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 |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
pOCC194 | 118882 | shuttle vector for baculovirus production, using FlashBac bacmid; for recombinant protein with C-terminal SNAP tag, cleavable with TEV, and C-terminal HIS6 tag, cleavable with 3C | Lemaitre et al BMC Biotechnol. 2019 Mar 29;19(1):20. doi: 10.1186/s12896-019-0512-z. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/232359/f6914861-5867-4da9-84e5-2fa050f931f2/addgene-plasmid-118882-sequence-232359.gbk | full | LOCUS sequence-232359- 5982 bp DNA circular SYN 26-AUG-2024
DEFINITION shuttle vector for baculovirus production, using FlashBac bacmid;
for recombinant protein with C-terminal SNAP tag, cleavable with
TEV, and C-terminal HIS6 tag, cleavable with 3C.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5982)
AUTHORS Lemaitre RP, Bogdanova A, Borgonovo B, Woodruff JB, Drechsel DN
TITLE FlexiBAC: a versatile, open-source baculovirus vector system for
protein expression, secretion, and proteolytic processing.
JOURNAL BMC Biotechnol. 2019 Mar 29;19(1):20. doi:
10.1186/s12896-019-0512-z.
PUBMED 30925874
REFERENCE 2 (bases 1 to 5982)
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: "BMC
Biotechnol."; date: "2019-03-29"; volume: "19(1):20. doi"; pages: "
10.1186/s12896-019-0512-z"
FEATURES Location/Qualifiers
source 1..5982
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 1..92
/gene="polh from Autographa californica"
/label=polyhedrin promoter
/note="promoter for the baculovirus polyhedrin gene"
primer_bind 17..34
/label=Polyhedrin-F
/note="Polyhedrin promoter, forward primer"
regulatory 134..143
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
regulatory 138..147
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 251..556
/codon_start=1
/gene="ccdB"
/product="CcdB, a bacterial toxin that poisons DNA gyrase"
/label=ccdB
/note="Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains."
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
primer_bind 435..454
/label=ccdB-fwd
/note="ccdB gene, forward primer"
CDS 612..632
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQG"
CDS 633..1178
/codon_start=1
/product="human O6-alkylguanine-DNA-alkyltransferase
variant that forms covalent bonds with benzylguanine
derivatives"
/label=SNAP-tag(R)
/note="fast-reacting variant, optimized for mammalian
expression"
/translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE
VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
VVKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLA
VKEWLLAHEGHRLGKPGLG"
CDS 1179..1202
/codon_start=1
/product="recognition and cleavage site for human
rhinovirus 3C and PreScission proteases"
/label=HRV 3C site
/translation="LEVLFQGP"
CDS 1212..1229
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
polyA_signal 1371..1505
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1408..1427)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 1462..1481
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
3'UTR 1578..1950
/label=polyhedrin 3' UTR
/note="3' UTR from the Autographa californica
nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
primer_bind complement(2302..2319)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2412..3000)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2492..2511)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3171..4031)
/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 3794..3813
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(4032..4136)
/gene="bla"
/label=AmpR promoter
primer_bind 4437..4459
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 4451..4468
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 4452..4468
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 4479..4498
/label=T7
/note="T7 promoter, forward primer"
promoter 4479..4497
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 5108..5232
/codon_start=3
/product="baculovirus late expression factor 2"
/label=lef2
/translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
CDS complement(5270..5875)
/codon_start=1
/product="baculovirus ORF603 protein"
/label=ORF603
/translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
ORIGIN
1 atcatggaga taattaaaat gataaccatc tcgcaaataa ataagtattt tactgttttc
61 gtaacagttt tgtaataaaa aaacctataa atattccgga ttattcatac cgtcccacca
121 tcgggcgcgg atcgccacca tggcggccgc aagcgctatg gccacaacct gttacattgc
181 acaagataaa aatatatcat cacgaacagt aaaactgtct gcttacataa acagtaatac
241 aaggggtgtt atgcagttta aggtttacac ctataaaaga gagagccgtt atcgtctgtt
301 tgtggatgta cagagtgata ttattgacac gcccgggcga cggatggtga tccccctggc
361 cagtgcccgt ctgctgtcag ataaagtctc ccgtgaactt tacccggtgg tgcatatcgg
421 ggatgaaagc tggcgcatga tgaccaccga tatggccagt gtgccggtct ccgttatcgg
481 ggaagaagtg gctgatctca gccaccgcga aaatgacatc aaaaacgcca ttaacctgat
541 gttctgggga atataaatgt caggctccct tatacacagc cagtggcgcg ccgggatcca
601 gcagcggcag agaaaacttg tatttccagg gcatggacaa agactgcgaa atgaagcgca
661 ccaccctgga tagccctctg ggcaagctgg aactgtctgg gtgcgaacag ggcctgcacc
721 gtatcatctt cctgggcaaa ggaacatctg ccgccgacgc cgtggaagtg cctgccccag
781 ccgccgtgct gggcggacca gagccactga tgcaggccac cgcctggctc aacgcctact
841 ttcaccagcc tgaggccatc gaggagttcc ctgtgccagc cctgcaccac ccagtgttcc
901 agcaggagag ctttacccgc caggtgctgt ggaaactgct gaaagtggtg aagttcggag
961 aggtcatcag ctacagccac ctggccgccc tggccggcaa tcccgccgcc accgccgccg
1021 tgaaaaccgc cctgagcgga aatcccgtgc ccattctgat cccctgccac cgggtggtgc
1081 agggcgacct ggacgtgggg ggctacgagg gcgggctcgc cgtgaaagag tggctgctgg
1141 cccacgaggg ccacagactg ggcaagcctg ggctgggtct ggaagttctg ttccaggggc
1201 cgggcagcag ccatcaccat caccatcaca gtggataata gaagcttgtc gagaagtact
1261 agaggatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
1321 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
1381 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
1441 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
1501 ggatctgatc actgcttgag cctaggagat ccgaaccaga taagtgaaag cctgcatccg
1561 cagatcttaa ttaataaaac acgatacatt gttattagta catttattaa gcgctagatt
1621 ctgtgcgttg ttgatttaca gacaattgtt gtacgtattt taataattca ttaaatttat
1681 aatctttagg gtggtatgtt agagcgaaaa tcaaatgatt ttcagcgtct ttatatctga
1741 atttaaatat taaatcctca atagatttgt aaaataggtt tcgattagtt tcaaacaagg
1801 gttgtttttc cgaaccgatg gctggactat ctaatggatt ttcgctcaac gccacaaaac
1861 ttgccaaatc ttgtagcagc aatctagctt tgtcgatatt cgtttgtgtt ttgttttgta
1921 ataaaggttc gacgtcgttc aaaatattat gcgcttttgt atttctttca tcactgtcgt
1981 tagtgtacaa ttgactcgac gtaaacacgt taaataacgc ttggacatat ttaacatcgg
2041 gcgtgttagc tttattaggc cgattatcgt cgtcgtccca accctcgtcg ttagaagttg
2101 cttccgaaga cgattttgcc atagccacac gacgcctatt aattgtgtcg gctaacacgt
2161 ccgcgatcaa atttgtagtt gagctttttg ggcctcatgg gccttccgct cactgcccgc
2221 tttccagtcg ggaaacctgt cgtgccagct gcattaacat ggtcatagct gtttccttgc
2281 gtattgggcg ctctccgctt cctcgctcac tgactcgctg cgctcggtcg ttcgggtaaa
2341 gcctggggtg cctaatgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
2401 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
2461 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
2521 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
2581 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
2641 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
2701 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
2761 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
2821 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
2881 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
2941 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
3001 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
3061 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
3121 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
3181 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
3241 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
3301 atgataccgc gagaaccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
3361 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
3421 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
3481 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
3541 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
3601 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
3661 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
3721 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
3781 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
3841 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
3901 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
3961 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
4021 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
4081 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
4141 tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
4201 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
4261 aatcaaaaga atagaccgag atagggttga gtggccgcta cagggcgctc ccattcgcca
4321 ttcaggctgc gcaactgttg ggaagggcgt ttcggtgcgg gcctcttcgc tattacgcca
4381 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
4441 gtcacgacgt tgtaaaacga cggccagtga gcgcgacgta atacgactca ctatagggcg
4501 aattggcgga aggccgtcaa ggccgtgacg ttaaaactat taagccatcc aatcgaccgt
4561 tagtcgaatc aggaccgctg gtgcgagaag ccgcgaagta tggcgaatgc atcgtataac
4621 gtgtggagtc cgctcattag agcgtcatgt ttagacaaga aagctacata tttaattgat
4681 cccgatgatt ttattgataa attgacccta actccataca cggtattcta caatggcggg
4741 gttttggtca aaatttccgg actgcgattg tacatgctgt taacggctcc gcccactatt
4801 aatgaaatta aaaattccaa ttttaaaaaa cgcagcaaga gaaacatttg tatgaaagaa
4861 tgcgtagaag gaaagaaaaa tgtcgtggac atgctgaaca acaagattaa tatgcctccg
4921 tgtataaaaa aaatattgaa cgatttgaaa gaaaacaatg taccgcgcgg cggtatgtac
4981 aggaagaggt ttatactaaa ctgttacatt gcaaacgtgg tttcgtgtgc caagtgtgaa
5041 aaccgatgtt taatcaaggc tctgacgcat ttctacaacc acgactccaa gtgtgtgggt
5101 gaagtcatgc atcttttaat caaatcccaa gatgtgtata aaccaccaaa ctgccaaaaa
5161 atgaaaactg tggacaagct ctgtccgttt gctggcaact gcaagggtct caatcctatt
5221 tgtaattatt gaataataaa acaattataa atgctaaatt tgttttttat taacgataca
5281 aaccaaacgc aacaagaaca tttgtagtat tatctataat tgaaaacgcg tagttataat
5341 cgctgaggta atatttaaaa tcattttcaa atgattcaca gttaatttgc gacaatataa
5401 ttttattttc acataaacta gacgccttgt cgtcttcttc ttcgtattcc ttctcttttt
5461 catttttctc ctcataaaaa ttaacatagt tattatcgta tccatatatg tatctatcgt
5521 atagagtaaa ttttttgttg tcataaatat atatgtcttt tttaatgggg tgtatagtac
5581 cgctgcgcat agtttttctg taatttacaa cagtgctatt ttctggtagt tcttcggagt
5641 gtgttgcttt aattattaaa tttatataat caatgaattt gggatcgtcg gttttgtaca
5701 atatgttgcc ggcatagtac gcagcttctt ctagttcaat tacaccattt tttagcagca
5761 ccggattaac ataactttcc aaaatgttgt acgaaccgtt aaacaaaaac agttcacctc
5821 ccttttctat actattgtct gcgagcagtt gtttgttgtt aaaaataaca gccattgtaa
5881 tgagacgcac aaactaatat cacaagctgc caccattttg ttcgcccagg actctagcta
5941 tagttctagt ggttggctac gtatactccg gaatattaat ag
//
| {
"Vector backbone": "pOET1",
"Backbone size w/o insert (bp)": "4900",
"Vector type": "Insect Expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DB3.1",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "FlexiBAC: a versatile, open-source baculovirus vector system for protein expression, secretion, and proteolytic processing",
"Publication": "BMC Biotechnol. 2019 Mar 29;19(1):20. doi: 10.1186/s12896-019-0512-z.",
"DOI": "10.1186/s12896-019-0512-z.",
"Authors": "Lemaitre RP, Bogdanova A, Borgonovo B, Woodruff JB, Drechsel DN /s---z",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/30925874",
"PubMed ID": "PubMed 30925874"
} | {
"Gene/Insert name": "NcoI-NotI-ccdB-AscI-TEV-SNAP-3C-HIS6-stop-HindIII cassette",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "polH",
"Tag / Fusion Protein": "SNAP tag, cleavable with TEV protease, HIS6, cleavable with 3C protease (C terminal on backbone)"
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "NotI (unknown if destroyed)",
"3′ cloning site": "AscI (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": ""
} |
||
Flag-Rab35 S22N inactive | 47429 | Peter McPherson | Allaire et al J Cell Sci. 2013 Feb 1;126(Pt 3):722-31. doi: 10.1242/jcs.112375. Epub 2012 Dec 21. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/68405/108cbc53-8256-4936-b2f5-baeacc29147f/addgene-plasmid-47429-sequence-68405.gbk | partial | LOCUS sequence-68405-e 972 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 972)
AUTHORS Allaire PD, Seyed Sadr M, Chaineau M, Seyed Sadr E, Konefal S,
Fotouhi M, Maret D, Ritter B, Del Maestro RF, McPherson PS
TITLE Interplay between Rab35 and Arf6 controls cargo recycling to
coordinate cell adhesion and migration.
JOURNAL J Cell Sci. 2013 Feb 1;126(Pt 3):722-31. doi: 10.1242/jcs.112375.
Epub 2012 Dec 21.
PUBMED 23264734
REFERENCE 2 (bases 1 to 972)
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 Cell
Sci."; date: "2013-02-1"; volume: "126(Pt 3):722-31. doi"; pages: "
10.1242/jcs.112375. Epub 2012 Dec 21"
FEATURES Location/Qualifiers
source 1..972
/mol_type="genomic DNA"
/organism="unspecified"
primer_bind 50..69
/label=T7
/note="T7 promoter, forward primer"
promoter 50..68
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory 82..91
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 91..114
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
promoter complement(787..805)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(788..805)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(825..842)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 831..942
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
ORIGIN
1 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
61 actataggga gacccaagct tgccaccatg gactacaaag acgatgacga caagggatcc
121 atggcccggg actacgacca cctcttcaag ctgctcatca tcggcgacag cggtgtgggc
181 aagaacagtt tactgttgcg ttttgcagac aacactttct caggcagcta catcaccacg
241 atcggagtgg atttcaagat ccggaccgtg gagatcaacg gggagaaggt gaagctgcag
301 atctgggaca cagcggggca ggagcgcttc cgcaccatca cctccacgta ttatcggggg
361 acccacgggg tcattgtggt ttacgacgtc accagtgccg agtcctttgt caacgtcaag
421 cggtggcttc acgaaatcaa ccagaactgt gatgatgtgt gccgaatatt agtgggtaat
481 aagaatgacg accctgagcg gaaggtggtg gagacggaag atgcctacaa attcgccggg
541 cagatgggca tccagttgtt cgagaccagc gccaaggaga atgtcaacgt ggaagagatg
601 ttcaactgca tcacggagct ggtcctccga gcaaagaaag acaacctggc aaaacagcag
661 cagcaacaac agaacgatgt ggtgaagctc acgaagaaca gtaaacgaaa gaaacgctgc
721 tgctaagaat tctgcagata tccatcacac tggcggccgc tcgagcatgc atctagaggg
781 ccctattcta tagtgtcacc taaatgctag agctcgctga tcagcctcga ctgtgccttc
841 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
901 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
961 tcattctatt ct
//
| {
"Vector backbone": "pcDNA3-Flag",
"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": "Interplay between Rab35 and Arf6 controls cargo recycling to coordinate cell adhesion and migration",
"Publication": "J Cell Sci. 2013 Feb 1;126(Pt 3):722-31. doi: 10.1242/jcs.112375. Epub 2012 Dec 21.",
"DOI": "10.1242/jcs.112375.",
"Authors": "Allaire PD, Seyed Sadr M, Chaineau M, Seyed Sadr E, Konefal S, Fotouhi M, Maret D, Ritter B, Del Maestro RF, McPherson PS /jcs",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/23264734",
"PubMed ID": "PubMed 23264734"
} | {
"Gene/Insert name": "Rab35 S22N",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "600",
"Mutation": "S22N inactive",
"Entrez Gene": "RAB35",
"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": "BGHrev"
} | {
"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": ""
} |
||
GluN2B/NR2B glutamate receptor scFv [N59/36] | 190563 | Mammalian Expression of GluN2B/NR2B glutamate receptor scFV. Derived from hybridoma N59/36. | James Trimmer | Mitchell et al Sci Rep. 2023 Sep 27;13(1):16200. doi: 10.1038/s41598-023-43233-4. | partial | {
"Vector backbone": "pCDNA3.1",
"Backbone size w/o insert (bp)": "5428",
"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": "High-volume hybridoma sequencing on the NeuroMabSeq platform enables efficient generation of recombinant monoclonal antibodies and scFvs for neuroscience research",
"Publication": "Sci Rep. 2023 Sep 27;13(1):16200. doi: 10.1038/s41598-023-43233-4.",
"DOI": "10.1038/s41598-023-43233-4.",
"Authors": "Mitchell KG, Gong B, Hunter SS, Burkart-Waco D, Gavira-O'Neill CE, Templeton KM, Goethel ME, Bzymek M, MacNiven LM, Murray KD, Settles ML, Froenicke L, Trimmer JS /s---",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/37758930",
"PubMed ID": "PubMed 37758930"
} | {
"Gene/Insert name": "GluN2B/NR2B glutamate receptor (Rattus norvegicus) recombinant scFV",
"Species": "M. musculus (mouse)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "Grin2b",
"Promoter": "CMV",
"Tag / Fusion Protein": "HA, Sortase, 6xHis (C terminal on insert)"
} | {
"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": ""
} |
|||
pDY0312 PhiBT Integrase | 179111 | PhiBT Integrase expression vector | Yarnall et al Nat Biotechnol. 2022 Nov 24. doi: 10.1038/s41587-022-01527-4. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/354284/4278c275-5bd8-4c3b-a3d2-761fcdfbb40b/addgene-plasmid-179111-sequence-354284.gbk | full | LOCUS sequence-354284- 6638 bp DNA circular SYN 25-AUG-2024
DEFINITION PhiBT Integrase expression vector.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6638)
AUTHORS Yarnall MTN, Ioannidi EI, Schmitt-Ulms C, Krajeski RN, Lim J,
Villiger L, Zhou W, Jiang K, Garushyants SK, Roberts N, Zhang L,
Vakulskas CA, Walker JA 2nd, Kadina AP, Zepeda AE, Holden K, Ma H,
Xie J, Gao G, Foquet L, Bial G, Donnelly SK, Miyata Y, Radiloff DR,
Henderson JM, Ujita A, Abudayyeh OO, Gootenberg JS
TITLE Drag-and-drop genome insertion of large sequences without
double-strand DNA cleavage using CRISPR-directed integrases.
JOURNAL Nat Biotechnol. 2022 Nov 24. doi: 10.1038/s41587-022-01527-4.
PUBMED 36424489
REFERENCE 2 (bases 1 to 6638)
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
Biotechnol. 2022 Nov 24. doi: 10.1038/s41587-022-01527-4."
FEATURES Location/Qualifiers
source 1..6638
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 6..385
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 387..664
/label=chicken beta-actin promoter
intron 665..1681
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
primer_bind 1605..1627
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 1694..1713
/label=T7
/note="T7 promoter, forward primer"
promoter 1694..1712
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 1732..1752
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
primer_bind complement(3557..3574)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 3563..3787
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(3859..3876)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(3859..3875)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3868..3890)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin complement(4016..4471)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4153..4174)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 4365..4384
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 4498..4602
/gene="bla"
/label=AmpR promoter
CDS 4603..5463
/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(4821..4840)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 5634..6222
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6123..6142
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 6376..6393
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 6510..6531
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 6546..6576
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6584..6600
/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 6589..6611
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 6608..6624
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 6608..6624
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
ORIGIN
1 gattcgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
61 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
121 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
181 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
241 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
301 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
361 cgtattagtc atcgctatta ccatggtcga ggtgagcccc acgttctgct tcactctccc
421 catctccccc ccctccccac ccccaatttt gtatttattt attttttaat tattttgtgc
481 agcgatgggg gcgggggggg ggggggggcg cgcgccrggs ggggsggggs ggggsgrggg
541 gsggggsggg gsgaggcgga gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa
601 gtttcctttt atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg
661 ggcgggagtc gctgcgcgct gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg
721 cccgccccgg ctctgactga ccgcgttact cccacaggtg agcgggcggg acggcccttc
781 tcctccgggc tgtaattagc gcttggttta atgacggctt gtttcttttc tgtggctgcg
841 tgaaagcctt gaggggctcc gggagggccc tttgtgcggg gggagcggct cggggggtgc
901 gtgcgtgtgt gtgtgcgtgg ggagcgccgc gtgcggctcc gcgctgcccg gcggctgtga
961 gcgctgcggg cgcggcgcgg ggctttgtgc gctccgcagt gtgcgcgagg ggagcgcggc
1021 cgggggcggt gccccgcggt gcgggggggg ctgcgagggg aacaaaggct gcgtgcgggg
1081 tgtgtgcgtg ggggggtgag cagggggtgt gggcgcgtcg gtcgggctgc aaccccccct
1141 gcacccccct ccccgagttg ctgagcacgg cccggcttcg ggtgcggggc tccgtacggg
1201 gcgtggcgcg gggctcgccg tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg
1261 ggcggggccg cctcgggccg gggagggctc gggggagggg cgcggcggcc cccggagcgc
1321 cggcggctgt cgaggcgcgg cgagccgcag ccattgcctt ttatggtaat cgtgcgagag
1381 ggcgcaggga cttcctttgt cccaaatctg tgcggagccg aaatctggga ggcgccgccg
1441 caccccctct agcgggcgcg gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg
1501 ggagggcctt cgtgcgtcgc cgcgccgccg tccccttctc cctctccagc ctcggggctg
1561 tccgcggggg gacggctgcc ttcggggggg acggggcagg gcggggttcg gcttctggcg
1621 tgtgaccggc ggctctagag cctctgctaa ccatgttcat gccttcttct ttttcctaca
1681 gatccttaat taataatacg actcactata gggggtcgac ccgccaccat gcccaagaag
1741 aaacggaaag tgatgagccc ctttatcgcc ccggacgtgc ccgagcacct cctggacact
1801 gtgcgcgtct ttctgtacgc ccgtcagagt aaaggacggt cagatggatc tgacgtgtcc
1861 accgaagcac agctcgctgc cggacgggcc cttgttgcct caagaaacgc acaaggggga
1921 gctagatggg tggtggcggg cgaattcgtg gatgtgggca gatcagggtg ggacccgaat
1981 gtgacacgcg ccgacttcga aagaatgatg ggcgaggtgc gcgccggtga gggagacgta
2041 gtggtggtta atgaactgag tcgccttacg aggaagggcg cccacgacgc tctggagatc
2101 gataacgaac tcaaaaaaca cggtgtgcgg ttcatgagcg tgctggaacc attcctggat
2161 accagcaccc caatcggtgt cgcgatcttt gccctgattg ccgcgctcgc taaacaggat
2221 tcagacctta aagctgagcg gctgaagggg gctaaagatg agatcgctgc cttggggggt
2281 gtgcacagct catctgcgcc attcggcatg agggcggtca gaaagaaagt ggataacctg
2341 gtcatatctg ttctggagcc tgatgaggac aacccggacc acgttgagct tgtggaacgg
2401 atggctaaga tgtctttcga aggcgtcagc gataacgcaa ttgccacaac atttgagaag
2461 gagaaaatcc cctctccggg gatggctgag agacgagcca cggagaagag gcttgcttct
2521 attaaggcac ggaggctcaa tggcgccgaa aagccgatca tgtggcgggc gcagacagtt
2581 agatggattc ttaaccatcc cgcgattggt ggattcgcat tcgagcgggt gaaacacgga
2641 aaagcccaca tcaacgtgat acgaagagat cccggcggca aaccccttac ccctcacact
2701 ggtatcctgt ctggatccaa gtggttggaa ctccaggaga agagaagcgg gaaaaatctc
2761 tccgaccgca aaccaggtgc cgaagtggaa cctacgctgc tttccgggtg gagatttctg
2821 ggatgtcgga tatgcggtgg gtcaatgggc cagtcccaag ggggccgtaa gaggaatggg
2881 gacttggctg agggcaatta catgtgtgca aacccaaagg ggcacggcgg tctgagcgtc
2941 aagaggtctg agcttgatga attcgtggca tcaaaagtct gggccaggtt gcgcacggct
3001 gacatggagg atgaacatga ccaagcatgg attgcagctg cagctgaacg gtttgctttg
3061 cagcacgacc tggcgggggt agctgacgag cgacgggagc aacaagctca cctggataac
3121 gttcggagat caataaaaga tctccaggcg gataggaagg caggtctcta cgtgggacgc
3181 gaagaactgg agacctggcg cagtaccgtc ctgcaatata ggagctacga ggctgagtgt
3241 actactaggt tggctgagct ggatgaaaaa atgaatggat ccacccgggt gccttcagaa
3301 tggtttagcg gcgaggaccc aaccgcggaa ggaggcatat gggcgagctg ggatgtctat
3361 gagcgccggg agtttctcag cttttttttg gactccgtaa tggttgacag gggcagacat
3421 cctgaaacca agaaatatat accattgaaa gaccgggtga ccttaaagtg ggcggagctg
3481 ttaaaggaag aggatgaagc aagcgaggcc acagaacggg agctggcagc tctttaggtc
3541 agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
3601 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
3661 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
3721 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
3781 tctatggctt ctgaggcgga aagaaccagc tggggctcga gatccactag ttctagcctc
3841 gaggctagag cggccgccac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
3901 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
3961 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggaaatt
4021 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
4081 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
4141 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
4201 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
4261 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
4321 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
4381 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
4441 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc
4501 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
4561 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
4621 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
4681 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
4741 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
4801 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
4861 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
4921 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
4981 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
5041 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
5101 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
5161 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
5221 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
5281 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
5341 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
5401 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
5461 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
5521 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
5581 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
5641 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
5701 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
5761 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
5821 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
5881 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
5941 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
6001 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
6061 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
6121 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
6181 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
6241 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
6301 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
6361 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
6421 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
6481 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
6541 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
6601 tttcacacag gaaacagcta tgaccatgag gcgcgccg
//
| {
"Vector backbone": "pCMV",
"Backbone size w/o insert (bp)": "",
"Vector type": "CRISPR",
"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": "Drag-and-drop genome insertion of large sequences without double-strand DNA cleavage using CRISPR-directed integrases",
"Publication": "Nat Biotechnol. 2022 Nov 24. doi: 10.1038/s41587-022-01527-4.",
"DOI": "10.1038/s41587-022-01527-4.",
"Authors": "Yarnall MTN, Ioannidi EI, Schmitt-Ulms C, Krajeski RN, Lim J, Villiger L, Zhou W, Jiang K, Garushyants SK, Roberts N, Zhang L, Vakulskas CA, Walker JA nd, Kadina AP, Zepeda AE, Holden K, Ma H, Xie J, Gao G, Foquet L, Bial G, Donnelly SK, Miyata Y, Radiloff DR, Henderson JM, Ujita A, Abudayyeh OO, Gootenberg JS /s---",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/36424489",
"PubMed ID": "PubMed 36424489"
} | {
"Gene/Insert name": "PhiBT Integrase",
"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": ""
} |
||
pDONR223_GSK3B_WT | 82126 | Gateway Donor vector containing GSK3B , 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/164661/b019215c-49fa-4a2f-a359-e822bcf1af06/addgene-plasmid-82126-sequence-164661.gbk | full | LOCUS sequence-164661- 4056 bp DNA circular SYN 25-AUG-2024
DEFINITION Gateway Donor vector containing GSK3B , part of the Target
Accelerator Plasmid Collection..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4056)
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 4056)
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:
"10.1158/2159-8290.CD-16-0160"; journalName: "Cancer Discov"; date:
"2016-07"; volume: "6"; issue: "7"; pages: "714-26"
FEATURES Location/Qualifiers
source 1..4056
/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 1488..1499
/codon_start=1
/product="WELQut protease recognition and cleavage site"
/label=WELQut site
/translation="WELQ"
protein_bind complement(1935..2034)
/gene="mutant version of attL"
/label=attL2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
primer_bind complement(2051..2070)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(2052..2070)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(2075..2091)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2075..2091)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
CDS 2522..3313
/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 3406..3994
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3895..3914
/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 agttggcacc atgtcagggc ggcccagaac cacctccttt gcggagagct gcaagccggt
721 gcagcagcct tcagcttttg gcagcatgaa agttagcaga gacaaggacg gcagcaaggt
781 gacaacagtg gtggcaactc ctgggcaggg tccagacagg ccacaagaag tcagctatac
841 agacactaaa gtgattggaa atggatcatt tggtgtggta tatcaagcca aactttgtga
901 ttcaggagaa ctggtcgcca tcaagaaagt attgcaggac aagagattta agaatcgaga
961 gctccagatc atgagaaagc tagatcactg taacatagtc cgattgcgtt atttcttcta
1021 ctccagtggt gagaagaaag atgaggtcta tcttaatctg gtgctggact atgttccgga
1081 aacagtatac agagttgcca gacactatag tcgagccaaa cagacgctcc ctgtgattta
1141 tgtcaagttg tatatgtatc agctgttccg aagtttagcc tatatccatt cctttggaat
1201 ctgccatcgg gatattaaac cgcagaacct cttgttggat cctgatactg ctgtattaaa
1261 actctgtgac tttggaagtg caaagcagct ggtccgagga gaacccaatg tttcgtatat
1321 ctgttctcgg tactataggg caccagagtt gatctttgga gccactgatt atacctctag
1381 tatagatgta tggtctgctg gctgtgtgtt ggctgagctg ttactaggac aaccaatatt
1441 tccaggggat agtggtgtgg atcagttggt agaaataatc aaggtcctgg gaactccaac
1501 aagggagcaa atcagagaaa tgaacccaaa ctacacagaa tttaaattcc ctcaaattaa
1561 ggcacatcct tggactaagg tcttccgacc ccgaactcca ccggaggcaa ttgcactgtg
1621 tagccgtctg ctggagtata caccaactgc ccgactaaca ccactggaag cttgtgcaca
1681 ttcatttttt gatgaattac gggacccaaa tgtcaaacta ccaaatgggc gagacacacc
1741 tgcactcttc aacttcacca ctcaagaact gtcaagtaat ccacctctgg ctaccatcct
1801 tattcctcct catgctcgga ttcaagcagc tgcttcaacc cccacaaatg ccacagcagc
1861 gtcagatgct aatactggag accgtggaca gaccaataat gctgcttctg catcagcttc
1921 caactccacc tgcttgccaa ctttcttgta caaagttggc attataagaa agcattgctt
1981 atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc
2041 agctgatatc ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg
2101 gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgcct
2161 cctctagacc agccaggaca gaaatgcctc gacttcgctg ctacccaagg ttgccgggtg
2221 acgcacaccg tggaaacgga tgaaggcacg aacccagtgg acataagcct gttcggttcg
2281 taagctgtaa tgcaagtagc gtatgcgctc acgcaactgg tccagaacct tgaccgaacg
2341 cagcggtggt aacggcgcag tggcggtttt catggcttgt tatgactgtt tttttggggt
2401 acagtctatg cctcgggcat ccaagcagca agcgcgttac gccgtgggtc gatgtttgat
2461 gttatggagc agcaacgatg ttacgcagca gggcagtcgc cctaaaacaa agttaaacat
2521 tatgagggaa gcggtgatcg ccgaagtatc gactcaacta tcagaggtag ttggcgtcat
2581 cgagcgccat ctcgaaccga cgttgctggc cgtacatttg tacggctccg cagtggatgg
2641 cggcctgaag ccacacagtg atattgattt gctggttacg gtgaccgtaa ggcttgatga
2701 aacaacgcgg cgagctttga tcaacgacct tttggaaact tcggcttccc ctggagagag
2761 cgagattctc cgcgctgtag aagtcaccat tgttgtgcac gacgacatca ttccgtggcg
2821 ttatccagct aagcgcgaac tgcaatttgg agaatggcag cgcaatgaca ttcttgcagg
2881 tatcttcgag ccagccacga tcgacattga tctggctatc ttgctgacaa aagcaagaga
2941 acatagcgtt gccttggtag gtccagcggc ggaggaactc tttgatccgg ttcctgaaca
3001 ggatctattt gaggcgctaa atgaaacctt aacgctatgg aactcgccgc ccgactgggc
3061 tggcgatgag cgaaatgtag tgcttacgtt gtcccgcatt tggtacagcg cagtaaccgg
3121 caaaatcgcg ccgaaggatg tcgctgccga ctgggcaatg gagcgcctgc cggcccagta
3181 tcagcccgtc atacttgaag ctagacaggc ttatcttgga caagaagaag atcgcttggc
3241 ctcgcgcgca gatcagttgg aagaatttgt ccactacgtg aaaggcgaga tcaccaaggt
3301 agtcggcaaa taaccctcga gccacccatg accaaaatcc cttaacgtga gttacgcgtc
3361 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
3421 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
3481 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
3541 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
3601 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
3661 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
3721 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
3781 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
3841 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
3901 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
3961 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
4021 gttcctggcc ttttgctggc cttttgctca catgtt
//
| {
"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": "GSK3B",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "WT",
"Entrez Gene": "GSK3B",
"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": ""
} |
||
pDLX4.1.0-gDNA | 132437 | CRISPR/Cas9 plasmid to create GFP fusion proteins | Kevin White | ENCODE collection: CRISPR constructs (unpublished) | https://media.addgene.org/snapgene-media/v2.0.0/sequences/266219/e19f65ad-f0f5-4759-8c73-a6db1a15db4e/addgene-plasmid-132437-sequence-266219.gbk | full | LOCUS sequence-266219- 8499 bp DNA circular SYN 26-AUG-2024
DEFINITION CRISPR/Cas9 plasmid to create GFP fusion proteins.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8499)
TITLE ENCODE collection: CRISPR constructs
REFERENCE 2 (bases 1 to 8499)
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..8499
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 1..249
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA
(Domitrovich & Kunkel, 2003)"
primer_bind 1..21
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 172..191
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
misc_RNA 271..346
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
enhancer 443..728
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer;
contains an 18-bp deletion relative to the standard CMV
enhancer"
intron 999..1226
/label=hybrid intron
/note="hybrid between chicken beta-actin (CBA) and minute
virus of mice (MMV) introns (Gray et al., 2011)"
regulatory 1238..1247
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 1247..1312
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
CDS 1319..1339
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 1364..5464
/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 1364..5464
/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 5465..5512
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
primer_bind complement(5540..5557)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 5546..5753
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
repeat_region 5762..5902
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 5762..5891
/label=AAV2 ITR
rep_origin 5977..6432
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(6064..6083)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 6274..6295
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(6449..6468)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 6568..6590
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(6628..6646)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 6714..6818
/gene="bla"
/label=AmpR promoter
CDS 6819..7679
/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(7037..7056)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 7850..8438
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 8339..8358
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
61 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
241 cgaaacaccg cctcagatga tgtgaatctg gttttagagc tagaaatagc aagttaaaat
301 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt ttgttttaga
361 gctagaaata gcaagttaaa ataaggctag tccgttttta gcgcgtgcgc caattctgca
421 gacaaatggc tctagaggta cccgttacat aacttacggt aaatggcccg cctggctgac
481 cgcccaacga cccccgccca ttgacgtcaa tagtaacgcc aatagggact ttccattgac
541 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
601 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattgtgccc
661 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
721 ttaccatggt cgaggtgagc cccacgttct gcttcactct ccccatctcc cccccctccc
781 cacccccaat tttgtattta tttatttttt aattattttg tgcagcgatg ggggcggggg
841 ggggggcgcc aggcggggcg gggcggggcg aggggcgggg cggggcgagg cggagaggtg
901 cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc
961 ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg agtcgctgcg cgctgccttc
1021 gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt
1081 tactcccaca ggtgagcggg cgggacggcc cttctcctcc gggctgtaat tagctgagca
1141 agaggtaagg gtttaaggga tggttggttg gtggggtatt aatgtttaat tacctggagc
1201 acctgcctga aatcactttt tttcaggttg gaccggtgcc accatggact ataaggacca
1261 cgacggagac tacaaggatc atgatattga ttacaaagac gatgacgata agatggcccc
1321 aaagaagaag cggaaggtcg gtatccacgg agtcccagca gccgacaaga agtacagcat
1381 cggcctggac atcggcacca actctgtggg ctgggccgtg atcaccgacg agtacaaggt
1441 gcccagcaag aaattcaagg tgctgggcaa caccgaccgg cacagcatca agaagaacct
1501 gatcggagcc ctgctgttcg acagcggcga aacagccgag gccacccggc tgaagagaac
1561 cgccagaaga agatacacca gacggaagaa ccggatctgc tatctgcaag agatcttcag
1621 caacgagatg gccaaggtgg acgacagctt cttccacaga ctggaagagt ccttcctggt
1681 ggaagaggat aagaagcacg agcggcaccc catcttcggc aacatcgtgg acgaggtggc
1741 ctaccacgag aagtacccca ccatctacca cctgagaaag aaactggtgg acagcaccga
1801 caaggccgac ctgcggctga tctatctggc cctggcccac atgatcaagt tccggggcca
1861 cttcctgatc gagggcgacc tgaaccccga caacagcgac gtggacaagc tgttcatcca
1921 gctggtgcag acctacaacc agctgttcga ggaaaacccc atcaacgcca gcggcgtgga
1981 cgccaaggcc atcctgtctg ccagactgag caagagcaga cggctggaaa atctgatcgc
2041 ccagctgccc ggcgagaaga agaatggcct gttcggaaac ctgattgccc tgagcctggg
2101 cctgaccccc aacttcaaga gcaacttcga cctggccgag gatgccaaac tgcagctgag
2161 caaggacacc tacgacgacg acctggacaa cctgctggcc cagatcggcg accagtacgc
2221 cgacctgttt ctggccgcca agaacctgtc cgacgccatc ctgctgagcg acatcctgag
2281 agtgaacacc gagatcacca aggcccccct gagcgcctct atgatcaaga gatacgacga
2341 gcaccaccag gacctgaccc tgctgaaagc tctcgtgcgg cagcagctgc ctgagaagta
2401 caaagagatt ttcttcgacc agagcaagaa cggctacgcc ggctacattg acggcggagc
2461 cagccaggaa gagttctaca agttcatcaa gcccatcctg gaaaagatgg acggcaccga
2521 ggaactgctc gtgaagctga acagagagga cctgctgcgg aagcagcgga ccttcgacaa
2581 cggcagcatc ccccaccaga tccacctggg agagctgcac gccattctgc ggcggcagga
2641 agatttttac ccattcctga aggacaaccg ggaaaagatc gagaagatcc tgaccttccg
2701 catcccctac tacgtgggcc ctctggccag gggaaacagc agattcgcct ggatgaccag
2761 aaagagcgag gaaaccatca ccccctggaa cttcgaggaa gtggtggaca agggcgcttc
2821 cgcccagagc ttcatcgagc ggatgaccaa cttcgataag aacctgccca acgagaaggt
2881 gctgcccaag cacagcctgc tgtacgagta cttcaccgtg tataacgagc tgaccaaagt
2941 gaaatacgtg accgagggaa tgagaaagcc cgccttcctg agcggcgagc agaaaaaggc
3001 catcgtggac ctgctgttca agaccaaccg gaaagtgacc gtgaagcagc tgaaagagga
3061 ctacttcaag aaaatcgagt gcttcgactc cgtggaaatc tccggcgtgg aagatcggtt
3121 caacgcctcc ctgggcacat accacgatct gctgaaaatt atcaaggaca aggacttcct
3181 ggacaatgag gaaaacgagg acattctgga agatatcgtg ctgaccctga cactgtttga
3241 ggacagagag atgatcgagg aacggctgaa aacctatgcc cacctgttcg acgacaaagt
3301 gatgaagcag ctgaagcggc ggagatacac cggctggggc aggctgagcc ggaagctgat
3361 caacggcatc cgggacaagc agtccggcaa gacaatcctg gatttcctga agtccgacgg
3421 cttcgccaac agaaacttca tgcagctgat ccacgacgac agcctgacct ttaaagagga
3481 catccagaaa gcccaggtgt ccggccaggg cgatagcctg cacgagcaca ttgccaatct
3541 ggccggcagc cccgccatta agaagggcat cctgcagaca gtgaaggtgg tggacgagct
3601 cgtgaaagtg atgggccggc acaagcccga gaacatcgtg atcgaaatgg ccagagagaa
3661 ccagaccacc cagaagggac agaagaacag ccgcgagaga atgaagcgga tcgaagaggg
3721 catcaaagag ctgggcagcc agatcctgaa agaacacccc gtggaaaaca cccagctgca
3781 gaacgagaag ctgtacctgt actacctgca gaatgggcgg gatatgtacg tggaccagga
3841 actggacatc aaccggctgt ccgactacga tgtggaccat atcgtgcctc agagctttct
3901 gaaggacgac tccatcgaca acaaggtgct gaccagaagc gacaagaacc ggggcaagag
3961 cgacaacgtg ccctccgaag aggtcgtgaa gaagatgaag aactactggc ggcagctgct
4021 gaacgccaag ctgattaccc agagaaagtt cgacaatctg accaaggccg agagaggcgg
4081 cctgagcgaa ctggataagg ccggcttcat caagagacag ctggtggaaa cccggcagat
4141 cacaaagcac gtggcacaga tcctggactc ccggatgaac actaagtacg acgagaatga
4201 caagctgatc cgggaagtga aagtgatcac cctgaagtcc aagctggtgt ccgatttccg
4261 gaaggatttc cagttttaca aagtgcgcga gatcaacaac taccaccacg cccacgacgc
4321 ctacctgaac gccgtcgtgg gaaccgccct gatcaaaaag taccctaagc tggaaagcga
4381 gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag atgatcgcca agagcgagca
4441 ggaaatcggc aaggctaccg ccaagtactt cttctacagc aacatcatga actttttcaa
4501 gaccgagatt accctggcca acggcgagat ccggaagcgg cctctgatcg agacaaacgg
4561 cgaaaccggg gagatcgtgt gggataaggg ccgggatttt gccaccgtgc ggaaagtgct
4621 gagcatgccc caagtgaata tcgtgaaaaa gaccgaggtg cagacaggcg gcttcagcaa
4681 agagtctatc ctgcccaaga ggaacagcga taagctgatc gccagaaaga aggactggga
4741 ccctaagaag tacggcggct tcgacagccc caccgtggcc tattctgtgc tggtggtggc
4801 caaagtggaa aagggcaagt ccaagaaact gaagagtgtg aaagagctgc tggggatcac
4861 catcatggaa agaagcagct tcgagaagaa tcccatcgac tttctggaag ccaagggcta
4921 caaagaagtg aaaaaggacc tgatcatcaa gctgcctaag tactccctgt tcgagctgga
4981 aaacggccgg aagagaatgc tggcctctgc cggcgaactg cagaagggaa acgaactggc
5041 cctgccctcc aaatatgtga acttcctgta cctggccagc cactatgaga agctgaaggg
5101 ctcccccgag gataatgagc agaaacagct gtttgtggaa cagcacaagc actacctgga
5161 cgagatcatc gagcagatca gcgagttctc caagagagtg atcctggccg acgctaatct
5221 ggacaaagtg ctgtccgcct acaacaagca ccgggataag cccatcagag agcaggccga
5281 gaatatcatc cacctgttta ccctgaccaa tctgggagcc cctgccgcct tcaagtactt
5341 tgacaccacc atcgaccgga agaggtacac cagcaccaaa gaggtgctgg acgccaccct
5401 gatccaccag agcatcaccg gcctgtacga gacacggatc gacctgtctc agctgggagg
5461 cgacaaaagg ccggcggcca cgaaaaaggc cggccaggca aaaaagaaaa agtaagaatt
5521 cctagagctc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
5581 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
5641 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
5701 gggcaggaca gcaaggggga ggattgggaa gagaatagca ggcatgctgg ggagcggccg
5761 caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
5821 gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag
5881 cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc
5941 ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa
6001 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccttagcgc
6061 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
6121 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
6181 aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
6241 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
6301 cactcaactc tatctcgggc tattcttttg atttataagg gattttgccg atttcggtct
6361 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
6421 cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
6481 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
6541 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
6601 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
6661 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
6721 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
6781 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
6841 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
6901 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
6961 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
7021 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
7081 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
7141 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
7201 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
7261 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
7321 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
7381 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
7441 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
7501 ttattgctga taaatctgga gccggtgagc gtggaagccg cggtatcatt gcagcactgg
7561 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
7621 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
7681 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
7741 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
7801 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
7861 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
7921 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
7981 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
8041 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
8101 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
8161 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
8221 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
8281 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
8341 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
8401 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
8461 tacggttcct ggccttttgc tggccttttg ctcacatgt
//
| {
"Vector backbone": "pX330",
"Backbone size w/o insert (bp)": "",
"Vector type": "CRISPR",
"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": "DLX4",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "DLX4",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "BbsI (destroyed during cloning)",
"3′ cloning site": "BbsI (destroyed during cloning)",
"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": ""
} |
|
pBZS-YP_009665506 | 198376 | Express YP_009665506 in bacterial cells. | Takashi Ito | Miura et al BMC Biotechnol. 2022 Nov 4;22(1):33. doi: 10.1186/s12896-022-00765-3. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/390269/8a345400-7a50-4ba1-916b-fe4000250ec6/addgene-plasmid-198376-sequence-390269.gbk | full | LOCUS sequence-390269- 5507 bp DNA circular SYN 27-AUG-2024
DEFINITION Express YP_009665506 in bacterial cells..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5507)
AUTHORS Miura F, Miura M, Shibata Y, Furuta Y, Miyamura K, Ino Y, Bayoumi
AMA, Oba U, Ito T
TITLE Identification, expression, and purification of DNA cytosine
5-methyltransferases with short recognition sequences.
