgagagcctg acctattgca tctcccgccg tgcacagggt
4801 gtcacgttgc aagacctgcc tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag
4861 gccatggatg cgatcgctgc ggccgatctt agccagacga gcgggttcgg cccattcgga
4921 ccgcaaggaa tcggtcaata cactacatgg cgtgatttca tatgcgcgat tgctgatccc
4981 catgtgtatc actggcaaac tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct
5041 ctcgatgagc tgatgctttg ggccgaggac tgccccgaag tccggcacct cgtgcacgcg
5101 gatttcggct ccaacaatgt cctgacggac aatggccgca taacagcggt cattgactgg
5161 agcgaggcga tgttcgggga ttcccaatac gaggtcgcca acatcttctt ctggaggccg
5221 tggttggctt gtatggagca gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca
5281 ggatcgccgc ggctccgggc gtatatgctc cgcattggtc ttgaccaact ctatcagagc
5341 ttggttgacg gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc
5401 cgatccggag ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg
5461 accgatggct gtgtagaagt actcgccgat agtggaaacc gacgccccag cactcgtccg
5521 agggcaaagg aatagcccgg gcggggcgcg tctggaacaa tcaacctctg gattacaaaa
5581 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
5641 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
5701 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
5761 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
5821 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
5881 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
5941 tgttgtcggg gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc
6001 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
6061 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc
6121 ggatctccct ttgggccgcc tccccgcctg gaattaattc tgcagtcgag acctagaaaa
6181 acatggagca atcacaagta gcaatacagc agctaccaat gctgattgtg cctggctaga
6241 agcacaagag gaggaggagg tgggttttcc agtcacacct caggtacctt taagaccaat
6301 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaagaggg gactggaagg
6361 gctaattcac tcccaacgaa gacaagatat ccttgatctg tggatctacc acacacaagg
6421 ctacttccct gattagcaga actacacacc agggccaggg gtcagatatc cactgacctt
6481 tggatggtgc tacaagctag taccagttga gccagataag gtagaagagg ccaataaagg
6541 agagaacacc agcttgttac accctgtgag cctgcatggg atggatgacc cggagagaga
6601 agtgttagag tggaggtttg acagccgcct agcatttcat cacgtggccc gagagctgca
6661 tccggagtac ttcaagaact gctgatatcg agcttgctac aagggacttt ccgctgggga
6721 ctttccaggg aggcgtggcc tgggcgggac tggggagtgg cgagccctca gatcctgcat
6781 ataagcagct gctttttgcc tgtactgggt ctctctggtt agaccagatc tgagcctggg
6841 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
6901 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
6961 tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt
7021 tataacttgc aaagaaatga atatcagaga gtgagaggcc ttgacattgc tagcgtttac
7081 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
7141 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
7201 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
7261 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
7321 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
7381 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
7441 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
7501 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
7561 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
7621 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
7681 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
7741 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
7801 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
7861 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
7921 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
7981 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
8041 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
8101 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
8161 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
8221 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
8281 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
8341 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
8401 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
8461 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
8521 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
8581 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
8641 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
8701 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
8761 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
8821 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
8881 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
8941 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
9001 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
9061 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
9121 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
9181 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
9241 gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatcaa cttgtttatt
9301 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
9361 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
9421 atcaactgga taactcaagc taaccaaaat catcccaaac ttcccacccc ataccctatt
9481 accactgcca attacctgtg gtttcattta ctctaaacct gtgattcctc tgaattattt
9541 tcattttaaa gaaattgtat ttgttaaata tgtactacaa acttagtagt
//
| {
"Vector backbone": "pLenti 2nd gen",
"Backbone size w/o insert (bp)": "",
"Vector type": "Mammalian Expression, Lentiviral",
"Selectable markers": "Hygromycin"
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "30°C",
"Growth Strain(s)": "NEB Stable",
"Copy number": "Unknown"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Signaling dynamics distinguish high- and low-priority neutrophil chemoattractant receptors",
"Publication": "Sci Signal. 2023 Oct 3;16(805):eadd1845. doi: 10.1126/scisignal.add1845. Epub 2023 Oct 3.",
"DOI": "10.1126/scisignal.add1845.",
"Authors": "Lundgren SM, Rocha-Gregg BL, Akdogan E, Mysore MN, Hayes S, Collins SR /scisignaladd",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/37788324",
"PubMed ID": "PubMed 37788324"
} | {
"Gene/Insert name": "3xHA-LTB4R(T308A)",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "1140",
"Mutation": "T308A",
"Entrez Gene": "LTB4R",
"Promoter": "PGK",
"Tag / Fusion Protein": "3xHA (N terminal on insert)"
} | {
"Cloning method": "Gibson Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "tgttccgcattctgcaagc",
"3′ sequencing primer": "cctcacattgccaaaagacg"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pMXs-Klf4-IP | 15920 | High | Shinya Yamanaka | Takahashi et al Cell. 2006 Aug 25. 126(4):663-76. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/169385/b922b9a3-0866-4a37-9dfe-db09523d662c/addgene-plasmid-15920-sequence-169385.gbk | full | LOCUS sequence-169385- 7409 bp DNA circular SYN 25-AUG-2024
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7409)
AUTHORS Takahashi K, Yamanaka S
TITLE Induction of pluripotent stem cells from mouse embryonic and adult
fibroblast cultures by defined factors.
JOURNAL Cell. 2006 Aug 25. 126(4):663-76.
PUBMED 16904174
REFERENCE 2 (bases 1 to 7409)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Cell. 2006
Aug 25. 126(4):663-76."
FEATURES Location/Qualifiers
source 1..7409
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 1..417
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
primer_bind 347..369
/label=pLXSN 5'
/note="Murine stem cell virus, forward primer. Also called
MSCV"
primer_bind 385..401
/label=pBABE 5'
/note="Psi packaging signal, forward primer for pBABE
vectors"
misc_feature 427..801
/label=pol region
/note="Moloney murine leukemia virus (MMLV) pol region
containing the splice acceptor site"
primer_bind 739..755
/label=pBMN 5'
/note="MMLV sequence, for inserts in pBMN retroviral
vector"
protein_bind 832..856
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
protein_bind complement(2336..2360)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
misc_feature 2408..2994
/label=IRES2
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(2575..2592)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 2802..2821
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 2995..3594
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind complement(2995..3014)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 3491..3511
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
LTR 3821..4290
/note="long terminal repeat from Moloney murine leukemia
virus"
primer_bind complement(4420..4437)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4591..5179)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4671..4690)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5343..6203)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 5966..5985
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6204..6308)
/gene="bla"
/label=AmpR promoter
LTR 6338..6931
/note="long terminal repeat from Moloney murine leukemia
virus"
misc_feature 6994..7351
/label=MMLV Psi
/note="packaging signal of Moloney murine leukemia virus
(MMLV)"
ORIGIN
1 ggccagactg ttaccactcc cttaagtttg accttaggtc actggaaaga tgtcgagcgg
61 atcgctcaca accagtcggt agatgtcaag aagagacgtt gggttacctt ctgctctgca
121 gaatggccaa cctttaacgt cggatggccg cgagacggca cctttaaccg agacctcatc
181 acccaggtta agatcaaggt cttttcacct ggcccgcatg gacacccaga ccaggtcccc
241 tacatcgtga cctgggaagc cttggctttt gacccccctc cctgggtcaa gccctttgta
301 caccctaagc ctccgcctcc tcttcctcca tccgccccgt ctctccccct tgaacctcct
361 cgttcgaccc cgcctcgatc ctccctttat ccagccctca ctccttctct aggcgccccc
421 atatggccat atgagatctt atatggggca cccccgcccc ttgtaaactt ccctgaccct
481 gacatgacaa gagttactaa cagcccctct ctccaagctc acttacaggc tctctactta
541 gtccagcacg aagtctggag acctctggcg gcagcctacc aagaacaact ggaccgaccg
601 gtggtacctc acccttaccg agtcggcgac acagtgtggg tccgccgaca ccagactaag
661 aacctagaac ctcgctggaa aggaccttac acagtcctgc tgaccacccc caccgccctc
721 aaagtagacg gcatcgcagc ttggatacac gccgcccacg tgaaggctgc cgaccccggg
781 ggtggaccat cctctagact gccggatcta gctagttaat taaggatcat cacaagtttg
841 tacaaaaaag caggctccgc ggccgccccc ttcaccatgg ctgtcagcga cgctctgctc
901 ccgtccttct ccacgttcgc gtccggcccg gcgggaaggg agaagacact gcgtccagca
961 ggtgccccga ctaaccgttg gcgtgaggaa ctctctcaca tgaagcgact tcccccactt
1021 cccggccgcc cctacgacct ggcggcgacg gtggccacag acctggagag tggcggagct
1081 ggtgcagctt gcagcagtaa caacccggcc ctcctagccc ggagggagac cgaggagttc
1141 aacgacctcc tggacctaga ctttatcctt tccaactcgc taacccacca ggaatcggtg
1201 gccgccaccg tgaccacctc ggcgtcagct tcatcctcgt cttccccagc gagcagcggc
1261 cctgccagcg cgccctccac ctgcagcttc agctatccga tccgggccgg gggtgacccg
1321 ggcgtggctg ccagcaacac aggtggaggg ctcctctaca gccgagaatc tgcgccacct
1381 cccacggccc ccttcaacct ggcggacatc aatgacgtga gcccctcggg cggcttcgtg
1441 gctgagctcc tgcggccgga gttggaccca gtatacattc cgccacagca gcctcagccg
1501 ccaggtggcg ggctgatggg caagtttgtg ctgaaggcgt ctctgaccac ccctggcagc
1561 gagtacagca gcccttcggt catcagtgtt agcaaaggaa gcccagacgg cagccacccc
1621 gtggtagtgg cgccctacag cggtggcccg ccgcgcatgt gccccaagat taagcaagag
1681 gcggtcccgt cctgcacggt cagccggtcc ctagaggccc atttgagcgc tggaccccag
1741 ctcagcaacg gccaccggcc caacacacac gacttccccc tggggcggca gctccccacc
1801 aggactaccc ctacactgag tcccgaggaa ctgctgaaca gcagggactg tcaccctggc
1861 ctgcctcttc ccccaggatt ccatccccat ccggggccca actaccctcc tttcctgcca
1921 gaccagatgc agtcacaagt cccctctctc cattatcaag agctcatgcc accgggttcc
1981 tgcctgccag aggagcccaa gccaaagagg ggaagaaggt cgtggccccg gaaaagaaca
2041 gccacccaca cttgtgacta tgcaggctgt ggcaaaacct ataccaagag ttctcatctc
2101 aaggcacacc tgcgaactca cacaggcgag aaaccttacc actgtgactg ggacggctgt
2161 gggtggaaat tcgcccgctc cgatgaactg accaggcact accgcaaaca cacagggcac
2221 cggccctttc agtgccagaa gtgcgacagg gccttttcca ggtcggacca ccttgcctta
2281 cacatgaaga ggcactttta aatcccacgt agtggatgta agggtgggcg cgccgaccca
2341 gctttcttgt acaaagtggt gattcgagcg gccgctcgat aagcttgata tcgaattccg
2401 cccccccccc ctctccctcc ccccccccta acgttactgg ccgaagccgc ttggaataag
2461 gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga
2521 gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg
2581 ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt
2641 gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca
2701 ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc
2761 agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat
2821 tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc
2881 ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga
2941 accacgggga cgtggttttc ctttgaaaaa cacgatgata atatggccac aaccatgacc
3001 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc
3061 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac
3121 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc
3181 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc
3241 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg
3301 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg
3361 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc
3421 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc
3481 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc
3541 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgacgcccg
3601 ccccacgacc cgcagcgccc gaccgaaagg agcgcacgac cccatgcatc gtcgacgata
3661 aaataaaaga ttttatttag tctccagaaa aaggggggaa tgaaagaccc cacctgtagg
3721 tttggcaagc tagcttaagt aacgccattt tgcaaggcat ggaaaaatac ataactgaga
3781 atagagaagt tcagatcaag gtcaggaaca gatggaacag ctgaatatgg gccaaacagg
3841 atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg aacagctgaa
3901 tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca
3961 gatggtcccc agatgcggtc cagccctcag cagtttctag agaaccatca gatgtttcca
4021 gggtgcccca aggacctgaa atgaccctgt gccttatttg aactaaccaa tcagttcgct
4081 tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc acaacccctc
4141 actcggggcg ccagtcctcc gattgactga gtcgcccggg tacccgtgta tccaataaac
4201 cctcttgcag ttgcatccga cttgtggtct cgctgttcct tgggagggtc tcctctgagt
4261 gattgactac ccgtcagcgg gggtctttca catgcagcat gtatcaaaat taatttggtt
4321 ttttttctta agtatttaca ttaaatggcc atagttgcat taatgaatcg gccaacgcgc
4381 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
4441 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
4501 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
4561 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
4621 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
4681 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
4741 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
4801 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
4861 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
4921 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
4981 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
5041 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
5101 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
5161 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
5221 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
5281 gatccttttg cggccggccg caaatcaatc taaagtatat atgagtaaac ttggtctgac
5341 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
5401 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
5461 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
5521 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
5581 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
5641 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
5701 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
5761 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
5821 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
5881 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
5941 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
6001 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
6061 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
6121 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
6181 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
6241 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
6301 gttccgcgca catttccccg aaaagtgcca cctgcataat gaaagacccc acctgtaggt
6361 ttggcaagct agcttaagta acgccatttt gcaaggcatg gaaaaataca taactgagaa
6421 tagaaaagtt cagatcaagg tcaggaacag atggaacagc tgaatatggg ccaaacagga
6481 tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatgga acagctgaat
6541 atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg ccaagaacag
6601 atggtcccca gatgcggtcc agccctcagc agtttctaga gaaccatcag atgtttccag
6661 ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga actaaccaat cagttcgctt
6721 ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata aaagagccca caacccctca
6781 ctcggcgcgc cagtcctccg attgactgag tcgcccgggt acccgtgtat ccaataaacc
6841 ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt gggagggtct cctctgagtg
6901 attgactacc cgtcagcggg ggtctttcat ttgggggctc gtccgggatc gggagacccc
6961 tgcccaggga ccaccgaccc accaccggga ggtaagctgg ccagcaactt atctgtgtct
7021 gtccgattgt ctagtgtcta tgactgattt tatgcgcctg cgtcggtact agttagctaa
7081 ctagctctgt atctggcgga cccgtggtgg aactgacgag ttcggaacac ccggccgcaa
7141 ccctgggaga cgtcccaggg acttcggggg ccgtttttgt ggcccgacct gagtccaaaa
7201 atcccgatcg ttttggactc tttggtgcac ccccctaata ggagggatat gtggttctgg
7261 taggagacga gaacctaaaa cagttcccgc ctccgtctga atttttgctt tcggtttggg
7321 accgaagccg cgccgcgcgt cttgtctgct gcagcatcgt tctgtgttgt ctctgtctga
7381 ctgtgtttct gtatttgtct gaaaattag
//
| {
"Vector backbone": "pMXs-IP",
"Backbone size w/o insert (bp)": "6100",
"Vector type": "Mammalian Expression, Retroviral",
"Selectable markers": "Puromycin"
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors",
"Publication": "Cell. 2006 Aug 25. 126(4):663-76.",
"DOI": "10.1016/j.cell.2006.07.024",
"Authors": "Takahashi K, Yamanaka S",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/16904174",
"PubMed ID": "PubMed 16904174"
} | {
"Gene/Insert name": "Kruppel-like factor 4 (gut)",
"Species": "M. musculus (mouse)",
"Insert Size (bp)": "1425",
"Mutation": "None",
"Entrez Gene": "Klf4",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Gateway Cloning",
"5′ cloning site": "Gateway (not destroyed)",
"3′ cloning site": "Gateway (not destroyed)",
"5′ sequencing primer": "pMX-S1811",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pDONR223-ZNF511 | 88081 | Donor Vector containing ZNF511 transcription factor, part of the Human TFome Collection | Marc Vidal | Ng et al Nat Biotechnol. 2021 Apr;39(4):510-519. doi: 10.1038/s41587-020-0742-6. Epub 2020 Nov 30. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/198735/82971aa1-e379-4949-81e7-203890e6e177/addgene-plasmid-88081-sequence-198735.gbk | full | LOCUS sequence-198735- 3546 bp DNA circular SYN 26-AUG-2024
DEFINITION Donor Vector containing ZNF511 transcription factor, part of the
Human TFome Collection.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3546)
AUTHORS Ng AHM, Khoshakhlagh P, Rojo Arias JE, Pasquini G, Wang K, Swiersy
A, Shipman SL, Appleton E, Kiaee K, Kohman RE, Vernet A, Dysart M,
Leeper K, Saylor W, Huang JY, Graveline A, Taipale J, Hill DE, Vidal
M, Melero-Martin JM, Busskamp V, Church GM
TITLE A comprehensive library of human transcription factors for cell fate
engineering.
JOURNAL Nat Biotechnol. 2021 Apr;39(4):510-519. doi:
10.1038/s41587-020-0742-6. Epub 2020 Nov 30.