JOURNAL BMC Biotechnol. 2022 Nov 4;22(1):33. doi:
10.1186/s12896-022-00765-3.
PUBMED 36333700
REFERENCE 2 (bases 1 to 5507)
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: "BMC
Biotechnol."; date: "2022-11-4"; volume: "22(1):33. doi"; pages: "
10.1186/s12896-022-00765-3"
FEATURES Location/Qualifiers
source 1..5507
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 1..285
/gene="araBAD"
/label=araBAD promoter
/note="promoter of the L-arabinose operon of E. coli; the
araC regulatory gene is transcribed in the opposite
direction (Guzman et al., 1995)"
primer_bind 208..227
/label=pBAD Forward
/note="For vectors with E. coli araBAD promoter, forward
primer"
CDS 331..348
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 352..384
/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 388..411
/codon_start=1
/product="Xpress(TM) epitope tag, including an enterokinase
recognition and cleavage site"
/label=Xpress(TM) tag
/translation="DLYDDDDK"
CDS 1846..1869
/codon_start=1
/product="peptide that binds Strep-Tactin(R), an engineered
form of streptavidin"
/label=Strep-Tag II
/translation="WSHPQFEK"
primer_bind complement(1908..1925)
/label=pBAD Reverse
/note="For vectors with E. coli araBAD promoter, reverse
primer"
terminator 2078..2164
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 2256..2283
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
promoter 2302..2393
/gene="bla"
/label=AmpR promoter
CDS 2394..3254
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(2612..2631)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 3425..4013
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3914..3933
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 4167..4184
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 4199..4339
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(4265..4284)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 4425..4447
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(4603..5481)
/codon_start=1
/gene="araC"
/product="L-arabinose regulatory protein"
/label=araC
/translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHFFRQQLGIS
VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
EKVNDVAVKLS"
ORIGIN
1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagccg
241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg ttttttgggc
301 taacaggagg aattaaccat ggggggttct catcatcatc atcatcatgg tatggctagc
361 atgactggtg gacagcaaat gggtcgggat ctgtacgacg atgacgataa gctcgaggga
421 tccatggttc tgcgtgcgct ggacctgttt agcggtatcg gtggcattac ccacggcctg
481 cgtgagatcg ttgaaccgat tgcgttcgtg gagaaaaacg acgaagcgcg tagctttctg
541 aagaaaaagt atccggaaat tccggttttc gacgatgtgt gcagctttga tgcgaccaaa
601 tggatcgaca agctggatat cattctggcg ggttggccgt gcaccggttt cagcaacgcg
661 ggtaccaaaa ccggcttcag ccacgaggcg agcggtctgt ttaccgaagt tatccgtatt
721 gtgaaagagt gccgtccgaa gtatgttttc ctggaaaaca gccacaccct gagcctgttt
781 gagaacgtga acgtggttgt gaaggcgttc gacgagctgg gttacgattg ccgttggatt
841 acctgccgtg cgacctgcgt tggtgcgctg caccagcgtc accgttggtt ttgcctggtt
901 gtgcgtcgtg acattgaact ggatatcggt attccgtacg tggaacgttt cgactggatg
961 accaacgagc cgccgcgtca gatccagaag agcaacaagg agaacatcct gcacattagc
1021 ctggcgggta acgcggttgt gccggatcaa gttcgttatg cgatggacat cctgtgcagc
1081 ttcgatacca acaaggaatt tagcctgagc agccgtgcga aaaacggttt cagcaagaac
1141 ggcgtgatct ttacctacga cattgagcac ccggcgcgtg aaccgctgaa catcgttctg
1201 accccgcgtg agaacgaaac cagctttgtg attagctgcg atccgaaaaa ggttctgacc
1261 aaaagcgtga tccagaagtt ctggccgacc ccgctgcaca gctgcaaata tgcgaccaag
1321 tgcccgcgta ccctgaccac ccgtgttagc aaaatgctga gcgcgtgcgt gggtttcagc
1381 gagggtggcg acaaaaacgg ctacctgagc gcgaagtggg tgctgtggct gatgggctac
1441 gattataagt acatgaccag cgaattccag catgatgaag cagtggataa caagtttaac
1501 aaagaacagc agaacgcgtt ttatgagatc ttgcacttac cgaatctgaa cgaggaacag
1561 cgcaatgcgt tcattcagtc ccttaaagac gatccatctc aaagcgcaaa tctgctggca
1621 gaagccaaga aactgaacga tgctcaagct ccgaaagtcg acaacaaatt caacaaagag
1681 caacagaatg cgttctacga aattctgcat ttaccgaacc tgaatgaaga acagcgtaac
1741 gcctttatcc aatcgctgaa agatgatcct agtcagtcag ccaatctgtt ggcggaagcg
1801 aagaaactca atgacgccca agctcccaaa gttgacgcga atagctggag ccatccgcag
1861 tttgaaaagc tttagggctg ttttggcgga tgagagaaga ttttcagcct gatacagatt
1921 aaatcagaac gcagaagcgg tctgataaaa cagaatttgc ctggcggcag tagcgcggtg
1981 gtcccacctg accccatgcc gaactcagaa gtgaaacgcc gtagcgccga tggtagtgtg
2041 gggtctcccc atgcgagagt agggaactgc caggcatcaa ataaaacgaa aggctcagtc
2101 gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac
2161 aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg
2221 acgcccgcca taaactgcca ggcatcaaat taagcagaag gccatcctga cggatggcct
2281 ttttgcgttt ctacaaactc ttttgtttat ttttctaaat acattcaaat atgtatccgc
2341 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
2401 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
2461 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
2521 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
2581 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg
2641 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
2701 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
2761 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
2821 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
2881 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
2941 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
3001 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
3061 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
3121 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
3181 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
3241 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
3301 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
3361 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
3421 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
3481 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
3541 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
3601 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
3661 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
3721 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
3781 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
3841 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
3901 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
3961 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
4021 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
4081 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
4141 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
4201 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc
4261 tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg
4321 tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
4381 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
4441 tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag cagatcaatt cgcgcgcgaa
4501 ggcgaagcgg catgcataat gtgcctgtca aatggacgaa gcagggattc tgcaaaccct
4561 atgctactcc gtcaagccgt caattgtctg attcgttacc aattatgaca acttgacggc
4621 tacatcattc actttttctt cacaaccggc acggaactcg ctcgggctgg ccccggtgca
4681 ttttttaaat acccgcgaga aatagagttg atcgtcaaaa ccaacattgc gaccgacggt
4741 ggcgataggc atccgggtgg tgctcaaaag cagcttcgcc tggctgatac gttggtcctc
4801 gcgccagctt aagacgctaa tccctaactg ctggcggaaa aaatgtgaca gacgcgacgg
4861 cgacaagcaa acatgctgtg cgacgctggc gatatcaaaa ttgctgtctg ccaggtgatc
4921 gctgatgtac tgacaagcct cgcgtacccg attatccatc ggtggatgga gcgactcgtt
4981 aatcgcttcc atgcgccgca gtaacaattg ctcaagcaga tttatcgcca gcagctccga
5041 atagcgccct tccccttgcc cggcgttaat gatttgccca aacaggtcgc tgaaatgcgg
5101 ctggtgcgct tcatccgggc gaaagaaccc cgtattggca aatattgacg gccagttaag
5161 ccattcatgc cagtaggcgc gcggacgaaa gtaaacccac tggtgatacc attcgcgagc
5221 ctccggatga cgaccgtagt gatgaatctc tcctggcggg aacagcaaaa tatcacccgg
5281 tcggcaaaca aattctcgtc cctgattttt caccaccccc tgaccgcgaa tggtgagatt
5341 gagaatataa cctttcattc ccagcggtcg gtcgataaaa aaatcgagat aaccgttggc
5401 ctcaatcggc gttaaacccg ccaccagatg ggcattaaac gagtatcccg gcagcagggg
5461 atcattttgc gcttcagcca tacttttcat actcccgcca ttcagag
//
| {
"Vector backbone": "pBZS",
"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)": "NEB T7 Express",
"Copy number": "Unknown"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Identification, expression, and purification of DNA cytosine 5-methyltransferases with short recognition sequences",
"Publication": "BMC Biotechnol. 2022 Nov 4;22(1):33. doi: 10.1186/s12896-022-00765-3.",
"DOI": "10.1186/s12896-022-00765-3.",
"Authors": "Miura F, Miura M, Shibata Y, Furuta Y, Miyamura K, Ino Y, Bayoumi AMA, Oba U, Ito T /s---",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/36333700",
"PubMed ID": "PubMed 36333700"
} | {
"Gene/Insert name": "YP_009665506",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "NYs-1_727R",
"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": ""
} |
|
pLPC-Flag-CHES1L-dC-NLS | 51840 | Flag-CHES1L mutant lacking the C-terminal, with remaining NLS | Gerardo Ferbeyre | Huot et al Mol Biol Cell. 2014 Mar;25(5):554-65. doi: 10.1091/mbc.E13-02-0110. Epub 2014 Jan 8. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/91891/48ce3b39-9303-437e-a842-7ab6d85b0d52/addgene-plasmid-51840-sequence-91891.gbk | partial | LOCUS sequence-91891-e 591 bp DNA linear UNA 26-AUG-2024
DEFINITION Flag-CHES1L mutant lacking the C-terminal, with remaining NLS.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE natural DNA sequence
ORGANISM unspecified
REFERENCE 1 (bases 1 to 591)
AUTHORS Huot G, Vernier M, Bourdeau V, Doucet L, Saint-Germain E,
Gaumont-Leclerc MF, Moro A, Ferbeyre G
TITLE CHES1/FOXN3 regulates cell proliferation by repressing PIM2 and
protein biosynthesis.
JOURNAL Mol Biol Cell. 2014 Mar;25(5):554-65. doi: 10.1091/mbc.E13-02-0110.
Epub 2014 Jan 8.
PUBMED 24403608
REFERENCE 2 (bases 1 to 591)
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:
"10.1091/mbc.E13-02-0110"; journalName: "Mol Biol Cell"; date:
"2014-03"; volume: "25"; issue: "5"; pages: "554-65"
FEATURES Location/Qualifiers
source 1..591
/mol_type="genomic DNA"
/organism="unspecified"
primer_bind 41..60
/label=T7
/note="T7 promoter, forward primer"
promoter 41..59
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 128..151
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
ORIGIN
1 ggctaactag agaacccact gcttactggc ttatcgaaat taatacgact cactataggg
61 agacccaagc ttggtaccga gctcggatcc actagtaacg gccgccagtg tgctggaatt
121 cgccatggac tacaaagacg atgacgacaa gggcatgggt ccagtcatgc ctcccagtaa
181 gaagccagaa agctcaggaa ttagtgtctc cagtggactg agtcagtgtt acgggggcag
241 cggtttctcc aaggcccttc aggaagacga tgacctcgac ttttctctgc ctgacatccg
301 attagaagag ggggccatgg aagatgaaga gctgaccaac ctgaactggc tgcacgagag
361 caagaacttg ctgaagagct ttggggagtc ggtcctcagg agtgtcagcc ccgtccagga
421 cctggacgat gacacccccc catcccctgc ccactctgac atgccctacg atgccaggca
481 gaaccccaac tgcaaacccc cctactcctt cagctgcctc atatttatgg ccatcgagga
541 ctctccaacc aagcgcctgc cagtgaagga tatctacaac tggatcttgg a
//
| {
"Vector backbone": "pLPC",
"Backbone size w/o insert (bp)": "",
"Vector type": "Retroviral",
"Selectable markers": "Puromycin"
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "Top10",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "CHES1/FOXN3 regulates cell proliferation by repressing PIM2 and protein biosynthesis",
"Publication": "Mol Biol Cell. 2014 Mar;25(5):554-65. doi: 10.1091/mbc.E13-02-0110. Epub 2014 Jan 8.",
"DOI": "10.1091/mbc.E13-02-0110.",
"Authors": "Huot G, Vernier M, Bourdeau V, Doucet L, Saint-Germain E, Gaumont-Leclerc MF, Moro A, Ferbeyre G /mbcE--",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/24403608",
"PubMed ID": "PubMed 24403608"
} | {
"Gene/Insert name": "CHES1",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "contains CHES1 amino acids 1-205",
"Entrez Gene": "FOXN3",
"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": ""
} |
|
UBCi | 25599 | Bacterial expression for structure determination; may not be full ORF | Cheryl Arrowsmith | Structural Genomics Consortium Clones (unpublished) | https://media.addgene.org/snapgene-media/v2.0.0/sequences/31585/f08436c9-6b42-4ef8-a895-38b9d6a00c6a/addgene-plasmid-25599-sequence-31585.gbk | partial | LOCUS sequence-31585-e 895 bp DNA linear UNA 26-AUG-2024
DEFINITION Bacterial expression for structure determination; may not be full
ORF.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE natural DNA sequence
ORGANISM unspecified
REFERENCE 1 (bases 1 to 895)
TITLE Structural Genomics Consortium Clones
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"
FEATURES Location/Qualifiers
source 1..895
/mol_type="genomic DNA"
/organism="unspecified"
RBS 11..33
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 52..69
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 79..96
/codon_start=1
/product="thrombin recognition and cleavage site"
/label=thrombin site
/translation="LVPRGS"
CDS 627..644
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(697..715)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 711..758
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
ORIGIN
1 tagaaataat tttgtttaac tttaagaagg agatatacca tgggcagcag ccatcatcat
61 catcatcaca gcagcggcct ggttccgcgt ggtagtggtg cagtgtctgg gtcagtgcaa
121 gcttcagata gacttatgaa agagctcagg gacatataca gatcacagag ttataaaaca
181 gggatttatt cagtggaact cataaatgac agtttatatg actggcatgt taaactgcag
241 aaggttgacc ctgatagtcc tttgcacagt gatcttcaga tcttaaaaga aaaagaaggc
301 atagaatata ttttgcttaa cttctctttt aaggataact ttccatttga tcctccattt
361 gttcgagtgg tgttacctgt tctctcagga gggtatgtat tgggtggagg agcattatgt
421 atggaacttc tcacaaaaca gggctggagc agtgcctact caatagaatc ggtcatcatg
481 caaataaatg ccaccttagt caaaggcaaa gccagagtgc agtttggagc aaataagaat
541 caatataatc tagcaagagc ccaacaatcc tataattcca ttgtacagat acatgagtga
601 tgacgaagct tgcggccgca ctcgagcacc accaccacca ccactgagat ccggctgcta
661 acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac
721 cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg
781 gattggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
841 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttc
//
| {
"Vector backbone": "pET28a-LIC",
"Backbone size w/o insert (bp)": "7328",
"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": "Industry MTA"
} | {
"Title": "",
"Publication": "",
"DOI": "",
"Authors": "",
"PubMed Link": "",
"PubMed ID": ""
} | {
"Gene/Insert name": "UBCi",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "501",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": "His (N terminal on insert)"
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "T7",
"3′ sequencing primer": "T7-term"
} | {
"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": ""
} |
|
CEPR2 X048_pECIA14 | 114899 | Prey vector CEPR2 X048_pECIA14 should be used with bait vector CEPR2 X048_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/276502/894494ed-be28-47e0-9b54-b32896920700/addgene-plasmid-114899-sequence-276502.gbk | full | LOCUS sequence-276502- 6983 bp DNA circular SYN 26-AUG-2024
DEFINITION Prey vector CEPR2 X048_pECIA14 should be used with bait vector CEPR2
X048_pECIA2..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6983)
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 6983)
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..6983
/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 2279..2302
/codon_start=1
/product="recognition and cleavage site for human
rhinovirus 3C and PreScission proteases"
/label=HRV 3C site
/translation="LEVLFQGP"
CDS 3977..4000
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS 4016..4033
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(4056..4073)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 4106..4240
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4130..4149)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 4184..4203
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind complement(4384..4400)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4384..4400)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4397..4419)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4408..4424
/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(4432..4462)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4477..4498
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4615..4632)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4786..5374)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4866..4885)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5545..6405)
/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 6168..6187
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6406..6510)
/gene="bla"
/label=AmpR promoter
primer_bind 6578..6596
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(6634..6656)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 6756..6775
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind join(6969..6983,1..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 tcgaccgttg agaaacaagc
541 tctgtttcgc ttcaaaaacc gcctcgacga ctctcacaac attctacaat cttggaagcc
601 ttccgattca ccatgcgtgt ttcgcggcat cacatgcgac ccgttatccg gcgaagtcat
661 cggtatttct ctaggaaacg taaatctctc cggcaccatt tctccatcaa tctcagctct
721 cacaaagctc tctacgctat cgcttccatc taacttcatc tccggaagaa tcccgccaga
781 gatagtcaac tgcaaaaacc ttaaagttct caatctcacc tccaacagac tctccggaac
841 aatcccaaac ctttctcctc taaagtccct agaaattctc gacatctccg ggaattttct
901 taacggagag tttcaaagct ggattgggaa tatgaatcag ttggtttctc tcggtcttgg
961 caacaatcac tacgaagaag gtataattcc tgagagtatt ggaggtttaa agaaacttac
1021 ttggctgttc ttagctcgct ccaacttgac cgggaagatt ccaaactcga tctttgatct
1081 gaatgctctc gacactttcg acatcgcgaa caacgcaatc tccgatgatt tcccaatctt
1141 aatctcaaga ttggtaaatc tcaccaagat tgagttgttc aacaacagct taacaggtaa
1201 aatccctccg gagataaaga acttgactcg tttacgcgaa ttcgacatct cttcgaatca
1261 gttgagcggg gttttacctg aagaactcgg agttttgaag gagctcagag ttttccactg
1321 tcatgaaaac aacttcaccg gcgaatttcc ttctggtttc ggcgatctaa gtcacctcac
1381 ttctctttcg atttatagaa acaacttctc cggtgaattt ccggttaaca tcggccggtt
1441 ctcgccgctc gacacggttg atatatctga aaacgagttt actggtccat ttccaagatt
1501 cttatgccaa aacaagaaac tacaattctt gctcgccttg cagaacgagt tttccggcga
1561 gattccgaga tcttacggcg agtgcaaatc tctattgaga ctaaggatta acaacaaccg
1621 tctaagtggt caagttgttg aaggattctg gtctctaccg cttgctaaga tgattgatct
1681 cagcgataac gaactcaccg gagaggtttc tcctcagatt ggactttcaa cagaactgag
1741 tcagctcatt ttacagaaca acagattttc cggcaagatt cctcgtgagc tcggaagatt
1801 gactaatata gagaggattt atctgagcaa caacaatctt tccggcgaaa ttccgatgga
1861 agttggagat ttgaaagagt tatcatctct gcatcttgaa aacaattcat taacaggatt
1921 tatccctaag gaattgaaga attgtgtcaa acttgttgat ctgaatctcg ctaagaactt
1981 cttgaccgga gaaatcccta acagtttatc tcagattgct tctttaaact cgttggattt
2041 ctcagggaat aggctaacag gtgaaatccc ggcgagtctt gtgaagttga agctgagttt
2101 cattgacttg tctggaaatc aactttcagg aagaatacca ccggatcttt tagcggtggg
2161 aggatccacc gcgttctcac gtaacgagaa gctctgtgtt gataaagaga acgctaaaac
2221 aaatcagaac ttggggttga gtatttgcag tggctaccaa aacgtgaaga ggaatagtct
2281 ggaggtgctg ttccagggac caagcggcaa tcccggcggc gatctggccc cccagatgct
2341 gcgcgagctg caggagacca atgccgccct gcaggacgtg cgcgagctgc tgcgccaccg
2401 cgtgaaggag atcaccttcc tgaagaacac cgtgatggag tgcgacgcct gcggcatgca
2461 gccctctaga ggctccggct ccatcatccc agttgaggag gagaacccgg acttctggaa
2521 ccgcgaggca gccgaggccc tgggtgccgc caagaagctg cagcctgcac agacagccgc
2581 caagaacctc atcatcttcc tgggcgatgg gatgggggtg tctacggtga cagctgccag
2641 gatcctaaaa gggcagaaga aggacaaact ggggcctgag atacccctgg ccatggaccg
2701 cttcccatat gtggctctgt ccaagacata caatgtagac aaacatgtgc cagacagtgg
2761 agccacagcc acggcctacc tgtgcggggt caagggcaac ttccagacca ttggcttgag
2821 tgcagccgcc cgctttaacc agtgcaacac gacacgcggc aacgaggtca tctccgtgat
2881 gaatcgggcc aagaaagcag ggaagtcagt gggagtggta accaccacac gagtgcagca
2941 cgcctcgcca gccggcacct acgcccacac ggtgaaccgc aactggtact cggacgccga
3001 cgtgcctgcc tcggcccgcc aggaggggtg ccaggacatc gctacgcagc tcatctccaa
3061 catggacatt gacgtgatcc taggtggagg ccgaaagtac atgtttcgca tgggaacccc
3121 agaccctgag tacccagatg actacagcca aggtgggacc aggctggacg ggaagaatct
3181 ggtgcaggaa tggctggcga agcgccaggg tgcccggtat gtgtggaacc gcactgagct
3241 catgcaggct tccctggacc cgtctgtgac ccatctcatg ggtctctttg agcctggaga
3301 catgaaatac gagatccacc gagactccac actggacccc tccctgatgg agatgacaga
3361 ggctgccctg cgcctgctga gcaggaaccc ccgcggcttc ttcctcttcg tggagggtgg
3421 tcgcatcgac catggtcatc atgaaagcag ggcttaccgg gcactgactg agacgatcat
3481 gttcgacgac gccattgaga gggcgggcca gctcaccagc gaggaggaca cgctgagcct
3541 cgtcactgcc gaccactccc acgtcttctc cttcggaggc taccccctgc gagggagctc
3601 catcttcggg ctggcccctg gcaaggcccg ggacaggaag gcctacacgg tcctcctata
3661 cggaaacggt ccaggctatg tgctcaagga cggcgcccgg ccggatgtta ccgagagcga
3721 gagcgggagc cccgagtatc ggcagcagtc agcagtgccc ctggacgaag agacccacgc
3781 aggcgaggac gtggcggtgt tcgcgcgcgg cccgcaggcg cacctggttc acggcgtgca
3841 ggagcagacc ttcatagcgc acgtcatggc cttcgccgcc tgcctggagc cctacaccgc
3901 ctgcgacctg gcgccccccg ccggcaccac cgacgccgcg cacccgggtt ctagaggaag
3961 cacctccggc gataacgatt acaaggatga tgatgataag gccagcggca ccggtcatca
4021 tcaccatcac cattgagttt aaacccgctg atcagcctcg actgtgcctt ctaaggcctg
4081 agctcgctga tcagcctcga tcgaggatcc agacatgata agatacattg atgagtttgg
4141 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
4201 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
4261 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
4321 caaatgtggt atggctgatt atgatcagtc gacctgcagg catgcaagct tggcgtaatc
4381 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
4441 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
4501 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
4561 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
4621 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
4681 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
4741 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
4801 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
4861 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
4921 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
4981 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
5041 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
5101 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
5161 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
5221 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
5281 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
5341 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
5401 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
5461 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
5521 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
5581 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
5641 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
5701 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
5761 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
5821 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
5881 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
5941 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
6001 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
6061 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
6121 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
6181 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
6241 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
6301 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
6361 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
6421 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
6481 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
6541 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
6601 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
6661 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
6721 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
6781 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
6841 gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
6901 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
6961 agggttttcc cagtcacgac gtt
//
| {
"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": "AT1G72180",
"Species": "A. thaliana (mustard weed)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "AT1G72180",
"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": ""
} |
|
T. aq RNA Polymerase | 215349 | Express T. aq RNA Polymerase subunits in E. coli cells | Julius Lucks | Hertz et al Nucleic Acids Res. 2024 May 8;52(8):4466-4482. doi: 10.1093/nar/gkae231. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/429510/d4fa703c-7e6a-4755-88bd-4bf183c8801c/addgene-plasmid-215349-sequence-429510.gbk | full | LOCUS sequence-429510- 14900 bp DNA circular SYN 27-AUG-2024
DEFINITION Express T. aq RNA Polymerase subunits in E. coli cells.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 14900)
AUTHORS Hertz LM, White EN, Kuznedelov K, Cheng L, Yu AM, Kakkaramadam R,
Severinov K, Chen A, Lucks JB
TITLE The effect of pseudoknot base pairing on cotranscriptional
structural switching of the fluoride riboswitch.
JOURNAL Nucleic Acids Res. 2024 May 8;52(8):4466-4482. doi:
10.1093/nar/gkae231.
PUBMED 38567721
REFERENCE 2 (bases 1 to 14900)
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"; doi: "10"; journalName:
"Nucleic Acids Res"; date: "2024-05-8- 8"; volume: "52"; issue: "8";
pages: "4466-4482"
FEATURES Location/Qualifiers
source 1..14900
/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"
CDS complement(560..1375)
/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(560..1375)
/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 1283..1302
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
primer_bind 1367..1386
/label=pENTR-R
/note="pENTR vectors, reverse primer"
rep_origin 1497..2085
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 1986..2005
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 2239..2256
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 2271..2413
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(2339..2358)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 2499..2521
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(2515..2706)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(3515..4597)
/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 4559..4578
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(4598..4675)
/gene="lacI"
/label=lacI promoter
primer_bind 4881..4900
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
promoter 4984..5002
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 5003..5027
/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)."
RBS 5042..5064
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
promoter 6070..6088
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 6089..6113
/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)."
RBS 6128..6150
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 9335..9346
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
promoter 9587..9605
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 9606..9630
/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)."
RBS 9645..9667
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 9686..9703
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 9713..9730
/codon_start=1
/product="thrombin recognition and cleavage site"
/label=thrombin site
/translation="LVPRGS"
promoter 14331..14349
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 14350..14374
/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)."
RBS 14389..14411
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 14744..14761
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(14814..14832)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 14828..14875
/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 aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat
601 tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa
661 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc
721 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga
781 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc
841 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac
901 cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac
961 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat
1021 tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag
1081 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca
1141 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac
1201 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg
1261 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca
1321 tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcataacac
1381 cccttgtatt actgtttatg taagcagaca gttttattgt tcatgaccaa aatcccttaa
1441 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
1501 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
1561 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
1621 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
1681 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
1741 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
1801 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
1861 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
1921 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
1981 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
2041 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
2101 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
2161 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
2221 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
2281 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatatggtgc actctcagta
2341 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg
2401 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
2461 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
2521 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc
2581 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag
2641 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt
2701 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa
2761 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg
2821 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg
2881 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc
2941 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta
3001 cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca
3061 gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc
3121 ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggggccgc
3181 catgccggcg ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa
3241 ggcttgagcg agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc
3301 gctccagcga aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac
3361 gagttgcatg ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca
3421 ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta
3481 atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
3541 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
3601 tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca
3661 ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa
3721 aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt
3781 atcccactac cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg
3841 cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca
3901 gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta
3961 tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg
4021 agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat
4081 gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct
4141 ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg
4201 catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat
4261 tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc
4321 tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca
4381 gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg
4441 ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt
4501 tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg
4561 catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct
4621 cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga
4681 tctcgacgct ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg
4741 ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc
4801 ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg
4861 cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg
4921 gcgccggtga tgccggccac gatgcgtccg gcgtagagga tcgagatctc gatcccgcga
4981 aattaatacg actcactata ggggaattgt gagcggataa caattcccct ctagaaataa
5041 ttttgtttaa ctttaagaag gagatatacc atgttggaga gcaagctgaa agccccggtc
5101 ttcacggcgc gcacccaggg gcgccactac ggcgagttcg tcctggagcc cctagaaagg
5161 gggtttgggg tcaccctggg gaaccctctc cgccgcatcc tcctctcctc catccccggg
5221 accgcggtca ccagcgtcta cattgaggac gtcctccacg agttctccac cattcccggg
5281 gtgaaggagg acgtggtgga gatcatcctg aacctgaagg agctggtggt gcgcttcctg
5341 gaccccaaga tggcgtccac caccctgatc ctcagggcgg agggccccaa ggaggtgagg
5401 gcgggcgact tcaccccctc cgccgacgtg gagatcatga accccgacct ccacatcgcc
5461 accctcgagg agggcgggaa gctctacatg gaggtgcggg tggaccgggg ggtggggtac
5521 gtgccggccg agcgccacgg catcaaggac cgcatcaacg ccatccctgt ggatgccatc
5581 ttctccccgg tgcgccgggt ggccttccag gtggaggaca cccgcctggg ccagcgcacg
5641 gacctggaca agctcacctt gaggatctgg accgacggct ccgtcacccc cctcgaggcc
5701 ctgaaccagg ccgtggccat cctgaaggag cacctgaact acttcgccaa ccccgaggcc
5761 tcccttctgc ccacccccga ggtgtcgaag ggggagaagc gggagagcgc cgaggaggac
5821 ctggacctgc ccttggagga gcttgggctt tccacccggg tcctccacag cctcaaggag
5881 gaggggattg agtccgtgcg cgcccttctg gccctcaacc tcaaggacct gaggaacatc
5941 cccggcatcg gggagcggag cctggaggag atccgccagg ccctggccaa gaagggcttc
6001 accctaaagg agtgaattat ggcggaaccg ggcattgaca agctcttcgg catctcgatc
6061 ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag
6121 aaataatttt gtttaacttt aagaaggaga tatacatatg gagattaagc ggttcggtcg
6181 catccgagag gtcattcccc ttccccccct tacggaaatc caggtggagt cctataagaa
6241 ggcgctccag gcggacgttc cccctgaaaa gcgggagaac gtgggcatcc aggccgcctt
6301 caaggagacc ttccctatag aggaggggga caagggcaag ggtggtctgg ttctggactt
6361 cctggagtac cgcatcggcg accccccctt ctcccaggac gagtgccggg agaaggacct
6421 cacctaccag gctccccttt acgcccggct ccagctcatc cacaaggaca ccgggctcat
6481 caaggaggac gaggtcttcc tgggccacct gcccctgatg accgaggacg gctccttcat
6541 catcaacggg gccgaccggg tcatcgtctc ccagatccac cgctcccccg gggtctactt
6601 cacccccgac cccgcccgcc ccggcaggta catcgccagc atcatccccc tgcccaagcg
6661 ggggccctgg attgacctcg aggtggaggc cagcggcgtg gtgaccatga aggtcaacaa
6721 gcgcaagttc cccctggtcc tcctcctcag ggtcctgggc tacgaccagg agaccctggt
6781 gcgggagctt tccgcctacg gggacctggt ccagggcctc ctggacgagg cggttttggc
6841 catgcgcccc gaggaggcca tggtccgcct gttcaccctc ctccgccccg gtgacccccc
6901 caagaaggac aaggccctgg cctacctctt tggccttctg gccgacccca agcgctacga
6961 cctgggagag gcgggccgct acaaggcaga ggagaagctg ggcgtgggcc tttcggggcg
7021 caccctggtc cggtttgagg acggcgagtt caaggacgag gtcttcctgc ccacgctccg
7081 ctacctcttc gccctaacgg ccggggttcc cggccacgag gtggacgaca ttgaccacct
7141 gggcaaccgg cgaatccgca ccgtgggcga gctcatggcg gaccagttcc gcgtgggcct
7201 ggccaggctg gcccgagggg tgcgggagcg catggtcatg ggctcccccg acaccctcac
7261 ccccgccaag ctggtcaaca gccgccccct ggaggccgcc ttgcgggagt ttttcagccg
7321 gagccagctc tcccagttca aggacgagac caaccccctc tcctccctga ggcacaaaag
7381 gcgcatttcc gccctgggcc ccgggggcct tacccgggag cgggcggggt ttgacgtgcg
7441 cgacgtgcac cgcacccact acgggcgcat ctgcccggtg gagacgccgg aaggggccaa
7501 catcggcctc atcacctccc tggccgccta cgcccgggtg gacgccttgg gcttcatccg
7561 caccccctac cgccgggtca agaacggcgt ggtcacggag gaggtggtct acatgaccgc
7621 ctccgaggag gaccgctaca ccatcgccca ggccaacacc cccctcgagg gcgaccgcat
7681 cgccaccgac cgggtggtgg ccaggcgcag gggggagccc gtcatcgtgg ctccggagga
7741 ggtggagttc atggacgtct cccccaagca ggtcttctct ctcaacacca acctcattcc
7801 cttcctggag cacgacgacg ccaaccgggc cctcatgggc tccaacatgc agacccaggc
7861 cgtgcccctc atccgggccc aggccccggt ggtcatgacg ggcctggagg agcgcgtggt
7921 gcgggactcc ctggccgccc tctacgccga ggaggacggg gaggtggtca aggtggacgg
7981 caccaggatc gccgtccgct atgaggacgg ccgcctggtg gagcaccctt tgcgccgcta
8041 tgcccgctcc aaccaaggca ccgccttcga ccagcggccc cgggtcaggg tgggccagcg
8101 ggtgaagaag ggggacctcc tggcggacgg ccccgcctcg gaagagggct ttctggccct
8161 ggggcagaac gtcctggtgg ccatcatgcc ctttgacggg tacaactttg aggacgccat
8221 cgtcatcagc gaggagctcc tcaagcggga cttctacacc tccatccaca ttgagcgcta
8281 cgagattgag gcccgggaca ccaagctggg ccccgagcgt attacccgtg acatccccca
8341 cctctccgag gccgcccttc gcgacctgga cgaggagggc atcgtccgca tcggggccga
8401 ggtcaagccc ggcgacatcc tggtggggcg caccagcttc aagggcgagc aggaaccttc
8461 ccccgaggag cgcctcctgc gctccatctt cggggagaag gccagggacg tgaaggacac
8521 ctccctgagg gtcccccccg gggaaggggg gatcgtggtg gggcgcctgc gccttaggcg
8581 gggcgatccc ggggtggagc tgaagcccgg ggtgcgggag gtggtccggg tcttcgtggc
8641 ccagaagcgc aagctccagg tgggggacaa gctggccaac cgccacggca acaagggcgt
8701 ggtggccaag atcctccccg tggaggacat gccccacctg cccgacggca cccccgtgga
8761 cgtgatcctg aaccccctgg gcgtgcccag ccggatgaac ctgggccaga tcctggagac
8821 ccacctgggc ctggcgggct acttcctggg ccagcgctac atctcccccg tctttgacgg
8881 ggccacggag cccgagatca aggagcttct ggccgaggcc ttcaacctct acttcggcaa
8941 gcgccagggc gagggcttcg gcgtggacaa gcgggagaag gaggtcctgg cccgggcgga
9001 gaagctgggc ctggtttccc cgggcaagag ccccgaggag caactcaaag agctcttcgt
9061 ccagggcaag gtggtcctct acgatggccg cacgggggag ccctttgagg ggcccatcgt
9121 ggtggggcag atgttcatca tgaagctcta ccacatggtg gaggacaaga tgcacgcccg
9181 ctccactggc ccctactccc tcatcaccca gcagcctctg ggcggcaagg cccagttcgg
9241 cggccagcgc ttcggggaga tggaggtgtg ggccctcgag gcctacgggg ccgcccacac
9301 cctgcaggag atgctgacca tcaagtccga cgacattgag ggcaggaacg ccgcctacga
9361 ggccatcatc aagggcgagg acgtgcccga gcccagcgtg cccgagtcct tccgcgtgct
9421 ggtgaaggag cttcaggccc tggccctgga cgtgcagacc ctggacgaga aggacaaccc
9481 cgtggacatc tttgaggggt tggcctccaa gcggtgaatt cggatcccgg tgatgccggc
9541 cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg cgaaattaat acgactcact
9601 ataggggaat tgtgagcgga taacaattcc cctctagaaa taattttgtt taactttaag
9661 aaggagatat accatgggca gcagccatca tcatcatcat cacagcagcg gcctggtgcc
9721 gcgcggcagc catatgaaaa aggaagtccg caaggtccgc atcgccctgg cctcccccga
9781 gaagatccgc tcctggagct acggggaggt ggagaagccc gagaccatca actaccgcac
9841 cctgaagccc gagcgggacg ggctctttga cgagcgcatc ttcggcccca tcaaggacta
9901 cgagtgcgcc tgcggcaagt acaagcgcca gcgctttgag ggcaaggtct gcgagcggtg
9961 cggggtggag gtgacgagga gcatcgtccg ccgctaccgc atgggccaca ttgagctggc
10021 cacccccgcc gcccacatct ggttcgtcaa ggacgtgccc tccaagatcg gcaccctcct
10081 ggacctctcc gccaccgagc tggagcaggt cctctacttc aacaagtaca tcgtcctgga
10141 ccccaagggg gccgtcctgg acggggtgcc ggtggaaaag cgccagctcc tcaccgacga
10201 ggagtaccgg gagctccgct acggcaagca ggagacctac cccctgcccg ccggggtgga
10261 tgccctggtc aaggacgggg aagaggtggt gaagggccag gagctggccc ccggcgtggt
10321 gagccgcatg gacggcgtgg ctctctaccg cttcccccgc cgggtgcggg tggactacct
10381 gaggaaggag cgggccgccc tcaggatccc cctttccgcc tgggtggaga aggaggccta
10441 ccggcccggg gaggtcctgg cggagctatc tgagccctac ctcttccggg ccgaggagag
10501 cggggtggtg gagctcaagg atctcgccga gggccacctc atctacctgc gccaggagga
10561 ggaggtggtg gcccgctact tcctccccgc gggcatgact cccctggtgg tggaggggga
10621 gatcgtggag gtgggccagc ctctggccga ggggaagggg ctattgcgcc tgccccgcca
10681 catgacggcc aaggaggtgg aggccgagga ggaaggggac agcgtccacc tcaccctgtt
10741 cctggagtgg accgagccca aggactacaa ggtggccccc cacatgaacg tcatcgtccc
10801 cgagggggcc aaggtgcagg cgggggagaa gatcgtggcc gccattgacc ccgaggagga
10861 ggtcatcgcc gaggccgagg gggtggtcca cctgcacgag cccgccagca tcctggtggt
10921 caaggcccgg gtctacccct ttgaggacga cgtggaggtg accaccgggg accgggtggc
10981 ccccggggac gtcctggccg acgggggcaa ggtcaagagc gagatctacg gccgggtgga
11041 ggtggacctg gtccgcaatg tggtgcgggt ggtggagtcc tacgacatag acgcccgcat
11101 gggggccgag gccatccagg agctcctcaa ggagctggac ctggagaagc tggagaggga
11161 gcttttggag gagatgaagc acccctcccg ggcccggcgg gccaaggcca ggaagcggct
11221 ggaggtggtg cgggccttcc tggactccgg caaccgcccg gagtggatga tcctggaggc
11281 cgtccccgtc ctgcccccgg acctccgccc catggtccag gtggacgggg ggcggttcgc
11341 caccagcgac ctcaacgacc tctaccgccg cctcatcaac cgcaacaacc gcttgaagaa
11401 gctcctggcc cagggggccc ccgagatcat catccgcaac gagaagcgca tgctccagga
11461 ggcggtggac gccgtcatag acaacggccg ccggggcagc cccgtcacca accccggctc
11521 cgagcgcccc ctccgcagcc tcaccgacat cctctccggc aagcagggcc gcttccgcca
11581 gaaccttctg ggcaaacggg tggactactc gggccggagc gtgatcgtgg tggggcccca
11641 gctcaagctc caccagtgcg gcctgcccaa gcgcatggcc ctggagctct tcaagccctt
11701 cctcctcaag aagatggagg agaagggcat cgcccccaac gtgaaggcgg ccaggcgcat
11761 gctggagcgc cagcgggaca tcaaggacga ggtgtgggac gccctcgagg aggtcatcca
11821 cggcaaggtg gtcctcctga accgggcccc taccctgcat cgcctaggga tccaggcctt
11881 ccagcccgtt ttggtggagg ggcagtccat ccagctccac cccctggtct gcgaggcctt
11941 caacgccgac tttgacgggg accagatggc cgtccacgtc cccctttcct ccttcgccca
12001 ggccgaggcc cggatccaga tgctctccgc ccacaacctt ctctccccgg cctcggggga
12061 gcccctggcc aagcccagcc gggacatcat cctgggcctt tactacatca cccaggtgcg
12121 aaaggagaag aagggagccg gcatggcctt cgctaccccc gaggaggctt tggcggctta
12181 cgaaaggggc gaggtggccc tgaacgcccc catcgtggtg gcgggcaggg agaccagcgt
12241 gggccgcttg aagttcgtct tcgccaaccc cgacgaggcc cttctggcgg tggcccacgg
12301 gcttttggac ctccaggacg tggtcaccgt ccgctacctg ggcaggcggc tggagacgag
12361 cccggggcgg atcctcttcg cccgcatcgt gggcgaggcc gtgggggacg aaaaggtggc
12421 ccaggagctc atccagatgg acgtccccca ggagaagaac tccctcaagg acctggtcta
12481 ccaggccttc ctgcgcctgg gcatggagaa gaccgcccgc ctcctggacg ccctcaagta
12541 ctacggtttc accctctcca ccaccagcgg catcaccatc ggcattgacg atgccgtgat
12601 ccccgaggag aagcagaggt acctggagga agccgaccgg aagctccgcc agattgagca
12661 ggcctacgag atgggcttcc tcaccgaccg ggagcggtac gaccaggtga tccagctttg
12721 gacggagacc acggagaagg tgacccaggc ggtcttcaag aactttgagg agaactaccc
12781 cttcaacccc ctctacgtca tggcccagtc cggggcccgg ggtaacccgc agcagatccg
12841 ccagctctgc ggcatgcggg gcctgatgca aaagccctcg ggggagacct ttgaggtgcc
12901 ggtgcgctcc tccttccgcg agggcctcac cgtcttggag tacttcatca gtagccacgg
12961 ggccaggaag ggcggggcgg acaccgcgct ccgcaccgcc gactccggct acctgacccg
13021 gaagctggtg gatgtggccc acgagatcgt ggtgcgggag gcggactgcg gcaccaccaa
13081 ctacatctcc gtccccctct tccagatgga cgaggtgacc cgcaccctgc gcctcaggaa
13141 gcgctcggac atagagtccg gcctctacgg ccgggtcctg gcccgggagg tggaggctct
13201 gggccgcagg ctggaggagg ggcgctacct ctccctcgag gacgtccact tcctcatcaa
13261 ggcggcggag gcgggggagg tgcgggaggt tcccgtgagg agccccctca cctgccagac
13321 ccgttacggg gtctgccaga agtgctacgg ctacgacctg tccatggccc gccccgtgtc
13381 catcggggag gccgtgggcg tagtggccgc cgagtccatc ggcgagcccg gcacccagct
13441 caccatgcgc accttccaca ccgggggcgt ggcggtgggc accgacatca cccagggtct
13501 tccccgggtc attgagctct ttgaggcccg ccgccccaag gccaaggcgg tcatctccga
13561 gattgacggc gtggtgcgca ttgaggaagg cgaggaccgg ctctcggtct tcgtggagtc
13621 cgagggcttc tccaaggagt acaagctccc caaggacgcc cgccttttgg tcaaggacgg
13681 ggactacgtg gaggcgggcc agcccctgac ccgcggggcc attgaccccc accagctttt
13741 ggaggccaag ggtcccgagg cggtggagcg ctacctggtg gacgagatcc agaaggtcta
13801 ccgggcccag ggcgtgaagc tccacgacaa gcacattgag atcgtggtgc ggcagatgct
13861 caagtacgtg gaggtcaccg accccgggga tagccgcctt ttggagggcc aggtcctgga
13921 gaagtgggac gtggaggccc tgaacgagcg cctgatcgcc gagggcaagg tgcccgtggc
13981 ctggaagccc ctcctcatgg gggtcaccaa gagcgccctc tccaccaaga gctggctctc
14041 cgccgccagc ttccagaaca ccacccacgt cctcactgag gcggccatcg ccggcaagaa
14101 ggacgagctc atcggcctca aggagaacgt gatcctgggt cgcctgatcc ctgcgggcac
14161 gggctccgac ttcgtccgct tcacccaggt ggtggaccag aggaccctga aggccattga
14221 ggaggcccgc aaggaggcgg tggaggccaa ggagaaggag gctccccgcc gccccgtgcg
14281 ccgggagcag ccggggaagg gcctttgaat tgatctcgat cccgcgaaat taatacgact
14341 cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt tgtttaactt
14401 taagaaggag atataccatg gcggaaccgg gcattgacaa gctcttcggc atggtggact
14461 ccaagtaccg gctcacggtg gtggtggcca aacgcgccca gcagcttctg cgccaccgct
14521 ttaagaacac ggttctggag ccggaggaga ggcccaagat gcgcaccctc gagggcctct
14581 acgacgaccc caacgccgtt acctgggcca tgaaggagct tttgacgggg cggcttttct
14641 tcggggagaa cctggtgccc gaggaccggc ttcagaagga gatggaaagg ctctacccca
14701 ccgaagagga ggcctagtcg acaagcttgc ggccgcactc gagcaccacc accaccacca
14761 ctgagatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga
14821 gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa
14881 aggaggaact atatccggat
//
| {
"Vector backbone": "pET28",
"Backbone size w/o insert (bp)": "",
"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",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "The effect of pseudoknot base pairing on cotranscriptional structural switching of the fluoride riboswitch",
"Publication": "Nucleic Acids Res. 2024 May 8;52(8):4466-4482. doi: 10.1093/nar/gkae231.",
"DOI": "10.1093/nar/gkae231.",
"Authors": "Hertz LM, White EN, Kuznedelov K, Cheng L, Yu AM, Kakkaramadam R, Severinov K, Chen A, Lucks JB /nar/gkae",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/38567721",
"PubMed ID": "PubMed 38567721"
} | {
"Gene/Insert name": "RNA polymerase",
"Species": "Thermus aquaticus",
"Insert Size (bp)": "9786",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": "6x His (C terminal on insert)"
} | {
"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": ""
} |
|
pHH0103_EPB41L3_FERM | 110011 | Protein expression and purification of EPB41L3_FERM | Sachdev Sidhu | Teyra et al Mol Syst Biol. 2020 Dec;16(12):e9310. doi: 10.15252/msb.20199310. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/215921/0b424ceb-65a5-4750-a0b6-760ff39bd929/addgene-plasmid-110011-sequence-215921.gbk | full | LOCUS sequence-215921- 7572 bp DNA circular SYN 26-AUG-2024
DEFINITION Protein expression and purification of EPB41L3_FERM.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7572)
AUTHORS Teyra J, Kelil A, Jain S, Helmy M, Jajodia R, Hooda Y, Gu J, D'Cruz
AA, Nicholson SE, Min J, Sudol M, Kim PM, Bader GD, Sidhu SS
TITLE Large-scale survey and database of high affinity ligands for peptide
recognition modules.
JOURNAL Mol Syst Biol. 2020 Dec;16(12):e9310. doi: 10.15252/msb.20199310.
PUBMED 33438817
REFERENCE 2 (bases 1 to 7572)
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 Syst
Biol."; date: "2020-12"; volume: "16(12):e9310. doi"; pages: "
10.15252/msb.20199310"
FEATURES Location/Qualifiers
source 1..7572
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 1..29
/label=tac promoter
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
protein_bind 37..53
/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 42..64
/label=M13/pUC Reverse
/note="In lacZ gene"
CDS 138..155
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 171..824
/codon_start=1
/product="glutathione S-transferase from Schistosoma
japonicum"
/label=GST
/translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
primer_bind 782..804
/label=pGEX 5'
/note="pGEX vectors, Glutathione-S-transferase, forward
primer"
CDS 831..854
/codon_start=1
/product="recognition and cleavage site for human
rhinovirus 3C and PreScission proteases"
/label=HRV 3C site
/translation="LEVLFQGP"
CDS 867..884
/codon_start=1
/product="thrombin recognition and cleavage site"
/label=thrombin site
/translation="LVPRGS"
terminator 1767..1814
/label=fd terminator
/note="central terminator from bacteriophage fd (Otsuka and
Kunisawa, 1982)"
rep_origin complement(3407..3862)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3544..3565)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 3756..3775
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind complement(3904..3926)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
misc_feature 4012..4152
/label=bom
/note="basis of mobility region from pBR322"
primer_bind 4067..4086
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(4167..4184)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4338..4925)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5096..5956)
/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 5719..5738
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5957..6061)
/gene="bla"
/label=AmpR promoter
primer_bind 6129..6147
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 6170..6247
/gene="lacI (mutant)"
/label=lacIq promoter
/note="In the lacIq allele, a single base change in the
promoter boosts expression of the lacI gene about 10-fold."
CDS 6248..7330
/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(6267..6286)
/label=LacI-R
/note="LacI, reverse primer"
ORIGIN
1 ttgacaatta atcatcggct cgtataatgt gtggaattgt gagcggataa caatttcaca
61 caggaaacag ccagtccgtt taggtgtttt cacgagcact tcaccaacaa ggaccataga
121 ttatgaaaat cgaagaacac catcaccatc accattccag cggtaagctt atgtccccta
181 tactaggtta ttggaaaatt aagggccttg tgcaacccac tcgacttctt ttggaatatc
241 ttgaagaaaa atatgaagag catttgtatg agcgcgatga aggtgataaa tggcgaaaca
301 aaaagtttga attgggtttg gagtttccca atcttcctta ttatattgat ggtgatgtta
361 aattaacaca gtctatggcc atcatacgtt atatagctga caagcacaac atgttgggtg
421 gttgtccaaa agagcgtgca gagatttcaa tgcttgaagg agcggttttg gatattagat
481 acggtgtttc gagaattgca tatagtaaag actttgaaac tctcaaagtt gattttctta
541 gcaagctacc tgaaatgctg aaaatgttcg aagatcgttt atgtcataaa acatatttaa
601 atggtgatca tgtaacccat cctgacttca tgttgtatga cgctcttgat gttgttttat
661 acatggaccc aatgtgcctg gatgcgttcc caaaattagt ttgttttaaa aaacgtattg
721 aagctatccc acaaattgat aagtacttga aatccagcaa gtatatagca tggcctttgc
781 agggctggca agccacgttt ggtggtggcg accatcctcc aaaatcggat ctggaagttc
841 tgttccaggg gcccctgtcc agcggtctgg ttccgcgtgg ttccggtacc gcggcccagc
901 cggccatgca gtgcaaagtt attctgctgg acggcagtga atacacctgt gatgtcgaaa
961 aacgttcccg cggtcaagtc ctgtttgata aagtgtgcga acatctgaac ctgctggaaa
1021 aagactattt cggcctgacg taccgcgatg cggaaaacca gaaaaattgg ctggatccgg
1081 ccaaagaaat caaaaaacaa gtccgtagcg gtgcgtggca cttttcgttc aatgtgaaat
1141 tttatccgcc ggacccggca cagctgtccg aagatattac ccgctattac ctgtgcctgc
1201 aactgcgcga tgacatcgtg agcggccgtc tgccgtgttc tttcgttacc ctggcgctgc
1261 tgggtagtta tacggtgcag tccgaactgg gcgattacga cccggatgaa tgtggtagcg
1321 actatatttc tgaatttcgc ttcgccccga accataccaa agaactggaa gataaagtga
1381 tcgaactgca taaaagccac cgtggcatga cgccggcaga agctgaaatg cactttctgg
1441 aaaatgcgaa aaaactgtca atgtacggcg ttgacctgca tcacgctaaa gattcggaag
1501 gtgttgaaat tatgctgggc gtctgcgcaa gcggtctgct gatttatcgt gatcgcctgc
1561 gtatcaaccg ctttgcttgg ccgaaagttc tgaaaatttc ttacaaacgc aacaacttct
1621 acatcaaaat ccgtccgggc gaatttgaac agttcgaatc aaccatcggt tttaaactgc
1681 cgaatcatcg cgcggccaaa cgtctgtgga aagtgtgtgt tgaacatcac acgtttttcc
1741 gtctgctgct ggcggccgca taataaaccg atacaattaa aggctccttt tggagccttt
1801 tttttggaga ttttcaacgt gaaaaaatta ttattcgcaa ttcctttagt tgttcctttc
1861 tattctcact ccgctgaaac tgttgaaagt tgtttagcaa aaccccatac agaaaattca
1921 tttactaacg tctggaaaga cgacaaaact ttagatcgtt acgctaacta tgagggctgt
1981 ctgtggaatg ctacaggcgt tgtagtttgt actggtgacg aaactcagtg tctagctaga
2041 gtggcggtgg ctctggttcc ggtgattttg attatgaaaa gatggcaaac gctaataagg
2101 gggctatgac cgaaaatgcc gatgaaaacg cgctacagtc tgacgctaaa ggcaaacttg
2161 attctgtcgc tactgattac ggtgctgcta tcgatggttt cattggtgac gtttccggcc
2221 ttgctaatgg taatggtgct actggtgatt ttgctggctc taattcccaa atggctcaag
2281 tcggtgacgg tgataattca cctttaatga ataatttccg tcaatattta ccttccctcc
2341 ctcaatcggt tgaatgtcgc ccttttgtct ttggcgctgg taaaccatat gaattttcta
2401 ttgattgtga caaaataaac ttattccgtg gtgtctttgc gtttctttta tatgttgcca
2461 cctttatgta tgtattttct acgtttgcta acatactgcg taataaggag tcttaatcat
2521 gccagttctt ttggctagcg ccgccctata ccttgtctgc ctccccgcgt tgcgtcgcgg
2581 tgcatggagc cgggccacct cgacctgaat ggaagccggc ggcacctcgc taacggattc
2641 accactccaa gaattggagc caatcaattc ttgcggagaa ctgtgaatgc gcaaaccaac
2701 ccttggcaga acatatccat cgcgtccgcc atctccagca gccgcacgcg gcgcatctcg
2761 ggcagcgttg ggtcctggcc acgggtgcgc atgatcgtgc tcctgtcgtt gaggacccgg
2821 ctaggctggc ggggttgcct tactggttag cagaatgaat caccgatacg cgagcgaacg
2881 tgaagcgact gctgctgcaa aacgtctgcg acctgagcaa caacatgaat ggtcttcggt
2941 ttccgtgttt cgtaaagtct ggaaacgcgg aagtcagcgc cctgcaccat tatgttccgg
3001 atctgcatcg caggatgctg ctggctaccc tgtggaacac ctacatctgt attaacgaag
3061 cgctggcatt gaccctgagt gatttttctc tggtcccgcc gcatccatac cgccagttgt
3121 ttaccctcac aacgttccag taaccgggca tgttcatcat cagtaacccg tatcgtgagc
3181 atcctctctc gtttcatcgg tatcattacc cccatgaaca gaaatccccc ttacacggag
3241 gcatcagtga ccaaacagga aaaaaccgcc cttaacatgg cccgctttat cagaagccag
3301 acattaacgc ttctggagaa actcaacgag ctggacgcgg atgaacaggc agacatctgt
3361 gaatcgcttc acgaccacgc tgatgagctt taccgcagga tccgcgaaat tgtaagcgtt
3421 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
3481 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
3541 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
3601 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
3661 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
3721 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
3781 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
3841 aatgcgccgc tacagggcgc gtcgcggatc ctgcctcgcg cgtttcggtg atgacggtga
3901 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
3961 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat
4021 gacccagtca cgtagcgata gcggagtgta tactggctta actatgcggc atcagagcag
4081 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
4141 taccgcatca ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
4201 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
4261 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
4321 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
4381 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggg tttccccctg
4441 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
4501 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
4561 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
4621 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
4681 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
4741 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
4801 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
4861 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
4921 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
4981 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
5041 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
5101 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
5161 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
5221 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
5281 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
5341 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
5401 ttgccattgc tgcaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
5461 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
5521 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
5581 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
5641 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
5701 gcccggcgtc aacacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
5761 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
5821 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
5881 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
5941 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
6001 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
6061 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
6121 cctataaaaa taggcgtatc acgaggccct ttcgtcttca agaattcccg acaccatcga
6181 atggtgcaaa acctttcgcg gtatggcatg atagcgcccg gaagagagtc aattcagggt
6241 ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag tatgccggtg tctcttatca
6301 gaccgtttcc cgcgtggtga accaggccag ccacgtttct gcgaaaacgc gggaaaaagt
6361 ggaagcggcg atggcggagc tgaattacat tcccaaccgc gtggcacaac aactggcggg
6421 caaacagtcg ttgctgattg gcgttgccac ctccagtctg gccctgcacg cgccgtcgca
6481 aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt gccagcgtgg tggtgtcgat
6541 ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg cacaatcttc tcgcgcaacg
6601 cgtcagtggg ctgatcatta actatccgct ggatgaccag gatgccattg ctgtggaagc
6661 tgcctgcact aatgttccgg cgttatttct tgatgtctct gaccagacac ccatcaacag
6721 tattattttc tcccatgaag acggtacgcg actgggcgtg gagcatctgg tcgcattggg
6781 tcaccagcaa atcgcgctgt tagcgggccc attaagttct gtctcggcgc gtctgcgtct
6841 ggctggctgg cataaatatc tcactcgcaa tcaaattcag ccgatagcgg aacgggaagg
6901 cgactggagt gccatgtccg gttttcaaca aaccatgcaa atgctgaatg agggcatcgt
6961 tcccactgcg atgctggttg ccaacgatca gatggcgctg ggcgcaatgc gcgccattac
7021 cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg ggatacgacg ataccgaaga
7081 cagctcatgt tatatcccgc cgttaaccac catcaaacag gattttcgcc tgctggggca
7141 aaccagcgtg gaccgcttgc tgcaactctc tcagggccag gcggtgaagg gcaatcagct
7201 gttgcccgtc tcactggtga aaagaaaaac caccctggcg cccaatacgc aaaccgcctc
7261 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
7321 cgggcagtga gcgcaacgca attaatgtaa gttagctcac tcattaggca caattctcat
7381 gtttgacagc ttatcatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc
7441 ggaagctgtg gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc
7501 gcactcccgt tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc
7561 tgaaatgagc tg
//
| {
"Vector backbone": "pHH0103",
"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": "Large-scale survey and database of high affinity ligands for peptide recognition modules",
"Publication": "Mol Syst Biol. 2020 Dec;16(12):e9310. doi: 10.15252/msb.20199310.",
"DOI": "10.15252/msb.20199310.",
"Authors": "Teyra J, Kelil A, Jain S, Helmy M, Jajodia R, Hooda Y, Gu J, D'Cruz AA, Nicholson SE, Min J, Sudol M, Kim PM, Bader GD, Sidhu SS /msb",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/33438817",
"PubMed ID": "PubMed 33438817"
} | {
"Gene/Insert name": "EPB41L3_FERM",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "EPB41L3",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "Sfi1 (unknown if destroyed)",
"3′ cloning site": "Not1 (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": ""
} |
|
pcDNA3.3_SARS2_BQ.1.1 | 194493 | Low | expressing SARS-CoV-2 BQ.1.1 spike protein for pseudovirus production | David Nemazee | Zhou et al Immunity. 2023 Mar 14;56(3):669-686.e7. doi: 10.1016/j.immuni.2023.02.005. Epub 2023 Feb 16. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/381631/2ae354c9-96e0-4415-b4b2-45fb5b25ce48/addgene-plasmid-194493-sequence-381631.gbk | full | LOCUS sequence-381631- 9191 bp DNA circular SYN 25-AUG-2024
DEFINITION expressing SARS-CoV-2 BQ.1.1 spike protein for pseudovirus
production.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9191)
AUTHORS Zhou P, Song G, Liu H, Yuan M, He WT, Beutler N, Zhu X, Tse LV,
Martinez DR, Schafer A, Anzanello F, Yong P, Peng L, Dueker K,
Musharrafieh R, Callaghan S, Capozzola T, Limbo O, Parren M, Garcia
E, Rawlings SA, Smith DM, Nemazee D, Jardine JG, Safonova Y, Briney
B, Rogers TF, Wilson IA, Baric RS, Gralinski LE, Burton DR, Andrabi
R
TITLE Broadly neutralizing anti-S2 antibodies protect against all three
human betacoronaviruses that cause deadly disease.
JOURNAL Immunity. 2023 Mar 14;56(3):669-686.e7. doi:
10.1016/j.immuni.2023.02.005. Epub 2023 Feb 16.
PUBMED 36889306
REFERENCE 2 (bases 1 to 9191)
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.immuni.2023.02.005"; journalName: "Immunity"; date:
"2023-03-14- 14"; volume: "56"; issue: "3"; pages: "669-686.e7"
FEATURES Location/Qualifiers
source 1..9191
/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"
primer_bind complement(4577..4596)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 4621..4669
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4871..5299
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4958..4977)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 5168..5189
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(5308..5328)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 5313..5642
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 5493..5628
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 5555..5574
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 5709..6503
/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(5763..5782)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 6373..6392
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 6679..6800
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(6716..6735)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 6770..6789
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(6849..6865)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6849..6865)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(6862..6884)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 6873..6889
/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(6897..6927)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6942..6963
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(7080..7097)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(7251..7839)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(7331..7350)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(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 8633..8652
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(8871..8975)
/gene="bla"
/label=AmpR promoter
primer_bind complement(9050..9069)
/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 actgagctcc ttaaggttaa ctctagagaa ttcggatcca tgtttgtgtt cctggtgctg
781 ctgccactgg tgtccagcca gtgtgtgaac ctgatcacca ggacccaatc ctacaccaac
841 tccttcacca ggggagtcta ctaccctgac aaggtgttca ggtcctctgt gctgcacagc
901 acccaggacc tgttcctgcc attcttcagc aatgtgacct ggttccatgc catctctggc
961 accaatggca ccaagaggtt tgacaaccct gtgctgccat tcaatgatgg agtctacttt
1021 gccagcacag agaagagcaa catcatcagg ggctggattt ttggcaccac cctggacagc
1081 aagacccagt ccctgctgat tgtgaacaat gccaccaatg tggtgattaa ggtgtgtgag
1141 ttccagttct gtaatgaccc attcctggac gtctactacc acaagaacaa caagtcctgg
1201 atggagtctg agttcagggt ctactcctct gccaacaact gtacctttga atatgtgagc
1261 caaccattcc tgatggactt ggagggcaag cagggcaact tcaagaacct gagggagttt
1321 gtgttcaaga acattgatgg ctacttcaag atttacagca aacacacacc aatcaacctg
1381 gggagggacc tgccacaggg cttctctgcc ttggaaccac tggtggacct gccaattggc
1441 atcaacatca ccaggttcca gaccctgctg gctctgcaca ggtcctacct gacacctgga
1501 gactcctcct ctggctggac agcaggagca gcagcctact atgtgggcta cctccaacca
1561 aggaccttcc tgctgaaata caatgagaat ggcaccatca cagatgctgt ggactgtgcc
1621 ctggacccac tgtctgagac caagtgtacc ctgaaatcct tcacagtgga gaagggcatc
1681 taccagacca gcaacttcag ggtccaacca acagagagca ttgtgaggtt tccaaacatc
1741 accaacctgt gtccatttga cgaggtgttc aatgccacca cgtttgcctc tgtctatgcc
1801 tggaacagga agaggattag caactgtgtg gctgactact ctgtgctcta caactttgcc
1861 cccttcttcg ccttcaagtg ttatggagtg agcccaacca aactgaatga cctgtgtttc
1921 accaatgtct atgctgactc ctttgtgatt aggggaaatg aggtgagtca gattgcccct
1981 ggacaaacag gcaacattgc tgactacaac tacaaactgc ctgatgactt cacaggctgt
2041 gtgattgcct ggaacagcaa caagctggac agcacggtgg gaggcaacta caactaccgc
2101 tacagactgt tcaggaagag caagctgaaa ccatttgaga gggacatcag cacagagatt
2161 taccaggctg gcaacaaacc atgtaatgga gtggcgggcg tcaactgtta ctttccactc
2221 cagtcctatg gcttcagacc aacctatgga gtgggccacc aaccatacag ggtggtggtg
2281 ctgtcctttg aactgctcca tgcccctgcc acagtgtgtg gaccaaagaa gagcaccaac
2341 ctggtgaaga acaagtgtgt gaacttcaac ttcaatggac tgacaggcac aggagtgctg
2401 acagagagca acaagaagtt cctgccattc caacagtttg gcagggacat tgctgacacc
2461 acagatgctg tgagggaccc acagaccttg gagattctgg acatcacacc atgttccttt
2521 ggaggagtgt ctgtgattac acctggcacc aacaccagca accaggtggc tgtgctctac
2581 cagggtgtga actgtactga ggtgcctgtg gctatccatg ctgaccaact tacaccaacc
2641 tggagggtct acagcacagg cagcaatgtg ttccagacca gggctggctg tctgattgga
2701 gcagagtatg tgaacaactc ctatgagtgt gacatcccaa ttggagcagg catctgtgcc
2761 tcctaccaga cccagaccaa gagccatagg agggcaaggt ctgtggcaag ccagagcatc
2821 attgcctaca caatgagtct gggagcagag aactctgtgg cttacagcaa caacagcatt
2881 gccatcccaa ccaacttcac catctctgtg accacagaaa ttctgcctgt gagtatgacc
2941 aagacctctg tggactgtac aatgtatatc tgtggagaca gcacagagtg tagcaacctg
3001 ctgctccaat atggctcctt ctgtacccaa cttaagaggg ctctgacagg cattgctgtg
3061 gaacaggaca agaacaccca ggaggtgttt gcccaggtga agcagattta caagacacct
3121 ccaatcaagt actttggagg cttcaacttc agccagattc tgcctgaccc aagcaagcca
3181 agcaagaggt ccttcattga ggacctgctg ttcaacaagg tgaccctggc tgatgctggc
3241 ttcatcaagc aatatggaga ctgtctggga gacattgctg caagggacct gatttgtgcc
3301 cagaagttca atggactgac cgtgctgcct ccactgctga cagatgagat gattgcccaa
3361 tacacctctg ccctgctggc tggcaccatc acctctggct ggacctttgg agcaggagca
3421 gccctccaaa tcccatttgc tatgcagatg gcttacaggt tcaatggcat tggagtgacc
3481 cagaatgtgc tctatgagaa ccagaaactg attgccaacc agttcaactc tgccattggc
3541 aagattcagg actccctgtc cagcacagcc tctgccctgg gcaaactcca agatgtggtg
3601 aaccataatg cccaggctct gaacaccctg gtgaagcaac tttccagcaa gtttggagcc
3661 atctcctctg tgctgaatga catcctgagc agactggaca aggtggaggc tgaggtccag
3721 attgacagac tgattacagg cagactccaa tccctccaaa cctatgtgac ccaacaactt
3781 atcagggctg ctgagattag ggcatctgcc aacctggctg ccaccaagat gagtgagtgt
3841 gtgctgggac aaagcaagag ggtggacttc tgtggcaagg gctaccacct gatgagtttt
3901 ccacagtctg cccctcatgg agtggtgttc ctgcatgtga cctatgtgcc tgcccaggag
3961 aagaacttca ccacagcccc tgccatctgc catgatggca aggctcactt tccaagggag
4021 ggagtgtttg tgagcaatgg cacccactgg tttgtgaccc agaggaactt ctatgaacca
4081 cagattatca ccacagacaa cacctttgtg tctggcaact gtgatgtggt gattggcatt
4141 gtgaacaaca cagtctatga cccactccaa cctgaactgg actccttcaa ggaggaactg
4201 gacaaatact tcaagaacca caccagccct gatgtggacc tgggagacat ctctggcatc
4261 aatgcctctg tggtgaacat ccagaaggag attgacagac tgaatgaggt ggctaagaac
4321 ctgaatgagt ccctgattga cctccaagaa ctgggcaaat atgaacaata catcaagtgg
4381 ccatggtaca tctggctggg cttcattgct ggactgattg ccattgtgat ggtgaccata
4441 atgctgtgtt gtatgacctc ctgttgttcc tgtctgaaag gctgttgttc ctgtggctcc
4501 tgttgtaagt gactcgagcc atggaagctt gatatctaac tgactgaacc ggtggtacca
4561 aacgggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
4621 cggcaataaa aagacagaat aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg
4681 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
4741 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
4801 gcagccaacg tcggggcggc aggccctgcc atagcagatc tgcgcagctg gggctctagg
4861 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
4921 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
4981 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
5041 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
5101 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
5161 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
5221 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
5281 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
5341 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
5401 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
5461 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
5521 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
5581 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
5641 aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat
5701 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
5761 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
5821 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
5881 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
5941 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
6001 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
6061 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
6121 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
6181 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
6241 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
6301 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
6361 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
6421 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
6481 gccttcttga cgagttcttc tgagcgggac tctggggttc gcgaaatgac cgaccaagcg
6541 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
6601 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
6661 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
6721 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
6781 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
6841 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
6901 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
6961 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
7021 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
7081 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
7141 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
7201 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
7261 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
7321 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
7381 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
7441 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
7501 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
7561 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
7621 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
7681 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
7741 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
7801 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
7861 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
7921 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
7981 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
8041 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
8101 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
8161 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
8221 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
8281 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
8341 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
8401 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
8461 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
8521 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
8581 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
8641 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
8701 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
8761 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
8821 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
8881 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
8941 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
9001 gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct
9061 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
9121 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga
9181 atctgcttag g
//
| {
"Vector backbone": "pcDNA3.3",
"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": "Broadly neutralizing anti-S2 antibodies protect against all three human betacoronaviruses that cause deadly disease",
"Publication": "Immunity. 2023 Mar 14;56(3):669-686.e7. doi: 10.1016/j.immuni.2023.02.005. Epub 2023 Feb 16.",
"DOI": "10.1016/j.immuni.2023.02.005.",
"Authors": "Zhou P, Song G, Liu H, Yuan M, He WT, Beutler N, Zhu X, Tse LV, Martinez DR, Schafer A, Anzanello F, Yong P, Peng L, Dueker K, Musharrafieh R, Callaghan S, Capozzola T, Limbo O, Parren M, Garcia E, Rawlings SA, Smith DM, Nemazee D, Jardine JG, Safonova Y, Briney B, Rogers TF, Wilson IA, Baric RS, Gralinski LE, Burton DR, Andrabi R /jimmuni",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/36889306",
"PubMed ID": "PubMed 36889306"
} | {
"Gene/Insert name": "Spike (BQ.1.1)",
"Species": "SARS-CoV-2",
"Insert Size (bp)": "3800",
"Mutation": "R346T, K444T, N460K and c-terminal 18aa del on the basis of BA.5",
"Entrez Gene": "S",
"Promoter": "CMV",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "BamHI (not destroyed)",
"3′ cloning site": "XhoI (not destroyed)",
"5′ sequencing primer": "CMV-F",
"3′ sequencing primer": "TK_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": ""
} |
FASTKD5 gRNA (BRDN0001147085) | 77621 | 3rd generation lentiviral gRNA plasmid targeting human FASTKD5 | Doench et al Nat Biotechnol. 2016 Jan 18. doi: 10.1038/nbt.3437. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/188118/c353e5a3-4e0a-485f-a770-9034856d2a1d/addgene-plasmid-77621-sequence-188118.gbk | full | LOCUS sequence-188118- 8323 bp DNA circular SYN 26-AUG-2024
DEFINITION 3rd generation lentiviral gRNA plasmid targeting human FASTKD5.