PUBMED 33257861
REFERENCE 2 (bases 1 to 3546)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1038/s41587-020-0742-6"; journalName: "Nat Biotechnol"; date:
"2021-04"; volume: "39"; issue: "4"; pages: "510-519"
FEATURES Location/Qualifiers
source 1..3546
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 1..17
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 33..132
/gene="mutant version of attL"
/label=attL1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
protein_bind complement(889..988)
/gene="mutant version of attL"
/label=attL2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
primer_bind complement(1005..1024)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(1006..1024)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(1029..1045)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1029..1045)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
CDS 1476..2267
/codon_start=1
/gene="aadA"
/product="aminoglycoside adenylyltransferase (Murphy,
1985)"
/label=SmR
/note="confers resistance to spectinomycin and
streptomycin"
/translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
rep_origin 2360..2948
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2849..2868
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 3102..3119
/label=L4440
/note="L4440 vector, forward primer"
terminator complement(3278..3305)
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator complement(3397..3483)
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
primer_bind join(3532..3546,1..8)
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind join(3546,1..17)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
ORIGIN
1 gtaaaacgac ggccagtctt aagctcgggc cccaaataat gattttattt tgactgatag
61 tgacctgttc gttgcaacaa attgatgagc aatgcttttt tataatgcca actttgtaca
121 aaaaagttgg catgcagttg ccccccgcgc tgtgcgcccg cctcgctgcg gggcccgggg
181 cggcggagcc actgcctgta gagcgggatc ccgcggctgg ggccgcgccc tttcgcttcg
241 ttgcgcgccc cgtgcgcttc ccgcgggagc accagttctt cgaggatggg gacgtgcagc
301 gccacctcta cctccaggac gtgatcatgc aggtggccga cgtgcctgag aagcccaggg
361 tgcccgcgtt tgcctgccag gtggccggct gctgccaggt gttcgatgcc ctggacgact
421 acgagcacca ctaccacacg ctgcacggaa atgtttgctc cttttgcaag cgggccttcc
481 cttccggaca cctgctggac gcccacatcc tggagtggca cgattcgctc ttccagatcc
541 tgtctgagag gcaggacatg tatcagtgct tggtagaagg ctgcacagag aagttcaaga
601 ccagcagaga ccggaaggat cacatggtga ggatgcacct gtaccccgcg gacttccggt
661 ttgataagcc aaagaaaagc agaagcccag cctcagcaga agccccaggg gacagtggag
721 agcggtcaga aggggaggcc atggaaatct gctctgagcc tgtggcagcc tcccctgcac
781 cggcaggtga gaggcggatc tacagacata gaataccctc taccatctgc tttggtcagg
841 gtgccgctcg aggatttaaa agcaacaaga agaaaaccaa acaatgctgc ccaactttct
901 tgtacaaagt tggcattata agaaagcatt gcttatcaat ttgttgcaac gaacaggtca
961 ctatcagtca aaataaaatc attatttgcc atccagctga tatcccctat agtgagtcgt
1021 attacatggt catagctgtt tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt
1081 acattgcaca agataaaaat atatcatcat gcctcctcta gaccagccag gacagaaatg
1141 cctcgacttc gctgctaccc aaggttgccg ggtgacgcac accgtggaaa cggatgaagg
1201 cacgaaccca gtggacataa gcctgttcgg ttcgtaagct gtaatgcaag tagcgtatgc
1261 gctcacgcaa ctggtccaga accttgaccg aacgcagcgg tggtaacggc gcagtggcgg
1321 ttttcatggc ttgttatgac tgtttttttg gggtacagtc tatgcctcgg gcatccaagc
1381 agcaagcgcg ttacgccgtg ggtcgatgtt tgatgttatg gagcagcaac gatgttacgc
1441 agcagggcag tcgccctaaa acaaagttaa acattatgag ggaagcggtg atcgccgaag
1501 tatcgactca actatcagag gtagttggcg tcatcgagcg ccatctcgaa ccgacgttgc
1561 tggccgtaca tttgtacggc tccgcagtgg atggcggcct gaagccacac agtgatattg
1621 atttgctggt tacggtgacc gtaaggcttg atgaaacaac gcggcgagct ttgatcaacg
1681 accttttgga aacttcggct tcccctggag agagcgagat tctccgcgct gtagaagtca
1741 ccattgttgt gcacgacgac atcattccgt ggcgttatcc agctaagcgc gaactgcaat
1801 ttggagaatg gcagcgcaat gacattcttg caggtatctt cgagccagcc acgatcgaca
1861 ttgatctggc tatcttgctg acaaaagcaa gagaacatag cgttgccttg gtaggtccag
1921 cggcggagga actctttgat ccggttcctg aacaggatct atttgaggcg ctaaatgaaa
1981 ccttaacgct atggaactcg ccgcccgact gggctggcga tgagcgaaat gtagtgctta
2041 cgttgtcccg catttggtac agcgcagtaa ccggcaaaat cgcgccgaag gatgtcgctg
2101 ccgactgggc aatggagcgc ctgccggccc agtatcagcc cgtcatactt gaagctagac
2161 aggcttatct tggacaagaa gaagatcgct tggcctcgcg cgcagatcag ttggaagaat
2221 ttgtccacta cgtgaaaggc gagatcacca aggtagtcgg caaataaccc tcgagccacc
2281 catgaccaaa atcccttaac gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag
2341 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
2401 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
2461 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
2521 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
2581 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
2641 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
2701 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
2761 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
2821 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
2881 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
2941 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
3001 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
3061 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
3121 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
3181 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata
3241 cgcgtaccgc tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc
3301 cttctgctta gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg
3361 ccgttgcttc acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt
3421 caccgacaaa caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat
3481 ttgatgcctg gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac
3541 gacgtt
//
| {
"Vector backbone": "pDONR223",
"Backbone size w/o insert (bp)": "3000",
"Vector type": "Gateway Donor Vector",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Spectinomycin, 50 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "A comprehensive library of human transcription factors for cell fate engineering",
"Publication": "Nat Biotechnol. 2021 Apr;39(4):510-519. doi: 10.1038/s41587-020-0742-6. Epub 2020 Nov 30.",
"DOI": "10.1038/s41587-020-0742-6.",
"Authors": "Ng AHM, Khoshakhlagh P, Rojo Arias JE, Pasquini G, Wang K, Swiersy A, Shipman SL, Appleton E, Kiaee K, Kohman RE, Vernet A, Dysart M, Leeper K, Saylor W, Huang JY, Graveline A, Taipale J, Hill DE, Vidal M, Melero-Martin JM, Busskamp V, Church GM /s---",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/33257861",
"PubMed ID": "PubMed 33257861"
} | {
"Gene/Insert name": "ZNF511",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "759",
"Mutation": "Last nucleotide of stop codon removed to allow for Gateway C-terminal fusions",
"Entrez Gene": "ZNF511",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Gateway Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "M13F(-21)",
"3′ sequencing primer": "M13R"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
TFORF1362 | 142380 | Lentiviral vector for overexpressing transcription factor ORFs with unique 24-bp barcodes. Barcodes facilitate identification of transcription factors in pooled screens. | Feng Zhang | Joung et al Cell. 2023 Jan 5;186(1):209-229.e26. doi: 10.1016/j.cell.2022.11.026. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/397170/0a85afb6-768a-4b76-afa2-936a8c85e665/addgene-plasmid-142380-sequence-397170.gbk | full | LOCUS sequence-397170- 9703 bp DNA circular SYN 26-AUG-2024
DEFINITION Lentiviral vector for overexpressing transcription factor ORFs with
unique 24-bp barcodes. Barcodes facilitate identification of
transcription factors in pooled screens..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9703)
AUTHORS Joung J, Ma S, Tay T, Geiger-Schuller KR, Kirchgatterer PC, Verdine
VK, Guo B, Arias-Garcia MA, Allen WE, Singh A, Kuksenko O, Abudayyeh
OO, Gootenberg JS, Fu Z, Macrae RK, Buenrostro JD, Regev A, Zhang F
TITLE A transcription factor atlas of directed differentiation.
JOURNAL Cell. 2023 Jan 5;186(1):209-229.e26. doi:
10.1016/j.cell.2022.11.026.
PUBMED 36608654
REFERENCE 2 (bases 1 to 9703)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1016/j.cell.2022.11"; journalName: "Cell"; date: "2023-01-5- 5";
volume: "186"; issue: "1"; pages: "209-229.e26"
FEATURES Location/Qualifiers
source 1..9703
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 5..231
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 232..412
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 459..584
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1077..1310
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1495..1539
/codon_start=1
/product="antigenic peptide corresponding to amino acids
655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
et al., 2013)"
/label=gp41 peptide
/note="recognized by the 2H10 single-chain llama nanobody"
/translation="KNEQELLELDKWASL"
promoter 1770..1980
/label=SV40 promoter
/note="SV40 early promoter"
promoter 1792..1988
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 1839..1974
/label=SV40 ori
/note="SV40 origin of replication"
CDS 2020..2622
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGAS"
primer_bind complement(2020..2039)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 2516..2536
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
misc_feature 2665..2782
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2920..4098
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 3151..4089
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 4046..4066
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
misc_feature 5560..6148
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(5613..5633)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(6031..6042)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 6220..6453
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 6531..6652
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(6568..6587)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 6622..6641
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 6692..6827
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(6847..6866)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(6848..6866)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(6876..6893)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(6876..6892)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6885..6907)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 7034..7489
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(7121..7140)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 7331..7352
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 7515..7619
/gene="bla"
/label=AmpR promoter
CDS 7620..8480
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(7838..7857)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 8651..9239
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 9140..9159
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 9393..9410
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 9527..9548
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 9563..9593
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 9601..9617
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
primer_bind 9606..9628
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 9625..9641
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 9625..9641
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 9660..9680
/label=T3
/note="T3 promoter, forward primer"
promoter 9662..9680
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
1 ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat gagttagcaa
61 catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt aaggtggtac
121 gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg aaccactgaa
181 ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa cgggtctctc
241 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
301 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
361 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
421 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
481 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
541 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
601 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
661 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
721 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
781 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
841 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
901 gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg
961 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta
1021 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga
1081 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg
1141 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg
1201 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc
1261 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg
1321 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat
1381 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac
1441 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat
1501 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca
1561 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga
1621 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg
1681 tttcagaccc acctcccaac cccgagggga cccatgcatt gcatctcaat tagtcagcaa
1741 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcgtctca
1801 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
1861 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
1921 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
1981 tttgcaaaaa gctttctaga ggtaccaccg gagcttacca tgaccgagta caagcccacg
2041 gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc
2101 gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc
2161 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg
2221 gacgacggcg ccgccgtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg
2281 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa
2341 cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc
2401 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga
2461 gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac
2521 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga
2581 ccgcgcacct ggtgcatgac ccgcaagccc ggtgccagtt aacgaattct agatcttgag
2641 acaaatggca gtattcatcc acaattttaa aagaaaaggg gggattgggg ggtacagtgc
2701 aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat tacaaaaaca
2761 aattacaaaa attcaaaatt ttcgggttta ttacagggac agcagagatc cactttggcg
2821 ccggctcgag gcctgcaggt gcaaagatgg ataaagtttt aaacagagag gaatctttgc
2881 agctaatgga ccttctaggt cttgaaagga gtgggaattg gctccggtgc ccgtcagtgg
2941 gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacc
3001 ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc
3061 ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt
3121 tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc ccgcgggcct
3181 ggcctcttta cgggttatgg cccttgcgtg ccttgaatta cttccacctg gctgcagtac
3241 gtgattcttg atcccgagct tcgggttgga agtgggtggg agagttcgag gccttgcgct
3301 taaggagccc cttcgcctcg tgcttgagtt gaggcctggc ctgggcgctg gggccgccgc
3361 gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc tctagccatt
3421 taaaattttt gatgacctgc tgcgacgctt tttttctggc aagatagtct tgtaaatgcg
3481 ggccaagatc tgcacactgg tatttcggtt tttggggccg cgggcggcga cggggcccgt
3541 gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc gagaatcgga
3601 cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg gcctcgcgcc gccgtgtatc
3661 gccccgccct gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc ggaaagatgg
3721 ccgcttcccg gccctgctgc agggagctca aaatggagga cgcggcgctc gggagagcgg
3781 gcgggtgagt cacccacaca aaggaaaagg gcctttccgt cctcagccgt cgcttcatgt
3841 gactccacgg agtaccgggc gccgtccagg cacctcgatt agttctcgag cttttggagt
3901 acgtcgtctt taggttgggg ggaggggttt tatgcgatgg agtttcccca cactgagtgg
3961 gtggagactg aagttaggcc agcttggcac ttgatgtaat tctccttgga atttgccctt
4021 tttgagtttg gatcttggtt cattctcaag cctcagacag tggttcaaag tttttttctt
4081 ccatttcagg tgtcgtgagg ctagcaccgc caccatgatg ttcacggaag accagggtgt
4141 agatgacagg ctgctctatg acattgtatt caagcacttc aaaagaaata aggtggagat
4201 ttcaaatgca ataaaaaaga catttccatt cctcgagggc ctccgtgatc gtgatctcat
4261 cacaaataaa atgtttgaag attctcaaga ttcttgtaga aacctggtcc ctgtacagag
4321 agtggtgtac aatgttctta gtgaactgga gaagacattt aacctgccag ttctggaagc
4381 actgttcagc gatgtcaaca tgcaggaata ccccgattta attcacattt ataaaggctt
4441 tgaaaatgta atccatgaca aattgcctct ccaagaaagt gaagaagaag agagggagga
4501 gaggtctggc ctccaactaa gtcttgaaca aggaactggt gaaaactctt ttcgaagcct
4561 gacttggcca ccttcgggtt ccccatctca tgctggtaca accccacctg aaaatggact
4621 ctcagagcac ccctgtgaaa cagaacagat aaatgcaaag agaaaagata caaccagtga
4681 caaagatgat tcgctaggaa gccaacaaac aaatgaacaa tgtgctcaaa aggctgagcc
4741 aacagagtcc tgcgaacaaa ttgctgtcca agtgaataat ggggatgctg gaagggagat
4801 gccctgcccg ttgccctgtg atgaagaaag cccagaggca gagctacaca accatggaat
4861 ccaaattaat tcctgttctg tgcgactggt ggatataaaa aaggaaaagc cattttctaa
4921 ttcaaaagtt gagtgccaag cccaagcaag aactcatcat aaccaggcat ctgacataat
4981 agtcatcagc agtgaggact ctgaaggatc cactgacgtt gatgagccct tagaagtctt
5041 catctcagca ccgagaagtg agcctgtgat caataatgac aaccctttag aatcaaatga
5101 tgaaaaggag ggccaagaag ccacttgctc acgaccccag attgtaccag agcccatgga
5161 tttcagaaaa ttatctacat tcagagaaag ttttaagaaa agagtgatag gacaagacca
5221 cgacttttca gaatccagtg aggaggaggc gcccgcagaa gcctcgagcg gggcactgag
5281 aagcaagcat ggtgagaagg ctcctatgac ttctagaagt acatctactt ggagaatacc
5341 cagcaggaag agacgtttca gcagtagtga cttttcagac ctgagtaatg gagaagagct
5401 tcaggaaacc tgcagctcat ccctaagaag agggtcaggt aaagaagatt agtaatgaac
5461 tagtccgtaa cttgaaagta tttcgatttc ttggctttat atatcttgtg gaaaggacga
5521 agtcaagaat tgtatcgact taatacgcgt taagtcgaca atcaacctct ggattacaaa
5581 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
5641 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
5701 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
5761 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
5821 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
5881 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
5941 gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt
6001 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
6061 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
6121 cggatctccc tttgggccgc ctccccgcgt cgactttaag accaatgact tacaaggcag
6181 ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc
6241 aacgaagaca agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag
6301 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
6361 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
6421 gaccctttta gtcagtgtgg aaaatctcta gcagtacgta tagtagttca tgtcatctta
6481 ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg aacttgttta
6541 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
6601 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
6661 ggctctagct atcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
6721 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
6781 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag ggacgtaccc
6841 aattcgccct atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt
6901 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
6961 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
7021 aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
7081 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
7141 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
7201 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
7261 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
7321 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
7381 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
7441 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt
7501 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
7561 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
7621 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
7681 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
7741 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
7801 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
7861 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
7921 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
7981 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
8041 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
8101 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
8161 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
8221 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
8281 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
8341 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
8401 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
8461 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
8521 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
8581 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
8641 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
8701 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
8761 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
8821 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
8881 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
8941 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
9001 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
9061 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
9121 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
9181 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
9241 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
9301 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
9361 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
9421 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
9481 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
9541 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
9601 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg
9661 caattaaccc tcactaaagg gaacaaaagc tggagctgca agc
//
| {
"Vector backbone": "pLX_TRC317",
"Backbone size w/o insert (bp)": "8264",
"Vector type": "Mammalian Expression, Lentiviral",
"Selectable markers": "Puromycin"
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "NEB Stable",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "A transcription factor atlas of directed differentiation",
"Publication": "Cell. 2023 Jan 5;186(1):209-229.e26. doi: 10.1016/j.cell.2022.11.026.",
"DOI": "10.1016/j.cell.2022.11.026.",
"Authors": "Joung J, Ma S, Tay T, Geiger-Schuller KR, Kirchgatterer PC, Verdine VK, Guo B, Arias-Garcia MA, Allen WE, Singh A, Kuksenko O, Abudayyeh OO, Gootenberg JS, Fu Z, Macrae RK, Buenrostro JD, Regev A, Zhang F /jcell",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/36608654",
"PubMed ID": "PubMed 36608654"
} | {
"Gene/Insert name": "SP100",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "1439",
"Mutation": "",
"Entrez Gene": "SP100",
"Promoter": "EF1a",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Unknown",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "GGGTGGAGACTGAAGTTAGGCCAG",
"3′ sequencing primer": "CACATAGCGTAAAAGGAGCAACATAG"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
KHBD00161 | 34314 | Bart Deplancke | Korneel et al Nat Methods. 2011 Oct 30;8(12):1065-70. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/41988/dc6cc583-266c-456c-adc7-6b990eddf939/addgene-plasmid-34314-sequence-41988.gbk | partial | LOCUS sequence-41988-e 719 bp DNA linear UNA 26-AUG-2024
DEFINITION natural linear DNA
ACCESSION .