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 cgacttaggc cgcaaagtac ctgttttaga 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": "FASTKD5",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "FASTKD5",
"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": ""
} |
||
pJK523 | 72442 | Produces Acetobacter aceti 1023 small hypothetical protein adjacent to purEK genes, with N-terminal His6 tag (H6AaOrfX) | T. J. Kappock | Additional purEK associated constructs (unpublished) | https://media.addgene.org/snapgene-media/v2.0.0/sequences/133863/f9793725-0271-4177-85eb-107b3225d276/addgene-plasmid-72442-sequence-133863.gbk | full | LOCUS sequence-133863- 5514 bp DNA circular SYN 26-AUG-2024
DEFINITION Produces Acetobacter aceti 1023 small hypothetical protein adjacent
to purEK genes, with N-terminal His6 tag (H6AaOrfX).
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5514)
AUTHORS Kappock
TITLE Additional purEK associated constructs
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 5514)
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..5514
/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(388..405)
/codon_start=1
/product="thrombin recognition and cleavage site"
/label=thrombin site
/translation="LVPRGS"
CDS complement(415..432)
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
RBS 451..473
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
protein_bind 488..512
/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(512..531)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(513..531)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(615..634)
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
promoter 840..917
/gene="lacI"
/label=lacI promoter
CDS 918..2000
/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(937..956)
/label=LacI-R
/note="LacI, reverse primer"
CDS 2809..3000
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
primer_bind complement(2994..3016)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
misc_feature 3102..3244
/label=bom
/note="basis of mobility region from pBR322"
primer_bind 3157..3176
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(3259..3276)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3430..4018)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3510..3529)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind complement(4129..4148)
/label=pENTR-R
/note="pENTR vectors, reverse primer"
CDS 4140..4955
/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 4140..4955
/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(4213..4232)
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
rep_origin complement(5048..5503)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5185..5206)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 5397..5416
/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 gcaagcttca
181 cgcgatgtca tccggaatca gtagactttc cactttcgta atttgatcct taagcagcag
241 tttgcgtttt ttcagacgtt gcagatacag ctgatcgata aacgtggttt catgggccat
301 acggttgatc acggtgtcca gatcgcggtg ttcgctacgc agttcatgca attgcgccag
361 cagcaggtcg cgatcacgca gcatatggct gccgcgcggc accaggccgc tgctgtgatg
421 atgatgatga tggctgctgc ccatggtata tctccttctt aaagttaaac aaaattattt
481 ctagagggga attgttatcc gctcacaatt cccctatagt gagtcgtatt aatttcgcgg
541 gatcgagatc tcgatcctct acgccggacg catcgtggcc ggcatcaccg gcgccacagg
601 tgcggttgct ggcgcctata tcgccgacat caccgatggg gaagatcggg ctcgccactt
661 cgggctcatg agcgcttgtt tcggcgtggg tatggtggca ggccccgtgg ccgggggact
721 gttgggcgcc atctccttgc atgcaccatt ccttgcggcg gcggtgctca acggcctcaa
781 cctactactg ggctgcttcc taatgcagga gtcgcataag ggagagcgtc gagatcccgg
841 acaccatcga atggcgcaaa acctttcgcg gtatggcatg atagcgcccg gaagagagtc
901 aattcagggt ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag tatgccggtg
961 tctcttatca gaccgtttcc cgcgtggtga accaggccag ccacgtttct gcgaaaacgc
1021 gggaaaaagt ggaagcggcg atggcggagc tgaattacat tcccaaccgc gtggcacaac
1081 aactggcggg caaacagtcg ttgctgattg gcgttgccac ctccagtctg gccctgcacg
1141 cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt gccagcgtgg
1201 tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg cacaatcttc
1261 tcgcgcaacg cgtcagtggg ctgatcatta actatccgct ggatgaccag gatgccattg
1321 ctgtggaagc tgcctgcact aatgttccgg cgttatttct tgatgtctct gaccagacac
1381 ccatcaacag tattattttc tcccatgaag acggtacgcg actgggcgtg gagcatctgg
1441 tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc attaagttct gtctcggcgc
1501 gtctgcgtct ggctggctgg cataaatatc tcactcgcaa tcaaattcag ccgatagcgg
1561 aacgggaagg cgactggagt gccatgtccg gttttcaaca aaccatgcaa atgctgaatg
1621 agggcatcgt tcccactgcg atgctggttg ccaacgatca gatggcgctg ggcgcaatgc
1681 gcgccattac cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg ggatacgacg
1741 ataccgaaga cagctcatgt tatatcccgc cgttaaccac catcaaacag gattttcgcc
1801 tgctggggca aaccagcgtg gaccgcttgc tgcaactctc tcagggccag gcggtgaagg
1861 gcaatcagct gttgcccgtc tcactggtga aaagaaaaac caccctggcg cccaatacgc
1921 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
1981 gactggaaag cgggcagtga gcgcaacgca attaatgtaa gttagctcac tcattaggca
2041 ccgggatctc gaccgatgcc cttgagagcc ttcaacccag tcagctcctt ccggtgggcg
2101 cggggcatga ctatcgtcgc cgcacttatg actgtcttct ttatcatgca actcgtagga
2161 caggtgccgg cagcgctctg ggtcattttc ggcgaggacc gctttcgctg gagcgcgacg
2221 atgatcggcc tgtcgcttgc ggtattcgga atcttgcacg ccctcgctca agccttcgtc
2281 actggtcccg ccaccaaacg tttcggcgag aagcaggcca ttatcgccgg catggcggcc
2341 ccacgggtgc gcatgatcgt gctcctgtcg ttgaggaccc ggctaggctg gcggggttgc
2401 cttactggtt agcagaatga atcaccgata cgcgagcgaa cgtgaagcga ctgctgctgc
2461 aaaacgtctg cgacctgagc aacaacatga atggtcttcg gtttccgtgt ttcgtaaagt
2521 ctggaaacgc ggaagtcagc gccctgcacc attatgttcc ggatctgcat cgcaggatgc
2581 tgctggctac cctgtggaac acctacatct gtattaacga agcgctggca ttgaccctga
2641 gtgatttttc tctggtcccg ccgcatccat accgccagtt gtttaccctc acaacgttcc
2701 agtaaccggg catgttcatc atcagtaacc cgtatcgtga gcatcctctc tcgtttcatc
2761 ggtatcatta cccccatgaa cagaaatccc ccttacacgg aggcatcagt gaccaaacag
2821 gaaaaaaccg cccttaacat ggcccgcttt atcagaagcc agacattaac gcttctggag
2881 aaactcaacg agctggacgc ggatgaacag gcagacatct gtgaatcgct tcacgaccac
2941 gctgatgagc tttaccgcag ctgcctcgcg cgtttcggtg atgacggtga aaacctctga
3001 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
3061 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gacccagtca
3121 cgtagcgata gcggagtgta tactggctta actatgcggc atcagagcag attgtactga
3181 gagtgcacca tatatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
3241 caggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
3301 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
3361 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
3421 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
3481 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
3541 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
3601 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
3661 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
3721 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
3781 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
3841 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
3901 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
3961 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
4021 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
4081 gattttggtc atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta
4141 tgagccatat tcaacgggaa acgtcttgct ctaggccgcg attaaattcc aacatggatg
4201 ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct
4261 atcgattgta tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg
4321 ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc
4381 ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga
4441 tccccgggaa aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg
4501 ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt
4561 ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg
4621 ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag
4681 aaatgcataa acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac
4741 ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg
4801 gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc
4861 cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat
4921 tgcagtttca tttgatgctc gatgagtttt tctaagaatt aattcatgag cggatacata
4981 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
5041 ccacctgaaa ttgtaaacgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
5101 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
5161 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
5221 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
5281 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
5341 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
5401 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
5461 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcccattc gcca
//
| {
"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": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "",
"Publication": "",
"DOI": "",
"Authors": "",
"PubMed Link": "",
"PubMed ID": ""
} | {
"Gene/Insert name": "hypothetical protein",
"Species": "Acetobacter aceti 1023",
"Insert Size (bp)": "",
"Mutation": "synthetic gene with ~40 silent substitutions",
"Entrez Gene": "",
"Promoter": "T7",
"Tag / Fusion Protein": "His6 (N terminal on backbone)"
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "NdeI (not destroyed)",
"3′ cloning site": "HindIII (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": ""
} |
|
3XFlag DVL2 (∂DIX) | 24805 | Low | Jeff Wrana | Narimatsu et al Cell. 2009 Apr 17. 137(2):295-307. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/30768/d3cfa012-7ab7-4cca-91ed-0d1b40914181/addgene-plasmid-24805-sequence-30768.gbk | partial | LOCUS sequence-30768-e 921 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 921)
AUTHORS Narimatsu M, Bose R, Pye M, Zhang L, Miller B, Ching P, Sakuma R,
Luga V, Roncari L, Attisano L, Wrana JL
TITLE Regulation of planar cell polarity by Smurf ubiquitin ligases.
JOURNAL Cell. 2009 Apr 17. 137(2):295-307.
PUBMED 19379695
REFERENCE 2 (bases 1 to 921)
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. 2009
Apr 17. 137(2):295-307."
FEATURES Location/Qualifiers
source 1..921
/mol_type="genomic DNA"
/organism="unspecified"
CDS 31..96
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
ORIGIN
1 gtgaaccgtc agaattgatc tggtaccatg gactacaaag accatgacgg tgattataaa
61 gatcatgaca tcgattacaa ggatgacgat gacaagcttg catgcctgca ggtcgactct
121 agatcctcag ataatcccca acccgagatg gcccctccag tccatgagcc tcgggcagaa
181 ctggcgcctc cagccccacc tttacctcct ttgccacccg agaggaccag cggcattggg
241 gactcaaggc ctccatcctt ccaccctaat gtgtccagca gccatgagaa tctggagcct
301 gagacagaaa ccgagtcagt agtgtcactg aggcgggagc ggcctcgcag gagagacagc
361 agtgagcatg gcgctggggg ccacaggact ggtggcccct caaggctgga gcgccacctg
421 gccggatacg agagctcctc taccctcatg accagcgagc tggagagtac cagcctgggg
481 gactcggacg aggaggacac catgagcagg ttcagcagct ccacggagca gagcagtgcc
541 tcccgcctcc ttaagcgcca ccggcggcga aggaagcaga ggccaccccg cctggagagg
601 acgtcatcct tcagcagcgt cacagattcc acaatgtctc tcaatatcat cacagtcacg
661 ctaaacatgg agaagtacaa cttcctgggt atctccattg ttggccagag caatgagcgg
721 ggagacggag gcatctacat tggctccatc atgaagggtg gggctgtggc ggccgacggg
781 cgcattgagc caggggacat gcttttgcag gtgaatgaca tgaactttga gaacatgagc
841 aacgatgacg ctgtgcgggt gctgagggac attgtgcaca agcctggccc cattgtgctg
901 actgtggcca agtgctggga t
//
| {
"Vector backbone": "pCMV5",
"Backbone size w/o insert (bp)": "4095",
"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": "Regulation of planar cell polarity by Smurf ubiquitin ligases",
"Publication": "Cell. 2009 Apr 17. 137(2):295-307.",
"DOI": "10.1016/j.cell.2009.02.025",
"Authors": "Narimatsu M, Bose R, Pye M, Zhang L, Miller B, Ching P, Sakuma R, Luga V, Roncari L, Attisano L, Wrana JL",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/19379695",
"PubMed ID": "PubMed 19379695"
} | {
"Gene/Insert name": "DVL2",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "Deletion of the N-terminal DIX domain",
"Entrez Gene": "DVL2",
"Promoter": "",
"Tag / Fusion Protein": "3XFlag (N terminal on insert)"
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "KpnI (not destroyed)",
"3′ cloning site": "EcoRI (not destroyed)",
"5′ sequencing primer": "CMV-F",
"3′ sequencing primer": "hGH-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": ""
} |
|
pET28-Kss1 | 52682 | Expression of Kss1 in bacteria | Benjamin Turk | Mok et al Sci Signal. 2010 Feb 16;3(109):ra12. doi: 10.1126/scisignal.2000482. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/82705/299922e1-a599-480f-ba3d-00365af80a15/addgene-plasmid-52682-sequence-82705.gbk | partial | LOCUS sequence-82705-e 868 bp DNA linear UNA 25-AUG-2024
DEFINITION Expression of Kss1 in bacteria.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE natural DNA sequence
ORGANISM unspecified
REFERENCE 1 (bases 1 to 868)
AUTHORS Mok J, Kim PM, Lam HY, Piccirillo S, Zhou X, Jeschke GR, Sheridan
DL, Parker SA, Desai V, Jwa M, Cameroni E, Niu H, Good M, Remenyi A,
Ma JL, Sheu YJ, Sassi HE, Sopko R, Chan CS, De Virgilio C,
Hollingsworth NM, Lim WA, Stern DF, Stillman B, Andrews BJ, Gerstein
MB, Snyder M, Turk BE
TITLE Deciphering protein kinase specificity through large-scale analysis
of yeast phosphorylation site motifs.
JOURNAL Sci Signal. 2010 Feb 16;3(109):ra12. doi: 10.1126/scisignal.2000482.
PUBMED 20159853
REFERENCE 2 (bases 1 to 868)
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: "Sci
Signal."; date: "2010-02-16"; volume: "3(109):ra12. doi"; pages: "
10.1126/scisignal.2000482"
FEATURES Location/Qualifiers
source 1..868
/mol_type="genomic DNA"
/organism="unspecified"
RBS 8..30
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 49..66
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 76..93
/codon_start=1
/product="thrombin recognition and cleavage site"
/label=thrombin site
/translation="LVPRGS"
CDS 97..129
/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"
ORIGIN
1 gaataatttt gtttaacttt aagaaggaga tataccatgg gcagcagcca tcatcatcat
61 catcacagca gcggcctggt gccgcgcggc agccatatgg ctagcatgac tggtggacag
121 caaatgggtc gcggatccaa gatggctaga accataactt ttgatatccc ttcccaatat
181 aaactcgtag atttaatagg tgagggagcg tacggaacag tatgttcagc aattcataag
241 ccttccggca taaaggtagc tatcaagaaa atacaaccgt ttagcaaaaa attgtttgtt
301 acaagaacta tacgtgagat caagctttta cggtatttcc atgaacacga aaacataata
361 agtatattgg ataaagtaag gccagtatcc atagacaaac taaacgctgt ttatttagtc
421 gaagagttga tggaaaccga tttacaaaaa gtaattaata atcagaatag cgggttttcc
481 actttaagtg atgaccatgt tcaatacttt acataccaaa tcctcagagc cttaaagtct
541 attcacagtg cacaagttat ccatagagac ataaagccat caaacctgtt actaaattcc
601 aattgtgatc tcaaagtctg cgattttgga ctagcgaggt gtttagctag cagtagcgat
661 tcaagagaaa cattggtagg attcatgacg gagtacgtcg caacgcgatg gtacagggca
721 cccgagataa tgctaacttt tcaagagtac acaactgcga tggatatatg gtcatgcgga
781 tgcattttgg ctgaaatggt ctccgggaag cctttgttcc caggcagaga ctatcatcat
841 caattatggc taattctaga agtcttgg
//
| {
"Vector backbone": "pET28a",
"Backbone size w/o insert (bp)": "5400",
"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",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Deciphering protein kinase specificity through large-scale analysis of yeast phosphorylation site motifs",
"Publication": "Sci Signal. 2010 Feb 16;3(109):ra12. doi: 10.1126/scisignal.2000482.",
"DOI": "10.1126/scisignal.2000482.",
"Authors": "Mok J, Kim PM, Lam HY, Piccirillo S, Zhou X, Jeschke GR, Sheridan DL, Parker SA, Desai V, Jwa M, Cameroni E, Niu H, Good M, Remenyi A, Ma JL, Sheu YJ, Sassi HE, Sopko R, Chan CS, De Virgilio C, Hollingsworth NM, Lim WA, Stern DF, Stillman B, Andrews BJ, Gerstein MB, Snyder M, Turk BE /scisignal",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/20159853",
"PubMed ID": "PubMed 20159853"
} | {
"Gene/Insert name": "KSS1",
"Species": "S. cerevisiae (budding yeast)",
"Insert Size (bp)": "1100",
"Mutation": "",
"Entrez Gene": "KSS1",
"Promoter": "T7",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "BamHI (not destroyed)",
"3′ cloning site": "SalI (not destroyed)",
"5′ sequencing primer": "T7",
"3′ sequencing primer": "T7 terminal"
} | {
"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": ""
} |
|
tdTomato--β-actin CDS2 | 183714 | tdTomato with fragment nucleotides 398-818 of beta-actin coding sequence inserted in 3'UTR | Jeffrey Gerst | Dasgupta et al Elife. 2023 May 30;12:e83584. doi: 10.7554/eLife.83584. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/361426/d903c024-25a8-40b0-8caa-1a6012a0c11e/addgene-plasmid-183714-sequence-361426.gbk | full | LOCUS sequence-361426- 9673 bp DNA circular SYN 27-AUG-2024
DEFINITION tdTomato with fragment nucleotides 398-818 of beta-actin coding
sequence inserted in 3'UTR.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9673)
AUTHORS Dasgupta S, Dayagi DY, Haimovich G, Wyler E, Olender T, Singer RH,
Landthaler M, Gerst JE
TITLE Global analysis of contact-dependent human-to-mouse intercellular
mRNA and lncRNA transfer in cell culture.
JOURNAL Elife. 2023 May 30;12:e83584. doi: 10.7554/eLife.83584.
PUBMED 37249209
REFERENCE 2 (bases 1 to 9673)
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: "Elife.";
date: "2023-05-30"; volume: "12:e83584. doi"; pages: "
10.7554/eLife.83584"
FEATURES Location/Qualifiers
source 1..9673
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 26..130
/gene="bla"
/label=AmpR promoter
CDS 131..991
/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(349..368)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 1162..1750
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 1651..1670
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 1904..1921
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 2038..2059
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2074..2104
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2112..2128
/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 2117..2139
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 2136..2152
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 2136..2152
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 2171..2191
/label=T3
/note="T3 promoter, forward primer"
promoter 2173..2191
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
enhancer 2292..2671
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 2672..2875
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 2826..2846
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
LTR 2890..3070
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 3117..3242
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 3735..3968
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 4153..4197
/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 4464..4581
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 4638..5849
/label=UbC promoter
/note="human ubiquitin C promoter"
primer_bind 5686..5705
/label=hUBCpro-F
/note="Human Ubiquitin C (UbC) promoter, forward primer"
regulatory 5883..5892
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 5889..7319
/codon_start=1
/product="tandem dimeric (pseudo-monomeric) derivative of
DsRed (Shaner et al., 2004)"
/label=tdTomato
/note="mammalian codon-optimized"
/translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
RSEGRHHLFLYGMDELYK"
misc_feature 7756..8344
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(7809..7829)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(8227..8238)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 8431..8664
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 8736..8857
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(8773..8792)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 8827..8846
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 8876..9011
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(9031..9050)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(9032..9050)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(9060..9077)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(9060..9076)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(9069..9091)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 9218..9646
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(9305..9324)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 9515..9536
/label=F1ori-F
/note="F1 origin, forward primer"
ORIGIN
1 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
61 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
121 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
181 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
241 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
301 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
361 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
421 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
481 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
541 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
601 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
661 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
721 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
781 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
841 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
901 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
961 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
1021 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
1081 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
1141 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
1201 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
1261 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
1321 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
1381 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
1441 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
1501 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
1561 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
1621 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
1681 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
1741 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
1801 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
1861 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
1921 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
1981 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
2041 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
2101 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
2161 cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag ctggagctgc aagcttggcc
2221 attgcatacg ttgtatccat atcataatat gtacatttat attggctcat gtccaacatt
2281 accgccatgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
2341 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
2401 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
2461 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
2521 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
2581 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
2641 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
2701 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
2761 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
2821 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctcgttt
2881 agtgaaccgg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
2941 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
3001 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
3061 atctctagca gtggcgcccg aacagggact tgaaagcgaa agggaaacca gaggagctct
3121 ctcgacgcag gactcggctt gctgaagcgc gcacggcaag aggcgagggg cggcgactgg
3181 tgagtacgcc aaaaattttg actagcggag gctagaagga gagagatggg tgcgagagcg
3241 tcagtattaa gcgggggaga attagatcgc gatgggaaaa aattcggtta aggccagggg
3301 gaaagaaaaa atataaatta aaacatatag tatgggcaag cagggagcta gaacgattcg
3361 cagttaatcc tggcctgtta gaaacatcag aaggctgtag acaaatactg ggacagctac
3421 aaccatccct tcagacagga tcagaagaac ttagatcatt atataataca gtagcaaccc
3481 tctattgtgt gcatcaaagg atagagataa aagacaccaa ggaagcttta gacaagatag
3541 aggaagagca aaacaaaagt aagaccaccg cacagcaagc ggccgctgat cttcagacct
3601 ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa
3661 attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa
3721 agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg
3781 ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag
3841 cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc
3901 tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa
3961 cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg
4021 aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag
4081 tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa
4141 aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg
4201 aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga
4261 ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag
4321 ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc
4381 gaaggaatag aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac
4441 ggatctcgac ggtatcggtt aacttttaaa agaaaagggg ggattggggg gtacagtgca
4501 ggggaaagaa tagtagacat aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa
4561 attacaaaaa ttcaaaattt tatcgatcac gagactagcc tcgagaagct tgattaaccc
4621 gtgtcggctc cagatctggc ctccgcgccg ggttttggcg cctcccgcgg gcgcccccct
4681 cctcacggcg agcgctgcca cgtcagacga agggcgcagc gagcgtcctg atccttccgc
4741 ccggacgctc aggacagcgg cccgctgctc ataagactcg gccttagaac cccagtatca
4801 gcagaaggac attttaggac gggacttggg tgactctagg gcactggttt tctttccaga
4861 gagcggaaca ggcgaggaaa agtagtccct tctcggcgat tctgcggagg gatctccgtg
4921 gggcggtgaa cgccgatgat tatataagga cgcgccgggt gtggcacagc tagttccgtc
4981 gcagccggga tttgggtcgc ggttcttgtt tgtggatcgc tgtgatcgtc acttggtgag
5041 tagcgggctg ctgggctggc cggggctttc gtggccgccg ggccgctcgg tgggacggaa
5101 gcgtgtggag agaccgccaa gggctgtagt ctgggtccgc gagcaaggtt gccctgaact
5161 gggggttggg gggagcgcag caaaatggcg gctgttcccg agtcttgaat ggaagacgct
5221 tgtgaggcgg gctgtgaggt cgttgaaaca aggtgggggg catggtgggc ggcaagaacc
5281 caaggtcttg aggccttcgc taatgcggga aagctcttat tcgggtgaga tgggctgggg
5341 caccatctgg ggaccctgac gtgaagtttg tcactgactg gagaactcgg tttgtcgtct
5401 gttgcggggg cggcagttat ggcggtgccg ttgggcagtg cacccgtacc tttgggagcg
5461 cgcgccctcg tcgtgtcgtg acgtcacccg ttctgttggc ttataatgca gggtggggcc
5521 acctgccggt aggtgtgcgg taggcttttc tccgtcgcag gacgcagggt tcgggcctag
5581 ggtaggctct cctgaatcga caggcgccgg acctctggtg aggggaggga taagtgaggc
5641 gtcagtttct ttggtcggtt ttatgtacct atcttcttaa gtagctgaag ctccggtttt
5701 gaactatgcg ctcggggttg gcgagtgtgt tttgtgaagt tttttaggca ccttttgaaa
5761 tgtaatcatt tgggtcaata tgtaattttc agtgttagac tagtaaattg tccgctaaat
5821 tctggccgtt tttggctttt ttgttagacg aagcttgggc tgcaggtcga gatccaccgg
5881 tcgccaccat ggtgagcaag ggcgaggagg tcatcaaaga gttcatgcgc ttcaaggtgc
5941 gcatggaggg ctccatgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
6001 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggcggcccc ctgcccttcg
6061 cctgggacat cctgtccccc cagttcatgt acggctccaa ggcgtacgtg aagcaccccg
6121 ccgacatccc cgattacaag aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga
6181 tgaacttcga ggacggcggt ctggtgaccg tgacccagga ctcctccctg caggacggca
6241 cgctgatcta caaggtgaag atgcgcggca ccaacttccc ccccgacggc cccgtaatgc
6301 agaagaagac catgggctgg gaggcctcca ccgagcgcct gtacccccgc gacggcgtgc
6361 tgaagggcga gatccaccag gccctgaagc tgaaggacgg cggccactac ctggtggagt
6421 tcaagaccat ctacatggcc aagaagcccg tgcaactgcc cggctactac tacgtggaca
6481 ccaagctgga catcacctcc cacaacgagg actacaccat cgtggaacag tacgagcgct
6541 ccgagggccg ccaccacctg ttcctggggc atggcaccgg cagcaccggc agcggcagct
6601 ccggcaccgc ctcctccgag gacaacaaca tggccgtcat caaagagttc atgcgcttca
6661 aggtgcgcat ggagggctcc atgaacggcc acgagttcga gatcgagggc gagggcgagg
6721 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggc ggccccctgc
6781 ccttcgcctg ggacatcctg tccccccagt tcatgtacgg ctccaaggcg tacgtgaagc
6841 accccgccga catccccgat tacaagaagc tgtccttccc cgagggcttc aagtgggagc
6901 gcgtgatgaa cttcgaggac ggcggtctgg tgaccgtgac ccaggactcc tccctgcagg
6961 acggcacgct gatctacaag gtgaagatgc gcggcaccaa cttccccccc gacggccccg
7021 taatgcagaa gaagaccatg ggctgggagg cctccaccga gcgcctgtac ccccgcgacg
7081 gcgtgctgaa gggcgagatc caccaggccc tgaagctgaa ggacggcggc cactacctgg
7141 tggagttcaa gaccatctac atggccaaga agcccgtgca actgcccggc tactactacg
7201 tggacaccaa gctggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg
7261 agcgctccga gggccgccac cacctgttcc tgtacggcat ggacgagctg tacaagtagc
7321 gtagccatcc aggctgtgct gtccctgtat gcctctggtc gtaccacagg cattgtgatg
7381 gactccggag acggggtcac ccacactgtg cccatctacg agggctatgc tctccctcac
7441 gccatcctgc gtctggacct ggctggccgg gacctgacag actacctcat gaagatcctg
7501 accgagcgtg gctacagctt caccaccaca gctgagaggg aaatcgtgcg tgacatcaaa
7561 gagaagctgt gctatgttgc tctagacttc gagcaggaga tggccactgc cgcatcctct
7621 tcctccctgg agaagagcta tgagctgcct gacggccagg tcatcactat tggcaacgag
7681 cggttccgat gccctgaggc tcttttccag ccttccttct tgggtatgga atcctgtggc
7741 agcggccgcg tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt
7801 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
7861 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
7921 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
7981 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
8041 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
8101 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc
8161 tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
8221 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
8281 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
8341 ccgcctggaa ttcgagctcg gtacctttaa gaccaatgac ttacaaggca gctgtagatc
8401 ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac
8461 aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc
8521 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
8581 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
8641 agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat
8701 aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg
8761 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
8821 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc yrscyatccc
8881 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
8941 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
9001 ttttggaggc ctagggacgt acccaattcg ccctatagtg agtcgtatta cgcgcgctca
9061 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
9121 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
9181 ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta
9241 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
9301 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
9361 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
9421 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
9481 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
9541 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
9601 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
9661 acgcttacaa ttt
//
| {
"Vector backbone": "pHAGE",
"Backbone size w/o insert (bp)": "7838",
"Vector type": "Lentiviral",
"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 ; Takara Bio Limited Use Label License (formerly Clontech)",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Global analysis of contact-dependent human-to-mouse intercellular mRNA and lncRNA transfer in cell culture",
"Publication": "Elife. 2023 May 30;12:e83584. doi: 10.7554/eLife.83584.",
"DOI": "10.7554/eLife.83584.",
"Authors": "Dasgupta S, Dayagi DY, Haimovich G, Wyler E, Olender T, Singer RH, Landthaler M, Gerst JE /eLife",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/37249209",
"PubMed ID": "PubMed 37249209"
} | {
"Gene/Insert name": "tdTomato with fragment (nucleotides 398-818) of mouse Actb coding dsequence",
"Species": "M. musculus (mouse)",
"Insert Size (bp)": "1852",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "UbiC",
"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": ""
} |
|
Eomes gRNA#1 | 170834 | Cas9-mediated knockout of Eomes in mammalian cells | Takeshi Imai | Aihara et al Cell Rep. 2021 Jun 22;35(12):109276. doi: 10.1016/j.celrep.2021.109276. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/341126/4e560247-42f1-48a0-9966-9d039b026226/addgene-plasmid-170834-sequence-341126.gbk | full | LOCUS sequence-341126- 2812 bp DNA circular SYN 26-AUG-2024
DEFINITION Cas9-mediated knockout of Eomes in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2812)
AUTHORS Aihara S, Fujimoto S, Sakaguchi R, Imai T
TITLE BMPR-2 gates activity-dependent stabilization of primary dendrites
during mitral cell remodeling.
JOURNAL Cell Rep. 2021 Jun 22;35(12):109276. doi:
10.1016/j.celrep.2021.109276.
PUBMED 34161760
REFERENCE 2 (bases 1 to 2812)
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: "2021-06-22"; volume: "35(12):109276. doi"; pages: "
10.1016/j.celrep.2021.109276"
FEATURES Location/Qualifiers
source 1..2812
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 72..320
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA
(Domitrovich & Kunkel, 2003)"
primer_bind 72..92
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 243..262
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
misc_RNA 341..416
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
protein_bind complement(428..452)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
misc_feature 658..714
/label=MCS
/note="pUC18/19 multiple cloning site"
primer_bind complement(727..743)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(727..743)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(740..762)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 751..767
/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(775..805)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 820..841
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(1129..1717)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1209..1228)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(1888..2748)
/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 2511..2530
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(join(2749..2812,1..41))
/gene="bla"
/label=AmpR promoter
ORIGIN
1 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
61 ccacctgacg tgagggccta tttcccatga ttccttcata tttgcatata cgatacaagg
121 ctgttagaga gataattaga attaatttga ctgtaaacac aaagatatta gtacaaaata
181 cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa
241 tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct ttatatatct
301 tgtggaaagg acgaaacacc gggcgcgcac ttctactcgc gttttagagc tagaaatagc
361 aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt
421 tttctagacc cagctttctt gtacaaagtt ggcattagat ccgctgcatt aatgaatcgg
481 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
541 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
601 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagaa
661 ttcgagctcg gtacccgggg atcctctaga gtcgacctgc aggcatgcaa gcttggcgta
721 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
781 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
841 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
901 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
961 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
1021 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
1081 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
1141 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
1201 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
1261 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
1321 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
1381 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
1441 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
1501 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
1561 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
1621 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
1681 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
1741 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
1801 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
1861 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
1921 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
1981 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
2041 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
2101 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
2161 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
2221 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
2281 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
2341 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
2401 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
2461 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
2521 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
2581 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
2641 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
2701 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
2761 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca ta
//
| {
"Vector backbone": "gRNA backbone",
"Backbone size w/o insert (bp)": "",
"Vector type": "Mammalian Expression, CRISPR",
"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": "BMPR-2 gates activity-dependent stabilization of primary dendrites during mitral cell remodeling",
"Publication": "Cell Rep. 2021 Jun 22;35(12):109276. doi: 10.1016/j.celrep.2021.109276.",
"DOI": "10.1016/j.celrep.2021.109276.",
"Authors": "Aihara S, Fujimoto S, Sakaguchi R, Imai T /jcelrep",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/34161760",
"PubMed ID": "PubMed 34161760"
} | {
"Gene/Insert name": "Eomes",
"Species": "M. musculus (mouse)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "Eomes",
"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": ""
} |
|
EP300 | 74658 | Bacterial expression for structure determination; may not be full ORF | Nicola Burgess-Brown | SGC 2016 plasmids (unpublished) | https://media.addgene.org/snapgene-media/v2.0.0/sequences/142516/dc0213d9-d4bd-415e-ba53-331826fc4db0/addgene-plasmid-74658-sequence-142516.gbk | partial | LOCUS sequence-142516- 799 bp DNA linear UNA 26-AUG-2024
DEFINITION Bacterial expression for structure determination; may not be full
ORF.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE natural DNA sequence
ORGANISM unspecified
REFERENCE 1 (bases 1 to 799)
TITLE SGC 2016 plasmids
REFERENCE 2 (bases 1 to 799)
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..799
/mol_type="genomic DNA"
/organism="unspecified"
CDS 26..43
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 68..88
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQS"
CDS 499..516
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(569..587)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 583..630
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
primer_bind complement(754..773)
/label=F1ori-R
/note="F1 origin, reverse primer"
ORIGIN
1 actttaagaa ggagatatac atatgcacca tcatcatcat cattcttctg gtgtagatct
61 gggtaccgag aacctgtact tccaatccat gattttcaaa ccggaagaac tgcgtcaggc
121 cctgatgccg accctggaag ccctgtatcg tcaggatccg gaaagcctgc cgtttcgcca
181 gccggtggat ccgcagctgc tgggtatccc ggattatttt gatattgtta aaagcccgat
241 ggatctgagc accattaaac gtaaactgga taccggccag tatcaggaac cgtggcagta
301 tgttgatgat atttggctga tgtttaataa tgcgtggctg tataatcgca aaaccagccg
361 cgtttataaa tattgtagca aactgagcga agtgtttgaa caggaaattg atccggtgat
421 gcagagcctg ggctgacagt aaaggtggat acggatccga attcgagctc cgtcgacaag
481 cttgcggccg cactcgagca ccaccaccac caccactgag atccggctgc taacaaagcc
541 cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg
601 gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc cggattggcg
661 aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
721 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
781 tcgccacgtt cgccggctt
//
| {
"Vector backbone": "pNIC28-Bsa4",
"Backbone size w/o insert (bp)": "",
"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": "",
"Publication": "",
"DOI": "",
"Authors": "",
"PubMed Link": "",
"PubMed ID": ""
} | {
"Gene/Insert name": "EP300",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "439",
"Mutation": "",
"Entrez Gene": "EP300",
"Promoter": "T7",
"Tag / Fusion Protein": "His-6-TEV (N terminal on backbone)"
} | {
"Cloning method": "Ligation Independent Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "T7",
"3′ sequencing primer": "T7term"
} | {
"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": ""
} |
|
pAWp7-5 | 72737 | pFUGW-UBCp-RFP-CMVp-GFP-[miR-128 + miR-132] | Timothy Lu | Wong et al Nat Biotechnol. 2015 Sep;33(9):952-61. doi: 10.1038/nbt.3326. Epub 2015 Aug 17. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/133520/d18282d1-9aa2-4eaa-86cf-23610e1e46ea/addgene-plasmid-72737-sequence-133520.gbk | full | LOCUS sequence-133520- 11961 bp DNA circular SYN 26-AUG-2024
DEFINITION pFUGW-UBCp-RFP-CMVp-GFP-[miR-128 + miR-132].
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11961)
AUTHORS Wong AS, Choi GC, Cheng AA, Purcell O, Lu TK
TITLE Massively parallel high-order combinatorial genetics in human cells.
JOURNAL Nat Biotechnol. 2015 Sep;33(9):952-61. doi: 10.1038/nbt.3326. Epub
2015 Aug 17.
PUBMED 26280411
REFERENCE 2 (bases 1 to 11961)
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.3326";
journalName: "Nat Biotechnol"; date: "2015-09"; volume: "33"; issue:
"9"; pages: "952-61"
FEATURES Location/Qualifiers
source 1..11961
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 20..608
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(73..93)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(491..502)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(611..627)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(612..628)
/label=pBluescriptKS
/note="For pBluescript vector"
LTR 1133..1313
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
primer_bind complement(1339..1356)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 1345..1569
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 1615..2043
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1702..1721)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 1912..1933
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2052..2072)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2057..2386
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2237..2372
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2299..2318
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter 2434..2481
/label=EM7 promoter
/note="synthetic bacterial promoter "
CDS 2500..2874
/codon_start=1
/gene="Sh ble from Streptoalloteichus hindustanus"
/product="antibiotic-binding protein"
/label=BleoR
/note="confers resistance to bleomycin, phleomycin, and
Zeocin(TM)"
/translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
EFALRDPAGNCVHFVAEEQD"
polyA_signal 3004..3125
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3041..3060)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3095..3114
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(3174..3190)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3174..3190)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3187..3209)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3198..3214
/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(3222..3252)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3267..3288
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(3405..3422)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3576..4164)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3656..3675)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4335..5195)
/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 4958..4977
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5196..5300)
/gene="bla"
/label=AmpR promoter
primer_bind complement(5375..5394)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 5566..5945
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 5947..6145
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
LTR 6163..6343
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 6390..6515
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 7008..7241
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 7426..7470
/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 7768..7885
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 7939..9150
/label=UbC promoter
/note="human ubiquitin C promoter"
primer_bind 8987..9006
/label=hUBCpro-F
/note="Human Ubiquitin C (UbC) promoter, forward primer"
CDS 9167..9862
/codon_start=1
/product="red fluorescent protein from Entacmaea
quadricolor"
/label=TurboRFP
/note="mammalian codon-optimized"
/translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMKIKV
VEGGPLPFAFDILATSFMYGSKAFINHTQGIPDFFKQSFPEGFTWERITTYEDGGVLTA
TQDTSFQNGCIIYNVKINGVNFPSNGPVMQKKTRGWEANTEMLYPADGGLRGHSQMALK
LVGGGYLHCSFKTTYRSKKPAKNLKMPGFHFVDHRLERIKEADKETYVEQHEMAVAKYC
DLPSKLGHR"
enhancer 9893..10272
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 10273..10476
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
regulatory 10495..10504
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 10501..11220
/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 10501..11217
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(10546..10567)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(10807..10826)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 11154..11175
/label=EGFP-C
/note="EGFP, forward primer"
ORIGIN
1 cgatatcaag cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg
61 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
121 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
181 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
241 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
301 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
361 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc
421 gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
481 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
541 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
601 ctccccgcat cgataccgtc gacctcgaga cctagaaaaa catggagcaa tcacaagtag
661 caatacagca gctaccaatg ctgattgtgc ctggctagaa gcacaagagg aggaggaggt
721 gggttttcca gtcacacctc aggtaccttt aagaccaatg acttacaagg cagctgtaga
781 tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag
841 acaagatatc cttgatctgt ggatctacca cacacaaggc tacttccctg attggcagaa
901 ctacacacca gggccaggga tcagatatcc actgaccttt ggatggtgct acaagctagt
961 accagttgag caagagaagg tagaagaagc caatgaagga gagaacaccc gcttgttaca
1021 ccctgtgagc ctgcatggga tggatgaccc ggagagagaa gtattagagt ggaggtttga
1081 cagccgccta gcatttcatc acatggcccg agagctgcat ccggactgta ctgggtctct
1141 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa
1201 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
1261 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagggcccg
1321 tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
1381 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
1441 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
1501 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
1561 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
1621 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
1681 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
1741 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
1801 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
1861 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
1921 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
1981 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
2041 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
2101 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
2161 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
2221 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
2281 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
2341 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
2401 ttgtatatcc attttcggat ctgatcagca cgtgttgaca attaatcatc ggcatagtat
2461 atcggcatag tataatacga caaggtgagg aactaaacca tggccaagtt gaccagtgcc
2521 gttccggtgc tcaccgcgcg cgacgtcgcc ggagcggtcg agttctggac cgaccggctc
2581 gggttctccc gggacttcgt ggaggacgac ttcgccggtg tggtccggga cgacgtgacc
2641 ctgttcatca gcgcggtcca ggaccaggtg gtgccggaca acaccctggc ctgggtgtgg
2701 gtgcgcggcc tggacgagct gtacgccgag tggtcggagg tcgtgtccac gaacttccgg
2761 gacgcctccg ggccggccat gaccgagatc ggcgagcagc cgtgggggcg ggagttcgcc
2821 ctgcgcgacc cggccggcaa ctgcgtgcac ttcgtggccg aggagcagga ctgacacgtg
2881 ctacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
2941 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
3001 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
3061 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
3121 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
3181 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
3241 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
3301 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
3361 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
3421 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
3481 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
3541 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
3601 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
3661 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
3721 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
3781 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
3841 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
3901 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
3961 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
4021 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
4081 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
4141 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
4201 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
4261 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
4321 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
4381 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
4441 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
4501 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
4561 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
4621 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
4681 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
4741 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
4801 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
4861 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
4921 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
4981 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
5041 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
5101 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
5161 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
5221 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
5281 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg
5341 ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
5401 taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa
5461 tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta
5521 ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga
5581 ctagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
5641 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
5701 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
5761 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
5821 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
5881 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
5941 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
6001 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
6061 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
6121 tacggtggga ggtctatata agcagcgcgt tttgcctgta ctgggtctct ctggttagac
6181 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
6241 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
6301 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg
6361 acttgaaagc gaaagggaaa ccagaggagc tctctcgacg caggactcgg cttgctgaag
6421 cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt ttgactagcg
6481 gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcggggg agaattagat
6541 cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa ttaaaacata
6601 tagtatgggc aagcagggag ctagaacgat tcgcagttaa tcctggcctg ttagaaacat
6661 cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca ggatcagaag
6721 aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa aggatagaga
6781 taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa agtaagacca
6841 ccgcacagca agcggccgct gatcttcaga cctggaggag gagatatgag ggacaattgg
6901 agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt agcacccacc
6961 aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg agctttgttc
7021 cttgggttct tgggagcagc aggaagcact atgggcgcag cgtcaatgac gctgacggta
7081 caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct gagggctatt
7141 gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct ccaggcaaga
7201 atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg gggttgctct
7261 ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa taaatctctg
7321 gaacagattt ggaatcacac gacctggatg gagtgggaca gagaaattaa caattacaca
7381 agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa
7441 ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac aaattggctg
7501 tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag aatagttttt
7561 gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc gtttcagacc
7621 cacctcccaa ccccgagggg acccgacagg cccgaaggaa tagaagaaga aggtggagag
7681 agagacagag acagatccat tcgattagtg aacggatcgg cactgcgtgc gccaattctg
7741 cagacaaatg gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag
7801 tgcaggggaa agaatagtag acataatagc aacagacata caaactaaag aattacaaaa
7861 acaaattaca aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg
7921 gttaattaag ggtgcagcgg cctccgcgcc gggttttggc gcctcccgcg ggcgcccccc
7981 tcctcacggc gagcgctgcc acgtcagacg aagggcgcag cgagcgtcct gatccttccg
8041 cccggacgct caggacagcg gcccgctgct cataagactc ggccttagaa ccccagtatc
8101 agcagaagga cattttagga cgggacttgg gtgactctag ggcactggtt ttctttccag
8161 agagcggaac aggcgaggaa aagtagtccc ttctcggcga ttctgcggag ggatctccgt
8221 ggggcggtga acgccgatga ttatataagg acgcgccggg tgtggcacag ctagttccgt
8281 cgcagccggg atttgggtcg cggttcttgt ttgtggatcg ctgtgatcgt cacttggtga
8341 gtagcgggct gctgggctgg ccggggcttt cgtggccgcc gggccgctcg gtgggacgga
8401 agcgtgtgga gagaccgcca agggctgtag tctgggtccg cgagcaaggt tgccctgaac
8461 tgggggttgg ggggagcgca gcaaaatggc ggctgttccc gagtcttgaa tggaagacgc
8521 ttgtgaggcg ggctgtgagg tcgttgaaac aaggtggggg gcatggtggg cggcaagaac
8581 ccaaggtctt gaggccttcg ctaatgcggg aaagctctta ttcgggtgag atgggctggg
8641 gcaccatctg gggaccctga cgtgaagttt gtcactgact ggagaactcg gtttgtcgtc
8701 tgttgcgggg gcggcagtta tggcggtgcc gttgggcagt gcacccgtac ctttgggagc
8761 gcgcgccctc gtcgtgtcgt gacgtcaccc gttctgttgg cttataatgc agggtggggc
8821 cacctgccgg taggtgtgcg gtaggctttt ctccgtcgca ggacgcaggg ttcgggccta
8881 gggtaggctc tcctgaatcg acaggcgccg gacctctggt gaggggaggg ataagtgagg
8941 cgtcagtttc tttggtcggt tttatgtacc tatcttctta agtagctgaa gctccggttt
9001 tgaactatgc gctcggggtt ggcgagtgtg ttttgtgaag ttttttaggc accttttgaa
9061 atgtaatcat ttgggtcaat atgtaatttt cagtgttaga ctagtaaatt gtccgctaaa
9121 ttctggccgt ttttggcttt tttgttagac agttaattaa gccaccatga gcgagctgat
9181 caaggagaac atgcacatga agctgtacat ggagggcacc gtgaacaacc accacttcaa
9241 gtgcacatcc gagggcgaag gcaagcccta cgagggcacc cagaccatga agatcaaggt
9301 ggtcgagggc ggccctctcc ccttcgcctt cgacatcctg gctaccagct tcatgtacgg
9361 cagcaaagcc ttcatcaacc acacccaggg catccccgac ttctttaagc agtccttccc
9421 tgagggcttc acatgggaga gaatcaccac atacgaagac gggggcgtgc tgaccgctac
9481 ccaggacacc agcttccaga acggctgcat catctacaac gtcaagatca acggggtgaa
9541 cttcccatcc aacggccctg tgatgcagaa gaaaacacgc ggctgggagg ccaacaccga
9601 gatgctgtac cccgctgacg gcggcctgag aggccacagc cagatggccc tgaagctcgt
9661 gggcgggggc tacctgcact gctccttcaa gaccacatac agatccaaga aacccgctaa
9721 gaacctcaag atgcccggct tccacttcgt ggaccacaga ctggaaagaa tcaaggaggc
9781 cgacaaagag acctacgtcg agcagcacga gatggctgtg gccaagtact gcgacctccc
9841 tagcaaactg gggcacagat aataacctgc aggcgggcca gatatacgcg ttgacattga
9901 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
9961 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
10021 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
10081 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
10141 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
10201 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
10261 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
10321 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
10381 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
10441 aggcgtgtac ggtgggaggt ctatataagc agcggtaacc cccgggtacc ggtcgccacc
10501 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
10561 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
10621 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
10681 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
10741 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
10801 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
10861 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
10921 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
10981 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
11041 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
11101 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
11161 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa
11221 agcggccgcg actctagaga tctctacagg aggatctcat gcatcggatc cgcgaaaagt
11281 ccggaattca tactgtgaag tacactgcat ataaggagtg tggtatagta taaagaaact
11341 ttctgcaggt agtaattata gtgaagattt taggtttaca aagccctagc tgttttctgt
11401 gtagctttta ttattcttat gactcttgac aagtttgtag cttcaccata tacatttaat
11461 attttgcaat aattggcctt gttcctgagc tgttggattc ggggccgtag cactgtctga
11521 gaggtttaca tttctcacag tgaaccggtc tctttttcag ctgcttcctg gcttcttttt
11581 actcaggttt ccactgcttt tttgcttttt ttaatgctgt atgaaggtgt taacatttgt
11641 ttatattttt cattaattgt aataccttta aatcatgcat catactcaga aatagggatt
11701 agaatttaag tgacatcttt ggcccaattc ctagccccgc agacactagc gctacccccg
11761 ccgcccgcga tgctgacgtc agcctgaaag ccccgccccc gcgtctccag ggcaaccgtg
11821 gctttcgatt gttactgtgg gaaccggagg taacagtcta cagccatggt cgccccgcag
11881 cacgcccacg ctccccacca ctcccgagtt ctgccagcct gggtttgggc agatacagag
11941 caagaggagg cggggcaatt g
//
| {
"Vector backbone": "pFUGW",
"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)": "Stbl3",
"Copy number": "Unknown"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Massively parallel high-order combinatorial genetics in human cells",
"Publication": "Nat Biotechnol. 2015 Sep;33(9):952-61. doi: 10.1038/nbt.3326. Epub 2015 Aug 17.",
"DOI": "10.1038/nbt.3326.",
"Authors": "Wong AS, Choi GC, Cheng AA, Purcell O, Lu TK /nbt",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/26280411",
"PubMed ID": "PubMed 26280411"
} | {
"Gene/Insert name": "UBCp-RFP-CMVp-GFP-[miR-128 + miR-132]",
"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": ""
} |
|
REST corepressor 2_R (OZ554) | 28079 | Keith Joung | Maeder et al Mol Cell. 2008 Jul 25. 31(2):294-301. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/34378/8670e901-c286-430a-a701-c1edec1ebd97/addgene-plasmid-28079-sequence-34378.gbk | partial | LOCUS sequence-34378-e 859 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 859)
AUTHORS Maeder ML, Thibodeau-Beganny S, Osiak A, Wright DA, Anthony RM,
Eichtinger M, Jiang T, Foley JE, Winfrey RJ, Townsend JA,
Unger-Wallace E, Sander JD, Muller-Lerch F, Fu F, Pearlberg J,
Goebel C, Dassie JP, Pruett-Miller SM, Porteus MH, Sgroi DC, Iafrate
AJ, Dobbs D, McCray PB, Cathomen T, Voytas DF, Joung JK
TITLE Rapid ""open-source"" engineering of customized zinc-finger
nucleases for highly efficient gene modification.
JOURNAL Mol Cell. 2008 Jul 25. 31(2):294-301.
PUBMED 18657511
REFERENCE 2 (bases 1 to 859)
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.
2008 Jul 25. 31(2):294-301."
FEATURES Location/Qualifiers
source 1..859
/mol_type="genomic DNA"
/organism="unspecified"
CDS 25..48
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
rep_origin 450..830
/direction=RIGHT
/label=M13 ori
/note="M13 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(537..556)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 747..768
/label=F1ori-F
/note="F1 origin, forward primer"
ORIGIN
1 aaaatccaag ctttggcggc cgctgactac aaagacgatg atgacaaatc tagacccggg
61 gagcgcccct tccagtgtcg catttgcatg cggaactttt cgcagaagca gcaccttatg
121 gtccataccc gtactcatac cggtgaaaaa ccgtttcagt gtcggatctg tatgcgaaat
181 ttctccgtga agcacaactt gcacaggcat ctacgtacgc acaccggcga gaagccattc
241 caatgccgaa tatgcatgcg caacttcagt aggcgctggc acctggcgct gcacctaaaa
301 acccacctga ggggatcctc tgcacagtag taactagaga ctagaaaaag gccgacaagt
361 cccgctccgc tgaagatcct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
421 acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc
481 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
541 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
601 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
661 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
721 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
781 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
841 aaaaatgagc tgatacata
//
| {
"Vector backbone": "MG414",
"Backbone size w/o insert (bp)": "5493",
"Vector type": "Zebrafish Targeting",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Kanamycin, 50 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "XL1 Blue",
"Copy number": "Low Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Rapid \"open-source\" engineering of customized zinc-finger nucleases for highly efficient gene modification",
"Publication": "Mol Cell. 2008 Jul 25. 31(2):294-301.",
"DOI": "10.1016/j.molcel.2008.06.016",
"Authors": "Maeder ML, Thibodeau-Beganny S, Osiak A, Wright DA, Anthony RM, Eichtinger M, Jiang T, Foley JE, Winfrey RJ, Townsend JA, Unger-Wallace E, Sander JD, Muller-Lerch F, Fu F, Pearlberg J, Goebel C, Dassie JP, Pruett-Miller SM, Porteus MH, Sgroi DC, Iafrate AJ, Dobbs D, McCray PB, Cathomen T, Voytas DF, Joung JK",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/18657511",
"PubMed ID": "PubMed 18657511"
} | {
"Gene/Insert name": "Zinc finger array targeting REST corepressor 2",
"Species": "D. rerio (zebrafish)",
"Insert Size (bp)": "270",
"Mutation": "",
"Entrez Gene": "rcor2",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "XbaI (not destroyed)",
"3′ cloning site": "BamHI (not destroyed)",
"5′ sequencing primer": "OK.61 GGGTAGTACGATGACGGAACCTGTC",
"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": ""
} |
||
MSCV-ASNS-OE-IRES-Thy1.1 | 192947 | Asparagine synthetase overexpression in murine cells | Ping-Chih Ho | Fernandez-Garcia et al Cell Rep. 2022 Nov 15;41(7):111639. doi: 10.1016/j.celrep.2022.111639. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/380517/fb8dee86-39df-4c4a-aaee-3a9c99a36100/addgene-plasmid-192947-sequence-380517.gbk | full | LOCUS sequence-380517- 7906 bp DNA circular SYN 26-AUG-2024
DEFINITION Asparagine synthetase overexpression in murine cells.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7906)
AUTHORS Fernandez-Garcia J, Franco F, Parik S, Altea-Manzano P, Pane AA,
Broekaert D, van Elsen J, Vermeire I, Schalley T, Planque M, van
Brussel T, Schepers R, Modave E, Karakach TK, Carmeliet P,
Lambrechts D, Ho PC, Fendt SM
TITLE CD8(+) T cell metabolic rewiring defined by scRNA-seq identifies a
critical role of ASNS expression dynamics in T cell differentiation.
JOURNAL Cell Rep. 2022 Nov 15;41(7):111639. doi:
10.1016/j.celrep.2022.111639.
PUBMED 36384124
REFERENCE 2 (bases 1 to 7906)
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: "2022-11-15"; volume: "41(7):111639. doi"; pages: "
10.1016/j.celrep.2022.111639"
FEATURES Location/Qualifiers
source 1..7906
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 211..233
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 361..380
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
LTR 580..1096
/label=5' LTR
/note="5' long terminal repeat from murine embryonic stem
cell virus"
misc_feature 1160..1501
/label=MESV Psi
/note="packaging signal of murine embryonic stem cell
virus"
CDS 1568..1984
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
primer_bind 1914..1936
/label=pLXSN 5'
/note="Murine stem cell virus, forward primer. Also called
MSCV"
primer_bind 1952..1968
/label=pBABE 5'
/note="Psi packaging signal, forward primer for pBABE
vectors"
misc_feature 3720..4286
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
misc_feature 3736..4288
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(3880..3897)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 4107..4126
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 4336..4824
/codon_start=1
/product="mouse GPI-anchored cell surface glycoprotein of
the immunoglobulin superfamily"
/label=Thy1.1
/note="This allele of Thy1 is also known as CD90.1."
/translation="MNPAISVALLLSVLQVSRGQKVTSLTACLVNQNLRLDCRHENNTK
DNSIQHEFSLTREKRKHVLSGTLGIPEHTYRSRVTLSNQPYIKVLTLANFTTKDEGDYF
CELRVSGANPMSSNKSISVYRDKLVKCGGISLLVQNTSWMLLLLLSLSLLQALDFISL"
LTR 4880..5394
/label=3' LTR
/note="3' long terminal repeat from murine embryonic stem
cell virus"
protein_bind 5556..5572
/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(5580..5610)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5625..5646
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(5763..5780)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5934..6522)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6014..6033)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6693..7553)
/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 7316..7335
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7554..7658)
/gene="bla"
/label=AmpR promoter
primer_bind 7726..7744
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(7782..7804)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind join(7904..7906,1..17)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
61 taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag
121 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
181 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcgc
241 aaggaatggt gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac
301 catacccacg ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc
361 ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac
421 gatgcgtccg gcgtagaggc gattagtcca atttgttaaa gacaggatat cagtggtcca
481 ggctctagtt ttgactcaac aatatcacca gctgaagcct atagagtacg agccatagat
541 aaaataaaag attttattta gtctccagaa aaagggggga atgaaagacc ccacctgtag
601 gtttggcaag ctagcttaag taacgccatt ttgcaaggca tggaaaatac ataactgaga
661 atagagaagt tcagatcaag gttaggaaca gagagacagc agaatatggg ccaaacagga
721 tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatggt ccccagatgc
781 ggtcccgccc tcagcagttt ctagagaacc atcagatgtt tccagggtgc cccaaggacc
841 tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc
901 gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg cgcgccagtc
961 ctccgataga ctgcgtcgcc cgggtacccg tattcccaat aaagcctctt gctgtttgca
1021 tccgaatcgt ggactcgctg atccttggga gggtctcctc agattgattg actgcccacc
1081 tcgggggtct ttcatttgga ggttccaccg agatttggag acccctgccc agggaccacc
1141 gacccccccg ccgggaggta agctggccag cggtcgtttc gtgtctgtct ctgtctttgt
1201 gcgtgtttgt gccggcatct aatgtttgcg cctgcgtctg tactagttag ctaactagct
1261 ctgtatctgg cggacccgtg gtggaactga cgagttctga acacccggcc gcaaccctgg
1321 gagacgtccc agggactttg ggggccgttt ttgtggcccg acctgaggaa gggagtcgat
1381 gtggaatccg accccgtcag gatatgtggt tctggtagga gacgagaacc taaaacagtt
1441 cccgcctccg tctgaatttt tgctttcggt ttggaaccga agccgcgcgt cttgtctgct
1501 gcagcgctgc agcatcgttc tgtgttgtct ctgtctgact gtgtttctgt atttgtctga
1561 aaattagggc cagactgtta ccactccctt aagtttgacc ttaggtcact ggaaagatgt
1621 cgagcggatc gctcacaacc agtcggtaga tgtcaagaag agacgttggg ttaccttctg
1681 ctctgcagaa tggccaacct ttaacgtcgg atggccgcga gacggcacct ttaaccgaga
1741 cctcatcacc caggttaaga tcaaggtctt ttcacctggc ccgcatggac acccagacca
1801 ggtcccctac atcgtgacct gggaagcctt ggcttttgac ccccctccct gggtcaagcc
1861 ctttgtacac cctaagcctc cgcctcctct tcctccatcc gccccgtctc tcccccttga
1921 acctcctcgt tcgaccccgc ctcgatcctc cctttatcca gccctcactc cttctctagg
1981 cgccggaatt agatctgcca ccatgtgtgg catctgggcc ctcttcggca gcgatgactg
2041 cctttccgtg cagtgtctga gtgcgatgaa gatcgcgcac agggggccag atgcatttcg
2101 ctttgagaat gtcaatggat acaccaactg ctgctttggc tttcaccgct tggctgtggt
2161 tgacccgctg tttggaatgc agccgataag agtgaggaaa tacccttatt tgtggctctg
2221 ttacaatggt gaaatctaca accacaaggc gctacagcaa cgttttgaat ttgaatatca
2281 gaccaatgtg gatggtgaga ttatcctcca cctctatgac aaaggaggca tcgagaaaac
2341 catctgtatg ctggacgggg tgtttgcatt catcttactg gacactgcca ataagaaagt
2401 atttctgggc agagacacct atggagtcag gcccttgttt aaagccatga cagaagatgg
2461 gtttctggct gtgtgttcag aagctaaagg ccttgtttcc ttaaaacact ccaccactcc
2521 cttcttaaaa gtggagccct tccttcctgg acactatgaa gttttggatt taaaaccaaa
2581 tggcaaagtt gcgtctgtgg aaatggtcaa ataccatcac tgtacggatg aaccattgca
2641 tgccatctat gacagcgtgg agaaactctt cccaggcttt gacctagaga ccgtgaagaa
2701 caatctgcgt atcctttttg acaacgctat caagaaacgc ttgatgacag accggaggat
2761 tggctgcctt ttatcagggg gcctggactc gagcttggtt gctgcctctc tgctgaagca
2821 actcaaggag gcccaagttc agtatcctct ccagacattt gctattggca tggaggacag
2881 ccccgatctc ctggccgcta gaaaggtggc aaattatatt ggaagcgagc atcatgaagt
2941 cctttttaac tctgaagaag gcattcaggc cctggatgaa gtcatatttt ccttggaaac
3001 ttatgatatt acgacagttc gggcatctgt gggcatgtat ttaatttcca agtatattcg
3061 gaagaacaca gacagcgtgg tgatcttctc cggagagggg tcagatgaac ttacacaggg
3121 ctatatatat ttccacaagg ctccttcccc tgagaaggcc gaggaggaga gtgagagact
3181 gctgaaggaa ctctacctgt ttgatgttct ccgggccgac cgcactactg ctgcacacgg
3241 tctcgaactg agagtcccct ttctggatca tcggttttct tcctattacc tgtctctgcc
3301 gccagatatg agaattccaa aaaatggcat agaaaaacat ctcctgagag agacttttga
3361 ggactgcaac ctgctaccca aagagattct ctggcgaccc aaagaagcct tcagtgatgg
3421 gatcacctca gtcaagaact cctggttcaa gattttgcag gactatgttg aacatcaggt
3481 tgatgatgag atgatgtctg cagcctccca gaagtttccc ttcaatactc ccaaaactaa
3541 ggaaggctac ttctaccgtc agatctttga acgccattac ccaggccggg ctgattggct
3601 gactcattat tggatgccta agtggatcaa tgctactgac ccttctgccc gcactctgac
3661 ccattataag tcagctgcca aagcttaggt cgacggtatc gataagcttg atatcgaatt
3721 ccgccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg
3781 tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg
3841 gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag
3901 gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt
3961 ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc
4021 aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga
4081 gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga
4141 aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tgcacatgct
4201 ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg
4261 ttttcctttg aaaaacacga tgataatatg gccacaacca tgggtctttt ctgcagtcac
4321 cgtcctcgag gcaccatgaa cccagccatc agcgtcgctc tcctgctctc agtcttgcag
4381 gtgtcccgag ggcagaaggt gaccagcctg acagcctgcc tggtgaacca aaaccttcgc
4441 ctggactgcc gccatgagaa taacaccaag gataactcca tccagcatga gttcagcctg
4501 acccgagaga agaggaagca cgtgctctca ggcacccttg ggatacccga gcacacgtac
4561 cgctcccgcg tcaccctctc caaccagccc tatatcaagg tccttaccct agccaacttc
4621 accaccaagg atgagggcga ctacttttgt gagcttcgcg tctcgggcgc gaatcccatg
4681 agctccaata aaagtatcag tgtgtataga gacaagctgg tcaagtgtgg cggcataagc
4741 ctgctggttc agaacacatc ctggatgctg ctgctgctgc tttccctctc cctcctccaa
4801 gccctggact tcatttctct gtgacgcgta gaattcgata aaataaaaga ttttatttag
4861 tctccagaaa aaggggggaa tgaaagaccc cacctgtagg tttggcaagc tagcttaagt
4921 aacgccattt tgcaaggcat ggaaaataca taactgagaa tagagaagtt cagatcaagg
4981 ttaggaacag agagacagca gaatatgggc caaacaggat atctgtggta agcagttcct
5041 gccccggctc agggccaaga acagatggtc cccagatgcg gtcccgccct cagcagtttc
5101 tagagaacca tcagatgttt ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat
5161 ttgaactaac caatcagttc gcttctcgct tctgttcgcg cgcttctgct ccccgagctc
5221 aataaaagag cccacaaccc ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc
5281 gggtacccgt gtatccaata aaccctcttg cagttgcatc cgacttgtgg tctcgctgtt
5341 ccttgggagg gtctcctctg agtgattgac tacccgtcag cgggggtctt tcatgggtaa
5401 cagtttcttg aagttggaga acaacattct gagggtagga gtcgaatatt aagtaatcct
5461 gactcaatta gccactgttt tgaatccaca tactccaata ctcctgaaat agttcattat
5521 ggacagcgca gaaagagctg gggagaattg tgaaattgtt atccgctcac aattccacac
5581 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
5641 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
5701 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
5761 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
5821 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
5881 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
5941 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
6001 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
6061 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
6121 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
6181 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
6241 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
6301 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
6361 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
6421 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
6481 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
6541 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
6601 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
6661 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
6721 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
6781 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
6841 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
6901 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
6961 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
7021 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
7081 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
7141 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
7201 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
7261 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
7321 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
7381 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
7441 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
7501 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
7561 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
7621 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
7681 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
7741 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
7801 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
7861 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcgg
//
| {
"Vector backbone": "MSCV-IRES-Thy1.1",
"Backbone size w/o insert (bp)": "6221",
"Vector type": "Mammalian Expression, Retroviral",
"Selectable markers": "Thy1.1"
} | {
"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": "CD8(+) T cell metabolic rewiring defined by scRNA-seq identifies a critical role of ASNS expression dynamics in T cell differentiation",
"Publication": "Cell Rep. 2022 Nov 15;41(7):111639. doi: 10.1016/j.celrep.2022.111639.",
"DOI": "10.1016/j.celrep.2022.111639.",
"Authors": "Fernandez-Garcia J, Franco F, Parik S, Altea-Manzano P, Pane AA, Broekaert D, van Elsen J, Vermeire I, Schalley T, Planque M, van Brussel T, Schepers R, Modave E, Karakach TK, Carmeliet P, Lambrechts D, Ho PC, Fendt SM /jcelrep",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/36384124",
"PubMed ID": "PubMed 36384124"
} | {
"Gene/Insert name": "Asparagine synthetase",
"Species": "M. musculus (mouse)",
"Insert Size (bp)": "1686",
"Mutation": "",
"Entrez Gene": "Asns",
"Promoter": "5'LTR",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "BglII (not destroyed)",
"3′ cloning site": "SalI (not destroyed)",
"5′ sequencing primer": "GCCCTTTGTACACCCTAAGCCTC",
"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": ""
} |
|
L754 | 1686 | (Empty Backbone) | Andrew Fire | Fire Lab C. Elegans Vector Kit 1999 (unpublished) | https://media.addgene.org/snapgene-media/v2.0.0/sequences/296/215fd7ca-bdcd-4812-9b70-90becf48efde/addgene-plasmid-1686-sequence-296.gbk | full | LOCUS sequence-296-exp 3410 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 3410)
AUTHORS Fire A et al
TITLE Fire Lab C. Elegans Vector Kit 1999
REFERENCE 2 (bases 1 to 3410)
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..3410
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(984..1006)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 1106..1125
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(1203..1221)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 1289..1393
/gene="bla"
/label=AmpR promoter
CDS 1394..2254
/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(1612..1631)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 2425..3013
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2914..2933
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 3167..3184
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 3301..3322
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 3337..3367
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3375..3391
/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 3380..3402
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind join(3399..3410,1..5)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind join(3399..3410,1..5)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
ORIGIN
1 atgaccatga ttacgccaag cttgcatgcc tgcaggtcga ctctagagga tccccgggat
61 tggccaaagg acccaaaggt atgtttcgaa tgatactaac ataacataga acattttcag
121 gaggaccctt ggctagcgtc gacggtacca tggtattgat atctgagctc cgcatcggcc
181 gctgtcatca gatcgccatc tcgcgcccgt gcctctgact tctaagtcca attactcttc
241 aacatcccta catgctcttt ctccctgtgc tcccaccccc tatttttgtt attatcaaaa
301 aacttctctt aatttctttg ttttttagct tcttttaagt cacctctaac aatgaaattg
361 tgtagattca aaaatagaat taattcgtaa taaaaagtcg aaaaaaattg tgctccctcc
421 ccccattaat aataattcta tcccaaaatc tacacaatgt tctgtgtaca cttcttatgt
481 tttttacttc tgataaattt ttttgaaaca tcatagaaaa aaccgcacac aaaatacctt
541 atcatatgtt acgtttcagt ttatgaccgc aatttttatt tcttcgcacg tctgggcctc
601 tcatgacgtc aaatcatgct catcgtgaaa aagttttgga gtatttttgg aatttttcaa
661 tcaagtgaaa gtttatgaaa ttaattttcc tgcttttgct ttttggggtt tcccctattg
721 tttgtcaaga tttcgaggac ggcgtttttc ttgctaaaat cacaagtatt gatgagcacg
781 atgcaagaaa gatcggaaga aggtttgggt ttgaggctca gtggaaggtg agtagaagtt
841 gataatttga aagtggagta gtgtctatgg ggtttttgcc ttaaatgaca gaatacattc
901 ccaatatacc aaacataact gtttcctact agtcggccgt acgggccctt tcgtctcgcg
961 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
1021 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
1081 gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
1141 atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gcggccttaa
1201 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
1261 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
1321 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
1381 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
1441 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
1501 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
1561 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
1621 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
1681 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
1741 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
1801 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
1861 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
1921 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
1981 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
2041 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
2101 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
2161 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
2221 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
2281 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
2341 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
2401 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
2461 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
2521 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
2581 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
2641 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
2701 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
2761 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagcattg
2821 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
2881 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
2941 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
3001 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
3061 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
3121 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
3181 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
3241 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
3301 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
3361 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
//
| {
"Vector backbone": "Unknown",
"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": "Unknown"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "",
"Publication": "",
"DOI": "",
"Authors": "",
"PubMed Link": "",
"PubMed ID": ""
} | {
"Gene/Insert name": "None",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": "IVSA, rf2128"
} | {
"Cloning method": "Restriction Enzyme",
"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": ""
} |
|
non-targeting control gRNA (BRDN0001146060) | 80244 | 3rd generation lentiviral gRNA plasmid, non-targeting control gRNA | Doench et al Nat Biotechnol. 2016 Jan 18. doi: 10.1038/nbt.3437. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/150966/d453ef5f-6c76-4742-ba3a-dc9931140d6f/addgene-plasmid-80244-sequence-150966.gbk | full | LOCUS sequence-150966- 8323 bp DNA circular SYN 26-AUG-2024
DEFINITION 3rd generation lentiviral gRNA plasmid, non-targeting control gRNA.
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 cggattcata ctaaacactc tagttttaga 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": "Non-targeting control gRNA",
"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": ""
} |
||
pJOG031 | 105422 | Synthetic biology | Johannes Stuttmann | Gantner et al PLoS One. 2018 May 30;13(5):e0197185. doi: 10.1371/journal.pone.0197185. eCollection 2018. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/207425/982f98df-a78d-4b7f-b01b-1a9b5d3d9a00/addgene-plasmid-105422-sequence-207425.gbk | full | LOCUS sequence-207425- 2915 bp DNA circular SYN 26-AUG-2024
DEFINITION Synthetic biology.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2915)
AUTHORS Gantner J, Ordon J, Ilse T, Kretschmer C, Gruetzner R, Lofke C,
Dagdas Y, Burstenbinder K, Marillonnet S, Stuttmann J
TITLE Peripheral infrastructure vectors and an extended set of plant parts
for the Modular Cloning system.
JOURNAL PLoS One. 2018 May 30;13(5):e0197185. doi:
10.1371/journal.pone.0197185. eCollection 2018.
PUBMED 29847550
REFERENCE 2 (bases 1 to 2915)
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 One.";
date: "2018-05-30"; volume: "13(5):e0197185. doi"; pages: "
10.1371/journal.pone.0197185. eCollection 2018"
FEATURES Location/Qualifiers
source 1..2915
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(24..42)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 110..214
/gene="bla"
/label=AmpR promoter
CDS 436..1227
/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 1398..1986
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 1887..1906
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 2140..2157
/label=L4440
/note="L4440 vector, forward primer"
ORIGIN
1 gtggctcttc agtggacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
61 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
121 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
181 ataaatgctt caataatatt gaaaaaggaa gagtatgcgc tcacgcaact ggtccagaac
241 cttgaccgaa cgcagcggtg gtaacggcgc agtggcggtt ttcatggctt gttatgactg
301 tttttttttg gggtacagtc tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg
361 ggtcgatgtt tgatgttatg gagcagcaac gatgttacgc agcagggcag tcgccctaaa
421 acaaagttaa acatcatgag ggaagcggtg atcgccgaag tatcgactca actatcagag
481 gtagttggcg tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca tttgtacggc
541 tccgcagtgg atggcggcct gaagccacac agcgatattg atttgctggt tacggtgacc
601 gtaaggcttg atgaaacaac gcggcgagct ttgatcaacg accttttgga aacttcggct
661 tcccctggag agagcgagat tctccgcgct gtagaagtca ccattgttgt gcacgacgac
721 atcattccgt ggcgttatcc agctaagcgc gaactgcaat ttggagaatg gcagcgcaat
781 gacattcttg caggtatctt cgagccagcc acgatcgaca ttgatctggc tatcttgctg
841 acaaaagcaa gagaacatag cgttgccttg gtaggtccag cggcggagga actctttgat
901 ccggttcctg aacaggatct atttgaggcg ctaaatgaaa ccttaacgct atggaactcg
961 ccgcccgact gggctggcga tgagcgaaat gtagtgctta cgttgtcccg catttggtac
1021 agcgcagtaa ccggcaaaat cgcgccgaag gatgtcgctg ccgactgggc aatggagcgc
1081 ctgccggccc agtatcagcc cgtcatactt gaagctagac aggcttatct tggacaagaa
1141 gaagatcgct tggcctcgcg cgcagatcag ttggaagaat ttgtccatta cgtaaaaggc
1201 gagatcacca aggtagtcgg caaataactg tcagaccaag tttactcata tatactttag
1261 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
1321 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
1381 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
1441 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
1501 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
1561 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
1621 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
1681 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
1741 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
1801 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
1861 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
1921 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
1981 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
2041 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
2101 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
2161 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatca
2221 ctctgtggtc tcaaatgact cgaatttcaa ccagttcagt aaattccagc ttcagttact
2281 cggcccctgc agaggaagcg caaaaccgcg tttcctcagc ccccaccaac tcctctccac
2341 ctggcaccac aaccgctgtc gcccaggcgt cagaggggca aaaaaagccg ggggcggtat
2401 taagcatgca tgcacagcga ctgcgccaat tgtgtgagag ttcatctcac cagtaccggc
2461 agaacgcgct attagctaaa tcttttgatg ctcaacgcct agacattaac actcaagcgg
2521 gcccttccaa tagccctcac ctgaacgctc tcaacaagct ccaacaacga gatttcaaac
2581 ctgcggctgg cgggctagaa atcccattta cacccaaatc cctattgggt ggaggtaaac
2641 gagtctatca aattggctca tcgtcacgcg acgtacaagt ctgcccacgt ggagctggtg
2701 ctgcattgag gcaagaaatc gaagataagc aactcatggt taacaatctc acggacgagc
2761 ttcaagacgc tatagatgag gccaacccag ccgaaatagc caatacttca caacagctgc
2821 gccatgcacg ggctgacctt gctgacctac agagacgctt tgcggtactc agaaacgaag
2881 atcgcagaat caaccaagct tcgtgagacc acgaa
//
| {
"Vector backbone": "pICH41308 (Engler et al., 2014)",
"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": "Peripheral infrastructure vectors and an extended set of plant parts for the Modular Cloning system",
"Publication": "PLoS One. 2018 May 30;13(5):e0197185. doi: 10.1371/journal.pone.0197185. eCollection 2018.",
"DOI": "10.1371/journal.pone.0197185.",
"Authors": "Gantner J, Ordon J, Ilse T, Kretschmer C, Gruetzner R, Lofke C, Dagdas Y, Burstenbinder K, Marillonnet S, Stuttmann J /journalpone",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/29847550",
"PubMed ID": "PubMed 29847550"
} | {
"Gene/Insert name": "AvrRps4 Pseudomonas syringae pv. pisi)",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "BsaI/ BpiI restriction sites removed",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "BpiI (destroyed during cloning)",
"3′ cloning site": "BpiI (destroyed during cloning)",
"5′ sequencing primer": "CTTTGAGTGAGCTGATACCG",
"3′ sequencing primer": "GGGTTCCGCGCACATTTC"
} | {
"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": ""
} |
|
AAV-mPGK-eGFP (beta-globin intron) | 162513 | AAV vector plasmid expressing enhanced green fluorescent protein (eGFP) under the mouse phosphoglycerate kinase (mPGK) promoter that is upstream of a beta-globin intron | Bart Nieuwenhuis | Nieuwenhuis et al Gene Ther. 2020 Jun 23. pii: 10.1038/s41434-020-0169-1. doi: 10.1038/s41434-020-0169-1. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/379932/03a5e28c-cb41-4a2a-8085-814e3edca075/addgene-plasmid-162513-sequence-379932.gbk | full | LOCUS sequence-379932- 5271 bp DNA circular SYN 26-AUG-2024
DEFINITION AAV vector plasmid expressing enhanced green fluorescent protein
(eGFP) under the mouse phosphoglycerate kinase (mPGK) promoter that
is upstream of a beta-globin intron.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5271)
AUTHORS Nieuwenhuis B, Haenzi B, Hilton S, Carnicer-Lombarte A, Hobo B,
Verhaagen J, Fawcett JW
TITLE Optimization of adeno-associated viral vector-mediated transduction
of the corticospinal tract: comparison of four promoters.
JOURNAL Gene Ther. 2020 Jun 23. pii: 10.1038/s41434-020-0169-1. doi:
10.1038/s41434-020-0169-1.
PUBMED 32576975
REFERENCE 2 (bases 1 to 5271)
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: "Gene Ther.
2020 Jun 23. pii: 10.1038/s41434-020-0169-1. doi:
10.1038/s41434-020-0169-1."