VERSION .
KEYWORDS .
SOURCE natural DNA sequence
ORGANISM unspecified
REFERENCE 1 (bases 1 to 719)
AUTHORS Korneel Hens, Jean-Daniel Feuz, Alina Isakova, Antonina Lagovitina,
Andreas Massouras, Julien Bryois, Patrick Callaerts, Susan E.
Celniker, Bart Deplancke
TITLE Automated protein-DNA interaction screening of Drosophila regulatory
elements
JOURNAL Nat Methods. 2011 Oct 30;8(12):1065-70.
PUBMED 22037703
REFERENCE 2 (bases 1 to 719)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat
Methods."; date: "2011-10-30"; volume: "8(12)"; pages: "1065-70"
FEATURES Location/Qualifiers
source 1..719
/mol_type="genomic DNA"
/organism="unspecified"
protein_bind 66..90
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
ORIGIN
1 ttttattttg actgatagtg acctgttcgt tgcaacaaat tgataagcaa tgctttctta
61 taatgccaac tttgtacaag aaagttggca agctctctaa catttccgga gagctgggaa
121 tggtttgctc cattttgttg cggtgggtta gcgtttgcag gtgagctttc agctggtgca
181 gcagagtgaa tgacttgcaa cagataccac agttgtaatt cttttgggcg gagtgcgaaa
241 ggatgtgatt tttgaggatg gctgtgtgcg taaaagtttt cccgcactcc tggcaggcat
301 aaggacgcgt gttggtgtgg acccgttcat ggcggacccg ttcgctgcga tcgtagaagt
361 ggcggtcgca gtacatacac ttgtatggac ggtcgcccgt atgacgacgg atgtgtgctc
421 gcaattcgcc catttgtcga aaagatttat tgcatatttc gcattggtgg ggcttgtact
481 ctgaatgggt catcatgtgg gccgtgaaaa cagatttctt gtaaaagacg tttccgcaca
541 ggatgcacat gaacttctcc tcttccttgc actttggctg gttccccctt tccttggcgt
601 tgatatcctt gcgtttgaat ttcagttcgg cgtccggttt gcgaggcctt cccggctttt
661 gtttgtccat tttgaagctg ggatgtggcg aaggagcgct tgacatgaga agttcctcg
//
| {
"Vector backbone": "pDONR221",
"Backbone size w/o insert (bp)": "2536",
"Vector type": "Gateway Donor Vector",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Kanamycin, 50 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Automated protein-DNA interaction screening of Drosophila regulatory elements",
"Publication": "Nat Methods. 2011 Oct 30;8(12):1065-70.",
"DOI": "10.1038/nmeth.1763",
"Authors": "Korneel Hens, Jean-Daniel Feuz, Alina Isakova, Antonina Lagovitina, Andreas Massouras, Julien Bryois, Patrick Callaerts, Susan E Celniker, Bart Deplancke",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/22037703",
"PubMed ID": "PubMed 22037703"
} | {
"Gene/Insert name": "CG14710",
"Species": "D. melanogaster (fly)",
"Insert Size (bp)": "1341",
"Mutation": "",
"Entrez Gene": "CG14710",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Gateway Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "M13F",
"3′ sequencing primer": "M13R"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
||
pD649-HAsp-IGFBP7-Fc(DAPA)-AviTag-6xHis | 156706 | Mammalian expression of secreted protein fused to Fc(DAPA)-Avi-6xHis. | Chris Garcia | Wojtowicz et al Cell. 2020 Aug 20;182(4):1027-1043.e17. doi: 10.1016/j.cell.2020.07.025. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/302521/779c6851-975f-4990-b4eb-8709915c6ccd/addgene-plasmid-156706-sequence-302521.gbk | full | LOCUS sequence-302521- 6874 bp DNA circular SYN 26-AUG-2024
DEFINITION Mammalian expression of secreted protein fused to
Fc(DAPA)-Avi-6xHis..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6874)
AUTHORS Wojtowicz WM, Vielmetter J, Fernandes RA, Siepe DH, Eastman CL,
Chisholm GB, Cox S, Klock H, Anderson PW, Rue SM, Miller JJ, Glaser
SM, Bragstad ML, Vance J, Lam AW, Lesley SA, Zinn K, Garcia KC
TITLE A Human IgSF Cell-Surface Interactome Reveals a Complex Network of
Protein-Protein Interactions.
JOURNAL Cell. 2020 Aug 20;182(4):1027-1043.e17. doi:
10.1016/j.cell.2020.07.025.
PUBMED 32822567
REFERENCE 2 (bases 1 to 6874)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1016/j.cell.2020.07"; journalName: "Cell"; date: "2020-08-20-
20"; volume: "182"; issue: "4"; pages: "1027-1043.e17"
FEATURES Location/Qualifiers
source 1..6874
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 124..143
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
primer_bind 149..168
/label=pENTR-R
/note="pENTR vectors, reverse primer"
rep_origin 254..842
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 743..762
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 996..1013
/label=L4440
/note="L4440 vector, forward primer"
primer_bind 1111..1128
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 1112..1128
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 1584..1600
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
enhancer 1767..2146
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 2147..2350
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 2301..2321
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 2347..2371
/label=LNCX
/note="Human CMV promoter, forward primer"
promoter 2581..2599
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 2581..2598
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind 2612..2631
/label=pMT2-F
/note="Synthetic intron, forward primer"
CDS 4216..4260
/codon_start=1
/product="peptide tag that allows for enzymatic
biotinylation"
/label=AviTag(TM)
/translation="GLNDIFEAQKIEWHE"
CDS 4261..4278
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(4418..4435)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 4424..4535
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(4653..4673)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 4658..4987
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 4838..4973
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4900..4919
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter 5035..5082
/label=EM7 promoter
/note="synthetic bacterial promoter "
CDS 5101..5700
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind complement(5101..5120)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 5597..5617
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
polyA_signal 5830..5951
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5867..5886)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 5921..5940
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
CDS complement(6004..6864)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 6627..6646
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(join(6865..6874,1..82))
/gene="bla"
/label=AmpR promoter
ORIGIN
1 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
61 tgtatttaga aaaataaaca aataggggtc agtgttacaa ccaattaacc aattctgaac
121 attatcgcga gcccatttat acctgaatat ggctcataac accccttgct catgaccaaa
181 atcccttaac gtgagttacg cgcgcgtcgt tccactgagc gtcagacccc gtagaaaaga
241 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
301 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
361 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
421 tagcccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
481 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
541 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
601 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
661 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
721 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
781 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
841 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
901 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
961 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
1021 aaggcgagag tagggaactg ccaggcatca aactaagcag aaggcccctg acggatggcc
1081 tttttgcgtt tctacaaact ctttctgtgt tgtaaaacga cggccagtct taagctcggg
1141 ccccctgggc ggttctgata acgagtaatc gttaatccgc aaataacgta aaaacccgct
1201 tcggcgggtt tttttatggg gggagtttag ggaaagagca tttgtcagaa tatttaaggg
1261 cgcctgtcac tttgcttgat atatgagaat tatttaacct tataaatgag aaaaaagcaa
1321 cgcactttaa ataagatacg ttgctttttc gattgatgaa cacctataat taaactattc
1381 atctattatt tatgattttt tgtatataca atatttctag tttgttaaag agaattaaga
1441 aaataaatct cgaaaataat aaagggaaaa tcagtttttg atatcaaaat tatacatgtc
1501 aacgataata caaaatataa tacaaactat aagatgttat cagtatttat tatcatttag
1561 aataaatttt gtgtcgccct taattgtgag cggataacaa ttacgagctt catgcacagt
1621 ggcgttgact caatattggc cattagccat attattcatt ggttatatag cataaatcaa
1681 tattggctat tggccattgc atacgttgta tctatatcat aatatgtaca tttatattgg
1741 ctcatgtcca atatgaccgc catgttggca ttgattattg actagttatt aatagtaatc
1801 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
1861 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
1921 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
1981 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
2041 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
2101 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
2161 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
2221 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
2281 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
2341 aagcagagct cgtttagtga accgtcagat cgcctggaga cgccatccac gctgttttga
2401 cctccataga agacaccggg accgatccag cctccgcggc cgggaacggt gcattggaac
2461 gcggattccc cgtgccaaga gtgacgtaag taccgcctat agagtctata ggcccacccc
2521 cttggcttcg ttagaacgcg gctacaatta atacataacc ttatgtatca tacacatacg
2581 atttaggtga cactatagaa taacatccac tttgcctttc tctccacagg tgtccactcc
2641 caggtccaac tgcacctcgg ttctatcgaa aacgcgcctg ctagcgccac catgaagacc
2701 atcattgccc tgagctacat cttctgcctg gtgttcgctg cggccgcctc tagctctgat
2761 acctgtggac cttgtgaacc cgccagctgt cctccattgc ctcctcttgg atgtctgctg
2821 ggcgagacaa gagatgcctg cggctgttgt cctatgtgtg ccagaggcga aggcgaacct
2881 tgtggcggag gcggagctgg aagaggatat tgtgcccctg gcatggaatg cgtgaagtcc
2941 agaaagcgga gaaagggcaa agctggtgct gctgctggtg gacctggtgt ttctggcgtg
3001 tgtgtgtgca agagcagata ccctgtgtgt ggcagcgacg gcacaacata tccaagcggc
3061 tgtcagctga gagctgcctc tcagagagcc gagagtagag gcgagaaggc catcacacag
3121 gtgtccaagg gcacatgtga acagggccct agcatcgtga cccctccaaa ggacatctgg
3181 aatgtgacag gcgcccaggt gtacctgagc tgcgaagtga tcggaatccc cacaccagtg
3241 ctgatctgga acaaagtgaa gcggggccac tacggcgtgc agagaacaga attgctgccc
3301 ggcgacagag acaacctggc cattcaaaca agaggcggac ccgagaagca cgaagtgaca
3361 ggatgggttc tcgtgtcccc tctgtctaaa gaggatgccg gcgagtacga gtgccacgcc
3421 tctaattctc aaggccaggc ctctgccagc gccaagatta cagtggtgga tgccctgcac
3481 gagatccccg tgaaaaaagg cgagggcgcc gaacttggcg cgccgggcgg cgacaagacc
3541 cacacctgcc ccccctgccc agctccagaa ctgctgggag ggccttccgt gttcctgttc
3601 ccccccaagc ccaaggacac cctgatgatc agcaggaccc ccgaggtgac ctgcgtggtg
3661 gtggccgtgt cccacgagga cccagaggtg aagttcaact ggtacgtgga cggcgtggag
3721 gtgcacaacg ccaagaccaa gcccagagag gagcagtaca acagcaccta cagggtggtg
3781 tccgtgctga ccgtgctgca ccaggactgg ctgaacggca aagaatacaa gtgcaaagtc
3841 tccaacaagg ccctggctgc cccaatcgaa aagacaatca gcaaggccaa gggccagcca
3901 cgggagcccc aggtgtacac cctgcccccc agccgggagg agatgaccaa gaaccaggtg
3961 tccctgacct gtctggtgaa gggcttctac cccagcgata tcgccgtgga gtgggagagc
4021 aacggccagc ccgagaacaa ctacaagacc acccccccag tgctggacag cgacggcagc
4081 ttcttcctgt acagcaagct gaccgtggac aagtccaggt ggcagcaggg caacgtgttc
4141 agctgcagcg tgatgcacga ggccctgcac aaccactaca cccagaagtc cctgagcctg
4201 agccccggcg gcagcggtct taatgatatt ttcgaggctc agaagattga gtggcatgaa
4261 caccaccacc atcaccacta aggccggcct gtagtcactg gcacaacgcg ttacccggta
4321 agccaatcgg gtatacacgg tcgtcatact gcagacaggg ttcttctact ttgcaagata
4381 gtcttgagta gtaaaataaa tagatagaga aaatcagcct cgactgtgcc ttctagttgc
4441 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
4501 actgtccttt cctaataaaa tgaggaaatt gcatcacaac actcaaccct atctcggtct
4561 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
4621 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
4681 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
4741 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
4801 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
4861 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
4921 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
4981 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcagc acgtgttgac
5041 aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc
5101 atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta
5161 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac
5221 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac
5281 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag
5341 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt
5401 tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag
5461 cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc
5521 agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg
5581 gagacatccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc
5641 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga
5701 cacgtgctac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
5761 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
5821 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
5881 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
5941 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
6001 tcattaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
6061 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
6121 ccccagcgct gcgatgatac cgcgagaacc acgctcaccg gctccggatt tatcagcaat
6181 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
6241 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
6301 caacgttgtt gccatcgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
6361 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
6421 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
6481 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
6541 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
6601 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
6661 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
6721 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
6781 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
6841 gacacggaaa tgttgaatac tcatattctt cctt
//
| {
"Vector backbone": "pD649",
"Backbone size w/o insert (bp)": "6106",
"Vector type": "Mammalian Expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "A Human IgSF Cell-Surface Interactome Reveals a Complex Network of Protein-Protein Interactions",
"Publication": "Cell. 2020 Aug 20;182(4):1027-1043.e17. doi: 10.1016/j.cell.2020.07.025.",
"DOI": "10.1016/j.cell.2020.07.025.",
"Authors": "Wojtowicz WM, Vielmetter J, Fernandes RA, Siepe DH, Eastman CL, Chisholm GB, Cox S, Klock H, Anderson PW, Rue SM, Miller JJ, Glaser SM, Bragstad ML, Vance J, Lam AW, Lesley SA, Zinn K, Garcia KC /jcell",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/32822567",
"PubMed ID": "PubMed 32822567"
} | {
"Gene/Insert name": "IGFBP7",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "768",
"Mutation": "Fc region of human IgG contains D265A and P329A mutations (DAPA)",
"Entrez Gene": "IGFBP7",
"Promoter": "CMV/SP6",
"Tag / Fusion Protein": "Fc(DAPA)-AviTag-6xHis (C terminal on backbone)"
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "GCGGCCGC (not destroyed)",
"3′ cloning site": "GGCGCGCC (not destroyed)",
"5′ sequencing primer": "ATTTAGGTGACACTATAG",
"3′ sequencing primer": "CACGTACCAGTTGAACTTCACC"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
mPA-GFP-MCHR1-N-10 | 57147 | Localization: Cilium, Excitation: 400 / 504, Emission: 515 / 517 | Michael Davidson | Davidson Photoactivatable Fluorescent Proteins (unpublished) | https://media.addgene.org/snapgene-media/v2.0.0/sequences/175698/ff3406e5-b78c-4951-acf6-5102b707f9d2/addgene-plasmid-57147-sequence-175698.gbk | full | LOCUS sequence-175698- 5746 bp DNA circular SYN 26-AUG-2024
DEFINITION Localization: Cilium, Excitation: 400 / 504, Emission: 515 / 517.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5746)
TITLE Davidson Photoactivatable Fluorescent Proteins
REFERENCE 2 (bases 1 to 5746)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..5746
/mol_type="other DNA"
/organism="synthetic DNA construct"
regulatory 1084..1093
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 1090..1806
/codon_start=1
/product="photoactivatable GFP variant (Patterson et al.,
2002)"
/label=PA-GFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(1135..1156)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1396..1415)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1743..1764
/label=EGFP-C
/note="EGFP, forward primer"
polyA_signal 1932..2053
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1969..1988)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2023..2042
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(2060..2515)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2197..2218)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2409..2428
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2542..2646
/gene="bla"
/label=AmpR promoter
promoter 2648..3005
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
enhancer 2648..2839
/label=SV40 enhancer
/note="enhancer for the SV40 early promoter (Herr, 1993)"
primer_bind complement(2671..2691)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 2856..2991
/label=SV40 ori
/note="SV40 origin of replication"
CDS 3040..3834
/codon_start=1
/product="aminoglycoside phosphotransferase"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
CDS 3040..3834
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(3094..3113)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3704..3723
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(4022..4041)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 4066..4113
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4442..5030
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4931..4950
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
enhancer 5205..5508
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 5509..5712
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 5663..5683
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory join(5741..5746,1..4)
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
ORIGIN
1 atggatctgc aagcctcgtt gctgtccact ggccccaatg ccagcaacat ctccgatggc
61 caggataatt tcacattggc ggggccacct cctcgcacaa ggagtgtctc ctacatcaac
121 atcatcatgc cttcagtgtt tggtaccatc tgtctcctgg gcattgtggg aaactccaca
181 gtcatttttg ccgtggtgaa gaaatccaag ctgcactggt gcagcaacgt ccctgacatc
241 ttcatcatca acctctctgt ggtggatctg cttttcctgc tgggcatgcc tttcatgatc
301 caccagctca tgggtaatgg tgtctggcac tttggggaaa ccatgtgcac cctcatcaca
361 gccatggacg ccaacagtca gttcaccagc acctacatcc tgactgctat ggccattgac
421 cgctacttgg ccaccgtcca tcccatctcc tccaccaagt tccggaagcc ctccatggcc
481 accctggtga tctgcctcct gtgggctctc tcgttcatta gcatcactcc tgtgtggctc
541 tatgccaggc ttatcccctt cccagggggt gctgtgggct gtggcatccg cctaccaaac
601 ccagatactg atctttactg gttcactctg tatcagtttt tcctggcctt cgcccttccg
661 tttgtggtca tcactgctgc gtacgtgaaa atactacagc gcatgacgtc ttcggtggcc
721 ccagcctctc aacgcagcat ccggcttcgg acaaagaggg tgacccgcac agccattgcc
781 atctgtctgg tcttctttgt gtgctgggcg ccctactacg tgctgcagct gacccagttg
841 tccatcagcc gcccgaccct cacattcgtc tacctgtaca atgcggctat cagcttgggc