FEATURES Location/Qualifiers
source 1..5271
/mol_type="other DNA"
/organism="synthetic DNA construct"
repeat_region 8..137
/label=AAV2 ITR (alternate)
/note="Functional equivalent of wild-type AAV2 ITR"
promoter 187..686
/label=PGK promoter
/note="mouse phosphoglycerate kinase 1 promoter"
primer_bind complement(222..245)
/label=MSCV-rev
/note="Murine stem cell virus, reverse primer"
primer_bind 619..638
/label=mPGK-F
/note="Mouse PGK promoter, forward primer"
intron 763..1141
/label=chimeric intron
/note="fusion between human cytomegalovirus intron A and
human beta-globin intron 2"
regulatory 1242..1251
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 1248..1967
/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 1248..1964
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(1293..1314)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1554..1573)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1901..1922
/label=EGFP-C
/note="EGFP, forward primer"
polyA_signal 2025..2501
/label=hGH poly(A) signal
/note="human growth hormone polyadenylation signal"
primer_bind complement(2224..2243)
/label=hGH-PA-R
/note="Human growth hormone terminator, reverse primer"
repeat_region 2541..2681
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 2541..2670
/label=AAV2 ITR
rep_origin 2756..3211
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2843..2862)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3053..3074
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(3228..3247)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 3347..3369
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(3407..3425)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 3493..3597
/gene="bla"
/label=AmpR promoter
CDS 3598..4458
/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(3816..3835)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 4629..5217
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5118..5137
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 cacatgtcct gcaggcagct gcgcgctcgc tcgctcactg aggccgcccg ggcgtcgggc
61 gacctttggt cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc
121 catcactagg ggttcctgcg gccgcacgcg cgatcacgag actagcctcg acgatggtcg
181 agtaccgggt aggggaggcg cttttcccaa ggcagtctgg agcatgcgct ttagcagccc
241 cgctgggcac ttggcgctac acaagtggcc tctggcctcg cacacattcc acatccaccg
301 gtaggcgcca accggctccg ttctttggtg gccccttcgc gccaccttct actcctcccc
361 tagtcaggaa gttccccccc gccccgcagc tcgcgtcgtg caggacgtga caaatggaag
421 tagcacgtct cactagtctc gtgcagatgg acagcaccgc tgagcaatgg aagcgggtag
481 gcctttgggg cagcggccaa tagcagcttt gctccttcgc tttctgggct cagaggctgg
541 gaaggggtgg gtccgggggc gggctcaggg gcgggctcag gggcggggcg ggcgcccgaa
601 ggtcctccgg aggcccggca ttctgcacgc ttcaaaagcg cacgtctgcc gcgctgttct
661 cctcttcctc atctccgggc ctttcgacct ctagcgggat cggattcgaa tcccggccgg
721 gaacggtgca ttggaacgcg gattccccgt gccaagagtg acgtaagtac cgcctataga
781 gtctataggc ccacaaaaaa tgctttcttc ttttaatata cttttttgtt tatcttattt
841 ctaatacttt ccctaatctc tttctttcag ggcaataatg atacaatgta tcatgcctct
901 ttgcaccatt ctaaagaata acagtgataa tttctgggtt aaggcaatag caatatttct
961 gcatataaat atttctgcat ataaattgta actgatgtaa gaggtttcat attgctaata
1021 gcagctacaa tccagctacc attctgcttt tattttatgg ttgggataag gctggattat
1081 tctgagtcca agctaggccc ttttgctaat catgttcata cctcttatct tcctcccaca
1141 gctcctgggc aacgtgctgg tctgtgtgct ggcccatcac tttggcaaag aattgggatt
1201 cgaacatcga ttgaattccg cgggcccggg atccaccggt cgccaccatg gtgagcaagg
1261 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
1321 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
1381 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
1441 tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct
1501 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
1561 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
1621 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
1681 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga
1741 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
1801 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
1861 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
1921 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaaagc ggccgctcta
1981 gaggatccaa gcttgcctcg agcagcgctg ctcgagagat ctacgggtgg catccctgtg
2041 acccctcccc agtgcctctc ctggccctgg aagttgccac tccagtgccc accagccttg
2101 tcctaataaa attaagttgc atcattttgt ctgactaggt gtccttctat aatattatgg
2161 ggtggagggg ggtggtatgg agcaaggggc aagttgggaa gacaacctgt agggcctgcg
2221 gggtctattg ggaaccaagc tggagtgcag tggcacaatc ttggctcact gcaatctccg
2281 cctcctgggt tcaagcgatt ctcctgcctc agcctcccga gttgttggga ttccaggcat
2341 gcatgaccag gctcagctaa tttttgtttt tttggtagag acggggtttc accatattgg
2401 ccaggctggt ctccaactcc taatctcagg tgatctaccc accttggcct cccaaattgc
2461 tgggattaca ggcgtgaacc actgctccct tccctgtcct tctgattttg taggtaacca
2521 cgtgcggacc gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg
2581 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg
2641 ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt
2701 tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg
2761 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
2821 cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
2881 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
2941 ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg
3001 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
3061 ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga tttataaggg
3121 attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
3181 aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct
3241 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
3301 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
3361 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
3421 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
3481 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
3541 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
3601 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
3661 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
3721 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
3781 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
3841 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
3901 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
3961 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
4021 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
4081 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
4141 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
4201 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
4261 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
4321 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
4381 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
4441 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
4501 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
4561 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
4621 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
4681 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
4741 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
4801 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
4861 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
4921 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
4981 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
5041 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
5101 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
5161 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
5221 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc t
//
| {
"Vector backbone": "Addgene plasmid #24593",
"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": "Optimization of adeno-associated viral vector-mediated transduction of the corticospinal tract: comparison of four promoters",
"Publication": "Gene Ther. 2020 Jun 23. pii: 10.1038/s41434-020-0169-1. doi: 10.1038/s41434-020-0169-1.",
"DOI": "10.1038/s41434-020-0169-1.",
"Authors": "Nieuwenhuis B, Haenzi B, Hilton S, Carnicer-Lombarte A, Hobo B, Verhaagen J, Fawcett JW /s---",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/32576975",
"PubMed ID": "PubMed 32576975"
} | {
"Gene/Insert name": "eGFP",
"Species": "",
"Insert Size (bp)": "720",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "mPGK",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "EcoRI (not destroyed)",
"3′ cloning site": "HindIII (not destroyed)",
"5′ sequencing primer": "Not applicable",
"3′ sequencing primer": "Not applicable"
} | {
"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": ""
} |
|
T4 Lysozyme T152S WT* | 18629 | Brian Matthews | T4 Lysozyme Plasmids - Tray 5 (unpublished) | https://media.addgene.org/snapgene-media/v2.0.0/sequences/24414/62b45f93-81db-4440-a1ef-f0881367076c/addgene-plasmid-18629-sequence-24414.gbk | partial | LOCUS sequence-24414-e 822 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 822)
TITLE T4 Lysozyme Plasmids - Tray 5
REFERENCE 2 (bases 1 to 822)
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..822
/mol_type="genomic DNA"
/organism="unspecified"
primer_bind complement(713..735)
/label=M13/pUC Reverse
/note="In lacZ gene"
ORIGIN
1 tantggaaaa agcccgctca ttaggcgggc tcagatctgc tcatgtttga cagcttatca
61 tcgataagct tcggcaataa attcttgata tgttttcata tgtttccttt aaatatttta
121 attagtaatt gtctattcaa gtaattgtga atatactatc acaattccaa gagaaagtaa
181 acagctttat agatttttat acgcgtccca agtgccagtt ctaaagcttg taatgactcg
241 ttttgcgcga ttaggtgttt gattatacca tctactttta gctaagttaa ctgctgcttc
301 atcccagcgt ttttgttgaa gcatacgtaa agagttagta aatcctgcca caccggtttc
361 tcccatttgg aaaaccatat taatcaatgc agcgcgacga accgcatcaa gagaatcata
421 aaccggtttt aatttagcat ttctcagaat tccgcgaaca gcagcatcaa catcctgatt
481 aaagagtttt tcagcctcat cttttgtaat tacaccatta gtattacgcc caatagcttt
541 atctaattca gatttagcag cattaagtga tggacttttt gtaagcaaat gaccgatgcc
601 aatagtgtaa tagccttctg tgtctttata gattttaagt ctaagacctt catctatacg
661 taacatttca aatatattca taatacctcc taagcatcga tggatcctgt ttcctgtgtg
721 aaattgttat ccgctcacan tccaacatta tacgagcgat gatantgtca caggggaggg
781 gagaancgac ccgtnccggg gaatngttcc cccnatcccc at
//
| {
"Vector backbone": "pHS1403",
"Backbone size w/o insert (bp)": "5000",
"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": "T4 Lysozyme",
"Species": "Bacteriophage T4",
"Insert Size (bp)": "600",
"Mutation": "T152S WT*",
"Entrez Gene": "e",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "BamHI (not destroyed)",
"3′ cloning site": "HindIII (not destroyed)",
"5′ sequencing primer": "na",
"3′ sequencing primer": "GGCGTATCACGAGGCCC"
} | {
"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": ""
} |
||
pLP-sfgfp | 209994 | Contains Level 0 Part: Coding sequences (sfGFP) for the construction of Level 1 plasmids | Stukenberg et al ACS Synth Biol. 2021 Aug 20;10(8):1904-1919. doi: 10.1021/acssynbio.1c00126. Epub 2021 Jul 13. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/419496/e4ff1ede-1115-41b4-bca9-7d07ff88c705/addgene-plasmid-209994-sequence-419496.gbk | full | LOCUS sequence-419496- 2823 bp DNA circular SYN 27-AUG-2024
DEFINITION Contains Level 0 Part: Coding sequences (sfGFP) for the construction
of Level 1 plasmids.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2823)
AUTHORS Stukenberg D, Hensel T, Hoff J, Daniel B, Inckemann R, Tedeschi JN,
Nousch F, Fritz G
TITLE The Marburg Collection: A Golden Gate DNA Assembly Framework for
Synthetic Biology Applications in Vibrio natriegens.
JOURNAL ACS Synth Biol. 2021 Aug 20;10(8):1904-1919. doi:
10.1021/acssynbio.1c00126. Epub 2021 Jul 13.
PUBMED 34255476
REFERENCE 2 (bases 1 to 2823)
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"; doi:
"10.1021/acssynbio.1c00126"; journalName: "ACS Synth Biol"; date:
"2021-08-20- 20"; volume: "10"; issue: "8"; pages: "1904-1919"
FEATURES Location/Qualifiers
source 1..2823
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 9..722
/codon_start=1
/product="GFP variant that folds robustly even when fused
to poorly folded proteins (Pedelacq et al., 2006)"
/label=superfolder GFP
/translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
FVTAAGITHGMDELYK"
terminator 767..824
/label=his operon terminator
/note="This putative transcriptin terminator from the E.
coli his operon has a 2-bp deletion introduced during
synthesis. Its efficiency has not been determined."
primer_bind complement(848..865)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(1019..1607)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1099..1118)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
terminator complement(1790..1884)
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
CDS complement(1905..2564)
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind 2479..2498
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
primer_bind 2726..2744
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
terminator complement(2748..2791)
/label=bacterial terminator
/note="putative bacterial transcription terminator"
misc_feature 2794..2815
/label=BioBrick prefix
/note="BioBrick prefix for parts that do not start with
""ATG"""
ORIGIN
1 ggtctcgaat gcgtaaaggt gaagaactgt ttaccggtgt tgttccaatt ctggttgaac
61 tggatggtga tgttaacggt cacaaatttt ctgttcgtgg tgaaggcgaa ggtgatgcaa
121 ccaacggtaa actgaccctg aaatttatct gtaccactgg taaactgcca gttccatggc
181 caactctggt taccactctg acctacggtg ttcaatgttt tgcacgttac ccagatcaca
241 tgaaacaaca cgattttttc aaaagcgcaa tgccagaagg ttacgttcaa gaacgtacca
301 tctcttttaa agatgacggc acctacaaaa cccgtgcgga agttaaattt gaaggtgata
361 ccctggttaa ccgcattgaa ctgaaaggca tcgattttaa agaagatggt aacatcctgg
421 gccacaaact ggaatacaac tttaactctc acaacgtgta catcaccgca gacaaacaaa
481 aaaacggtat caaagcgaac ttcaagatcc gtcacaacgt tgaagatggt tctgttcaac
541 tggcagatca ctaccaacaa aacaccccaa ttggtgatgg tccagttctg ctgccagata
601 accactacct gtctacccaa agcgttctgt ctaaagatcc aaacgaaaaa cgtgatcaca
661 tggtgctgct ggaatttgtt accgcagcag gtattaccca cggtatggat gaactgtaca
721 aagcagcttt gagaccactc atcgcgacta gtagcggccg ctgcagtccg gcaaaaaagg
781 gcaaggtgtc accaccctgc cctttttctt taaaaccgaa aagattactt cgcgttatgc
841 aggcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
901 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
961 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
1021 tccacaggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
1081 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
1141 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
1201 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
1261 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
1321 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
1381 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
1441 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
1501 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
1561 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
1621 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
1681 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
1741 caatctaaag tatatatgag taaacttggt ctgacagctc gaggcttgga ttctcaccaa
1801 taaaaaacgc ccggcggcaa ccgagcgttc tgaacaaatc cagatggagt tctgaggtca
1861 ttactggatc tatcaacagg agtccaagcg agctcgatat caaattacgc cccgccctgc
1921 cactcgtcgc agtactgttg taattcatta agcattctgc cgacatggaa gccatcacaa
1981 acggcatgat gaacctgaat cgccagcggc atcagcacct tgtcgccttg cgtataatat
2041 ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa
2101 ctggtgaaac tcacccaggg attggctgat acgaaaaaca tattctcaat aaacccttta
2161 gggaaatagg ccaggttttc accgtaacac gccacatctt gcgaatatat gtgtagaaac
2221 tgccggaaat cgtcgtggta ttcactccag agtgatgaaa acgtttcagt ttgctcatgg
2281 aaaacggtgt aacaagggtg aacactatcc catatcacca gctcaccgtc tttcattgcc
2341 atacgaaatt ccggatgagc attcatcagg cgggcaagaa tgtgaataaa ggccggataa
2401 aacttgtgct tatttttctt tacggtcttt aaaaaggccg taatatccag ctgaacggtc
2461 tggttatagg tacattgagc aactgactga aatgcctcaa aatgttcttt acgatgccat
2521 tgggatatat caacggtggt atatccagtg atttttttct ccattttagc ttccttagct
2581 cctgaaaatc tcgataactc aaaaaatacg cccggtagtg atcttatttc attatggtga
2641 aagttggaac ctcttacgtg cccgatcaac tcgagtgcca cctgacgtct aagaaaccat
2701 tattatcatg acattaacct ataaaaatag gcgtatcacg aggcagaatt tcagataaaa
2761 aaaatcctta gctttcgcta aggatgattt ctggaattcg cggccgcttc tagagcgatg
2821 agt
//
| {
"Vector backbone": "pLP1",
"Backbone size w/o insert (bp)": "",
"Vector type": "Bacterial Expression",
"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": "The Marburg Collection: A Golden Gate DNA Assembly Framework for Synthetic Biology Applications in Vibrio natriegens",
"Publication": "ACS Synth Biol. 2021 Aug 20;10(8):1904-1919. doi: 10.1021/acssynbio.1c00126. Epub 2021 Jul 13.",
"DOI": "10.1021/acssynbio.1c00126.",
"Authors": "Stukenberg D, Hensel T, Hoff J, Daniel B, Inckemann R, Tedeschi JN, Nousch F, Fritz G /acssynbioc",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/34255476",
"PubMed ID": "PubMed 34255476"
} | {
"Gene/Insert name": "sfGFP",
"Species": "Aequorea victoria",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "BsmBI (destroyed during cloning)",
"3′ cloning site": "BsmBI (destroyed during cloning)",
"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": ""
} |
||
pCAG-EHC | 112619 | expression plasmid with CAG promoter driving multicistronic cassette H2BEmerald_p2A_HygroR_p2A_CreERt2 | Stefan Heller | Hartman et al Dev Biol. 2018 Sep 1. pii: S0012-1606(18)30499-8. doi: 10.1016/j.ydbio.2018.08.013. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/226490/e802b99e-5c0d-4462-8817-788409f3840d/addgene-plasmid-112619-sequence-226490.gbk | full | LOCUS sequence-226490- 9852 bp DNA circular SYN 26-AUG-2024
DEFINITION expression plasmid with CAG promoter driving multicistronic cassette
H2BEmerald_p2A_HygroR_p2A_CreERt2.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9852)
AUTHORS Hartman BH, Bscke R, Ellwanger DC, Keymeulen S, Scheibinger M,
Heller S
TITLE Fbxo2(VHC) mouse and embryonic stem cell reporter lines delineate in
vitro-generated inner ear sensory epithelia cells and enable otic
lineage selection and Cre-recombination.
JOURNAL Dev Biol. 2018 Sep 1. pii: S0012-1606(18)30499-8. doi:
10.1016/j.ydbio.2018.08.013.
PUBMED 30179592
REFERENCE 2 (bases 1 to 9852)
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: "Dev Biol.
2018 Sep 1. pii: S0012-1606(18)30499-8. doi:
10.1016/j.ydbio.2018.08.013."
FEATURES Location/Qualifiers
source 1..9852
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 28..405
/codon_start=1
/gene="HIST1H2BJ"
/product="human histone H2B"
/label=H2B
/translation="MPEPAKSAPAPKKGSKKAVTKAQKKDGKKRKRSRKESYSIYVYKV
LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
GELAKHAVSEGTKAVTKYTSAK"
CDS 412..1128
/codon_start=1
/product="brightly fluorescent monomeric variant of GFP"
/label=mEmerald
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(718..737)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1065..1086
/label=EGFP-C
/note="EGFP, forward primer"
CDS 1138..1194
/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 1195..2217
/codon_start=1
/product="hygromycin B phosphotransferase"
/label=HygR
/note="confers resistance to hygromycin"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
E"
CDS 1195..2217
/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"
CDS 2230..2286
/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 2287..3315
/codon_start=1
/product="site-specific recombinase"
/label=Cre
/note="Cre is a site-specific recombinase from
bacteriophage P1. Recombination occurs at loxP sequences."
/translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
DGD"
CDS 3328..4263
/codon_start=1
/product="mutated ligand-binding domain of the human
estrogen receptor (Feil et al., 1997)"
/label=ERT2
/translation="AGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPIL
YSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEI
LMIGLVWRSMEHPVKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEF
VCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRL
AQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEET
DQSHLATAGSTSSHSLQKYYITGEAEGFPAT"
CDS 3376..4269
/codon_start=1
/gene="ESR1"
/product="estrogen nuclear receptor alpha ligand-binding
domain"
/label=ER-LBD
/translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPVKL
LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
GMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
QKYYITGEAEGFPATA"
primer_bind complement(4357..4376)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_signal 4422..4477
/label=beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(4476..4495)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
primer_bind complement(4838..4854)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4838..4854)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4851..4873)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4862..4878
/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(4886..4916)
/label=lac promoter
/note="promoter for the E. coli lac operon"
rep_origin complement(4964..5419)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5101..5122)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 5313..5332
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 5446..5550
/gene="bla"
/label=AmpR promoter
promoter 5552..5909
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
enhancer 5552..5743
/label=SV40 enhancer
/note="enhancer for the SV40 early promoter (Herr, 1993)"
primer_bind complement(5575..5595)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 5760..5895
/label=SV40 ori
/note="SV40 origin of replication"
CDS 5944..6738
/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 5944..6738
/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(5998..6017)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 6608..6627
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(6926..6945)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 6970..7017
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 7346..7934
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 7835..7854
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
enhancer 8108..8487
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 8490..8765
/label=chicken beta-actin promoter
intron 8766..9774
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
primer_bind 9698..9720
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 9782..9801
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
ORIGIN
1 ggcaaagaat tcatgaggcc ggccaccatg ccagagccag cgaagtctgc tcccgccccg
61 aaaaagggct ccaagaaggc ggtgactaag gcgcagaaga aagacggcaa gaagcgcaag
121 cgcagccgca aggagagcta ttccatctat gtgtacaagg ttctgaagca ggtccaccct
181 gacaccggca tttcgtccaa ggccatgggc atcatgaatt cgtttgtgaa cgacattttc
241 gagcgcatcg caggtgaggc ttcccgcctg gcgcattaca acaagcgctc gaccatcacc
301 tccagggaga tccagacggc cgtgcgcctg ctgctgcctg gggagttggc caagcacgcc
361 gtgtccgagg gtactaaggc cgtcaccaag tacaccagcg ctaaatccac catggtgagc
421 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
481 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
541 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
601 accttgacct acggcgtgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac
661 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
721 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
781 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
841 tacaactaca acagccacaa ggtctatatc accgccgaca agcagaagaa cggcatcaag
901 gtgaacttca agacccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac
961 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
1021 acccagtcca agctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
1081 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaaggg aagcggagct
1141 actaacttca gcctgctgaa gcaggctgga gacgtggagg agaaccctgg acctatgaaa
1201 aagcctgaac tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt cgacagcgtc
1261 tccgacctga tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga
1321 gggcgtggat atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa agatcgttat
1381 gtttatcggc actttgcatc ggccgcgctc ccgattccgg aagtgcttga cattggggaa
1441 ttcagcgaga gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac gttgcaagac
1501 ctgcctgaaa ccgaactgcc cgctgttctg cagccggtcg cggaggccat ggatgcgatc
1561 gctgcggccg atcttagcca gacgagcggg ttcggcccat tcggaccgca aggaatcggt
1621 caatacacta catggcgtga tttcatatgc gcgattgctg atccccatgt gtatcactgg
1681 caaactgtga tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg
1741 ctttgggccg aggactgccc cgaagtccgg cacctcgtgc acgcggattt cggctccaac
1801 aatgtcctga cggacaatgg ccgcataaca gcggtcattg actggagcga ggcgatgttc
1861 ggggattccc aatacgaggt cgccaacatc ttcttctgga ggccgtggtt ggcttgtatg
1921 gagcagcaga cgcgctactt cgagcggagg catccggagc ttgcaggatc gccgcggctc
1981 cgggcgtata tgctccgcat tggtcttgac caactctatc agagcttggt tgacggcaat
2041 ttcgatgatg cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg
2101 actgtcgggc gtacacaaat cgcccgcaga agcgcggccg tctggaccga tggctgtgta
2161 gaagtactcg ccgatagtgg aaaccgacgc cccagcactc gtccgagggc aaaggaacgc
2221 ggctccggcg caacaaattt ctctctcctc aaacaagcag gcgatgttga agaaaatccc
2281 gggcccatgt ccaatttact gaccgtacac caaaatttgc ctgcattacc ggtcgatgca
2341 acgagtgatg aggttcgcaa gaacctgatg gacatgttca gggatcgcca ggcgttttct
2401 gagcatacct ggaaaatgct tctgtccgtt tgccggtcgt gggcggcatg gtgcaagttg
2461 aataaccgga aatggtttcc cgcagaacct gaagatgttc gcgattatct tctatatctt
2521 caggcgcgcg gtctggcagt aaaaactatc cagcaacatt tgggccagct aaacatgctt
2581 catcgtcggt ccgggctgcc acgaccaagt gacagcaatg ctgtttcact ggttatgcgg
2641 cggatccgaa aagaaaacgt tgatgccggt gaacgtgcaa aacaggctct agcgttcgaa
2701 cgcactgatt tcgaccaggt tcgttcactc atggaaaata gcgatcgctg ccaggatata
2761 cgtaatctgg catttctggg gattgcttat aacaccctgt tacgtatagc cgaaattgcc
2821 aggatcaggg ttaaagatat ctcacgtact gacggtggga gaatgttaat ccatattggc
2881 agaacgaaaa cgctggttag caccgcaggt gtagagaagg cacttagcct gggggtaact
2941 aaactggtcg agcgatggat ttccgtctct ggtgtagctg atgatccgaa taactacctg
3001 ttttgccggg tcagaaaaaa tggtgttgcc gcgccatctg ccaccagcca gctatcaact
3061 cgcgccctgg aagggatttt tgaagcaact catcgattga tttacggcgc taaggatgac
3121 tctggtcaga gatacctggc ctggtctgga cacagtgccc gtgtcggagc cgcgcgagat
3181 atggcccgcg ctggagtttc aataccggag atcatgcaag ctggtggctg gaccaatgta
3241 aatattgtca tgaactatat ccgtaacctg gatagtgaaa caggggcaat ggtgcgcctg
3301 ctggaagatg gcgatctcga gccatctgct ggagacatga gagctgccaa cctttggcca
3361 agcccgctca tgatcaaacg ctctaagaag aacagcctgg ccttgtccct gacggccgac
3421 cagatggtca gtgccttgtt ggatgctgag ccccccatac tctattccga gtatgatcct
3481 accagaccct tcagtgaagc ttcgatgatg ggcttactga ccaacctggc agacagggag
3541 ctggttcaca tgatcaactg ggcgaagagg gtgccaggct ttgtggattt gaccctccat
3601 gatcaggtcc accttctaga atgtgcctgg ctagagatcc tgatgattgg tctcgtctgg
3661 cgctccatgg agcacccagt gaagctactg tttgctccta acttgctctt ggacaggaac
3721 cagggaaaat gtgtagaggg catggtggag atcttcgaca tgctgctggc tacatcatct
3781 cggttccgca tgatgaatct gcagggagag gagtttgtgt gcctcaaatc tattattttg
3841 cttaattctg gagtgtacac atttctgtcc agcaccctga agtctctgga agagaaggac
3901 catatccacc gagtcctgga caagatcaca gacactttga tccacctgat ggccaaggca
3961 ggcctgaccc tgcagcagca gcaccagcgg ctggcccagc tcctcctcat cctctcccac
4021 atcaggcaca tgagtaacaa aggcatggag catctgtaca gcatgaagtg caagaacgtg
4081 gtgcccctct atgacctgct gctggaggcg gcggacgccc accgcctaca tgcgcccact
4141 agccgtggag gggcatccgt ggaggagacg gaccaaagcc acttggccac tgcgggctct
4201 acttcatcgc attccttgca aaagtattac atcacggggg aggcagaggg tttccctgcc
4261 acagcttgat agcggccgca ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg
4321 tgtggccaat gccctggctc acaaatacca ctgagatctt tttccctctg ccaaaaatta
4381 tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga aatttatttt
4441 cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc
4501 aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat atgcccatat
4561 gctggctgcc atgaacaaag gttggctata aagaggtcat cagtatatga aacagccccc
4621 tgctgtccat tccttattcc atagaaaagc cttgacttga ggttagattt tttttatatt
4681 ttgttttgtg ttattttttt ctttaacatc cctaaaattt tccttacatg ttttactagc
4741 cagatttttc ctcctctcct gactactccc agtcatagct gtccctcttc tcttatggag
4801 atccctcgac ctgcagccca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
4861 attgttatcc gctcacaatt ccacacaaca tacgagccgg atgcataaag tgtaaagcct
4921 ggggtgccta atgagtgaaa ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat
4981 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
5041 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
5101 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
5161 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
5221 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
5281 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
5341 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
5401 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
5461 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
5521 atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg
5581 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
5641 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
5701 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
5761 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
5821 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
5881 gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt
5941 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
6001 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
6061 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
6121 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
6181 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
6241 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
6301 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
6361 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
6421 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc
6481 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
6541 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
6601 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
6661 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
6721 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
6781 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
6841 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
6901 tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
6961 gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata
7021 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
7081 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
7141 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata
7201 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
7261 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
7321 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
7381 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
7441 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
7501 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
7561 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
7621 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
7681 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
7741 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
7801 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
7861 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
7921 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
7981 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
8041 ccatgcatta gttattacta gcgctaccgg actcagatct cgagctcaag cttcgaattc
8101 tgcagtcgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
8161 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
8221 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
8281 atagggactt tccattgacg tcaatgggtg gactatttac ggtaaactgc ccacttggca
8341 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
8401 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
8461 tacgtattag tcatcgctat taccatgggt cgaggtgagc cccacgttct gcttcactct
8521 ccccatctcc cccccctccc cacccccaat tttgtattta tttatttttt aattattttg
8581 tgcagcgatg ggggcggggg gggggggggc gcgcgccagg cggggcgggg cggggcgagg
8641 ggcggggcgg ggcgaggcgg agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa
8701 agtttccttt tatggcgagg cggcggcggc ggcggcccta taaaaagcga agcgcgcggc
8761 gggcgggagt cgctgcgttg ccttcgcccc gtgccccgct ccgcgccgcc tcgcgccgcc
8821 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
8881 ctccgggctg taattagcgc ttggtttaat gacggctcgt ttcttttctg tggctgcgtg
8941 aaagccttaa agggctccgg gagggccctt tgtgcggggg ggagcggctc ggggggtgcg
9001 tgcgtgtgtg tgtgcgtggg gagcgccgcg tgcggcccgc gctgcccggc ggctgtgagc
9061 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcgtgtg cgcgagggga gcgcggccgg
9121 gggcggtgcc ccgcggtgcg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg
9181 tgcgtggggg ggtgagcagg gggtgtgggc gcggcggtcg ggctgtaacc cccccctgca
9241 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtgcggggcg
9301 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
9361 ggggccgcct cgggccgggg agggctcggg ggaggggcgc ggcggccccg gagcgccggc
9421 ggctgtcgag gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg
9481 cagggacttc ctttgtccca aatctggcgg agccgaaatc tgggaggcgc cgccgcaccc
9541 cctctagcgg gcgcgggcga agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg
9601 ccttcgtgcg tcgccgcgcc gccgtcccct tctccatctc cagcctcggg gctgccgcag
9661 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
9721 ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg
9781 ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattcct cgaggatgat
9841 ccaccggtcg cc
//
| {
"Vector backbone": "promoterless backbone from pd2EGFP-1",
"Backbone size w/o insert (bp)": "3110",
"Vector type": "Mammalian Expression",
"Selectable markers": "Hygromycin"
} | {
"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": "Fbxo2(VHC) mouse and embryonic stem cell reporter lines delineate in vitro-generated inner ear sensory epithelia cells and enable otic lineage selection and Cre-recombination",
"Publication": "Dev Biol. 2018 Sep 1. pii: S0012-1606(18)30499-8. doi: 10.1016/j.ydbio.2018.08.013.",
"DOI": "10.1016/j.ydbio.2018.08.013.",
"Authors": "Hartman BH, Bscke R, Ellwanger DC, Keymeulen S, Scheibinger M, Heller S /jydbio",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/30179592",
"PubMed ID": "PubMed 30179592"
} | {
"Gene/Insert name": "H2B-Emerald",
"Species": "H. sapiens (human); Aequorea victoria",
"Insert Size (bp)": "1101",
"Mutation": "",
"Entrez Gene": "H2BC3",
"Promoter": "CAG",
"Tag / Fusion Protein": "Venus (C terminal on insert)"
} | {
"Cloning method": "Gibson Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "AAAGAATTCATGAGGCCGGCC",
"3′ sequencing primer": "TGAAGTTAGTAGCTCCGCTTCC"
} | {
"Gene/Insert name": "Hygrophosphotransferase",
"Species": "Coccidioides posadasii C735 delta SOWgp",
"Insert Size (bp)": "1023",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "CAG",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Gibson Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "atgaaaaagcctgaactcaccgc",
"3′ sequencing primer": "ttcctttgccctcggacga"
} | {
"Gene/Insert name": "Cre-ERT2 fusion protein",
"Species": "Bacteriophage P1",
"Insert Size (bp)": "1983",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "CAG",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Gibson Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "ATGTCCAATTTACTGACCGTACA",
"3′ sequencing primer": "TCAAGCTGTGGCAGGGAAACC"
} |
|
pET28a-PACT | 50352 | bacterial expression of PACT | Kumiko Ui-Tei | Takahashi et al PLoS One. 2013 May 2;8(5):e63434. doi: 10.1371/journal.pone.0063434. Print 2013. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/75135/cabac27a-8f01-4682-a836-6d2281d82e94/addgene-plasmid-50352-sequence-75135.gbk | full | LOCUS sequence-75135-e 6349 bp DNA circular SYN 25-AUG-2024
DEFINITION bacterial expression of PACT.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6349)
AUTHORS Takahashi T, Miyakawa T, Zenno S, Nishi K, Tanokura M, Ui-Tei K
TITLE Distinguishable in vitro binding mode of monomeric TRBP and dimeric
PACT with siRNA.
JOURNAL PLoS One. 2013 May 2;8(5):e63434. doi: 10.1371/journal.pone.0063434.
Print 2013.
PUBMED 23658827
REFERENCE 2 (bases 1 to 6349)
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: "PLoS One.";
date: "2013-05-2"; volume: "8(5):e63434. doi"; pages: "
10.1371/journal.pone.0063434. Print 2013"
FEATURES Location/Qualifiers
source 1..6349
/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"
CDS complement(560..1375)
/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(560..1375)
/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 1283..1302
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
primer_bind 1367..1386
/label=pENTR-R
/note="pENTR vectors, reverse primer"
rep_origin 1497..2085
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 1986..2005
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 2239..2256
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 2271..2413
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(2339..2358)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 2499..2521
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(2515..2706)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(3515..4597)
/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 4559..4578
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(4598..4675)
/gene="lacI"
/label=lacI promoter
primer_bind 4881..4900
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind 4984..5003
/label=T7
/note="T7 promoter, forward primer"
promoter 4984..5002
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 5003..5027
/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)."