901 tatgccaaca gctgcctcaa tccctttgtg tacatagtac tctgtgagac ctttcgaaaa
961 cgcttggtgc tgtcggtgaa gcccgcggcc caggggcagc ttcgcacggt cagcaatgct
1021 cagacagctg acgaggagag gacagaaagc aaaggcaccg gaggtagcgg ggatccaccg
1081 gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
1141 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
1201 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
1261 tggcccaccc tcgtgaccac cttcagctac ggcgtgcagt gcttcagccg ctaccccgac
1321 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
1381 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
1441 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
1501 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
1561 cagaagaacg gcatcaaggc caacttcaag atccgccaca acatcgagga cggcagcgtg
1621 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
1681 gacaaccact acctgagcca ccagtccaag ctgagcaaag accccaacga gaagcgcgat
1741 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
1801 tacaagtaaa gcggccgcga ctctagatca taatcagcca taccacattt gtagaggttt
1861 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
1921 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
1981 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
2041 tcaatgtatc ttaaggcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
2101 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
2161 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
2221 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
2281 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
2341 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
2401 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
2461 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt
2521 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
2581 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
2641 aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt
2701 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
2761 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
2821 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
2881 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
2941 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
3001 gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
3061 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
3121 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
3181 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta
3241 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
3301 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
3361 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
3421 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
3481 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
3541 gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc
3601 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
3661 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
3721 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
3781 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
3841 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
3901 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
3961 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa
4021 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac
4081 agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct
4141 ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct
4201 tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg
4261 cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc
4321 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
4381 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
4441 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
4501 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
4561 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
4621 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
4681 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
4741 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
4801 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
4861 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
4921 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
4981 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
5041 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
5101 cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta ttaatagtaa
5161 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
5221 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
5281 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
5341 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
5401 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
5461 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
5521 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
5581 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
5641 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
5701 ataagcagag ctggtttagt gaaccgtcag atccgctagc gccacc
//
| {
"Vector backbone": "mPA-GFP-N1",
"Backbone size w/o insert (bp)": "4750",
"Vector type": "Mammalian Expression",
"Selectable markers": "Neomycin (select with G418)"
} | {
"Bacterial Resistance(s)": "Kanamycin, 50 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Industry MTA"
} | {
"Title": "",
"Publication": "",
"DOI": "",
"Authors": "",
"PubMed Link": "",
"PubMed ID": ""
} | {
"Gene/Insert name": "MCHR1",
"Species": "M. musculus (mouse)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "CMV",
"Tag / Fusion Protein": "mPA-GFP (C terminal on backbone)"
} | {
"Cloning method": "Unknown",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "CMV-F",
"3′ sequencing primer": "SV40pA-R"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
mRuby2-Actin-7 | 55888 | Localization: Actin, Excitation: 559, Emission: 600 | Michael Davidson | Lam et al Nat Methods. 2012 Sep 9. doi: 10.1038/nmeth.2171. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/108820/1413c98a-2d7a-4d68-b67f-da64b8316e7b/addgene-plasmid-55888-sequence-108820.gbk | partial | LOCUS sequence-108820- 895 bp DNA linear UNA 26-AUG-2024
DEFINITION Localization: Actin, Excitation: 559, Emission: 600.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE natural DNA sequence
ORGANISM unspecified
REFERENCE 1 (bases 1 to 895)
AUTHORS Lam AJ, St-Pierre F, Gong Y, Marshall JD, Cranfill PJ, Baird MA,
McKeown MR, Wiedenmann J, Davidson MW, Schnitzer MJ, Tsien RY, Lin
MZ
TITLE Improving FRET dynamic range with bright green and red fluorescent
proteins.
JOURNAL Nat Methods. 2012 Sep 9. doi: 10.1038/nmeth.2171.
PUBMED 22961245
REFERENCE 2 (bases 1 to 895)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat
Methods. 2012 Sep 9. doi: 10.1038/nmeth.2171."
FEATURES Location/Qualifiers
source 1..895
/mol_type="genomic DNA"
/organism="unspecified"
ORIGIN
1 gagatgatgt atccagcaga tggtggtctg aggggataca ctcatatggc actgaaagtt
61 gatggtggtg gccatctgtc ttgctctttc gtaacaactt acaggtcaaa aaagaccgtc
121 gggaacatca agatgcccgg tatccatgcc gttgatcacc gcctggaaag gttagaggaa
181 agtgacaatg aaatgttcgt agtacaacgc gaacacgcag ttgccaagtt cgccgggctt
241 ggtggtggga tggacgagct gtacaagtcc ggactcagat ctcgagctat ggatgatgat
301 atcgccgcgc tcgtcgtcga caacggctcc ggcatgtgca aggccggctt cgcgggcgac
361 gatgcccccc gggccgtctt cccctccatc gtggggcgcc ccaggcacca gggcgtgatg
421 gtgggcatgg gtcagaagga ttcctatgtg ggcgacgagg cccagagcaa gagaggcatc
481 ctcaccctga agtaccccat cgagcacggc atcgtcacca actgggacga catggagaaa
541 atctggcacc acaccttcta caatgagctg cgtgtggctc ccgaggagca ccccgtgctg
601 ctgaccgagg cccccctgaa ccccaaggcc aaccgcgaga agatgaccca gatcatgttt
661 gagaccttca acaccccagc catgtacgtt gctatccagg ctgtgctatc cctgtacgcc
721 tctggccgta ccactggcat cgtgatggac tccggtgacg gggtcaccca cactgtgccc
781 atctacgagg ggtatgccct cccccatgcc atcctgcgtc tggacctggc tggccgggac
841 ctgactgact acctcatgaa gatcctcacc gagcgcggct acagcttcac cacca
//
| {
"Vector backbone": "mRuby2",
"Backbone size w/o insert (bp)": "4750",
"Vector type": "Mammalian Expression",
"Selectable markers": "Neomycin (select with G418)"
} | {
"Bacterial Resistance(s)": "Kanamycin, 50 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Industry MTA"
} | {
"Title": "Improving FRET dynamic range with bright green and red fluorescent proteins",
"Publication": "Nat Methods. 2012 Sep 9. doi: 10.1038/nmeth.2171.",
"DOI": "10.1038/nmeth.2171.",
"Authors": "Lam AJ, St-Pierre F, Gong Y, Marshall JD, Cranfill PJ, Baird MA, McKeown MR, Wiedenmann J, Davidson MW, Schnitzer MJ, Tsien RY, Lin MZ /nmeth",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/22961245",
"PubMed ID": "PubMed 22961245"
} | {
"Gene/Insert name": "Actin",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "ACTB",
"Promoter": "CMV",
"Tag / Fusion Protein": "mRuby2"
} | {
"Cloning method": "Unknown",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "CMV-F",
"3′ sequencing primer": "SV40pA-R"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pDONR221-SLC44A5_STOP | 161321 | Gateway entry clone with codon-optimized ORF sequence for subcloning into custom expression vectors. Contains a STOP codon at the end of the codon-optimized ORF sequence. | Superti-Furga et al Nat Rev Drug Discov. 2020 Jul;19(7):429-430. doi: 10.1038/d41573-020-00056-6. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/373518/177ec0fd-2f75-41f3-90f8-7bbc53acc484/addgene-plasmid-161321-sequence-373518.gbk | full | LOCUS sequence-373518- 4710 bp DNA circular SYN 26-AUG-2024
DEFINITION Gateway entry clone with codon-optimized ORF sequence for subcloning
into custom expression vectors. Contains a STOP codon at the end of
the codon-optimized ORF sequence. .
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4710)
AUTHORS Superti-Furga G, Lackner D, Wiedmer T, Ingles-Prieto A, Barbosa B,
Girardi E, Goldmann U, Gurtl B, Klavins K, Klimek C, Lindinger S,
Lineiro-Retes E, Muller AC, Onstein S, Redinger G, Reil D, Sedlyarov
V, Wolf G, Crawford M, Everley R, ... Steppan CM
TITLE The RESOLUTE consortium: unlocking SLC transporters for drug
discovery.
JOURNAL Nat Rev Drug Discov. 2020 Jul;19(7):429-430. doi:
10.1038/d41573-020-00056-6.
PUBMED 32265506
REFERENCE 2 (bases 1 to 4710)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10"; journalName:
"Nat Rev Drug Discov"; date: "2020-07"; volume: "19"; issue: "7";
pages: "429-430"
FEATURES Location/Qualifiers
source 1..4710
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind 1..100
/gene="mutant version of attL"
/label=attL1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
protein_bind complement(2267..2366)
/gene="mutant version of attL"
/label=attL2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
primer_bind complement(2383..2402)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(2384..2402)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(2407..2423)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2407..2423)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(2525..2544)
/label=pENTR-R
/note="pENTR vectors, reverse primer"
CDS 2536..3345
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind complement(2603..2622)
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
rep_origin 3492..4080
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3981..4000
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 4234..4251
/label=L4440
/note="L4440 vector, forward primer"
terminator complement(4410..4437)
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator complement(4529..4615)
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
primer_bind 4664..4686
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 4678..4695
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 4679..4695
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
ORIGIN
1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacacat tgatgagcaa
61 tgctttttta taatgccaac tttgtacaaa aaagcaggct ccaccatgaa tgataccgag
121 aagcctgccg acacaccatc cgaggaggag gattttggcg accccagaac ctacgatcct
181 gacttcaagg gcccagtggc caacaggtct tgcacagatg tgctgtgctg tatgatcttt
241 ctgctgtgca tcatcggcta catcgtgctg ggactggtgg catgggtgca cggcgatcca
301 aggagagccg cctatcccac agacagccag ggacacttct gcggacagaa gggaacccct
361 aacgagaata agacaatcct gttctacttc aacctgctgc gctgcacctc tcctagcgtg
421 ctgctgaatc tgcagtgtcc aaccacacag atctgcgtga gcaagtgtcc agagaagttt
481 ctgacctacg tggagatgca gctgctgtat acaaaggata agtcctactg ggaggactat
541 cggcagttct gtaagaccac agccaagcca gtgaagtctc tgacccagct gctgctggac
601 gatgactgcc ccacagccat cttccccagc aagccttttc tgcagcggtg cttccctgat
661 ttttccacca agaacggcac cctgacaatc ggctctaaga tgatgttcca ggacggcaat
721 ggaggaacac ggtccgtggt ggagctggga atcgcagcaa acggcatcaa taagctgctg
781 gatgccaagt ctctgggcct gaaggtgttc gaggactacg ccagaacctg gtattggatc
841 ctgatcggcc tgacaatcgc catggtgctg tcttggatct tcctgatcct gctgaggttt
901 atcgccggct gcctgttctg ggtgtttatg atcggcgtga tcggcatcat cggctacggc
961 atctggcact gttaccagca gtataccaac ctgcaggaga gacccagctc cgtgctgacc
1021 atctacgata tcggcatcca gacaaatatc tccatgtatt ttgagctgca gcagacctgg
1081 ttcaccttca tgatcatcct gtgcatcatc gaagtgatcg tgatcctgat gctgatcttc
1141 ctgaggaatc gcatccgggt ggccatcatc ctgctgaagg agggcagcaa ggccatcggc
1201 tacgtgcctt ccaccctggt gtatcccgcc ctgaccttca tcctgctgag catctgcatc
1261 tgttactggg tggtgaccgc cgtgttcctg gccacatccg gagtgccagt gtataaagtg
1321 atcgcaccag gaggacactg catccacgag aaccagacct gtgaccccga gatctttaat
1381 accacagaga tcgcaaaggc atgccctggc gccctgtgca acttcgcctt ttacggcggc
1441 aagagcctgt accaccagta tatccccacc ttccacgtgt ataacctgtt cgtgtttctg
1501 tggctgatca attttgtgat cgccctggga cagtgcgcac tggcaggagc cttcgccacc
1561 tactattggg ccatgaagaa gcctgatgac atcccaaggt accccctgtt caccgccttt
1621 ggcagagcca tcaggtatca cacaggctcc ctggcctttg gctctctgat catcgccctg
1681 atccagatgt tcaagatcgt gctggagtac ctggaccacc gcctgaagcg gacccagaac
1741 acactgagca agttcctgca gtgctgtctg cgctgctgtt tttggtgcct ggagaatgcc
1801 atcaagttcc tgaaccggaa tgcctacatc atgatcgcca tctatggcag aaacttctgt
1861 aggtccgcca aggatgcctt taacctgctg atgagaaatg tgctgaaggt ggccgtgacc
1921 gacgaggtga catacttcgt gctgtttctg ggcaagctgc tggtggcagg atctatcggc
1981 gtgctggcct ttctgttctt tacccagagg ctgcccgtga tcgcacaggg acctgccagc
2041 ctgaactact attgggtgcc actgctgaca gtgatcttcg gctcttacct gatcgcccac
2101 ggcttcttta gcgtgtatgc catgtgcgtg gagaccatct tcatctgttt tctggaggat
2161 ctggagagga acgacggatc caccgcaagg ccatacttcg tgacacctaa tctgcacggc
2221 atcctgatca agaagcagct ggtgccccag aagcagaagg agtaagaccc agctttcttg
2281 tacaaagttg gcattataag aaagcattgc ttatcaattt gttgcaacga acaggtcact
2341 atcagtcaaa ataaaatcat tatttgccat ccagctgata tcccctatag tgagtcgtat
2401 tacatggtca tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac
2461 attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt
2521 aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg attaaattcc
2581 aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt
2641 gcgacaatct atcgcttgta tgggaagccc gatgcgccag agttgtttct gaaacatggc
2701 aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa
2761 tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc
2821 accactgcga tccccggaaa aacagcattc caggtattag aagaatatcc tgattcaggt
2881 gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt
2941 aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat
3001 aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa
3061 gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt cactcatggt
3121 gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt
3181 ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt
3241 gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat
3301 atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcaga attggttaat
3361 tggttgtaac actggcagag cattacgctg acttgacggg acggcgcaag ctcatgacca
3421 aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc
3481 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
3541 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
3601 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
3661 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
3721 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
3781 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
3841 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
3901 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
3961 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
4021 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
4081 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
4141 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
4201 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
4261 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
4321 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tacgcgtacc
4381 gctagccagg aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg gccttctgct
4441 tagtttgatg cctggcagtt tatggcgggc gtcctgcccg ccaccctccg ggccgttgct
4501 tcacaacgtt caaatccgct cccggcggat ttgtcctact caggagagcg ttcaccgaca
4561 aacaacagat aaaacgaaag gcccagtctt ccgactgagc ctttcgtttt atttgatgcc
4621 tggcagttcc ctactctcgc gttaacgcta gcatggatgt tttcccagtc acgacgttgt
4681 aaaacgacgg ccagtcttaa gctcgggccc
//
| {
"Vector backbone": "pDONR221",
"Backbone size w/o insert (bp)": "2550",
"Vector type": "Mammalian Expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Kanamycin, 50 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "NEB Stable",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Industry MTA"
} | {
"Title": "The RESOLUTE consortium: unlocking SLC transporters for drug discovery",
"Publication": "Nat Rev Drug Discov. 2020 Jul;19(7):429-430. doi: 10.1038/d41573-020-00056-6.",
"DOI": "10.1038/d41573-020-00056-6.",
"Authors": "Superti-Furga G, Lackner D, Wiedmer T, Ingles-Prieto A, Barbosa B, Girardi E, Goldmann U, Gurtl B, Klavins K, Klimek C, Lindinger S, Lineiro-Retes E, Muller AC, Onstein S, Redinger G, Reil D, Sedlyarov V, Wolf G, Crawford M, Everley R, Steppan CM /d---",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/32265506",
"PubMed ID": "PubMed 32265506"
} | {
"Gene/Insert name": "SLC44A5",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "2160",
"Mutation": "",
"Entrez Gene": "SLC44A5",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Gateway Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "M13-fwd",
"3′ sequencing primer": "M13-rev"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
||
pFA6a-13Myc-His3MX6 | 41602 | Low | John Pringle | Longtine et al Yeast. 1998 Jul;14(10):953-61. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/55636/4066ba80-fe93-44c2-8bf7-4bc5f1abb1a7/addgene-plasmid-41602-sequence-55636.gbk | full | LOCUS sequence-55636-e 4590 bp DNA circular SYN 25-AUG-2024
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4590)
AUTHORS Longtine MS, McKenzie A 3rd, Demarini DJ, Shah NG, Wach A, Brachat
A, Philippsen P, Pringle JR
TITLE Additional modules for versatile and economical PCR-based gene
deletion and modification in Saccharomyces cerevisiae.
JOURNAL Yeast. 1998 Jul;14(10):953-61.
PUBMED 9717241
REFERENCE 2 (bases 1 to 4590)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Yeast.";
date: "1998-07"; volume: "14(10)"; pages: "953-61"
FEATURES Location/Qualifiers
source 1..4590
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 64..561
/codon_start=1
/product="13 tandem Myc epitope tags"
/label=13xMyc
/translation="EQKLISEEDLNGEQKLISEEDLNGLDGEQKLISEEDLNGEQKLIS
EEDLNGEQKLISEEDLNGLDGEQKLISEEDLNGEQKLISEEDLNGEQKLISEEDLNGLD
GEQKLISEEDLNGEQKLISEEDLNGEQKLISEEDLNGLDGEQKLISEEDLNGEQKLISE
EDL"
terminator 616..803
/gene="S. cerevisiae ADH1"
/label=ADH1 terminator
/note="transcription terminator for the S. cerevisiae
alcohol dehydrogenase 1 (ADH1) gene"
primer_bind complement(671..695)
/label=pHybLex-R
/note="ADH terminator, reverse primer"
promoter 878..1221
/label=TEF promoter
/note="Ashbya gossypii TEF promoter"
CDS 1222..1920
/codon_start=1
/product="S. kluyveri imidazoleglycerol phosphate
dehydratase"
/label=k-HIS3
/translation="MAEPAQKKQKQTVQERKAFISRITNETKIQIAISLNGGYIQIKDS
ILPAKKDDDVASQATQSQVIDIHTGVGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHT
TEDCGIALGQAFKEAMGAVRGVKRFGTGFAPLDEALSRAVVDLSNRPFAVIDLGLKREM
IGDLSTEMIPHFLESFAEAARITLHVDCLRGFNDHHRSESAFKALAVAIREAISSNGTN
DVPSTKGVLM"
terminator 1924..2121
/label=TEF terminator
/note="Ashbya gossypii TEF terminator"
primer_bind complement(2225..2244)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(2226..2244)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(2331..2348)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2502..3090)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2582..2601)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3261..4121)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 3884..3903
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(4122..4226)
/gene="bla"
/label=AmpR promoter
primer_bind 4294..4312
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(4350..4372)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 4472..4491
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
promoter 4572..4590
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 4572..4589
/label=SP6
/note="SP6 promoter, forward primer"
ORIGIN
1 acgcggccgc cagctgaagc ttcgtacgct gcaggtcgac ggatccccgg gttaattaac
61 ggtgaacaaa agctaatctc cgaggaagac ttgaacggtg aacaaaaatt aatctcagaa
121 gaagacttga acggactcga cggtgaacaa aagttgattt ctgaagaaga tttgaacggt
181 gaacaaaagc taatctccga ggaagacttg aacggtgaac aaaaattaat ctcagaagaa
241 gacttgaacg gactcgacgg tgaacaaaag ttgatttctg aagaagattt gaacggtgaa
301 caaaagctaa tctccgagga agacttgaac ggtgaacaaa aattaatctc agaagaagac
361 ttgaacggac tcgacggtga acaaaagttg atttctgaag aagatttgaa cggtgaacaa
421 aagctaatct ccgaggaaga cttgaacggt gaacaaaaat taatctcaga agaagacttg
481 aacggactcg acggtgaaca aaagttgatt tctgaagaag atttgaacgg tgaacaaaag
541 ctaatctccg aggaagactt gaacggtgaa caaaaattaa tcaatcacta gtgaattcgc
601 gccacttcta aataagcgaa tttcttatga tttatgattt ttattattaa ataagttata
661 aaaaaaataa gtgtatacaa attttaaagt gactcttagg ttttaaaacg aaaattctta
721 ttcttgagta actctttcct gtaggtcagg ttgctttctc aggtatagta tgaggtcgct
781 cttattgacc acacctctac cggcagatcc gctagggata acagggtaat atagatctgt
841 ttagcttgcc tcgtccccgc cgggtcaccc ggccagcgac atggaggccc agaataccct
901 ccttgacagt cttgacgtgc gcagctcagg ggcatgatgt gactgtcgcc cgtacattta
961 gcccatacat ccccatgtat aatcatttgc atccatacat tttgatggcc gcacggcgcg
1021 aagcaaaaat tacggctcct cgctgcagac ctgcgagcag ggaaacgctc ccctcacaga
1081 cgcgttgaat tgtccccacg ccgcgcccct gtagagaaat ataaaaggtt aggatttgcc
1141 actgaggttc ttctttcata tacttccttt taaaatcttg ctaggataca gttctcacat
1201 cacatccgaa cataaacaac catggcagaa ccagcccaaa aaaagcaaaa acaaactgtt
1261 caggagcgca aggcgtttat ctcccgtatc actaatgaaa ctaaaattca aatcgctatt
1321 tcgctgaatg gtggttatat tcaaataaaa gattcgattc ttcctgcaaa gaaggatgac
1381 gatgtagctt cccaagctac tcagtcacag gtcatcgata ttcacacagg tgttggcttt
1441 ttggatcata tgatccatgc gttggcaaaa cactctggtt ggtctcttat tgttgaatgt
1501 atcggtgacc tgcacattga cgatcaccat actaccgaag attgcggtat cgcattaggg
1561 caagcgttca aagaagcaat gggtgctgtc cgtggtgtaa aaagattcgg tactgggttc
1621 gcaccattgg atgaggcgct atcacgtgcc gtagtcgatt tatctaatag accatttgct
1681 gtaatcgacc ttggattgaa gagagagatg attggtgatt tatccactga aatgattcca
1741 cactttttgg aaagtttcgc ggaggcggcc agaattactt tgcatgttga ttgtctgaga
1801 ggtttcaacg atcaccacag aagtgagagt gcgttcaagg ctttggctgt tgccataaga
1861 gaagctattt ctagcaatgg caccaatgac gttccctcaa ccaaaggtgt tttgatgtga
1921 agtactgaca ataaaaagat tcttgttttc aagaacttgt catttgtata gtttttttat
1981 attgtagttg ttctatttta atcaaatgtt agcgtgattt atattttttt tcgcctcgac
2041 atcatctgcc cagatgcgaa gttaagtgcg cagaaagtaa tatcatgcgt caatcgtatg
2101 tgaatgctgg tcgctatact gctgtcgatt cgatactaac gccgccatcc agtttaaacg
2161 agctcgaatt catcgatgat atcagatcca ctagtggcct atgcggccgc ggatctgccg
2221 gtctccctat agtgagtcgt attaatttcg ataagccagg ttaacctgca ttaatgaatc
2281 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
2341 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
2401 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
2461 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
2521 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
2581 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
2641 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc
2701 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
2761 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
2821 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
2881 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
2941 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
3001 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
3061 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
3121 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
3181 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
3241 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
3301 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
3361 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
3421 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
3481 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
3541 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
3601 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
3661 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
3721 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
3781 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
3841 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
3901 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
3961 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
4021 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
4081 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
4141 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
4201 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
4261 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
4321 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
4381 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
4441 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
4501 caccatatgg acatattgtc gttagaacgc ggctacaatt aatacataac cttatgtatc
4561 atacacatac gatttaggtg acactataga
//
| {
"Vector backbone": "pFA6a",
"Backbone size w/o insert (bp)": "2300",
"Vector type": "Yeast genomic targeting",
"Selectable markers": "HIS3"
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "Unknown"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Additional modules for versatile and economical PCR-based gene deletion and modification in Saccharomyces cerevisiae",
"Publication": "Yeast. 1998 Jul;14(10):953-61.",
"DOI": "10.1002/(SICI)1097-0061(199807)14:10",
"Authors": "Longtine MS, McKenzie A rd, Demarini DJ, Shah NG, Wach A, Brachat A, Philippsen P, Pringle JR <::AID-YEA>CO;-U",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/9717241",
"PubMed ID": "PubMed 9717241"
} | {
"Gene/Insert name": "His3",
"Species": "S. kluyveri",
"Insert Size (bp)": "2169",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "PacI (not destroyed)",
"3′ cloning site": "PmeI (not destroyed)",
"5′ sequencing primer": "Sp6_primer",
"3′ sequencing primer": "T7_promoter"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
GPR158-Tango | 66332 | Low | Expression of G protein-coupled receptors for PRESTO-Tango: parallel receptorome expression and screening via transcriptional output, with transcriptional activation following arrestin translocation | Bryan Roth | Kroeze et al Nat Struct Mol Biol. 2015 May;22(5):362-9. doi: 10.1038/nsmb.3014. Epub 2015 Apr 20. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/119882/58df4dd9-88d6-4d1b-8cdd-d3d2630fb232/addgene-plasmid-66332-sequence-119882.gbk | partial | LOCUS sequence-119882- 833 bp DNA linear UNA 25-AUG-2024
DEFINITION Expression of G protein-coupled receptors for PRESTO-Tango: parallel
receptorome expression and screening via transcriptional output,
with transcriptional activation following arrestin translocation.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE natural DNA sequence
ORGANISM unspecified
REFERENCE 1 (bases 1 to 833)
AUTHORS Kroeze WK, Sassano MF, Huang XP, Lansu K, McCorvy JD, Giguere PM,
Sciaky N, Roth BL
TITLE PRESTO-Tango as an open-source resource for interrogation of the
druggable human GPCRome.
JOURNAL Nat Struct Mol Biol. 2015 May;22(5):362-9. doi: 10.1038/nsmb.3014.
Epub 2015 Apr 20.
PUBMED 25895059
REFERENCE 2 (bases 1 to 833)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10.1038/nsmb.3014";
journalName: "Nat Struct Mol Biol"; date: "2015-05"; volume: "22";
issue: "5"; pages: "362-9"
FEATURES Location/Qualifiers
source 1..833
/mol_type="genomic DNA"
/organism="unspecified"
primer_bind 48..67
/label=T7
/note="T7 promoter, forward primer"
promoter 48..66
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
1 gctctctggc taactagaga acccactgct tactggctta tcgaaattaa tacgactcac
61 tatagggaga cccaagctgg ctagcgttta aacttaagct tggtaccgag ctcggatcca
121 ctagtccagt gtggtggaat tctgcagata tccagcacag tggcggccgc gccaccatga
181 agacgatcat cgccctgagc tacatcttct gcctggtatt cgccgactac aaggacgatg
241 atgacgccag catcgatatg ggcgctatgg cataccctct gctgctgtgc ctgcttcttg
301 ctcaactcgg gctgggcgcc gtaggcgcca gtagagatcc tcagggcaga cctgacagtc
361 ccagggaacg cacccctaag ggcaaaccgc acgcacagca acccggtcgg gcttctgcta
421 gcgactcctc tgccccatgg tctaggagta cagatggcac aatcctcgct cagaaacttg
481 cagaagaagt gccgatggac gtggcctcct atttgtatac gggtgactca caccagctga
541 agcgagccaa ttgtagtggc aggtacgaac tcgcaggctt gccaggcaag tggccagcac
601 tcgcatccgc gcatccctca ctgcaccggg ctctggatac cctgacccat gcaactaact
661 ttcttaatgt tatgcttcag tctaacaaga gtcgagagca gaaccttcaa gacgacctgg
721 actggtatca ggccctcgtg tggtcactcc tggagggcga gcctagcatt tccagggctg
781 caatcacctt ttccactgat tccctgagcg ccccagctcc gcaggtcttc ttg
//
| {
"Vector backbone": "empty Tango",
"Backbone size w/o insert (bp)": "6632",
"Vector type": "Mammalian Expression",
"Selectable markers": "Neomycin (select with G418)"
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "PRESTO-Tango as an open-source resource for interrogation of the druggable human GPCRome",
"Publication": "Nat Struct Mol Biol. 2015 May;22(5):362-9. doi: 10.1038/nsmb.3014. Epub 2015 Apr 20.",
"DOI": "10.1038/nsmb.3014.",
"Authors": "Kroeze WK, Sassano MF, Huang XP, Lansu K, McCorvy JD, Giguere PM, Sciaky N, Roth BL /nsmb",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/25895059",
"PubMed ID": "PubMed 25895059"
} | {
"Gene/Insert name": "GPR158",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "3645",
"Mutation": "",
"Entrez Gene": "GPR158",
"Promoter": "CMV",
"Tag / Fusion Protein": "FLAG (N terminal on backbone)"
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "NotI (not destroyed)",
"3′ cloning site": "XhoI (unknown if destroyed)",
"5′ sequencing primer": "CMV-F",
"3′ sequencing primer": "Tet-R"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
pmCherry-histone H3 | 179699 | visualization of nucleus/chromosome | Kenji Sugimoto | Yasui et al Mutat Res. 2010 Oct 13;692(1-2):12-8. doi: 10.1016/j.mrfmmm.2010.07.009. Epub 2010 Aug 5. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/356505/e7947980-ec0b-4cb3-b2bc-980710905eac/addgene-plasmid-179699-sequence-356505.gbk | full | LOCUS sequence-356505- 5109 bp DNA circular SYN 25-AUG-2024
DEFINITION visualization of nucleus/chromosome.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5109)
AUTHORS Yasui M, Koyama N, Koizumi T, Senda-Murata K, Takashima Y, Hayashi
M, Sugimoto K, Honma M
TITLE Live cell imaging of micronucleus formation and development.
JOURNAL Mutat Res. 2010 Oct 13;692(1-2):12-8. doi:
10.1016/j.mrfmmm.2010.07.009. Epub 2010 Aug 5.
PUBMED 20691709
REFERENCE 2 (bases 1 to 5109)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Mutat
Res."; date: "2010-10-13"; volume: "692(1-2):12-8. doi"; pages: "
10.1016/j.mrfmmm.2010.07.009. Epub 2010 Aug 5"
FEATURES Location/Qualifiers
source 1..5109
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 61..364
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 365..568
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 519..539
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 598..607
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 604..1311
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
primer_bind complement(749..767)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 1017..1036
/label=mCherry-F
/note="mCherry, forward primer"
polyA_signal 1897..2018
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1934..1953)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 1988..2007
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(2025..2480)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2162..2183)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2374..2393
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2507..2611
/gene="bla"
/label=AmpR promoter
promoter 2613..2970
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
enhancer 2613..2804
/label=SV40 enhancer
/note="enhancer for the SV40 early promoter (Herr, 1993)"
primer_bind complement(2636..2656)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 2821..2956
/label=SV40 ori
/note="SV40 origin of replication"
CDS 3005..3799
/codon_start=1
/product="aminoglycoside phosphotransferase"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
CDS 3005..3799
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(3059..3078)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3669..3688
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(3987..4006)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 4031..4078
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4407..4995
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4896..4915
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagccgcc
601 accatggtga gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc
661 aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag
721 ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg
781 cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag
841 caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag
901 cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag
961 gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc
1021 gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac
1081 ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac
1141 gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac
1201 gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac
1261 gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa gagatctcga
1321 gctcaagctt cgaattcggc tcgtactaag cagaccgctc gcaagtccac cggcggcaag
1381 gccccgcgca agcagctggc caccaaggct gcccgcaaga gcgccccggc caccggcggc
1441 gtgaagaagc ctcaccgcta ccgtcccggc accgtggcgc tgcgcgagat ccggcgctac
1501 cagaagtcga ccgagctgct gatccgcaag ctgccgttcc agcgcctggt gcgcgagatc
1561 gcgcaggact tcaagaccga cctgcgcttc cagagctcgg ctgtcatggc tctgcaggag
1621 gcgagcgagg cctaccttgt gggtctgttt gaggacacca acctgtgcgc catccacgcc
1681 aagcgtgtca ccatcatgcc caaggacatc cagctcgccc gccgcatccg cggggagagg
1741 gcttaactcg acggtaccgc gggcccggga tccaccggat ctagataact gatcataatc
1801 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
1861 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
1921 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
1981 tctagttgtg gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa
2041 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
2101 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
2161 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
2221 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
2281 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
2341 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
2401 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
2461 tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
2521 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
2581 aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat
2641 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
2701 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
2761 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
2821 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
2881 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
2941 ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt
3001 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
3061 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
3121 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
3181 actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
3241 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
3301 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
3361 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
3421 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
3481 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc
3541 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
3601 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
3661 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
3721 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
3781 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
3841 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
3901 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
3961 ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
4021 cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat
4081 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
4141 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
4201 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat
4261 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
4321 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
4381 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
4441 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
4501 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
4561 gtgtagccgt agttaggcca ccacttcaag aactctgtat caccgcctac atacctcgct
4621 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
4681 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
4741 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
4801 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
4861 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
4921 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
4981 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
5041 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
5101 gccatgcat
//
| {
"Vector backbone": "pECFP-C1",
"Backbone size w/o insert (bp)": "",
"Vector type": "Mammalian Expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Kanamycin, 50 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA ; Takara Bio Limited Use Label License (formerly Clontech)",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Live cell imaging of micronucleus formation and development",
"Publication": "Mutat Res. 2010 Oct 13;692(1-2):12-8. doi: 10.1016/j.mrfmmm.2010.07.009. Epub 2010 Aug 5.",
"DOI": "10.1016/j.mrfmmm.2010.07.009.",
"Authors": "Yasui M, Koyama N, Koizumi T, Senda-Murata K, Takashima Y, Hayashi M, Sugimoto K, Honma M /jmrfmmm",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/20691709",
"PubMed ID": "PubMed 20691709"
} | {
"Gene/Insert name": "Histone H3",
"Species": "M. musculus (mouse)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "Hist1h3h",
"Promoter": "",
"Tag / Fusion Protein": "mCherry (N terminal on insert)"
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "unknown (unknown if destroyed)",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pcDNA3.1(+)/AUU-nLuc-3XFLAG-CL1/PEST | 127319 | Express 3xFLAG tagged highly destabilized nanoLuciferase (nLuc) protein from AUU start codon | Jeremy Wilusz | Kearse et al Genes Dev. 2019 Jun 6. pii: gad.324715.119. doi: 10.1101/gad.324715.119. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/247731/75436918-ef5c-4efe-8543-c35860ac1170/addgene-plasmid-127319-sequence-247731.gbk | full | LOCUS sequence-247731- 6157 bp DNA circular SYN 26-AUG-2024
DEFINITION Express 3xFLAG tagged highly destabilized nanoLuciferase (nLuc)
protein from AUU start codon.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6157)
AUTHORS Kearse MG, Goldman DH, Choi J, Nwaezeapu C, Liang D, Green KM,
Goldstrohm AC, Todd PK, Green R, Wilusz JE
TITLE Ribosome queuing enables non-AUG translation to be resistant to
multiple protein synthesis inhibitors.