RBS 5042..5064
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 5083..5100
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 5110..5127
/codon_start=1
/product="thrombin recognition and cleavage site"
/label=thrombin site
/translation="LVPRGS"
CDS 6139..6168
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 6193..6210
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(6263..6281)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 6277..6324
/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 aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat
601 tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa
661 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc
721 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga
781 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc
841 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac
901 cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac
961 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat
1021 tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag
1081 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca
1141 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac
1201 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg
1261 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca
1321 tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcataacac
1381 cccttgtatt actgtttatg taagcagaca gttttattgt tcatgaccaa aatcccttaa
1441 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
1501 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
1561 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
1621 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
1681 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
1741 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
1801 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
1861 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
1921 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
1981 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
2041 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
2101 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
2161 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
2221 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
2281 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatatggtgc actctcagta
2341 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg
2401 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
2461 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
2521 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc
2581 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag
2641 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt
2701 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa
2761 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg
2821 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg
2881 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc
2941 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta
3001 cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca
3061 gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc
3121 ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggggccgc
3181 catgccggcg ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa
3241 ggcttgagcg agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc
3301 gctccagcga aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac
3361 gagttgcatg ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca
3421 ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta
3481 atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
3541 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
3601 tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca
3661 ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa
3721 aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt
3781 atcccactac cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg
3841 cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca
3901 gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta
3961 tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg
4021 agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat
4081 gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct
4141 ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg
4201 catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat
4261 tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc
4321 tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca
4381 gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg
4441 ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt
4501 tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg
4561 catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct
4621 cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga
4681 tctcgacgct ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg
4741 ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc
4801 ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg
4861 cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg
4921 gcgccggtga tgccggccac gatgcgtccg gcgtagagga tcgagatctc gatcccgcga
4981 aattaatacg actcactata ggggaattgt gagcggataa caattcccct ctagaaataa
5041 ttttgtttaa ctttaagaag gagatatacc atgggcagca gccatcatca tcatcatcac
5101 agcagcggcc tggtgccgcg cggcagccat atggctagcg gatccatgtc ccagagcagg
5161 caccgcgccg aggccccgcc gctggagcgc gaggacagtg ggaccttcag tttggggaag
5221 atgataacag ctaagccagg gaaaacaccg attcaggtat tacacgaata cggcatgaag
5281 accaagaaca tcccagttta tgaatgtgaa agatctgatg tgcaaataca cgtgcccact
5341 ttcaccttca gagtaaccgt tggtgacata acctgcacag gtgaaggtac aagtaagaag
5401 ctggcgaaac atagagctgc agaggctgcc ataaacattt tgaaagccaa tgcaagtatt
5461 tgctttgcag ttcctgaccc cttaatgcct gacccttcca agcaaccaaa gaaccagctt
5521 aatcctattg gttcattaca ggaattggct attcatcatg gctggagact tcctgaatat
5581 accctttccc aggagggagg acctgctcat aagagagaat atactacaat ttgcaggcta
5641 gagtcattta tggaaactgg aaagggggca tcaaaaaagc aagccaaaag gaatgctgct
5701 gagaaatttc ttgccaaatt tagtaatatt tctccagaga accacatttc tttaacaaat
5761 gtagtaggac attctttagg atgtacttgg cattccttga ggaattctcc tggtgaaaag
5821 atcaacttac tgaaaagaag cctccttagt attccaaata cagattacat ccagctgctt
5881 agtgaaattg ccaaggaaca aggttttaat ataacatatt tggatataga tgaactgagc
5941 gccaatggac aatatcaatg tcttgctgaa ctgtccacca gccccatcac agtctgtcat
6001 ggctccggta tctcctgtgg caatgcacaa agtgatgcag ctcacaatgc tttgcagtat
6061 ttaaagataa tagcagaaag aaaggtcgac gcggccgctg caagctcctc gagcatgcat
6121 ctagagggcc cgcggttcga acaaaaactc atctcagaag aggatctgtg aaagcttgcg
6181 gccgcactcg agcaccacca ccaccaccac tgagatccgg ctgctaacaa agcccgaaag
6241 gaagctgagt tggctgctgc caccgctgag caataactag cataacccct tggggcctct
6301 aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggat
//
| {
"Vector backbone": "pET28a",
"Backbone size w/o insert (bp)": "",
"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": "Distinguishable in vitro binding mode of monomeric TRBP and dimeric PACT with siRNA",
"Publication": "PLoS One. 2013 May 2;8(5):e63434. doi: 10.1371/journal.pone.0063434. Print 2013.",
"DOI": "10.1371/journal.pone.0063434.",
"Authors": "Takahashi T, Miyakawa T, Zenno S, Nishi K, Tanokura M, Ui-Tei K /journalpone",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/23658827",
"PubMed ID": "PubMed 23658827"
} | {
"Gene/Insert name": "PACT",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "PRKRA",
"Promoter": "T7",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "BamHI (not destroyed)",
"3′ cloning site": "SalI (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": ""
} |
|
LCB7_crtYB | 140232 | Expression of gene coding lycopene bifunctional β-cyclase for production beta-carotene from lycopene | Kang Zhou | Engineering E.coli to produce lycopene and its derivations from isopentenol (unpublished) | https://media.addgene.org/snapgene-media/v2.0.0/sequences/288146/3fecffcb-e946-47a1-b191-5f6e008864dc/addgene-plasmid-140232-sequence-288146.gbk | full | LOCUS sequence-288146- 4859 bp DNA circular SYN 26-AUG-2024
DEFINITION Expression of gene coding lycopene bifunctional beta-cyclase for
production beta-carotene from lycopene.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4859)
TITLE Engineering E.coli to produce lycopene and its derivations from
isopentenol
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 4859)
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..4859
/mol_type="other DNA"
/organism="synthetic DNA construct"
RBS 378..400
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
primer_bind complement(2492..2510)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 2506..2553
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS 2651..2668
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 2840..3631
/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"
primer_bind complement(3958..3975)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4092..4637)
/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(4172..4191)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 tagcaatcta atctaagttt tctagaaaaa tgagcttttc attctgactg caacgggcaa
61 tatgtctctg tgtggattaa aaaaagagtg tctgatagca gcttctgaac tggttacctg
121 ccgtgagtaa attaaaattt tattgactta ggtcactaaa tactttaacc aatataggca
181 tagcgcacag acagataaaa attacagagt acacaacatc catgaaacgc attagcacca
241 ccattaccac caccatcacc attaccacag gtaacggtgc gggctgacgc gtacaggaaa
301 cacagaaaaa agcccgcacc tgacagtgcg ggcttttttt tcgaccaaag gtaacgaggt
361 aacaacctag aaataatttt gtttaacttt aagaaggaga tatacatatg actgctttgg
421 cctactacca aatccatttg atctacacct tgccaatttt gggtttgttg ggtttattga
481 cctctccaat tttgaccaag ttcgacatct acaagatctc catcttggtt ttcattgctt
541 tctctgctac tactccatgg gattcttgga ttattagaaa tggtgcttgg acttacccat
601 ctgctgaatc tggtcaaggt gtttttggta cttttttgga tgtcccatac gaagagtacg
661 ctttcttcgt tattcaaacc gttattaccg gtttggttta cgttttggct accagacatt
721 tgttgccatc tttggctttg ccaaagacta gatcttctgc tttgtcattg gctttgaagg
781 ctttgattcc attgccaatc atctacttgt ttaccgctca tccatctcca tcaccagatc
841 cattggttac tgatcattac ttctacatga gagccctgtc tttgttgatt actccaccaa
901 ctatgttgtt ggctgcttta tctggtgaat acgcttttga ttggaaatcc ggtagagcta
961 agtctaccat tgctgctatt atgattccaa ccgtttactt gatctgggtt gattatgttg
1021 ctgttggtca agattcctgg tctattaacg acgaaaaaat cgtcggttgg agattaggtg
1081 gtgttttgcc aattgaagaa gctatgttct tcctgttgac caacttgatg atcgtcttag
1141 gtttgtctgc ttgtgatcat actcaagcct tgtatttgtt gcatggtaga actatctacg
1201 gcaacaaaaa gatgccatct tctttcccat tgatcacacc accagttttg tccttgtttt
1261 tctcatctag accatactct tctcagccaa agagagattt ggaattggct gttaagttgc
1321 tggaagaaaa gtccagatct tttttcgttg cttctgctgg ttttccatcc gaagttagag
1381 aaagattggt tggcttgtat gctttctgta gagttaccga tgatttgatc gattccccag
1441 aagtttcttc taatccacat gctaccatcg atatggtttc tgatttcttg actttgttgt
1501 tcggtccacc attgcatcca tcacaaccag ataagatttt gtcctctcca ttattgccac
1561 catcacatcc ttctagacca actggtatgt atccattgcc tccaccacca tctttgtctc
1621 cagctgaatt ggttcaattc ttgactgaaa gagttccagt ccaataccat tttgctttca
1681 gattattggc caagttgcaa ggtttgatcc caagatatcc attggatgaa ttgctgagag
1741 gttacaccac tgatttgata ttcccattgt ctaccgaagc tgttcaagct agaaaaactc
1801 caattgaaac taccgccgat ttgttggatt atggtttgtg tgttgctggt tccgttgctg
1861 aattattggt ttatgtttct tgggcttctg ccccatctca agttccagct actattgaag
1921 aaagagaagc tgttttggtt gcctctagag aaatgggtac tgctttacag ttggttaaca
1981 ttgccagaga tattaagggt gatgctactg aaggtagatt ctacttgcca ttgtcattct
2041 tcggtttgag ggatgaatct aagttggcta ttccaactga ttggactgaa cctagaccac
2101 aagatttcga taagctgttg tctttgtcac catcttcaac tttgccatcc tctaatgcct
2161 ctgaatcctt tagatttgag tggaaaactt actccttgcc tttggttgct tatgctgaag
2221 atttggctaa acactcctac aagggtattg atagattgcc aactgaagtt caagctggta
2281 tgagagctgc ttgtgcttct tatttgttaa tcggtagaga aatcaaggtc gtttggaaag
2341 gtgatgttgg tgaaagaaga actgttgcag gttggagaag agttagaaag gttttgtctg
2401 ttgttatgtc tggttgggaa ggtcaatgat gggctgaagg gtttaagggc tgctaacaaa
2461 gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt
2521 ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggatat
2581 cccgcaagag gcccggcagt accggcataa ccaagcctat gcctacagca tccagggtga
2641 cggtgcctaa catcatcatc accatcacta aaagtcgacc tgcagaagct tagatctatt
2701 accctgttat ccctactcga gttcatgtgc agctccataa gcaaaagggg atgataagtt
2761 tatcaccacc gactatttgc aacagtgccg ttgatcgtgc tatgatcgac tgatgtcatc
2821 agcggtggag tgcaatgtca tgagggaagc ggtgatcgcc gaagtatcga ctcaactatc
2881 agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg ttgctggccg tacatttgta
2941 cggctccgca gtggatggcg gcctgaagcc acacagtgat attgatttgc tggttacggt
3001 gaccgtaagg cttgatgaaa caacgcggcg agctttgatc aacgaccttt tggaaacttc
3061 ggcttcccct ggagagagcg agattctccg cgctgtagaa gtcaccattg ttgtgcacga
3121 cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg caatttggag aatggcagcg
3181 caatgacatt cttgcaggta tcttcgagcc agccacgatc gacattgatc tggctatctt
3241 gctgacaaaa gcaagagaac atagcgttgc cttggtaggt ccagcggcgg aggaactctt
3301 tgatccggtt cctgaacagg atctatttga ggcgctaaat gaaaccttaa cgctatggaa
3361 ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg cttacgttgt cccgcatttg
3421 gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc gctgccgact gggcaatgga
3481 gcgcctgccg gcccagtatc agcccgtcat acttgaagct agacaggctt atcttggaca
3541 agaagaagat cgcttggcct cgcgcgcaga tcagttggaa gaatttgtcc actacgtgaa
3601 aggcgagatc accaaggtag tcggcaaata agatgccgct cgccagtcga ttggctgagc
3661 tcataagttc ctattccgaa gttccgcgaa cgcgtaaagg atctaggtga agatcctttt
3721 tgataatctc atgaccaaaa tcccttaacg ttcacacaac atagccacgg gtgttcagct
3781 actgacgggg tggtgcgtaa cggcaaaagc accgccggac atcagcgcta gcggagtgta
3841 tactggctta ctatgttggc actgatgagg gtgtcagtga agtgcttcat gtggcaggag
3901 aaaaaaggct gcaccggtgc gtcagcagaa tatgtgatac aggatatatt ccgcttcctc
3961 gctcactgac tcgctacgct cggtcgttcg actgcggcga gcggaaatgg cttacgaacg
4021 gggcggagat ttcctggaag atgccaggaa gatacttaac agggaagtga gagggccgcg
4081 gcaaagccgt ttttccatag gctccgcccc cctgacaagc atcacgaaat ctgacgctca
4141 aatcagtggt gacgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggcggc
4201 tccctcgtgc gctctcctgt tcctgccttt cggtttaccg gtgtcattcc gctgttatgg
4261 ccgcgtttgt ctcattccac gcctgacact cagttccggg taggcagttc gctccaagct
4321 ggactgtatg cacgaacccc ccgttcagtc cgaccgctgc gccttatccg gtaactatcg
4381 tcttgagtcc aacccggaaa gacatgcaaa agcaccactg gcagcagcca ctggtaattg
4441 atttagagga gttagtcttg aagtcatgcg ccggttaagg ctaaactgaa aggacaagtt
4501 ttggtgactg cgctcctcca agccagttac ctcggttcaa agagttggta gctcagagaa
4561 ccttcgaaaa accgccctgc aaggcggttt tttcgttttc agagcaagag attacgcgca
4621 gaccaaaacg atctcaagaa gatcatctta ttaatcagat aaaatatttc tagatttcag
4681 tgcaatttat ctcttcaaat gtagcacctg aagtcagccc catacgatat aagttgtaat
4741 tctcatgttt gacagcttat cacccagtcc tgctcgcttc gctacttggc catacccacg
4801 ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtc
//
| {
"Vector backbone": "LCB7",
"Backbone size w/o insert (bp)": "",
"Vector type": "Bacterial Expression",
"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": "",
"Publication": "",
"DOI": "",
"Authors": "",
"PubMed Link": "",
"PubMed ID": ""
} | {
"Gene/Insert name": "phytoene-beta carotene synthase",
"Species": "",
"Insert Size (bp)": "2016",
"Mutation": "K307E",
"Entrez Gene": "",
"Promoter": "pthrC3",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Ligation Independent Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "TagcaatctaatctaagttttctagaaaaatG",
"3′ sequencing primer": "CttttagtgatggtgatgatgatgttA"
} | {
"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": ""
} |
|
pH9-ilvA-acp (DE2) | 135163 | Expressing synthetic overlapping gene, ilvA-acpP | Harris Wang | Blazejewski et al Science. 2019 Aug 9;365(6453):595-598. doi: 10.1126/science.aav5477. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/267932/468ecab8-b8a5-4cf3-b786-829d6a88a043/addgene-plasmid-135163-sequence-267932.gbk | full | LOCUS sequence-267932- 4694 bp DNA circular SYN 26-AUG-2024
DEFINITION Expressing synthetic overlapping gene, ilvA-acpP.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4694)
AUTHORS Blazejewski T, Ho HI, Wang HH
TITLE Synthetic sequence entanglement augments stability and containment
of genetic information in cells.
JOURNAL Science. 2019 Aug 9;365(6453):595-598. doi: 10.1126/science.aav5477.
PUBMED 31395784
REFERENCE 2 (bases 1 to 4694)
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: "10.1126/science";
journalName: "Science"; date: "2019-08-9- 9"; volume: "365"; issue:
"6453"; pages: "595-598"
FEATURES Location/Qualifiers
source 1..4694
/mol_type="other DNA"
/organism="synthetic DNA construct"
terminator 1584..1655
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 1671..1698
/label=T7Te terminator
/note="phage T7 early transcription terminator"
primer_bind complement(1726..1743)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(1860..2405)
/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(1940..1959)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
terminator complement(2519..2613)
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
terminator complement(2678..2772)
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
CDS complement(2803..3594)
/codon_start=1
/gene="aadA"
/product="aminoglycoside adenylyltransferase (Murphy,
1985)"
/label=SmR
/note="confers resistance to spectinomycin and
streptomycin"
/translation="MREAVITEVSTQLSEVVGAIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
CDS complement(3962..4588)
/codon_start=1
/gene="tetR"
/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="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
primer_bind 4493..4512
/label=Tet-R
/note="Tetracycline resistance gene, reverse primer"
promoter 4604..4659
/gene="tetR"
/label=tetR/tetA promoters
/note="overlapping promoters for bacterial tetR and tetA"
protein_bind 4640..4658
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
RBS 4677..4688
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
ORIGIN
1 atggctgact cgcaacccct gtccggtgct ccggaaggtg ccgaatattt aagagcagtg
61 ctgcgcgcgc cggtttacga ggcggcgcag gttacgccgc tacaaaaaat ggaaaaactg
121 tcgtcgcgtc ttgataacgt cattctggtg aagcgcgaag atcgccagcc agtgcacagc
181 tttaagctgc gcggcgcata cgccatgatg gcgggcctga cggaagaaca gaaagcgcac
241 ggcgtgatca ctgcttctgc gggtaaccac gcgcagggcg tcgcgttttc ttctgcgcgg
301 ttaggcgtga aggccctgat cgttatgcca accgccaccg ccgacatcaa agtcgacgcg
361 gtgcgcggct tcggcggcga agtgctgctc cacggcgcga actttgatga agcgaaagcc
421 aaagcgatcg aactgtcaca gcagcagggg ttcacctggg tgccgccgtt cgaccatccg
481 atggtgattg ccgggcaagg cacgctggcg ctggaactgc tccagcagga cgcccatctc
541 gaccgcgtat ttgtgccagt cggcggcggc ggtctggctg ctggcgtggc ggtgctgatc
601 aaacaactga tgccgcaaat caaagtgatc gccgtagaag cggaagactc cgcctgcctg
661 aaagcagcgc tggatgcggg tcatccggtt gatctgccgc gcgtagggct atttgctgaa
721 ggcgtagcgg taaaacgcat cggtgacgaa accttccgtt tatgccagga gtatctcgac
781 gacatcatca ccgtcgatag cgatgcgatc tgtgcggcga tgaaggattt attcgaagat
841 gtgcgcgcgg tggcggaacc ctctggcgcg ctggcgctgg cgggaatgaa aaaatatatc
901 gccctgcaca acattcgcgg cgaacggctg gcgcatattc tttccggtgc caacgtgaac
961 ttccacggcc tgcgctacgt ctcagaacgc tgcgaactgg gcgaacagcg tgaagcgttg
1021 ttggctgtca ccatccctga agagaaagga tcattcctta agttttgcca attgttggga
1081 gggaggagtg ttacggatgt caactataga agagcgggtg aagaagataa taggagagca
1141 gttgggagtc aagcaagaag aagtgacgaa caacgcgagc ttcgtagaag acttaggagc
1201 agacagttta gatacagtgg aattagtgat ggctctagag gaagagttcg atacagaaat
1261 accggacgag gaggcagaaa agataacaac agttcaagcc gctatagact atataaacgg
1321 acatcaagct tagtcccgga atcaccgggc gcgctgctgc gcttcctcaa cacgctgggt
1381 acgtactgga acatttcttt gttccactat cgcagccatg gcaccgacta cgggcgcgta
1441 ctggcggcgt tcgaacttgg cgaccatgaa ccggatttcg aaacccggct gaatgagctg
1501 ggctacgatt gccacgacga aaccaataac ccggcgttca ggttcttttt ggcgggttag
1561 ctcgagtaag gatctccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct
1621 ttcgttttat ctgttgtttg tcggtgaacg ctctctacta gagtcacact ggctcacctt
1681 cgggtgggcc tttctgcgtt tatacctagg gatatattcc gcttcctcgc tcactgactc
1741 gctacgctcg gtcgttcgac tgcggcgagc ggaaatggct tacgaacggg gcggagattt
1801 cctggaagat gccaggaaga tacttaacag ggaagtgaga gggccgcggc aaagccgttt
1861 ttccataggc tccgcccccc tgacaagcat cacgaaatct gacgctcaaa tcagtggtgg
1921 cgaaacccga caggactata aagataccag gcgtttcccc ctggcggctc cctcgtgcgc
1981 tctcctgttc ctgcctttcg gtttaccggt gtcattccgc tgttatggcc gcgtttgtct
2041 cattccacgc ctgacactca gttccgggta ggcagttcgc tccaagctgg actgtatgca
2101 cgaacccccc gttcagtccg acagctgcgc cttatccggt aactatcgtc ttgagtccaa
2161 cccggaaaga catgcaaaag cacaactggc agcagccact ggtaattgat ttagaggagt
2221 tagtcttgaa gtcatgcgcc ggttaaggct aaactgaaag gacaagtttt ggtgactgcg
2281 ctcctccaag ccagttacct cggttcaaag agttggtagc tcagagaacc ttcgaaaaac
2341 cgccctgcaa ggcggttttt tcgttttcag agcaagagat tacgcgcaga ccaaaacgat
2401 ctcaagaaga tcatcttatt aatcagataa aatatttcta gatttcagtg caatttatct
2461 cttcaaatgt agcacctgaa gtcagcccca tacgatataa gttgttacta gtgcttggat
2521 tctcaccaat aaaaaacgcc cggcggcaac cgagcgttct gaacaaatcc agatggagtt
2581 ctgaggtcat tactggatct atcaacagga gtccaagcga gctcgatatc aaattacgcc
2641 ccgccctgcc actcatcgca gtactactag tgcttggatt ctcaccaata aaaaacgccc
2701 ggcggcaacc gagcgttctg aacaaatcca gatggagttc tgaggtcatt actggatcta
2761 tcaacaggag tccaagcgag ctctcgaacc ccagagtccc gcttatttgc cgactacctt
2821 ggtgatctcg cctttcacgt agtggacaaa ttcttccaac tgatctgcgc gcgaggccaa
2881 gcgatcttct tcttgtccaa gataagcctg tctagcttca agtatgacgg gctgatactg
2941 ggccggcagg cgctccattg cccagtcggc agcgacatcc ttcggcgcga ttttgccggt
3001 tactgcgctg taccaaatgc gggacaacgt aagcactaca tttcgctcat cgccagccca
3061 gtcgggcggc gagttccata gcgttaaggt ttcatttagc gcctcaaata gatcctgttc
3121 aggaaccgga tcaaagagtt cctccgccgc tggacctacc aaggcaacgc tatgttctct
3181 tgcttttgtc agcaagatag ccagatcaat gtcgatcgtg gctggctcga agatacctgc
3241 aagaatgtca ttgcgctgcc attctccaaa ttgcagttcg cgcttagctg gataacgcca
3301 cggaatgatg tcgtcgtgca caacaatggt gacttctaca gcgcggagaa tctcgctctc
3361 tccaggggaa gccgaagttt ccaaaaggtc gttgatcaaa gctcgccgcg ttgtttcatc
3421 aagccttacg gtcaccgtaa ctagcaaatc aatatcactg tgtggcttca ggccgccatc
3481 cactgcggag ccgtacaaat gtacggccag caacgtcggt tcgagatggc gctcgatggc
3541 gccaactacc tctgatagtt gagtcgatac ttcggtgatc accgcttccc tcataatgtt
3601 taactttgtt ttagggcgac tgccctgctg cgtaacatcg ttgctgctcc ataacatcaa
3661 acatcgaccc atggcgtaac gcgcttgctg cttggatgcc cgaggcatag actgtacccc
3721 aaaaaaacag tcatagcaag ccatgaaaac cgccactgca ccgttaccac cgctgcgttc
3781 ggtcaaggtt ctggaccagt tgcgtgagcg catgcgaaac gatcctcatc ctgtctcttg
3841 atcagatctt gatcccctgc gccatcagat ccttggcggc aagaaagcca tccagtttac
3901 tttgcagggc ttcccaacct taccagaggg cgccccagct ggcaattccg acgtcgacgt
3961 cttaagaccc actttcacat ttaagttgtt tttctaatcc gcatatgatc aattcaaggc
4021 cgaataagaa ggctggctct gcaccttggt gatcaaataa ttcgatagct tgtcgtaata
4081 atggcggcat actatcagta gtaggtgttt ccctttcttc tttagcgact tgatgctctt
4141 gatcttccaa tacgcaacct aaagtaaaat gccccacagc gctgagtgca tataatgcat
4201 tctctagtga aaaaccttgt tggcataaaa aggctaattg attttcgaga gtttcatact
4261 gtttttctgt aggccgtgta cctaaatgta cttttgctcc atcgcgatga cttagtaaag
4321 cacatctaaa acttttagcg ttattacgta aaaaatcttg ccagctttcc ccttctaaag
4381 ggcaaaagtg agtatggtgc ctatctaaca tctcaatggc taaggcgtcg agcaaagccc
4441 gcttattttt tacatgccaa tacaatgtag gctgctctac acctagcttc tgggcgagtt
4501 tacgggttgt taaaccttcg attcctacct cattaagcag ctctaatgcg ctgttaatca
4561 ctttactttt atctaatcta gacatcatta attcctaatt tttgttgaca ctctatcgtt
4621 gatagagtta ttttaccact ccctatcagt gatagagaaa agaattcaaa agatctaaag
4681 aggagaaagg atct
//
| {
"Vector backbone": "pH9",
"Backbone size w/o insert (bp)": "3000",
"Vector type": "Synthetic Biology",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Spectinomycin, 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": "Synthetic sequence entanglement augments stability and containment of genetic information in cells",
"Publication": "Science. 2019 Aug 9;365(6453):595-598. doi: 10.1126/science.aav5477.",
"DOI": "10.1126/science.aav5477.",
"Authors": "Blazejewski T, Ho HI, Wang HH /scienceaav",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/31395784",
"PubMed ID": "PubMed 31395784"
} | {
"Gene/Insert name": "ilvA-acp",
"Species": "Synthetic",
"Insert Size (bp)": "1600",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "TetO",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Gibson Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "ACCTCATTAAGCAGCTCTAATGCGCTG",
"3′ sequencing primer": "TGTGACTCTAGTAGAGAGCGTTCAC"
} | {
"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": ""
} |
|
pAc5.1B-lambdaN-HA-DmeIF4Ga_578-650-CG32016-E_T | 147588 | Insect Expression of DmeIF4Ga_578-650-CG32016-E | Elisa Izaurralde | Igreja et al Nat Commun. 2014 Sep 2;5:4790. doi: 10.1038/ncomms5790. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/290035/119246e2-8426-4f44-baf6-3be2134e2692/addgene-plasmid-147588-sequence-290035.gbk | full | LOCUS sequence-290035- 7196 bp DNA circular SYN 26-AUG-2024
DEFINITION Insect Expression of DmeIF4Ga_578-650-CG32016-E.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7196)
AUTHORS Igreja C, Peter D, Weiler C, Izaurralde E
TITLE 4E-BPs require non-canonical 4E-binding motifs and a lateral surface
of eIF4E to repress translation.
JOURNAL Nat Commun. 2014 Sep 2;5:4790. doi: 10.1038/ncomms5790.
PUBMED 25179781
REFERENCE 2 (bases 1 to 7196)
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
Commun."; date: "2014-09-2"; volume: "5:4790. doi"; pages: "
10.1038/ncomms5790"
FEATURES Location/Qualifiers
source 1..7196
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 56..2568
/label=Ac5 promoter
/note="Drosophila melanogaster actin 5C promoter"
CDS 2588..2653
/codon_start=1
/product="N-terminal RNA-binding domain from the lambda
bacteriophage antiterminator protein N (Baron-Benhamou et
al., 2004)"
/label=lambda N peptide
/translation="MDAQTRRRERRAEKQAQWKAAN"
CDS 2678..2704
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 4481..4522
/codon_start=1
/product="epitope tag from simian virus 5"
/label=V5 tag
/translation="GKPIPNPLLGLDST"
CDS 4532..4549
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(4572..4589)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 4622..4756
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4646..4665)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 4700..4719
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
rep_origin complement(5378..5966)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5458..5477)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6137..6997)
/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 6760..6779
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6998..7102)
/gene="bla"
/label=AmpR promoter
primer_bind 7170..7188
/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 taccaacatg gacgcacaaa cacgacgacg tgagcgtcgc gctgagaaac aagctcaatg
2641 gaaagctgca aacggtggat ctcctccaca tatggcttac ccatacgatg ttccagatta
2701 cgctcctcca agctttccag agactaccct taacgataaa caggactcta ctgacctaaa
2761 agtgaaagtg tctgcgaaaa tttcatctat cataaactat aatgagggtc aatggtcacc
2821 aaacaatcca agcggaaaaa aacaatacga ccgagaacag ttattacagt tacgcgaggt
2881 taaggcctca cgtatacaac cagaggtaaa aaatgtgtct attcttcctc aaccattaat
2941 taatatgata aaagtggata tgaggactga tttaataaat ccaatagcag ggtcattatg
3001 tcagacagtg caaatggaaa aacctattgc tagagacact gaggcattca aaaaactttt
3061 gcaacagttg ggttctcaag ccaggcagca tcatccatgc aacgacgatt gtcgtacaat
3121 aaacctctca aatatagcaa atcatgtaca ccttgagtca aaattacatc aaaaaataaa
3181 tgatggtcat ttgcagcaac cagagttaag tgttaacgtg ccaaccatgc caactagtag
3241 tcatgtattt ttgcaaaagc gtttggaaat ccaacattta attcaaagac ttcgttgtgg
3301 agacgtttca catgattttt tggaaaaaga gttggataat cccagcaccc cagcagccac
3361 aaaggatgtc attgccacag tgttgagtga atattctcat tctaaaagaa atccggtggt
3421 cacaggcgat cccaatattt ttacccaaca atcgtttctt cagcctcaat cagtccatca
3481 gcactactct caagaattgc attcgcaaaa tactgctaac catacgatta atcagctaat
3541 ttcccacggt aactctccaa ctccgttggc atttactcct acttcagttt taagaaaaat
3601 gacagccgat aaggatacgc aatcaccaag cacttattgt cagaacccac aatatcatgt
3661 acaccaacaa aatgccaagc aagtcggtac tcgtgagaac gtacttgagc cacagctgac
3721 cgcaacaatg gctgtacagc cccgaatgat attaggcggt ggaaactttg ccattggcca
3781 aaacaatcaa catttgtctc ccaacatgtc acaaagtcgt aatcaacaag tgttaaagtg
3841 gacttctggc aatatgcaaa tggtccacgg aaaaacattt ggccgacata ttctgaaagg
3901 aggtcttaat tcaatgccgc attccaattc tgcgcttccc tttactgccc ataaaattga
3961 aatgcaaccc attcatcaac cacatctaca gcagcagcaa catagattta aagcagttca
4021 atctgtggaa tcaaatctga ataccgaaag tgttcaccaa aacattacat ctcctgttgg
4081 atggcaccaa ctttacatgc aacatcagca acaacatcat cacacacgac agcagcttag
4141 tcaaagagtg atttatggag aaatgcaccg tcaatcaaat ccacaaatgt ctccgccagt
4201 cccaggtttt tctgacagct ccgattccgg taatgtgatc aaagctaaca gcctcacatc
4261 tcctagctac cagagagatg agaggatttc atcaccaacc aatcaattgg cgcaatggtt
4321 ttcccctgag cttcttgcaa aagcctctgc tggaaaactg ccacttttaa atgtgaacca
4381 agcactcagc ttggaagaat ttgaacgaag cattcaacat tcctcaggcg tcgtgcataa
4441 ctaaggcggc cgctcgagtc tagagggccc gcggttcgaa ggtaagccta tccctaaccc
4501 tctcctcggt ctcgattcta cgcgtaccgg tcatcatcac catcaccatt gagtttaaac
4561 ccgctgatca gcctcgactg tgccttctaa ggcctgagct cgctgatcag cctcgatcga
4621 ggatccagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
4681 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
4741 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
4801 ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtatgg ctgattatga
4861 tcagtcgacc gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg
4921 gcatgactat cgtcgccgca cttatgactg tcttctttat catgcaactc gtaggacagg
4981 tgccggcagc gctctgggtc attttcggcg aggaccgctt tcgctggagc gcgacgatga
5041 tcggcctgtc gcttgcggta ttcggaatct tgcacgccct cgctcaagcc ttcgtcactg
5101 gtcccgccac caaacgtttc ggcgagaagc aggccattat cgccggcatg gcggccgacg
5161 cgctgggcta cgtcttgctg gcgttcgcga cgcgaggctg gatggccttc cccattatga
5221 ttcttctcgc ttccggcggc atcgggatgc ccgcgttgca ggccatgctg tccaggcagg
5281 tagatgacga ccatcaggga cagcttcaag gatcgctcgc ggctcttacc agccagcaaa
5341 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
5401 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
5461 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
5521 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac
5581 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
5641 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
5701 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
5761 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
5821 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
5881 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
5941 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
6001 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
6061 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
6121 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
6181 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
6241 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
6301 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
6361 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
6421 ttaatagttt gcgcaacgtt gttgccattg ctgcaggcat cgtggtgtca cgctcgtcgt
6481 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
6541 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
6601 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
6661 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
6721 tgcggcgacc gagttgctct tgcccggcgt caacacggga taataccgcg ccacatagca
6781 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
6841 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
6901 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
6961 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
7021 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
7081 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
7141 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgt
//
| {
"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": "4E-BPs require non-canonical 4E-binding motifs and a lateral surface of eIF4E to repress translation",
"Publication": "Nat Commun. 2014 Sep 2;5:4790. doi: 10.1038/ncomms5790.",
"DOI": "10.1038/ncomms5790.",
"Authors": "Igreja C, Peter D, Weiler C, Izaurralde E /ncomms",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/25179781",
"PubMed ID": "PubMed 25179781"
} | {
"Gene/Insert name": "DmeIF4Ga_578-650-CG32016-E",
"Species": "D. melanogaster (fly)",
"Insert Size (bp)": "",
"Mutation": "CG32016-E 5 silent and three no silent mutations H117R, N148S and P315H compared to the sequence given by NCBI:NM_166800.1",
"Entrez Gene": "eIF4G1",
"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": ""
} |
|
pVRa14_1324 | 49665 | Christopher Voigt | Rhodius et al Mol Syst Biol. 2013 Oct 29;9:702. doi: 10.1038/msb.2013.58. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/74438/369fdd2b-f021-450e-97cb-4c5aed7bba9f/addgene-plasmid-49665-sequence-74438.gbk | full | LOCUS sequence-74438-e 6125 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 6125)
AUTHORS Rhodius VA, Segall-Shapiro TH, Sharon BD, Ghodasara A, Orlova E,
Tabakh H, Burkhardt DH, Clancy K, Peterson TC, Gross CA, Voigt CA
TITLE Design of orthogonal genetic switches based on a crosstalk map of
sigmas, anti-sigmas, and promoters.
JOURNAL Mol Syst Biol. 2013 Oct 29;9:702. doi: 10.1038/msb.2013.58.
PUBMED 24169405
REFERENCE 2 (bases 1 to 6125)
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 Syst
Biol."; date: "2013-10-29"; volume: "9:702. doi"; pages: "
10.1038/msb.2013.58"
FEATURES Location/Qualifiers
source 1..6125
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 700..717
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(770..788)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 784..831
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
rep_origin 868..1323
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(955..974)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 1165..1186
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 1350..1454
/gene="bla"
/label=AmpR promoter
CDS 1455..2315
/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(1673..1692)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 2486..3074
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2975..2994
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 3228..3245
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 3260..3402
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(3328..3347)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 3488..3510
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(3504..3695)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(4504..5586)
/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 5548..5567
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(5587..5664)
/gene="lacI"
/label=lacI promoter
primer_bind 5870..5889
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind 5973..5992
/label=T7
/note="T7 promoter, forward primer"
promoter 5973..5991
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 5992..6016
/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)."
RBS 6031..6053
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 6072..6089
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 6099..6122
/codon_start=1
/product="recognition and cleavage site for human
rhinovirus 3C and PreScission proteases"
/label=HRV 3C site
/translation="LEVLFQGP"
ORIGIN
1 atgaatgata ccgcagcaga tcatagccat gcagatggtc cgaccaccgg tcatgcacat
61 accgcaacct ttagcagtga tgccgatggt caggcatgga cccctccgac ctgggaagaa
121 attgttagca cccatagcgg tcgtgtttat cgtctggcat atcgtctgac cggtaatcag
181 catgatgcag aagatctgac ccaagaagtt tttgttcgcg tttttcgtag cctgagcacc
241 tatacaccgg gtacatttga aggttggctg catcgtatta ccaccaacct gtttctggat
301 atggttcgtc gtaaacagcg tattcgtttt gatgcactgg gtgatgatgc agcagaacgt
361 ctgccgagcc gtgaaccgac tccgcagcag ctgtttcatg atgcacattt tgatgcggat
421 gttcagcagg cactggatac cctggcaccg gaatttcgtg cagcagttgt tctgtgtgat
481 attgaaggtc tgtcctatga agaaatcgca gcaaccctgg gtgttaaact gggcaccgtt
541 cgtagccgta ttcatcgtgg tcgtagccag ctgcgtaaag cactggcaca tcgtagcccg
601 aaagcacgtg cagatcgtcg tagtctggca gccggtgttc cggctctggg tggtggtggt
661 acaagcgcat aaggatccaa gcttgcggcc gcactcgagc accaccacca ccaccactga
721 gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa
781 taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga
841 ggaactatat ccggattggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg
901 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
961 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
1021 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
1081 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
1141 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
1201 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
1261 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa
1321 tttcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
1381 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
1441 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
1501 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
1561 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
1621 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
1681 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
1741 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
1801 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
1861 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
1921 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
1981 gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac
2041 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
2101 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
2161 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
2221 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
2281 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
2341 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
2401 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
2461 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
2521 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
2581 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
2641 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
2701 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
2761 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
2821 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
2881 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
2941 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
3001 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
3061 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
3121 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
3181 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
3241 gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
3301 cacaccgcat atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
3361 gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca
3421 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
3481 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg
3541 cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca
3601 tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg
3661 gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt
3721 ctgttcatgg gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact
3781 gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg
3841 cggcgggacc agagaaaaat cactcagggt caatgccagc gcttcgttaa tacagatgta
3901 ggtgttccac agggtagcca gcagcatcct gcgatgcaga tccggaacat aatggtgcag
3961 ggcgctgact tccgcgtttc cagactttac gaaacacgga aaccgaagac cattcatgtt
4021 gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt
4081 gattcattct gctaaccagt aaggcaaccc cgccagccta gccgggtcct caacgacagg
4141 agcacgatca tgcgcacccg tggggccgcc atgccggcga taatggcctg cttctcgccg
4201 aaacgtttgg tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa gattccgaat
4261 accgcaagcg acaggccgat catcgtcgcg ctccagcgaa agcggtcctc gccgaaaatg
4321 acccagagcg ctgccggcac ctgtcctacg agttgcatga taaagaagac agtcataagt
4381 gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc
4441 aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct aacttacatt aattgcgttg
4501 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
4561 caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag
4621 tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg
4681 gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat
4741 ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatatccg caccaacgcg
4801 cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag
4861 catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat
4921 ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt
4981 atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc
5041 gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg
5101 ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac
5161 attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat
5221 cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc
5281 gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat
5341 cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag
5401 caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc
5461 catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac
5521 gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg
5581 tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa
5641 ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc tcccttatgc gactcctgca
5701 ttaggaagca gcccagtagt aggttgaggc cgttgagcac cgccgccgca aggaatggtg
5761 catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc
5821 cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg
5881 cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg
5941 cgtagaggat cgagatctcg atcccgcgaa attaatacga ctcactatag gggaattgtg
6001 agcggataac aattcccctc tagaaataat tttgtttaac tttaagaagg agatatacca
6061 tgggcagcag ccatcatcat catcatcaca gcagcggcct ggaagttctg ttccaggggc
6121 cccat
//
| {
"Vector backbone": "pVRa",
"Backbone size w/o insert (bp)": "",
"Vector type": "Bacterial Expression, Synthetic Biology",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH10B",
"Copy number": "Low Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Design of orthogonal genetic switches based on a crosstalk map of sigmas, anti-sigmas, and promoters",
"Publication": "Mol Syst Biol. 2013 Oct 29;9:702. doi: 10.1038/msb.2013.58.",
"DOI": "10.1038/msb.2013.58.",
"Authors": "Rhodius VA, Segall-Shapiro TH, Sharon BD, Ghodasara A, Orlova E, Tabakh H, Burkhardt DH, Clancy K, Peterson TC, Gross CA, Voigt CA /msb",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/24169405",
"PubMed ID": "PubMed 24169405"
} | {
"Gene/Insert name": "ECF14_1324",
"Species": "Streptomyces coelicolor A3(2)",
"Insert Size (bp)": "",
"Mutation": "Codon optimized for E.coli",
"Entrez Gene": "",
"Promoter": "pT7-lacO",
"Tag / Fusion Protein": "His6-PreScission (N terminal on backbone)"
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "NdeI (not destroyed)",
"3′ cloning site": "HindIII (not destroyed)",
"5′ sequencing primer": "CGATATAGGCGCCAGCAAC",
"3′ sequencing primer": "CAGCAGCCAACTCAGCTTC"
} | {
"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-mPCSK4 | 122675 | Expresses mouse PCSK4 in mammalian cells | Liming Pei | Li et al Mol Cell Biol. 2018 Oct 15;38(21). pii: MCB.00249-18. doi: 10.1128/MCB.00249-18. Print 2018 Nov 1. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/244737/5869b5cd-c10c-4a36-af36-ae2935c548bd/addgene-plasmid-122675-sequence-244737.gbk | full | LOCUS sequence-244737- 7480 bp DNA circular SYN 26-AUG-2024
DEFINITION Expresses mouse PCSK4 in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7480)
AUTHORS Li JJ, Liu J, Lupino K, Liu X, Zhang L, Pei L
TITLE Growth Differentiation Factor 15 Maturation Requires Proteolytic
Cleavage by PCSK3, -5, and -6.
JOURNAL Mol Cell Biol. 2018 Oct 15;38(21). pii: MCB.00249-18. doi:
10.1128/MCB.00249-18. Print 2018 Nov 1.