JOURNAL Genes Dev. 2019 Jun 6. pii: gad.324715.119. doi:
10.1101/gad.324715.119.
PUBMED 31171704
REFERENCE 2 (bases 1 to 6157)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Genes Dev.
2019 Jun 6. pii: gad.324715.119. doi: 10.1101/gad.324715.119."
FEATURES Location/Qualifiers
source 1..6157
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 61..440
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 441..644
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 595..615
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 689..708
/label=T7
/note="T7 promoter, forward primer"
promoter 689..707
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 778..1287
/codon_start=1
/product="NanoLuc(R) luciferase"
/label=Nluc
/note="human codon-optimized"
/translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
LCERILA"
CDS 1288..1353
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
CDS 1360..1407
/codon_start=1
/product="non-ORF yeast peptide conferring
ubiquitin-dependent degradation"
/label=hCL1
/note="human codon-optimized"
/translation="ACKNWFSSLSHFVIHL"
CDS 1411..1530
/codon_start=1
/product="PEST degradation sequence from mouse ornithine
decarboxylase"
/label=hPEST
/note="human codon-optimized"
/translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
primer_bind complement(1574..1591)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 1580..1804
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 1850..2278
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1937..1956)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2147..2168
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2287..2307)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2292..2621
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2472..2607
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2534..2553
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2688..3482
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(2742..2761)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3352..3371
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 3656..3777
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3693..3712)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3747..3766
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(3826..3842)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3826..3842)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3839..3861)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3850..3866
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(3874..3904)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3919..3940
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4057..4074)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4228..4816)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4308..4327)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4987..5847)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 5610..5629
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5848..5952)
/gene="bla"
/label=AmpR promoter
primer_bind complement(6027..6046)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt
61 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
121 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
181 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
241 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
301 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
361 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
421 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
481 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
541 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
601 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
661 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
721 gctagcgttt aaacttaagc ttggcaatcc ggtactgttg gtaaataagc caccattgtc
781 ttcacactcg aagatttcgt tggggactgg cgacagacag ccggctacaa cctggaccaa
841 gtccttgaac agggaggtgt gtccagtttg tttcagaatc tcggggtgtc cgtaactccg
901 atccaaagga ttgtcctgag cggtgaaaat gggctgaaga tcgacatcca tgtcatcatc
961 ccgtatgaag gtctgagcgg cgaccaaatg ggccagatcg aaaaaatttt taaggtggtg
1021 taccctgtgg atgatcatca ctttaaggtg atcctgcact atggcacact ggtaatcgac
1081 ggggttacgc cgaacatgat cgactatttc ggacggccgt atgaaggcat cgccgtgttc
1141 gacggcaaaa agatcactgt aacagggacc ctgtggaacg gcaacaaaat tatcgacgag
1201 cgcctgatca accccgacgg ctccctgctg ttccgagtaa ccatcaacgg agtgaccggc
1261 tggcggctgt gcgaacgcat tctggcggac tacaaagacc atgacggtga ttataaagat
1321 catgacatcg attacaagga tgacgatgac aagaattctg cttgcaagaa ctggttcagt
1381 agcttaagcc actttgtgat ccaccttaac agccacggct ttccgcctga ggttgaagag
1441 caagccgccg gtacattgcc tatgtcctgc gcacaagaaa gcggtatgga ccggcaccca
1501 gccgcttgtg cttcagctcg catcaacgtc taaggccgcg actctagagg gcccgtttaa
1561 acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
1621 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
1681 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
1741 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
1801 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt
1861 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
1921 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
1981 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
2041 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
2101 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
2161 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
2221 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
2281 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
2341 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
2401 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
2461 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
2521 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
2581 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
2641 tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag
2701 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
2761 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
2821 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
2881 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
2941 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
3001 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
3061 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
3121 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
3181 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg
3241 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
3301 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
3361 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
3421 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
3481 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
3541 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
3601 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
3661 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
3721 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
3781 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
3841 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
3901 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
3961 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
4021 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
4081 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
4141 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
4201 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
4261 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
4321 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
4381 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
4441 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
4501 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
4561 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
4621 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
4681 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
4741 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
4801 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
4861 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
4921 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
4981 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
5041 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
5101 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
5161 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
5221 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
5281 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
5341 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
5401 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
5461 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
5521 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
5581 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
5641 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
5701 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
5761 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
5821 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
5881 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
5941 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct
6001 cccgatcccc tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
6061 tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct
6121 acaacaaggc aaggcttgac cgacaattgc atgaaga
//
| {
"Vector backbone": "pcDNA3.1(+)",
"Backbone size w/o insert (bp)": "",
"Vector type": "Mammalian Expression",
"Selectable markers": "Neomycin (select with G418)"
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA ; NanoLuc Luciferase Limited Use Label License",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Ribosome queuing enables non-AUG translation to be resistant to multiple protein synthesis inhibitors",
"Publication": "Genes Dev. 2019 Jun 6. pii: gad.324715.119. doi: 10.1101/gad.324715.119.",
"DOI": "10.1101/gad.324715.119.",
"Authors": "Kearse MG, Goldman DH, Choi J, Nwaezeapu C, Liang D, Green KM, Goldstrohm AC, Todd PK, Green R, Wilusz JE /gad",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/31171704",
"PubMed ID": "PubMed 31171704"
} | {
"Gene/Insert name": "nanoLuciferase",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "CMV",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "HindIII (not destroyed)",
"3′ cloning site": "ApaI (not destroyed)",
"5′ sequencing primer": "CGCAAATGGGCGGTAGGCGTG",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pDONR223_ABCB9_WT_V5 | 82946 | Gateway Donor vector containing ABCB9, used as an over expression control for gene expression analysis. Part of the Target Accelerator Plasmid Collection. | Berger et al Cancer Cell. 2016 Aug 8;30(2):214-28. doi: 10.1016/j.ccell.2016.06.022. Epub 2016 Jul 28. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/162610/cf37f074-f6f4-43b4-9622-d1c30a51b565/addgene-plasmid-82946-sequence-162610.gbk | full | LOCUS sequence-162610- 4578 bp DNA circular SYN 26-AUG-2024
DEFINITION Gateway Donor vector containing ABCB9, used as an over expression
control for gene expression analysis. Part of the Target Accelerator
Plasmid Collection..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4578)
AUTHORS Berger AH, Brooks AN, Wu X, Shrestha Y, Chouinard C, Piccioni F,
Bagul M, Kamburov A, Imielinski M, Hogstrom L, Zhu C, Yang X, Pantel
S, Sakai R, Watson J, Kaplan N, Campbell JD, Singh S, Root DE,
Narayan R, Natoli T, Lahr DL, Tirosh I, Tamayo P, Getz G, Wong B,
Doench J, Subramanian A, Golub TR, Meyerson M, Boehm JS
TITLE High-throughput Phenotyping of Lung Cancer Somatic Mutations.
JOURNAL Cancer Cell. 2016 Aug 8;30(2):214-28. doi:
10.1016/j.ccell.2016.06.022. Epub 2016 Jul 28.
PUBMED 27478040
REFERENCE 2 (bases 1 to 4578)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1016/j.ccell.2016.06.022"; journalName: "Cancer Cell"; date:
"2016-08-8- 8"; volume: "30"; issue: "2"; pages: "214-28"
FEATURES Location/Qualifiers
source 1..4578
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 43..65
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 57..74
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 58..74
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 90..189
/gene="mutant version of attL"
/label=attL1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
CDS 374..388
/codon_start=1
/product="tag for protein cleavage using Ni2+ ion (Dang et
al., 2019)"
/label=SNAC tag
/translation="GSHHW"
protein_bind complement(1978..2077)
/gene="mutant version of attL"
/label=attL2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
primer_bind complement(2094..2113)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(2095..2113)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(2118..2134)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2118..2134)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
CDS 2565..3356
/codon_start=1
/gene="aadA"
/product="aminoglycoside adenylyltransferase (Murphy,
1985)"
/label=SmR
/note="confers resistance to spectinomycin and
streptomycin"
/translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
rep_origin 3449..4037
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3938..3957
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 4191..4208
/label=L4440
/note="L4440 vector, forward primer"
terminator complement(4367..4394)
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator complement(4486..4572)
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
ORIGIN
1 ggcagttccc tactctcgcg ttaacgctag catggatgtt ttcccagtca cgacgttgta
61 aaacgacggc cagtcttaag ctcgggcccc aaataatgat tttattttga ctgatagtga
121 cctgttcgtt gcaacaaatt gatgagcaat gcttttttat aatgccaact ttgtacaaaa
181 aagttggcat gcggctgtgg aaggcggtgg tggtgacttt ggccttcatg agtgtggaca
241 tctgcgtgac cacggccatc tatgtcttca gccacctgga ccgcagcctc ctggaggaca
301 tccgccactt caacatcttt gactcggtgc tggatctctg ggcagcctgc ctgtaccgca
361 gctgcctgct gctgggagcc accattggtg tggccaagaa cagtgcgctg gggccccggc
421 ggctgcgggc ctcgtggctg gtcatcaccc tcgtgtgcct cttcgtgggc atctatgcca
481 tggtgaagct gctgctcttc tcagaggtgc gcaggcccat ccgggacccc tggttttggg
541 ccctgttcgt gtggacgtac atttcactcg gcgcatcctt cctgctctgg tggctgctgt
601 ccaccgtgcg gccaggcacc caggccctgg agccaggggc ggccaccgag gctgagggct
661 tccctgggag cggccggcca ccgcccgagc aggcgtctgg ggccacgctg cagaagctgc
721 tctcctacac caagcccgac gtggccttcc tcgtggccgc ctccttcttc ctcatcgtgg
781 cagctctggg agagaccttc ctgccctact acacgggccg cgccattgat ggcatcgtca
841 tccagaaaag catggatcag ttcagcacgg ctgtcgtcat cgtgtgcctg ctggccattg
901 gcagctcatt tgccgcaggt attcggggcg gcatttttac cctcatattt gccagactga
961 acattcgcct tcgaaactgt ctcttccgct cactggtgtc ccaggagaca agcttctttg
1021 atgagaaccg cacaggggac ctcatctccc gcctgacctc ggacaccacc atggtcagcg
1081 acctggtctc ccagaacatc aatgtcttcc tgcggaacac agtcaaggtc acgggcgtgg
1141 tggtcttcat gttcagcctc tcatggcagc tctccttggt caccttcatg ggcttcccca
1201 tcatcatgat ggtgtccaac atctacggca agtactacaa gaggctctcc aaagaggtcc
1261 agaatgccct ggccagagcg agcaacacgg cggaggagac catcagtgcc atgaagactg
1321 tccggagctt cgccaatgag gaggaggagg cagaggtgta cctgcggaag ctgcagcagg
1381 tgtacaagct gaacaggaag gaggcagctg cctacatgta ctacgtctgg ggcagcgggc
1441 tcacactgct ggtggtccag gtcagcatcc tctactacgg gggccacctt gtcatctcag
1501 gccagatgac cagcggcaac ctcatcgcct tcatcatcta cgagtttgtc ctgggagatt
1561 gtatggagtc cgtgggctcc gtctacagtg gcctgatgca gggagtgggg gctgctgaga
1621 aggtgttcga gttcatcgac cggcagccga ccatggtgca cgatggcagc ttggcccccg
1681 accacctgga gggccgggtg gactttgaga atgtgacctt cacctaccgc actcggcccc
1741 acacccaggt cctgcagaat gtctccttca gcctgtcccc cggcaaggtg acggccctgg
1801 tggggccctc gggcagtggg aagagctcct gtgtcaacat cctggagaac ttctaccccc
1861 tggagggggg ccgggtgctg ctggacggca agcccatcag cgcctacgac cacaagtact
1921 tgcaccgtgt ggtatgtgca cgggcctggg ccacacttct ccgccctttc tgcatctacc
1981 caactttctt gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg
2041 aacaggtcac tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata
2101 gtgagtcgta ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc
2161 tctgatgtta cattgcacaa gataaaaata tatcatcatg cctcctctag accagccagg
2221 acagaaatgc ctcgacttcg ctgctaccca aggttgccgg gtgacgcaca ccgtggaaac
2281 ggatgaaggc acgaacccag tggacataag cctgttcggt tcgtaagctg taatgcaagt
2341 agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga acgcagcggt ggtaacggcg
2401 cagtggcggt tttcatggct tgttatgact gtttttttgg ggtacagtct atgcctcggg
2461 catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg agcagcaacg
2521 atgttacgca gcagggcagt cgccctaaaa caaagttaaa cattatgagg gaagcggtga
2581 tcgccgaagt atcgactcaa ctatcagagg tagttggcgt catcgagcgc catctcgaac
2641 cgacgttgct ggccgtacat ttgtacggct ccgcagtgga tggcggcctg aagccacaca
2701 gtgatattga tttgctggtt acggtgaccg taaggcttga tgaaacaacg cggcgagctt
2761 tgatcaacga ccttttggaa acttcggctt cccctggaga gagcgagatt ctccgcgctg
2821 tagaagtcac cattgttgtg cacgacgaca tcattccgtg gcgttatcca gctaagcgcg
2881 aactgcaatt tggagaatgg cagcgcaatg acattcttgc aggtatcttc gagccagcca
2941 cgatcgacat tgatctggct atcttgctga caaaagcaag agaacatagc gttgccttgg
3001 taggtccagc ggcggaggaa ctctttgatc cggttcctga acaggatcta tttgaggcgc
3061 taaatgaaac cttaacgcta tggaactcgc cgcccgactg ggctggcgat gagcgaaatg
3121 tagtgcttac gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc gcgccgaagg
3181 atgtcgctgc cgactgggca atggagcgcc tgccggccca gtatcagccc gtcatacttg
3241 aagctagaca ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt
3301 tggaagaatt tgtccactac gtgaaaggcg agatcaccaa ggtagtcggc aaataaccct
3361 cgagccaccc atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag
3421 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
3481 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
3541 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
3601 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
3661 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
3721 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
3781 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
3841 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
3901 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
3961 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
4021 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
4081 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
4141 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
4201 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
4261 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
4321 caattaatac gcgtaccgct agccaggaag agtttgtaga aacgcaaaaa ggccatccgt
4381 caggatggcc ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca
4441 ccctccgggc cgttgcttca caacgttcaa atccgctccc ggcggatttg tcctactcag
4501 gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtcttccg actgagcctt
4561 tcgttttatt tgatgcct
//
| {
"Vector backbone": "pDONR223",
"Backbone size w/o insert (bp)": "",
"Vector type": "Gateway Entry vector",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Spectinomycin, 50 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "High-throughput Phenotyping of Lung Cancer Somatic Mutations",
"Publication": "Cancer Cell. 2016 Aug 8;30(2):214-28. doi: 10.1016/j.ccell.2016.06.022. Epub 2016 Jul 28.",
"DOI": "10.1016/j.ccell.2016.06.022.",
"Authors": "Berger AH, Brooks AN, Wu X, Shrestha Y, Chouinard C, Piccioni F, Bagul M, Kamburov A, Imielinski M, Hogstrom L, Zhu C, Yang X, Pantel S, Sakai R, Watson J, Kaplan N, Campbell JD, Singh S, Root DE, Narayan R, Natoli T, Lahr DL, Tirosh I, Tamayo P, Getz G, Wong B, Doench J, Subramanian A, Golub TR, Meyerson M, Boehm JS /jccell",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/27478040",
"PubMed ID": "PubMed 27478040"
} | {
"Gene/Insert name": "ABCB9",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "AA 582- 590 VCARAWATL; AA 592-595 RPFC, AA 597-681 deleted compared to GenBank",
"Entrez Gene": "ABCB9",
"Promoter": "None",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Gateway Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "M13pUC-fwd",
"3′ sequencing primer": "M13-Rev"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
||
pCR8/GW/TOPO-ETS1 | 98621 | Gateway entry TOPO TA cloning vector for ETS1 | Richard Harvey | Bouveret et al Elife. 2015 Jul 6;4. doi: 10.7554/eLife.06942. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/182584/6764afb9-ef62-4e45-b74e-2889f58104e0/addgene-plasmid-98621-sequence-182584.gbk | full | LOCUS sequence-182584- 4140 bp DNA circular SYN 26-AUG-2024
DEFINITION Gateway entry TOPO TA cloning vector for ETS1.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4140)
AUTHORS Bouveret R, Waardenberg AJ, Schonrock N, Ramialison M, Doan T, de
Jong D, Bondue A, Kaur G, Mohamed S, Fonoudi H, Chen CM, Wouters MA,
Bhattacharya S, Plachta N, Dunwoodie SL, Chapman G, Blanpain C,
Harvey RP
TITLE NKX2-5 mutations causative for congenital heart disease retain
functionality and are directed to hundreds of targets.
JOURNAL Elife. 2015 Jul 6;4. doi: 10.7554/eLife.06942.
PUBMED 26146939
REFERENCE 2 (bases 1 to 4140)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Elife.";
date: "2015-07-6"; volume: "4. doi"; pages: " 10.7554/eLife.06942"
FEATURES Location/Qualifiers
source 1..4140
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 1..17
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 33..132
/gene="mutant version of attL"
/label=attL1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
protein_bind complement(1483..1582)
/gene="mutant version of attL"
/label=attL2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
primer_bind complement(1599..1618)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(1600..1618)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(1623..1639)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1623..1639)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
CDS 2070..2861
/codon_start=1
/gene="aadA"
/product="aminoglycoside adenylyltransferase (Murphy,
1985)"
/label=SmR
/note="confers resistance to spectinomycin and
streptomycin"
/translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
rep_origin 2954..3542
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3443..3462
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 3696..3713
/label=L4440
/note="L4440 vector, forward primer"
terminator complement(3872..3899)
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator complement(3991..4077)
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
primer_bind join(4126..4140,1..8)
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind join(4140,1..17)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
ORIGIN
1 gtaaaacgac ggccagtctt aagctcgggc cccaaataat gattttattt tgactgatag
61 tgacctgttc gttgcaacaa attgatgagc aatgcttttt tataatgcca actttgtaca
121 aaaaagcagg ctccgaattc gcccttacca tgaaggcggc cgtcgatctc aagccgactc
181 tcaccatcat caagacagaa aaagtggatc tcgagctttt cccttccccg gacatggaat
241 gtgcagatgt cccgctgtta actccgagca gcaaagaaat gatgtcccaa gccttgaaag
301 ctactttcag tggtttcaca aaagaacagc agcggctggg aatccccaaa gacccccggc
361 agtggacaga aacccacgtc cgggactggg tgatgtgggc tgtgaatgag ttcagcctga
421 aaggtgtgga cttccagaag ttctgtatga gtggagcagc actgtgtgcc ctgggtaaag
481 aatgcttcct cgagctggct ccagactttg ttggggatat cctgtgggag catctagaga
541 tcctgcagaa agaggatgtg aaaccatatc aggttaatgg agccaaccct acctacccag
601 aatcctgtta cacctcggat tacttcatca gctacggtat cgagcatgct cagtgtgttc
661 ctccctcaga gttctcagag cccagcttca tcacagagtc ctatcagacg ctgcatccta
721 tcagctcaga agaactcctg tccctcaagt atgagaacga ctacccttct gtcattctcc
781 aggaccctct ccagacagac accttgcaga cagactactt tgccatcaag caagaggtgt
841 taactccaga caacatgtgc ctggggagag ccagtcgtgg taaactcggg ggccaggact
901 cttttgagag cgtagagagc tacgatagtt gtgaccgcct cacccagtcc tggagcagcc
961 agtcatcctt caacagcctg cagcgggtcc cctcctatga cagcttcgac tacgaggatt
1021 atcccgctgc cctgcccaac cacaagccca agggcacctt caaggactat gtgcgtgacc
1081 gtgctgacct caacaaggac aagcctgtca ttcctgctgc tgccctggct ggctacacag
1141 gaagtgggcc gatccagctg tggcagtttc ttctggaatt actcactgat aagtcttgtc
1201 agtcctttat cagctggaca ggagatggct gggaattcaa gctttctgac ccagatgagg
1261 tggccaggag atggggaaag aggaaaaaca aacctaagat gaattatgag aaactgagcc
1321 gtggccttcg ctactattat gacaaaaata tcatccacaa gacggcgggc aagcgctacg
1381 tataccgctt tgtgtgcgac ctgcagagcc tgctgggata cacccctgag gagctgcacg
1441 ccatgctgga cgtaaagccg gatgctgaca agggcgaatt cgacccagct ttcttgtaca
1501 aagttggcat tataaaaaat aattgctcat caatttgttg caacgaacag gtcactatca
1561 gtcaaaataa aatcattatt tgccatccag ctgatatccc ctatagtgag tcgtattaca
1621 tggtcatagc tgtttcctgg cagctctggc ccgtgtctca aaatctctga tgttacattg
1681 cacaagataa aaatatatca tcatgcctcc tctagaccag ccaggacaga aatgcctcga
1741 cttcgctgct acccaaggtt gccgggtgac gcacaccgtg gaaacggatg aaggcacgaa
1801 cccagtggac ataagcctgt tcggttcgta agctgtaatg caagtagcgt atgcgctcac
1861 gcaactggtc cagaaccttg accgaacgca gcggtggtaa cggcgcagtg gcggttttca
1921 tggcttgtta tgactgtttt tttggggtac agtctatgcc tcgggcatcc aagcagcaag
1981 cgcgttacgc cgtgggtcga tgtttgatgt tatggagcag caacgatgtt acgcagcagg
2041 gcagtcgccc taaaacaaag ttaaacatta tgagggaagc ggtgatcgcc gaagtatcga
2101 ctcaactatc agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg ttgctggccg
2161 tacatttgta cggctccgca gtggatggcg gcctgaagcc acacagtgat attgatttgc
2221 tggttacggt gaccgtaagg cttgatgaaa caacgcggcg agctttgatc aacgaccttt
2281 tggaaacttc ggcttcccct ggagagagcg agattctccg cgctgtagaa gtcaccattg
2341 ttgtgcacga cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg caatttggag
2401 aatggcagcg caatgacatt cttgcaggta tcttcgagcc agccacgatc gacattgatc
2461 tggctatctt gctgacaaaa gcaagagaac atagcgttgc cttggtaggt ccagcggcgg
2521 aggaactctt tgatccggtt cctgaacagg atctatttga ggcgctaaat gaaaccttaa
2581 cgctatggaa ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg cttacgttgt
2641 cccgcatttg gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc gctgccgact
2701 gggcaatgga gcgcctgccg gcccagtatc agcccgtcat acttgaagct agacaggctt
2761 atcttggaca agaagaagat cgcttggcct cgcgcgcaga tcagttggaa gaatttgtcc
2821 actacgtgaa aggcgagatc accaaggtag tcggcaaata accctcgagc cacccatgac
2881 caaaatccct taacgtgagt tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga
2941 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
3001 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
3061 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
3121 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
3181 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
3241 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
3301 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
3361 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
3421 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
3481 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
3541 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
3601 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
3661 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
3721 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
3781 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatacgcgta
3841 ccgctagcca ggaagagttt gtagaaacgc aaaaaggcca tccgtcagga tggccttctg
3901 cttagtttga tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc cgggccgttg
3961 cttcacaacg ttcaaatccg ctcccggcgg atttgtccta ctcaggagag cgttcaccga
4021 caaacaacag ataaaacgaa aggcccagtc ttccgactga gcctttcgtt ttatttgatg
4081 cctggcagtt ccctactctc gcgttaacgc tagcatggat gttttcccag tcacgacgtt
//
| {
"Vector backbone": "pCR8/GW/TOPO",
"Backbone size w/o insert (bp)": "2817",
"Vector type": "TOPO TA cloning vector",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Spectinomycin, 50 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "NEB Stable",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "NKX2-5 mutations causative for congenital heart disease retain functionality and are directed to hundreds of targets",
"Publication": "Elife. 2015 Jul 6;4. doi: 10.7554/eLife.06942.",
"DOI": "10.7554/eLife.06942.",
"Authors": "Bouveret R, Waardenberg AJ, Schonrock N, Ramialison M, Doan T, de Jong D, Bondue A, Kaur G, Mohamed S, Fonoudi H, Chen CM, Wouters MA, Bhattacharya S, Plachta N, Dunwoodie SL, Chapman G, Blanpain C, Harvey RP /eLife",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/26146939",
"PubMed ID": "PubMed 26146939"
} | {
"Gene/Insert name": "ETS1",
"Species": "M. musculus (mouse)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "Ets1",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "TOPO Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
JGI_018341 | 158848 | Rare earth element alcohol dehydrogenases | Joint Genome Institute | Huang et al ISME J. 2019 Aug;13(8):2005-2017. doi: 10.1038/s41396-019-0414-z. Epub 2019 Apr 5. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/316580/761f5c02-8c4a-45ec-ad24-94d7d75b8533/addgene-plasmid-158848-sequence-316580.gbk | full | LOCUS sequence-316580- 9477 bp DNA circular SYN 26-AUG-2024
DEFINITION Rare earth element alcohol dehydrogenases.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9477)
AUTHORS Huang J, Yu Z, Groom J, Cheng JF, Tarver A, Yoshikuni Y,
Chistoserdova L
TITLE Rare earth element alcohol dehydrogenases widely occur among
globally distributed, numerically abundant and environmentally
important microbes.
JOURNAL ISME J. 2019 Aug;13(8):2005-2017. doi: 10.1038/s41396-019-0414-z.
Epub 2019 Apr 5.
PUBMED 30952993
REFERENCE 2 (bases 1 to 9477)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1038/s41396-019-0414-z"; journalName: "ISME J"; date: "2019-08";
volume: "13"; issue: "8"; pages: "2005-2017"
FEATURES Location/Qualifiers
source 1..9477
/mol_type="other DNA"
/organism="synthetic DNA construct"
RBS 325..347
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 2181..2201
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQG"
CDS 2202..2225
/codon_start=1
/product="8xHis affinity tag"
/label=8xHis
/translation="HHHHHHHH"
misc_feature 2251..2307
/label=MCS
/note="pUC18/19 multiple cloning site"
primer_bind complement(2308..2325)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(2308..2324)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2317..2339)
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind complement(2533..2552)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 2652..2674
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(2712..2730)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
CDS 3516..4715
/codon_start=1
/gene="tetA"
/product="tetracycline efflux protein"
/label=TcR
/note="confers resistance to tetracycline"
/translation="MKPNIPLIVILSTVALDAVGIGLIMPVLPGLLRDLVHSNDVTAHY
GILLALYALVQFACAPVLGALSDRFGRRPILLVSLAGATVDYAIMATAPFLWVLYIGRI
VAGITGATGAVAGAYIADITDGDERARHFGFMSACFGFGMVAGPVLGGLMGGFSPHAPF
FAAAALNGLNFLTGCFLLPESHKGERRPLRREALNPLASFRWARGMTVVAALMAVFFIM
QLVGQVPAALWVIFGEDRFHWDATTIGISLAAFGILHSLAQAMITGPVAARLGERRALM
LGMIADGTGYILLAFATRGWMAFPIMVLLASGGIGMPALQAMLSRQVDEERQGQLQGSL
AALTSLTSIVGPLLFTAIYAASITTWNGWAWIAGAALYLLCLPALRRGLWSGAGQRADR
"
CDS complement(4946..6094)
/codon_start=1
/product="trans-acting replication protein that binds to
and activates oriV"
/label=trfA
/translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
CDS complement(6366..6737)
/codon_start=1
/gene="traJ"
/product="oriT-recognizing protein"
/label=traJ
/translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSL*AYLLAV
GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
KQDELGKVMMGVVRPRAEP"
oriT 6770..6879
/note="incP origin of transfer"
rep_origin 7032..7663
/label=oriV
/note="incP origin of replication"
rep_origin 8126..8714
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 8615..8634
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 8868..8885
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 9002..9023
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 9038..9068
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 9076..9092
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
primer_bind 9081..9103
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 9100..9116
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 9100..9116
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
ORIGIN
1 gcaacgcaaa atgctgaatt tagcaggcaa gtttcctgat aaagggccct gtgagtgcga
61 cattcgacgc gcgaaagcga gttgacggcg cccgaaattt tcttagagtt cgttcaagct
121 tcggtttcgg ggagccgcta agggatgcgg acccagccgg gtaagtcgcc gggaactatg
181 gaataatcct aatttctgtg cggcaggcgc gagctgccgg agcgaatatc atccctgtcg
241 caaacagcag gtcaatcaac ggtgggtagc gggctgctat ccagcggcgg ataagacttg
301 tcggaggaat cctctagaaa taattttgtt taactttaag aaggagatat accatgtcgc
361 tgaagcgcgc cactggccag accggcctcc agaagaagcg cctttcgcgc gcggtggcga
421 ccgccatcgc cgggtttgcc ctggcctcag cgtcgggcgc tgccctcgcc aacgccgagc
481 tgatcaagct gtcgcaggac gatcgccaat gggtcatgca gggcaaggac tatgaccact
541 cgcactacag cggtctgaag cagatcaatc ggtcgaacgt caagaacctc aaggcggcct
601 ggaccttctc caccggcgtc ctccacggcc atgagggcgg cccgttggtg gtgaacggca
661 tcatgtacat ccatacgccc ttcccgaaca tcacgtacgc catcgatctc gataacccca
721 acaagatcct ctgggagcac aagccaaagc aggatgccag cgtccgggcc gtggcctgct
781 gcgacgtggt taaccgcggc ctggcgtacg gcgacggtaa gatcttcaag acccagctcg
841 acggccacat cgtcgctctc gatgctaaga cggggaagga gctctggaag atggagaact
901 cggaccccaa ggtgggcagc acgctgactc aggcgccgat ggttgtcgag gacatggtga
961 tcgtggggtc cagtggcgcg gagctcggcg tccgcgggta tgtcaccgcg taccgaatca
1021 aggacggcaa gcaggtgtgg agggcctacg ccacggggcc cgacgaagac atactgctgg
1081 cggacgattt caacatggcc aaccctcact acggacagaa gggcctgggc acatctacgt
1141 gggaggggga cgcctggaag atcggcggcg gcaccaattg gggctggtat gcctacgatc
1201 cggagcttaa gctgatgtac tatggcagcg gcaaccccgc cccctggaac gagaccatgc
1261 ggccgggcga caacaagtgg accatggcta tctggggccg tgacgtcaag acgggcaagg
1321 cgaagttcgc cttccagaag acgccccacg acgagtggga ttacgccggg gtcaattgga
1381 tcggactgtc cgaccaggtc gtcgatggca agaagcagaa gctcctgacc catccagacc
1441 gcaatggcat cgtgtacacc ctgaatcgcg aaaacggcaa cctcgtgcgc gccgacaaaa
1501 tcgatccgtc ggtgaacgtg tttaagggct acgatatgaa gaagggcgtc ccgatccgcg
1561 accctgagta ttccacccgt atggatcacc tcgccaaggg catctgcccc tcagccatgg
1621 ggtaccacaa ccagggacac gattctatcg acaagaagcg cgagctggtc atgctcggga
1681 caaacatgat ctgcatggac tgggagccct tcatgctccc ctaccgggcg gggcagttct
1741 ttgtgggcgc gacgctgaat atgtacgcga ccccgggcaa caacggtaac atggggcagg
1801 tgaaggccta tgacgtgaag acgagcaagt tcaagtgggt caaggacgaa aagttctccg
1861 tgtggggcgg caccacctcc accgctggcg acctggtctt ctacggcacc ctcgacgggc
1921 tgatcaaggc cgtggacgcc gacaatggga agaccttgtg ggagttcaag ctcccctcgg
1981 gcgtcatcgg ccatcccgtc acatacgagc acaaaggcaa gcagtacgtc gcgatctatt
2041 acggtgtcgg gggctggccg ggcgtcggcc tggtcttcga cctcaccgat cccaccgcgg
2101 gtctgggcgc cgtgggggcg ttcaaggagc tgcagcagta cacgaagatg ggcggtggcg
2161 tgatggtgtt ctccctcgcc gaaaacctct attttcaggg acatcatcac catcaccatc
2221 accactgaca gctatgacca tgattacgcc aagcttgcat gcctgcaggt cgactctaga
2281 ggatccccgg gtaccgagct cgaattcact ggccgtcgtt ttacaacgtc gtgactggga
2341 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
2401 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
2461 atggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg
2521 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
2581 aacacccgct gacgcgccct gacgggcttg tatgctcccg gcatccgctt acagacaagc
2641 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
2701 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
2761 ttcttagcac cctttctcgg tccttcaacg ttcctgacaa cgagcctcct tttcgccaat
2821 ccatcgacaa tcaccgcgag tccctgctcg aacgctgcgt ccggaccggc ttcgtcgaag
2881 gcgtctatcg cggcccgcaa cagcggcgag agcggagcct gttcaacggt gccgccgcgc
2941 tcgccggcat cgctgtcgcc ggcctgctcc tcaagcacgg ccccaacagt gaagtagctg
3001 attgtcatca gcgcattgac ggcgtccccg gccgaaaaac ccgcctcgca gaggaagcga
3061 agctgcgcgt cggccgtttc catctgcggt gcgcccggtc gcgtgccggc atggatgcgc
3121 gcgccatcgc ggtaggcgag cagcgcctgc ctgaagctgc gggcattccc gatcagaaat
3181 gagcgccagt cgtcgtcggc tctcggcacc gaatgcgtat gattctccgc cagcatggct
3241 tcggccagtg cgtcgagcag cgcccgcttg ttcctgaagt gccagtaaag cgccggctgc
3301 tgaaccccca accgttccgc cagtttgcgt gtcgtcagac cgtctacgcc gacctcgttc
3361 aacaggtcca gggcggcacg gatcactgta ttcggctgca actttgtcat gcttgacact
3421 ttatcactga taaacataat atgtccacca acttatcagt gataaagaat ccgcgcgttc
3481 aatcggacca gcggaggctg gtccggaggc cagacgtgaa acccaacata cccctgatcg
3541 taattctgag cactgtcgcg ctcgacgctg tcggcatcgg cctgattatg ccggtgctgc