PUBMED 30104250
REFERENCE 2 (bases 1 to 7480)
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."; date: "2018-10-15"; volume: "38(21). pii: MCB.00249-18.
doi"; pages: " 10.1128/MCB.00249-18. Print 2018 Nov 1"
FEATURES Location/Qualifiers
source 1..7480
/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 2978..3019
/codon_start=1
/product="epitope tag from simian virus 5"
/label=V5 tag
/translation="GKPIPNPLLGLDST"
CDS 3029..3046
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(3069..3086)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 3075..3299
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 3345..3773
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3432..3451)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3642..3663
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(3782..3802)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 3787..4116
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3967..4102
/label=SV40 ori
/note="SV40 origin of replication"
CDS 4183..4977
/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(4237..4256)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4847..4866
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 5153..5274
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5190..5209)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 5244..5263
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(5323..5339)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5323..5339)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(5336..5358)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 5347..5363
/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(5371..5401)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5416..5437
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(5554..5571)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5725..6313)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5805..5824)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6484..7344)
/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 7107..7126
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7345..7449)
/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 gctggctagt
901 taagcttggt accgagctcg gatcctatgc ggccctccca gacagagctg tggctgggtc
961 tgactttgac tttggccctc ctggctgtga ggtgggcctc agcccaggcc cccatctatg
1021 tcagcagctg ggcagtgcgg gtgaccaaag gttaccagga agctgagcgc ctggcccgta
1081 aatttggctt cgtcaacctg ggacagatct tccctgacga ccagtatttc catctgcggc
1141 accggggtgt ggcccagcag tccctgactc cacactgggg ccatcgtcta cgactgaaga
1201 aagatcccaa ggtgcgctgg tttgagcagc agactttgag gcgccgggtg aaacgctccc
1261 tggtggtgcc cacagacccc tggttttcca agcagtggta catgaacaag gagatacaac
1321 aagatctcaa catcctaaag gcttggaacc agggactgac cggccgggga gtggtgattt
1381 ccatcttgga cgatggcatt gagaaagacc acccggacct ctgggctaat tatgaccctc
1441 tggccagcta tgacttcaac gactatgacc cagaccccca gcctcgctac acacccaacg
1501 atgagaaccg acatggaacg cgctgtgctg gggaggtgtc tgccacagct aacaacggtt
1561 tctgtggtgc cggtgtggcc ttcaatgcta gaattggagg cgtgcgcatg ttggatggag
1621 ccatcactga catcgtggag gctcagtccc tcagcctgca gccgcaacac atacacatct
1681 atagcgccag ctggggcccg gaggatgatg gtcgcacggt ggacggacct ggcctcctca
1741 cacaggaggc cttcaggcgt ggtgtaacca agggccgcca agggctgggt acactgttca
1801 tctgggcctc gggaaacggt ggcctccatt acgacaactg caattgtgac ggctacacca
1861 acagcatcca cacactgtca gtgggcagca ccacgcggca gggtcgagtg ccctggtaca
1921 gcgaggcctg tgcctccacg tttaccacca ccttcagcag cggtgtggtc accgacccac
1981 agatcgtcac cacggacctg caccaccagt gcaccgacaa gcacacgggc acctcggcat
2041 ctgccccgct ggctgctggc atgatcgccc tggccctgga ggccaacccg ctcctgacct
2101 ggagggacct acagcacctg gtggtccgcg cgtccaggcc ggcgcagctg caggcggagg
2161 actggaggat caacggcgtg gggcgccaag tgagccacca ttatggctat gggctgctgg
2221 acgcggggtt gctggtggac ctggctcgcg tgtggctgcc cactaagcct cagaagaaat
2281 gtgccattcg ggtggtgcac acccccaccc ccatcctgcc tcggatgctg gtgccgaaga
2341 acgtgaccgc gtgctcagat ggctcgcgcc gccgcctcat ccgctcgctc gagcacgtgc
2401 aggtccagct ttcgctctcc tacagccgcc gcggggacct ggagatcttc ctcaccagcc
2461 ccatgggcac gcgctccacg ctcgtggcca tcagaccgtt ggatatcagc ggccaaggct
2521 acaacaactg gatcttcatg tccactcact actgggatga ggacccgcag ggcctgtgga
2581 ccctgggcct ggagaataag ggctactatt ttaacacagg aactctgtac tactacaccc
2641 tgctgctgta tgggacggcc gaggacatga cagcacggcc ccaggccccc caggtgacca
2701 gccgcgcgcg cgcatgtgtg cagagggaca cagaggggct gtgccaggaa agtcacagtc
2761 ccctctccat cctggcagga ctctgcctca tctccagcca gcagtggtgg tggctctaca
2821 gccacccaca gcagccagtg accgaaggac aggccagctg tcaccctcct gttacacctg
2881 ctgcagcagc ctgagcagcg actacactgc ctgtcccctg tcccttgtgt tgggagagct
2941 ccgcggccgc tcgagtctag agggcccgcg gttcgaaggt aagcctatcc ctaaccctct
3001 cctcggtctc gattctacgc gtaccggtca tcatcaccat caccattgag tttaaacccg
3061 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
3121 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
3181 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
3241 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
3301 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
3361 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
3421 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
3481 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
3541 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
3601 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
3661 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
3721 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
3781 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
3841 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
3901 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
3961 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
4021 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
4081 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
4141 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
4201 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
4261 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
4321 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
4381 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
4441 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
4501 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
4561 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
4621 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
4681 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
4741 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
4801 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
4861 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
4921 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
4981 ggactctggg gttcgcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
5041 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
5101 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
5161 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
5221 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
5281 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
5341 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
5401 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
5461 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
5521 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
5581 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
5641 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
5701 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
5761 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
5821 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
5881 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
5941 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
6001 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
6061 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
6121 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
6181 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
6241 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
6301 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
6361 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
6421 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
6481 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
6541 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
6601 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
6661 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
6721 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
6781 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
6841 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
6901 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
6961 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
7021 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
7081 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
7141 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
7201 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
7261 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
7321 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
7381 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
7441 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
| {
"Vector backbone": "pcDNA3.1",
"Backbone size w/o insert (bp)": "5428",
"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": "Growth Differentiation Factor 15 Maturation Requires Proteolytic Cleavage by PCSK3, -5, and -6",
"Publication": "Mol Cell Biol. 2018 Oct 15;38(21). pii: MCB.00249-18. doi: 10.1128/MCB.00249-18. Print 2018 Nov 1.",
"DOI": "10.1128/MCB.00249-18.",
"Authors": "Li JJ, Liu J, Lupino K, Liu X, Zhang L, Pei L /MCB-",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/30104250",
"PubMed ID": "PubMed 30104250"
} | {
"Gene/Insert name": "Mus musculus proprotein convertase subtilisin/kexin type 4 (Pcsk4)",
"Species": "M. musculus (mouse)",
"Insert Size (bp)": "1967",
"Mutation": "",
"Entrez Gene": "Pcsk4",
"Promoter": "T7",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "BamHI (not destroyed)",
"3′ cloning site": "NotI (not destroyed)",
"5′ sequencing primer": "GTA ATA CGA CTC ACT ATA GGG",
"3′ sequencing primer": "TAG AAG GCA CAG TCG AGG"
} | {
"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": ""
} |
|
yqhD-RFP | 63845 | This plasmid is designed as a bacterial oxidative stress sensor | Howard Salis | Salis Lab Plasmids (unpublished) | https://media.addgene.org/snapgene-media/v2.0.0/sequences/106568/a311552b-1fb4-446c-a4f8-854918764cef/addgene-plasmid-63845-sequence-106568.gbk | full | LOCUS sequence-106568- 5735 bp DNA circular SYN 25-AUG-2024
DEFINITION This plasmid is designed as a bacterial oxidative stress sensor.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5735)
TITLE Salis Lab Plasmids
REFERENCE 2 (bases 1 to 5735)
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"
FEATURES Location/Qualifiers
source 1..5735
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(130..147)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(264..809)
/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(344..363)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 885..904
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
CDS 1407..2078
/codon_start=1
/product="monomeric derivative of DsRed (Campbell et al.,
2002)"
/label=mRFP1
/translation="SSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAKLK
VTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGVVT
VTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKMRL
KLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHSTG
A"
promoter 2153..2187
/label=BBa_J23102
/note="Anderson constitutive promoter
(https://parts.igem.org/Promoters/Catalog/Anderson).
Measured strength relative to BBa_J223100: 0.86"
terminator 3022..3108
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
primer_bind complement(3191..3208)
/label=pBAD Reverse
/note="For vectors with E. coli araBAD promoter, reverse
primer"
terminator 3361..3447
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 3539..3566
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
promoter 3585..3676
/gene="bla"
/label=AmpR promoter
CDS 3677..4537
/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(3895..3914)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 4579..5034
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4666..4685)
/label=F1ori-R
/note="F1 origin, reverse primer"
ORIGIN
1 tagcggagtg tatactggct tactatgttg gcactgatga gggtgtcagt gaagtgcttc
61 atgtggcagg agaaaaaagg ctgcaccggt gcgtcagcag aatatgtgat acaggatata
121 ttccgcttcc tcgctcactg actcgctacg ctcggtcgtt cgactgcggc gagcggaaat
181 ggcttacgaa cggggcggag atttcctgga agatgccagg aagatactta acagggaagt
241 gagagggccg cggcaaagcc gtttttccat aggctccgcc cccctgacaa gcatcacgaa
301 atctgacgct caaatcagtg gtggcgaaac ccgacaggac tataaagata ccaggcgttt
361 ccccctggcg gctccctcgt gcgctctcct gttcctgcct ttcggtttac cggtgtcatt
421 ccgctgttat ggccgcgttt gtctcattcc acgcctgaca ctcagttccg ggtaggcagt
481 tcgctccaag ctggactgta tgcacgaacc ccccgttcag tccgaccgct gcgccttatc
541 cggtaactat cgtcttgagt ccaacccgga aagacatgca aaagcaccac tggcagcagc
601 cactggtaat tgatttagag gagttagtct tgaagtcatg cgccggttaa ggctaaactg
661 aaaggacaag ttttggtgac tgcgctcctc caagccagtt acctcggttc aaagagttgg
721 tagctcagag aaccttcgaa aaaccgccct gcaaggcggt tttttcgttt tcagagcaag
781 agattacgcg cagaccaaaa cgatctcaag aagatcatct tattaatcag ataaaatatt
841 tctaggatca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata
901 taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag
961 aggatctgct catgtttgac agcttatcat cgatgcataa tgtgcctgtc aaatggacga
1021 agcagggatt ctgcaaaccc tatgctactc cctcgagccg gatccagcgt aatttcttag
1081 gaataatgcg gcaatttgat tgtgcgcaat tttgtagcat ttctccagca ctctggagaa
1141 ataggcaaga cattggcaga aatgagcatt gagagccagg gcgctggcga tcacaatgaa
1201 aaacatcagg cagatcgttc tctgccctca tattggccca gcaaagggag caagtaatga
1261 acaactttaa tctgcacacc ccaacccgca ttctgtttgg taaaggcgca atcgctggtt
1321 tacgcgaaca aattcctcac gatgctcgcg tattgattac ctacggcggc ggcagcgtga
1381 aaaaaaccgg gaattctaaa aatctgagct ctgaagacgt tatcaaagag ttcatgcgtt
1441 tcaaagttcg tatggaaggt tccgttaacg gtcacgagtt cgaaatcgaa ggtgaaggtg
1501 aaggtcgtcc gtacgaaggt acccagaccg ctaaactgaa agttaccaaa ggtggtccgc
1561 tgccgttcgc ttgggacatc ctgtccccgc agttccagta cggttccaaa gcttacgtta
1621 aacacccggc tgacatcccg gactacctga aactgtcctt cccggaaggt ttcaaatggg
1681 aacgtgttat gaacttcgaa gacggtggtg ttgttaccgt tacccaggac tcctccctgc
1741 aagacggtga gttcatctac aaagttaaac tgcgtggtac caacttcccg tccgacggtc
1801 cggttatgca gaaaaaaacc atgggttggg aagcttccac cgaacgtatg tacccggaag
1861 acggtgctct gaaaggtgaa atcaaaatgc gtctgaaact gaaagacggt ggtcactacg
1921 acgctgaagt taaaaccacc tacatggcta aaaaaccggt tcagctgccg ggtgcttaca
1981 aaaccgacat caaactggac atcacctccc acaacgaaga ctacaccatc gttgaacagt
2041 acgaacgtgc tgaaggtcgt cactccaccg gtgcttaata aggccggccg tgctagtgta
2101 gaaatagccg ccagttccgc tggcggcatt ttaactttct ttagtactag tattgacagc
2161 tagctcagtc ctaggtactg tgctagcacc ccaatcgacg tcatactaga atcgggtcac
2221 ggaggtacaa taataatgcg taaactcgag gaacttttca cgggggtcgt cccaattctg
2281 gtagaattag acggtgatgt aaacgggcac aagtttagcg tcagcggaga gggcgagggg
2341 gacgccacct acggtaaact tacccttaag tttatttgca cgacgggcaa attacccgtt
2401 ccgtggccaa cgctcgtcac tactttcggc tacggcgttc agtgcttcgc gagatatccg
2461 gatcacatga agcagcacga tttcttcaag tcggccatgc cagagggtta cgttcaggag
2521 agaactatat ttttcaagga cgacggcaat tacaagacac gcgccgaagt caagttcgaa
2581 ggcgataccc tggttaaccg tatcgagtta aaaggtatcg attttaaaga agacggaaac
2641 attctcggtc acaaactcga atacaactat aacagccata atgtctacat tatggctgac
2701 aaacagaaga acggcattaa ggttaacttc aaaattcgcc acaacatcga agacggtagc
2761 gtccaactag cagaccatta tcaacaaaat actcctatcg gcgacggccc agtcctttta
2821 ccagacaacc attaccttag cacacaaagc gcccttagca aagatcccaa cgagaagcgc
2881 gaccacatgg tccttctcga atttgtcaca gcggcaggga tcacccacgg catggacgaa
2941 ctctacaaat aataattaat taaaaaaaaa aaccccgccc ctgacagggc ggggtttttt
3001 tttctaatat gcggccgcca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt
3061 cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgccctag
3121 agcatgcgcg ttcggctgcg gcgagcggta tcaggcttgg ctgttttggc ggatgagaga
3181 agattttcag cctgatacag attaaatcag aacgcagaag cggtctgata aaacagaatt
3241 tgcctggcgg cagtagcgcg gtggtcccac ctgaccccat gccgaactca gaagtgaaac
3301 gccgtagcgc cgatggtagt gtggggtctc cccatgcgag agtagggaac tgccaggcat
3361 caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg
3421 gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa
3481 cggcccggag ggtggcgggc aggacgcccg ccataaactg ccaggcatca aattaagcag
3541 aaggccatcc tgacggatgg cctttttgcg tttctacaaa ctcttttgtt tatttttcta
3601 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
3661 ttgaaaaagg aagactatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
3721 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
3781 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
3841 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
3901 tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
3961 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
4021 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
4081 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
4141 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
4201 gcgtgacacc acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga
4261 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
4321 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
4381 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
4441 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
4501 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
4561 tatactttag attgatttac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
4621 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
4681 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
4741 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg
4801 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
4861 ccacgttctt taatagtgga ctcttgttcc aaacttgaac aacactcaac cctatctcgg
4921 gctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
4981 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca atttaaaagg
5041 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
5101 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
5161 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
5221 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
5281 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
5341 ccgcctacat acctcgctct gctaatcctg ttaccagtca ggcatttgag aagcacacgg
5401 tcacactgct tccggtagtc aataaaccgg taaaccagca atagacataa gcggctattt
5461 aacgaccctg ccctgaaccg acgaccgggt cgaatttgct ttcgaatttc tgccattcat
5521 ccgcttatta tcacttattc aggcgtagca ccaggcgttt aagggcacca ataactgcct
5581 taaaaaaatt acgccccgcc ctgccactca tcgcagtact gttgtaattc attaagcatt
5641 ctgccgacat ggaagccatc acagacggca tgatgaacct gaatcgccag cggcatcagc
5701 accttgtcgc cttgcgtata atatttgccc atggc
//
| {
"Vector backbone": "P15A",
"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)": "DH10B",
"Copy number": "Low 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": "yqhD, mRFP1 and GFPmut3b",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "Gene is translationally fused to RFP",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": "mRFP1 (C terminal on insert)"
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "unknown (unknown if destroyed)",
"3′ cloning site": "unknown (unknown if destroyed)",
"5′ sequencing primer": "RNAde-BB-3 (tacggacgaccttcaccttc)",
"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": ""
} |
|
pMDJ22 - pEXP[Psmu-2 | Cre Recombinase (PATCs) | gpd-2 tagRFP-T smu-1 UTR] | 159814 | Optimized enzyme for genome editing in C. elegans | Christian Froekjaer-Jensen | Aljohani et al Nat Commun. 2020 Dec 9;11(1):6300. doi: 10.1038/s41467-020-19898-0. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/317354/b2d35e20-5ad5-463d-80c9-578e9ac73512/addgene-plasmid-159814-sequence-317354.gbk | full | LOCUS sequence-317354- 10042 bp DNA circular SYN 25-AUG-2024
DEFINITION Optimized enzyme for genome editing in C. elegans.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10042)
AUTHORS Aljohani MD, El Mouridi S, Priyadarshini M, Vargas-Velazquez AM,
Frokjaer-Jensen C
TITLE Engineering rules that minimize germline silencing of transgenes in
simple extrachromosomal arrays in C. elegans.
JOURNAL Nat Commun. 2020 Dec 9;11(1):6300. doi: 10.1038/s41467-020-19898-0.
PUBMED 33298957
REFERENCE 2 (bases 1 to 10042)
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
Commun."; date: "2020-12-9"; volume: "11(1):6300. doi"; pages: "
10.1038/s41467-020-19898-0"
FEATURES Location/Qualifiers
source 1..10042
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 1..17
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 1..17
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
protein_bind 37..57
/gene="mutant version of attB"
/label=attB4
/bound_moiety="BP Clonase(TM)"
/note="core recombination site for the Gateway(R) BP
reaction"
protein_bind 2121..2145
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
protein_bind complement(4393..4417)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
protein_bind complement(7655..7675)
/gene="mutant version of attB"
/label=attB3
/bound_moiety="BP Clonase(TM)"
/note="core recombination site for the Gateway(R) BP
reaction"
primer_bind complement(7683..7700)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(7683..7699)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(7692..7714)
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 8027..8049
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(8087..8105)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 8173..8277
/gene="bla"
/label=AmpR promoter
CDS 8278..9138
/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 complement(8496..8515)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 9309..9897
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 9798..9817
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind join(10024..10042,1..4)
/label=M13/pUC Reverse
/note="In lacZ gene"
ORIGIN
1 caggaaacag ctatgaccat gattacgcca agctatcaac tttgtataga aaagttgtgc
61 accaatttga gtttccatgg ggaatcaagc ggataccgat tgagttgaag aaccctaaat
121 aaaattttat atattttacc ttccaacttc gaaaaaatca ctcaccattt tagcatgttt
181 ccctcaaaaa ccgtggcaaa ttgctcttgg aaaataatag acacatcagt tggtcgagca
241 ttttttggag gaattttaaa actgtgatga tggttgaagg tgactatatt gtctgcgagt
301 tgatactctg gaatattgat ttttttgaag atttcatagt cttcttggat ttcaaaaatt
361 gaaatatttc agctgtatta aaagagaata aataaaatga tattgcctag gggtgagcgg
421 caaacgattt tttccggcaa atcggcaaat cgacaaattg ccggaattgg aaatttccag
481 caaatcggca tatcggcaat ttgccggaat ttaaaatttc cggcaaaccg gcaaaccggc
541 aaattgccga tttgccgaac ttcggtaaat ttgcaaatcg gtaatttgcc gaaaattttc
601 ggcaaattgt ggttttgcac ttattttttg gaatttcaat ttcaaatggc aaaattaaac
661 gcatcatatg aatttttcta tatctatttt gagaagttct atgaaaatat gtagaaaaaa
721 cgggaaaaat tcaataatag acagttttaa gtgcttccgt cttataaaaa atccctctaa
781 acatttccgg caaatctggt attcggcaaa tcggcaagtc ggcaatttgc cgtaaattaa
841 aatttccggc aaaacggcaa accggcaaat tgccggaatt ggaaatttcc ggcaaatcgg
901 caaattgctt gcccattttt tgattcttag tcggcgtttt acacaaaaga taaattccaa
961 taaaagacag aactttgctt tttgccagaa attaaaaaat tggggcaaac caatccaaat
1021 atgaagtgta aacttgggtt tgcaccaggg ctgtgcggaa accggcaatt gccgaaattg
1081 ccgaacccta aaaactttgg caaccggcaa cttttgctgg ctggaacttc ggcaactttt
1141 cggcaaacgg caacttggtt gccggttgcg caaagctctg gtttgcgttt agtagcaagg
1201 cttcctgaaa gaattccgga ggaaaactcc ccctaactca ccttttactc tatgcatctg
1261 cttaattttg tgcaattgta gtgcaaatag taggaaatga attttttttc cgccattttt
1321 tccttttcct atttctgctt ttcaatttcc acgtggagct cttttaaaat gaaaatttaa
1381 actctagata tttcacgttt taaaaaaaaa aaattttaat tttctatcgg ttttactttt
1441 gacgaaaagg gttcacagac attggaaata atcatcattt actatatata aagcgcttat
1501 tctgtgtgtc catagtttgt agtctctgta gtctttgtag tctgtgaagt tttggcttct
1561 ggagggatag tgagttgggg tcagtgtagg gatatagtcg gggtactgta gtggtacaat
1621 agtggtacgg taggaatact gtagggttac ggtagtttca taaaaaatta attttcagcc
1681 ccagaagtcg ggggccgcgc cgaaggtgcc tagacctaag cctaggcctc agcctaagcc
1741 taattacatc aaaaaagcta tgacttttgg atcattttga cttcttttct agaacgcata
1801 atgaaataat tcttaatttc atcggctttt tctcatcgtt gcttagaagt attgtcttga
1861 aaattgatat ttttagaaag tacacatatt tctcaatcct tttatttaaa tttctgaaag
1921 aaactttgcc acaacggtga gaaaagggtg cccgaaaaat gtgcatttaa atgtgaaaaa
1981 tatccgtggc cgcggcctaa gaacccccgt cggccatgtg gtgcaagtgc gctctgttgc
2041 ggatttttat atttaattgc attttcgcat attcgaacga ttttcctgat tttccagccg
2101 aaattgtcca ttaaaaagtg gcaagtttgt acaaaaaagc aggctaaaaa tgtccaacct
2161 cctcaccgtc caccaaaacc tcccagccct cccagtcgac gccacctccg acgaggtccg
2221 taaggtattt tcctgcattt ttcaactggg aaaatgaaag aaaatcgata atttcagaac
2281 ctcatggaca tgttccgtga ccgtcaagcc ttctccgagc acacctggaa gatgctcctc
2341 tccgtctgcc gttcctgggc cgcctggtgc aagctcaaca accgtaagtg gttcccagcc
2401 gagccaggta agttttctta tgggaaagaa ggaaaaaacc gagattttac ttgaaaaatt
2461 gaatttttcg cgggattttc accaaaaatt gttgaatatt cattatttca cgctgtaaaa
2521 caaaaaaaaa aaaaatcaaa aactacgttg aaatcgcgtt tttaagcgaa ttttcttcag
2581 aattgccaga ttttaacccc aaattttgca gtttttaaat aaaatttcac cttttcggct
2641 caaattgtag attttcctga aaatttagta caaaaaacaa tttcctcgta aatttttcaa
2701 atagattttc agaggacgtc cgtgactacc tcctctacct ccaagcccgt ggactcgccg
2761 tcaagaccat ccaacaacac ctcggacaac tcaacatgct ccaccgtcgt tccggactcc
2821 cacgtccatc cgactccaac gccgtctccc tcgtcatgcg tcgtatccgt aaggagaacg
2881 tcgacgccgg agagcgtgcc aagcaagccc tcgccttcga gcgtaccgac ttcgaccaag
2941 tccgttccct catggagaac tccgaccgtt gccaaggtaa gttatgtcta cctgcctgcc
3001 taccgcctaa attttttgtg aagtttcttc aaaaaatcca gaaaaaaaaa caattttcat
3061 acgatttttt cccttaaaat tgtgaatttt catgcttttt agccccaaaa gtcattattt
3121 gagaaaaaat ttcatacaaa aaagttttga gaaatacaca attttttaaa tgtaattttc
3181 aaattttcaa ttttcaacta gaaaattcac aaaacttgta aattttggac caaaacattt
3241 atacaattac tttttttgaa tctaataact acaataacta caaatttaac attttcagac
3301 atccgtaacc tcgccttcct cggaatcgcc tacaacaccc tcctccgtat cgccgagatc
3361 gcccgtatcc gtgtcaagga catctcccgt accgacggag gacgtatgct catccacatc
3421 ggacgtacca agaccctcgt ctccaccgcc ggagtcgaga aggccctctc cctcggagtc
3481 accaagctcg tcgagcgttg gatctccgtc tccggagtcg ccgacgaccc aaacaactac
3541 ctcttctgcc gtgtccgtaa ggtaagttct ttttttgaaa agtgcagttt gtagtctaat
3601 tttcatttat tttcttttta aaaaacgcat caatttttaa atattttggg acaaaatccg
3661 aaaactgtac taatttgtag tttgtaaaat taaaaaaaaa acgcaaaaaa atgttttcaa
3721 aaatgcttag aataaaaata aaatttctaa aaaattgaac aaaataaaca tttaaaaatt
3781 caaaaagttt gaaaaaatgc aaagtttttt aatgcaaaaa atatttaatg tttaaacaaa
3841 tttaaaaaat gttgtaagaa tatgtttaga gacagttttt aaatgttaaa ttgcaaaaaa
3901 aaaaagcaaa aaaaaatttg aaaaatgtaa cagaaaattt taattaaaaa agtcgtgttt
3961 ttttcaaacc tgcaattctt tgtaatttta tttataaaga gaaacaaatc aaattgtata
4021 ttacgaaaaa cgatactaaa attcgaaaat ttgcggtttt ttgcgtaaaa aatacggttc
4081 cgtaattttc agaacggagt cgccgcccca tccgccacct cccaactctc cacccgtgcc
4141 ctcgagggaa tcttcgaggc cacccaccgt ctcatctacg gagccaagga cgactccgga
4201 caacgttacc tcgcctggtc cggacactcc gcccgtgtcg gagccgcccg tgacatggcc
4261 cgtgccggag tctccatccc agagatcatg caagccggag gatggaccaa cgtcaacatc
4321 gtcatgaact acatccgtaa cctcgactcc gagacgggag ccatggtccg tctcctcgag
4381 gacggagact agacccagct ttcttgtaca aagtggccta agctgtctca tcctactttc
4441 acctagttaa ctgcttgtct taaaatctat gcttctcttt agtatctaaa attttcctag
4501 aagcttacaa gtatataaat ggtctcttct caataaaggt tgtatattta ttcatcttat
4561 tgaatctgcc atttcctcgt ttttgcgagt ttatatacct tccaattttc tttctattgt
4621 attttcaact tctaatttta attcagggaa actgcttcaa cgcatcatgg gttgtaccat
4681 gtcacaggaa gagcgtgccg ctcttgaaag atcacgaatg tcctcagtct ccaaggtaac
4741 acttagtttt tgttgctctg tttaaaaatt aattttaagg gagaggagct catcaaggag
4801 aacatgcaca tgaagctcta catggaggga accgtcaaca accaccactt caagtgcacc
4861 tccgagggag agggaaagcc atacgaggga acccaaacca tgcgtatcaa ggtcgtcgag
4921 ggaggaccac tcccattcgc cttcgacatc ctcgccacct ccttcatgta cggatcgcgt
4981 accttcatca accacaccca aggtaagttt tcttatggga aagaaggaaa aaaccgagat
5041 tttacttgaa aaattgaatt tttcgcggga ttttcaccaa aaattgttga atattcatta
5101 tttcacgctg taaaacaaaa aaaaaaaaaa tcaaaaacta cgttgaaatc gcgtttttaa
5161 gcgaattttc ttcagaattg ccagatttta accccaaatt ttgcagtttt taaataaaat
5221 ttcacctttt cggctcaaat tgtagatttt cctgaaaatt tagtacaaaa aacaatttcc
5281 tcgtaaattt ttcaaataga ttttcaggaa tcccagactt cttcaagcaa tccttcccag
5341 agggattcac ctgggagcgt gtcaccacct acgaggacgg aggagtcctc accgccaccc
5401 aagacacctc cctccaagac ggatgcctca tctacaacgt caaggtaagt tatgtctacc
5461 tgcctgccta ccgcctaaat tttttgtgaa gtttcttcaa aaaatccaga aaaaaaaaca
5521 attttcatac gattttttcc cttaaaattg tgaattttca tgctttttag ccccaaaagt
5581 cattatttga gaaaaaattt catacaaaaa agttttgaga aatacacaat tttttaaatg
5641 taattttcaa attttcaatt ttcaactaga aaattcacaa aacttgtaaa ttttggacca
5701 aaacatttat acaattactt tttttgaatc taataactac aataactaca aatttaacat
5761 tttcagatcc gtggagtcaa cttcccatcc aacggaccag tcatgcaaaa gaagaccctc
5821 ggatgggagg ccaacaccga gatgctctac ccagccgacg gaggactgga gggacgtacc
5881 gacatggccc tcaagctcgt cggaggagga cacctcatct gcaacttcaa gaccacctac
5941 cgttccaaga agccagccaa ggtaagttct ttttttgaaa agtgcagttt gtagtctaat
6001 tttcatttat tttcttttta aaaaacgcat caatttttaa atattttggg acaaaatccg
6061 aaaactgtac taatttgtag tttgtaaaat taaaaaaaaa acgcaaaaaa atgttttcaa
6121 aaatgcttag aataaaaata aaatttctaa aaaattgaac aaaataaaca tttaaaaatt
6181 caaaaagttt gaaaaaatgc aaagtttttt aatgcaaaaa atatttaatg tttaaacaaa
6241 tttaaaaaat gttgtaagaa tatgtttaga gacagttttt aaatgttaaa ttgcaaaaaa
6301 aaaaagcaaa aaaaaatttg aaaaatgtaa cagaaaattt taattaaaaa agtcgtgttt
6361 ttttcaaacc tgcaattctt tgtaatttta tttataaaga gaaacaaatc aaattgtata
6421 ttacgaaaaa cgatactaaa attcgaaaat ttgcggtttt ttgcgtaaaa aatacggttc
6481 cgtaattttc agaacctcaa gatgccagga gtctactacg tcgaccaccg tctggagcgt
6541 atcaaggagg ccgacaagga gacgtacgtc gagcaacacg aggtcgccgt cgcccgttac
6601 tgcgacctcc catccaagct cggacacaag ctcaacggaa tggacgagct ctacaagtcg
6661 ggataatttt ccccagattt ccccacgatt ttttttgaca tttttatgaa aattttctct
6721 aaaagttacc cctattttgt ttagactgat ctcttatttt atccaatgtt tcaaattttt
6781 gaaattgctc gaaaattcca attttcacat atttttgttg attttttcgc cattttcccc
6841 tgataaatta acttattttt gcaatttttt ctttcgaaaa aatattggtt tctcatcacc
6901 atttatcctt ttttttatgg agttaaaaat tgttaaaaaa atttaatttt ttttctaatt
6961 ttagagtcaa aacgggttcg aatctagagc gtagctgtac gattttttgg cgatttttgc
7021 ggttttctgg tgagtttttt taaagttatt tttttgaaaa aaattcttga aaataaaagg
7081 aaattgtttt aaaaaaaaat ttcaaagaaa aaattttgaa aaaattgaaa catttttttt
7141 attttgacca aaaatattga aaattaatta acattttttt ttggaaagtt aatttttaat
7201 acaaattttt taaatttatt ttacaaaaaa aattgaattt ctgtgaaaaa tcagttttcc
7261 agtacataat gatttgacca aaaaaaattg aaaaaaatga ttttttaaaa ttaaaaatac
7321 cataaaatgc tgaaaattca gaaaaataag taaattttaa aaaattctgg aaaaacatct
7381 aaaaaccccc caaatataat attttggacc cttttatttt ctgctcgaca tctcaaatat
7441 cctccaattt ttcgcttttt tgattaattg aaagcatagt gggggcaggt gttataggac
7501 acccttccct ccactttttt ggagcatttt tcaaagctct tccgagagca gaagcttctc
7561 aatttttcca ccccaaaagt ttgaaattct gccaaaaaat ctcgcatttt aacaattttt
7621 cgagatgttt cacgtccttt tcattgcgat ttcccaactt tattatacat agttgataat
7681 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
7741 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
7801 cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc
7861 cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtgc aatctgctct
7921 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
7981 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
8041 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
8101 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
8161 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
8221 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
8281 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
8341 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
8401 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
8461 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
8521 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
8581 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
8641 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
8701 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
8761 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
8821 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
8881 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
8941 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tggctctcgc
9001 ggtatcattg cagcactggg gtcagatggt aagccctccc gtatcgtagt tatctacacg
9061 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
9121 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
9181 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
9241 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
9301 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
9361 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
9421 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
9481 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
9541 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
9601 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
9661 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
9721 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
9781 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
9841 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
9901 gccagcaacg cggccttttt acggttcctg gtcttttgct ggccttttgc tcacatgttc
9961 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
10021 accagcggat aacaatttca ca
//
| {
"Vector backbone": "pDEST R4-R3",
"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 ; Evrogen Limited Use Label License for FPs",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Engineering rules that minimize germline silencing of transgenes in simple extrachromosomal arrays in C. elegans",
"Publication": "Nat Commun. 2020 Dec 9;11(1):6300. doi: 10.1038/s41467-020-19898-0.",
"DOI": "10.1038/s41467-020-19898-0.",
"Authors": "Aljohani MD, El Mouridi S, Priyadarshini M, Vargas-Velazquez AM, Frokjaer-Jensen C /s---",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/33298957",
"PubMed ID": "PubMed 33298957"
} | {
"Gene/Insert name": "pEXP[Psmu-2 | Cre Recombinase (PATCs) | gpd-2 tagRFP-T smu-1 UTR]",
"Species": "C. elegans (nematode)",
"Insert Size (bp)": "",
"Mutation": "Not applicable",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Gateway 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": ""
} |
|
FKBP_cSbMVp | 118968 | Expresses C-terminus of soybean mosaic virus protease (SbMVp) fused to the FK-506 binding protein (FKBP) | Roman Jerala | Fink et al Nat Chem Biol. 2019 Feb;15(2):115-122. doi: 10.1038/s41589-018-0181-6. Epub 2018 Dec 10. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/232050/3178011f-4816-47e3-a35a-9f006d948a1d/addgene-plasmid-118968-sequence-232050.gbk | full | LOCUS sequence-232050- 6199 bp DNA circular SYN 26-AUG-2024
DEFINITION Expresses C-terminus of soybean mosaic virus protease (SbMVp) fused
to the FK-506 binding protein (FKBP).
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6199)
AUTHORS Fink T, Lonzaric J, Praznik A, Plaper T, Merljak E, Leben K, Jerala
N, Lebar T, Strmsek Z, Lapenta F, Bencina M, Jerala R
TITLE Design of fast proteolysis-based signaling and logic circuits in
mammalian cells.
JOURNAL Nat Chem Biol. 2019 Feb;15(2):115-122. doi:
10.1038/s41589-018-0181-6. Epub 2018 Dec 10.
PUBMED 30531965
REFERENCE 2 (bases 1 to 6199)
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/s41589-018-0181-6"; journalName: "Nat Chem Biol"; date:
"2019-02"; volume: "15"; issue: "2"; pages: "115-122"
FEATURES Location/Qualifiers
source 1..6199
/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 958..967
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 964..1287
/codon_start=1
/product="human FK506-binding protein FKBP12"
/label=FKBP
/translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
LKLE"
CDS 1675..1701
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
promoter complement(1752..1770)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(1753..1770)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(1790..1807)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 1796..2020
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2066..2494
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2153..2172)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2363..2384
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2503..2523)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2508..2837
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2688..2823
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2750..2769
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2904..3698
/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(2958..2977)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3568..3587
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 3872..3993
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3909..3928)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3963..3982
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4042..4058)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4042..4058)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4055..4077)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4066..4082
/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(4090..4120)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4135..4156
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4273..4290)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4444..5032)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4524..4543)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5203..6063)
/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 5826..5845
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6064..6168)
/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 gcttggtacc
901 gagctcggat ccactagtaa cggccgccag tgtgctggaa ttcgcggccg ctctagagcc
961 accatgggag tgcaagtgga aaccatctcc ccaggagacg ggcgcacctt ccccaagcgc
1021 ggccagacct gcgtggtgca ctacaccggg atgcttgaag atggaaagaa atttgattcc
1081 tcccgggaca gaaacaagcc ctttaagttt atgctaggca agcaggaggt gatccgaggc
1141 tgggaagaag gggttgccca gatgagtgtg ggtcagagag ccaaactgac tatatctcca
1201 gattatgcct atggtgccac tgggcaccca ggcatcatcc caccacatgc cactctcgtc
1261 ttcgatgtgg agcttctaaa actggaagga tccggaagta agagcctgcg cgccaccgtg
1321 agcgagagca gcatgatcct gcccgagggc aagggcagct tctggataca ctggatcacc
1381 acccaggacg gcttctgcgg cctgcccctg gtgagcgtga acgacggcca catcgtgggc
1441 atccacggcc tgaccagcaa cgacagcgag aagaacttct tcgtgcccct gaccgacggc
1501 ttcgagaagg agtacctgga gaacgccgac aacctgagct gggacaagca ctggttctgg
1561 gagcccagca agatcgcctg gggcagcctg aacctggtgg aggagcagcc caaggaggag
1621 ttcaagatca gcaagctggt gagcgacctg ttcggcaaca ccgtgaccgt gcagtaccca
1681 tacgatgttc cagattacgc ttaaactagt gcggccgcct gcagctcgag catgcatcta
1741 gagggcccta ttctatagtg tcacctaaat gctagagctc gctgatcagc ctcgactgtg
1801 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
1861 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
1921 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
1981 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
2041 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
2101 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
2161 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
2221 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
2281 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
2341 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
2401 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
2461 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
2521 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
2581 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
2641 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
2701 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
2761 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
2821 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
2881 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
2941 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
3001 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
3061 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
3121 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
3181 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
3241 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
3301 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
3361 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
3421 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
3481 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
3541 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
3601 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
3661 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
3721 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
3781 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
3841 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
3901 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
3961 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
4021 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
4081 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
4141 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
4201 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
4261 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
4321 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
4381 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
4441 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
4501 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
4561 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
4621 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
4681 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
4741 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
4801 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
4861 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
4921 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
4981 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
5041 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
5101 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
5161 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
5221 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
5281 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
5341 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
5401 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
5461 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
5521 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
5581 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
5641 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
5701 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
5761 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
5821 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
5881 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
5941 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
6001 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
6061 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
6121 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
6181 aaaagtgcca cctgacgtc
//
| {
"Vector backbone": "pcDNA3",
"Backbone size w/o insert (bp)": "5443",
"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": "Design of fast proteolysis-based signaling and logic circuits in mammalian cells",
"Publication": "Nat Chem Biol. 2019 Feb;15(2):115-122. doi: 10.1038/s41589-018-0181-6. Epub 2018 Dec 10.",
"DOI": "10.1038/s41589-018-0181-6.",
"Authors": "Fink T, Lonzaric J, Praznik A, Plaper T, Merljak E, Leben K, Jerala N, Lebar T, Strmsek Z, Lapenta F, Bencina M, Jerala R /s---",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/30531965",
"PubMed ID": "PubMed 30531965"
} | {
"Gene/Insert name": "split cSbMVp in fusion with FKBP",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "CMV",
"Tag / Fusion Protein": "HA tag (C terminal on insert)"
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "EcoRI (unknown if destroyed)",
"3′ cloning site": "XhoI (unknown if destroyed)",
"5′ sequencing primer": "CGCAAATGGGCGGTAGGCGTG",
"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": ""
} |
|
pAG305GAL-ccdB-TAP | 14257 | (Empty Backbone) | Susan Lindquist | Alberti et al Yeast. 2007 Oct . 24(10):913-9. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/7920/74038d84-29f5-4111-8f6d-611e3f3f5185/addgene-plasmid-14257-sequence-7920.gbk | full | LOCUS sequence-7920-ex 8787 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 8787)
AUTHORS Alberti S, Gitler AD, Lindquist S
TITLE A suite of Gateway cloning vectors for high-throughput genetic
analysis in Saccharomyces cerevisiae.
JOURNAL Yeast. 2007 Oct . 24(10):913-9.
PUBMED 17583893
REFERENCE 2 (bases 1 to 8787)
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: "Yeast. 2007
Oct . 24(10):913-9."
FEATURES Location/Qualifiers
source 1..8787
/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"
CDS complement(663..1757)
/codon_start=1
/gene="S. cerevisiae LEU2"
/product="3-isopropylmalate dehydrogenase, required for
leucine biosynthesis"
/label=LEU2
/note="yeast auxotrophic marker"
/translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
DLGGSNSTTEVGDAVAEEVKKILA"
promoter complement(1770..2174)
/gene="S. cerevisiae LEU2"
/label=LEU2 promoter
rep_origin complement(2474..2929)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2611..2632)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2823..2842
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3059..3081
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 3073..3090
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 3074..3090
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 3097..3116
/label=T7
/note="T7 promoter, forward primer"
promoter 3097..3115
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 3135..3576
/gene="S. cerevisiae GAL1"
/label=GAL1 promoter
/note="inducible promoter, regulated by Gal4"
protein_bind 