3601 cgggcctcct gcgcgatctg gttcactcga acgacgtcac cgcccactat ggcattctgc
3661 tggcgctgta tgcgttggtg caatttgcct gcgcacctgt gctgggcgcg ctgtcggatc
3721 gtttcgggcg gcggccaatc ttgctcgtct cgctggccgg cgccactgtc gactacgcca
3781 tcatggcgac agcgcctttc ctttgggttc tctatatcgg gcggatcgtg gccggcatca
3841 ccggggcgac tggggcggta gccggcgctt atattgccga tatcactgat ggcgatgagc
3901 gcgcgcggca cttcggcttc atgagcgcct gtttcgggtt cgggatggtc gcgggacctg
3961 tgctcggtgg gctgatgggc ggtttctccc cccacgctcc gttcttcgcc gcggcagcct
4021 tgaacggcct caatttcctg acgggctgtt tccttttgcc ggagtcgcac aaaggcgaac
4081 gccggccgtt acgccgggag gctctcaacc cgctcgcttc gttccggtgg gcccggggca
4141 tgaccgtcgt cgccgccctg atggcggtct tcttcatcat gcaacttgtc ggacaggtgc
4201 cggccgcgct ttgggtcatt ttcggcgagg atcgctttca ctgggacgcg accacgatcg
4261 gcatttcgct tgccgcattt ggcattctgc attcactcgc ccaggcaatg atcaccggcc
4321 ctgtagccgc ccggctcggc gaaaggcggg cactcatgct cggaatgatt gccgacggca
4381 caggctacat cctgcttgcc ttcgcgacac ggggatggat ggcgttcccg atcatggtcc
4441 tgcttgcttc gggtggcatc ggaatgccgg cgctgcaagc aatgttgtcc aggcaggtgg
4501 atgaggaacg tcaggggcag ctgcaaggct cactggcggc gctcaccagc ctgacctcga
4561 tcgtcggacc cctcctcttc acggcgatct atgcggcttc tataacaacg tggaacgggt
4621 gggcatggat tgcaggcgct gccctctact tgctctgcct gccggcgctg cgtcgcgggc
4681 tttggagcgg cgcagggcaa cgagccgatc gctgatcgtg gaaacgatag gcctatgcca
4741 tgcgggtcaa ggcgacttcc ggcaagctat acgcgcccta gaattgtcaa ttttaatcct
4801 ctgtttatcg gcagttcgta gagcgcgccg tgcgtcccga gcgatactga gcgaagcaag
4861 tgcgtcgagc agtgcccgct tgttcctgaa atgccagtaa agcgctggct gctgaacccc
4921 cagccggaac tgaccccaca aggccctagc gtttgcaatg caccaggtca tcattgaccc
4981 aggcgtgttc caccaggccg ctgcctcgca actcttcgca ggcttcgccg acctgctcgc
5041 gccacttctt cacgcgggtg gaatccgatc cgcacatgag gcggaaggtt tccagcttga
5101 gcgggtacgg ctcccggtgc gagctgaaat agtcgaacat ccgtcgggcc gtcggcgaca
5161 gcttgcggta cttctcccat atgaatttcg tgtagtggtc gccagcaaac agcacgacga
5221 tttcctcgtc gatcaggacc tggcaacggg acgttttctt gccacggtcc aggacgcgga
5281 agcggtgcag cagcgacacc gattccaggt gcccaacgcg gtcggacgtg aagcccatcg
5341 ccgtcgcctg taggcgcgac aggcattcct cggccttcgt gtaataccgg ccattgatcg
5401 accagcccag gtcctggcaa agctcgtaga acgtgaaggt gatcggctcg ccgatagggg
5461 tgcgcttcgc gtactccaac acctgctgcc acaccagttc gtcatcgtcg gcccgcagct
5521 cgacgccggt gtaggtgatc ttcacgtcct tgttgacgtg gaaaatgacc ttgttttgca
5581 gcgcctcgcg cgggattttc ttgttgcgcg tggtgaacag ggcagagcgg gccgtgtcgt
5641 ttggcatcgc tcgcatcgtg tccggccacg gcgcaatatc gaacaaggaa agctgcattt
5701 ccttgatctg ctgcttcgtg tgtttcagca acgcggcctg cttggcctcg ctgacctgtt
5761 ttgccaggtc ctcgccggcg gtttttcgct tcttggtcgt catagttcct cgcgtgtcga
5821 tggtcatcga cttcgccaaa cctgccgcct cctgttcgag acgacgcgaa cgctccacgg
5881 cggccgatgg cgcgggcagg gcagggggag ccagttgcac gctgtcgcgc tcgatcttgg
5941 ccgtagcttg ctggaccatc gagccgacgg actggaaggt ttcgcggggc gcacgcatga
6001 cggtgcggct tgcgatggtt tcggcatcct cggcggaaaa ccccgcgtcg atcagttctt
6061 gcctgtatgc cttccggtca aacgtccgat tcattcaccc tccttgcggg attgccccga
6121 ctcacgccgg ggcaatgtgc ccttattcct gatttgaccc gcctggtgcc ttggtgtcca
6181 gataatccac cttatcggca atgaagtcgg tcccgtagac cgtctggccg tccttctcgt
6241 acttggtatt ccgaatcttg ccctgcacga ataccagctc cgcgaagtcg ctcttcttga
6301 tggagcgcat ggggacgtgc ttggcaatca cgcgcacccc ccggccgttt tagcggctaa
6361 aaaagtcatg gctctgccct cgggcggacc acgcccatca tgaccttgcc aagctcgtcc
6421 tgcttctctt cgatcttcgc cagcagggcg aggatcgtgg catcaccgaa ccgcgccgtg
6481 cgcgggtcgt cggtgagcca gagtttcagc aggccgccca ggcggcccag gtcgccattg
6541 atgcgggcca gctcgcggac gtgctcatag tccacgacgc ccgtgatttt gtagccctgg
6601 ccgacggcca gcaggtaggc ctacaggctc atgccggccg ccgccgcctt ttcctcaatc
6661 gctcttcgtt cgtctggaag gcagtacacc ttgataggtg ggctgccctt cctggttggc
6721 ttggtttcat cagccatccg cttgccctca tctgttacgc cggcggtagc cggccagcct
6781 cgcagagcag gattcccgtt gagcaccgcc aggtgcgaat aagggacagt gaagaaggaa
6841 cacccgctcg cgggtgggcc tacttcacct atcctgcccg gctgacgccg ttggatacac
6901 caaggaaagt ctacacgaac cctttggcaa aatcctgtat atcgtgcgaa aaaggatgga
6961 tataccgaaa aaatcgctat aatgaccccg aagcagggtt atgcagcgga aaagatccgt
7021 cgaccctttc cgacgctcac cgggctggtt gccctcgccg ctgggctggc ggccgtctat
7081 ggccctgcaa acgcgccaga aacgccgtcg aagccgtgtg cgagacaccg cggccgccgg
7141 cgttgtggat acctcgcgga aaacttggcc ctcactgaca gatgaggggc ggacgttgac
7201 acttgagggg ccgactcacc cggcgcggcg ttgacagatg aggggcaggc tcgatttcgg
7261 ccggcgacgt ggagctggcc agcctcgcaa atcggcgaaa acgcctgatt ttacgcgagt
7321 ttcccacaga tgatgtggac aagcctgggg ataagtgccc tgcggtattg acacttgagg
7381 ggcgcgacta ctgacagatg aggggcgcga tccttgacac ttgaggggca gagtgctgac
7441 agatgagggg cgcacctatt gacatttgag gggctgtcca caggcagaaa atccagcatt
7501 tgcaagggtt tccgcccgtt tttcggccac cgctaacctg tcttttaacc tgcttttaaa
7561 ccaatattta taaaccttgt ttttaaccag ggctgcgccc tgtgcgcgtg accgcgcacg
7621 ccgaaggggg gtgccccccc ttctcgaacc ctcccggccc gctaacgcgg gcctcccatc
7681 cccccagggg ctgcgcccct cggccgcgaa cggcctcacc ccaaaaatgg cagccaagct
7741 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
7801 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
7861 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
7921 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
7981 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
8041 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
8101 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
8161 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
8221 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
8281 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
8341 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
8401 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
8461 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
8521 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
8581 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
8641 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
8701 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
8761 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
8821 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
8881 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
8941 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
9001 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
9061 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
9121 attacgccaa gctagcttcc cgcttggtcg ggccgcttcg cgagggcccg ttgacgacaa
9181 cggtgcgatg ggtcccggcc ccggtcaaga cgatgccaat acgttgcgac actacgcctt
9241 ggcactttta gaattgcctt atcgtcctga taagaaatgt ccgaccagct aaagacatcg
9301 cgtccaatca aagcctagaa aatataggcg aagggacgct aataagtctt tcataagacc
9361 gcgcaaatct aaaaatatcc ttagattcac gatgcggcac ttcggatgac ttccgagcga
9421 gcctggaacc tcagaaaaac gtctgagaga taccgcggat ctcacacagg aaatttc
//
| {
"Vector backbone": "pCM80_PxoxF",
"Backbone size w/o insert (bp)": "",
"Vector type": "Synthetic Biology",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Tetracycline, 10 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "Top10",
"Copy number": "Low Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Rare earth element alcohol dehydrogenases widely occur among globally distributed, numerically abundant and environmentally important microbes",
"Publication": "ISME J. 2019 Aug;13(8):2005-2017. doi: 10.1038/s41396-019-0414-z. Epub 2019 Apr 5.",
"DOI": "10.1038/s41396-019-0414-z.",
"Authors": "Huang J, Yu Z, Groom J, Cheng JF, Tarver A, Yoshikuni Y, Chistoserdova L /s---z",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/30952993",
"PubMed ID": "PubMed 30952993"
} | {
"Gene/Insert name": "FAM1_1436",
"Species": "Synthetic",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Gibson Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
FJZ 663 | 108689 | in vitro mRNA transcription | Jon Lorsch | Yourik et al Elife. 2017 Nov 30;6. doi: 10.7554/eLife.31476. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/208356/76acce34-ae4c-47a3-baee-c172d858fac0/addgene-plasmid-108689-sequence-208356.gbk | full | LOCUS sequence-208356- 3236 bp DNA circular SYN 26-AUG-2024
DEFINITION in vitro mRNA transcription.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3236)
AUTHORS Yourik P, Aitken CE, Zhou F, Gupta N, Hinnebusch AG, Lorsch JR
TITLE Yeast eIF4A enhances recruitment of mRNAs regardless of their
structural complexity.
JOURNAL Elife. 2017 Nov 30;6. doi: 10.7554/eLife.31476.
PUBMED 29192585
REFERENCE 2 (bases 1 to 3236)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Elife.";
date: "2017-11-30"; volume: "6. doi"; pages: " 10.7554/eLife.31476"
FEATURES Location/Qualifiers
source 1..3236
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 1..19
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter complement(426..444)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(465..481)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(465..481)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(478..500)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 489..505
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(513..543)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 558..579
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(696..713)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(867..1455)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(947..966)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(1626..2486)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 2249..2268
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(2487..2591)
/gene="bla"
/label=AmpR promoter
rep_origin complement(2617..3072)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2754..2775)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2966..2985
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3199..3221
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 3213..3230
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 3214..3230
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
ORIGIN
1 taatacgact cactatagga aagaattgcc atcttggcaa ttcaacaaca acaacaacaa
61 caacaacaac aacaacaaca acaacaacaa caacaaaaaa tgagagccaa gtggagaaag
121 aagagaacta gaagacttaa gagaaagaga cggaaggtga gagccagatc caaataagcg
181 gattatgagt aaataactct aattttgttt taaattcttt caagagtatc gtaatgtcat
241 tgatgaatta acatgttagt ttctattcta cctcataatg gatctaaatt gcatactaat
301 ctcacggtgg ggtgtaaacc attgcctact atttatatag tgctttatat atgtctcaca
361 tagtttaatc aattgtccgt ttttttggtc gacctcgagg gggggcccgg taccagcttt
421 tgttcccttt agtgagggtt aatttcgagc ttggcgtaat catggtcata gctgtttcct
481 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
541 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
601 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
661 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
721 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
781 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
841 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
901 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
961 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
1021 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
1081 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
1141 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
1201 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
1261 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
1321 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
1381 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
1441 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
1501 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
1561 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
1621 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
1681 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
1741 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
1801 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
1861 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
1921 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
1981 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
2041 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
2101 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
2161 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
2221 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
2281 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
2341 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
2401 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
2461 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
2521 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
2581 ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt
2641 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
2701 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
2761 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
2821 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
2881 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
2941 agccggcgga cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
3001 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
3061 tacagggcgc gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
3121 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
3181 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattg
//
| {
"Vector backbone": "pBluescript II KS (+)",
"Backbone size w/o insert (bp)": "2960",
"Vector type": "Bacterial Expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Yeast eIF4A enhances recruitment of mRNAs regardless of their structural complexity",
"Publication": "Elife. 2017 Nov 30;6. doi: 10.7554/eLife.31476.",
"DOI": "10.7554/eLife.31476.",
"Authors": "Yourik P, Aitken CE, Zhou F, Gupta N, Hinnebusch AG, Lorsch JR /eLife",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/29192585",
"PubMed ID": "PubMed 29192585"
} | {
"Gene/Insert name": "RPL41A",
"Species": "S. cerevisiae (budding yeast)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "RPL41A",
"Promoter": "T7",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "unknown (unknown if destroyed)",
"3′ cloning site": "unknown (unknown if destroyed)",
"5′ sequencing primer": "M13",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
C2-66 pComb3xss anti-cFos nanobody | 145833 | Bacterial expression of anti-cFos (Human) recombinant llama nanobody for use as an immunolabel | James Trimmer | Dong et al Elife. 2019 Sep 30;8. pii: 48750. doi: 10.7554/eLife.48750. | partial | {
"Vector backbone": "pComb3xss",
"Backbone size w/o insert (bp)": "",
"Vector type": "Bacterial Expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "A toolbox of nanobodies developed and validated for use as intrabodies and nanoscale immunolabels in brain neurons",
"Publication": "Elife. 2019 Sep 30;8. pii: 48750. doi: 10.7554/eLife.48750.",
"DOI": "10.7554/eLife.48750.",
"Authors": "Dong JX, Lee Y, Kirmiz M, Palacio S, Dumitras C, Moreno CM, Sando R, Santana LF, Sudhof TC, Gong B, Murray KD, Trimmer JS /eLife",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/31566565",
"PubMed ID": "PubMed 31566565"
} | {
"Gene/Insert name": "anti-cFos (Human) recombinant llama nanobody C2-66",
"Species": "L. glama (llama)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "FOS",
"Promoter": "lac",
"Tag / Fusion Protein": "6xHis-HA (C terminal on backbone)"
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "SfiI (not destroyed)",
"3′ cloning site": "SfiI (not destroyed)",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |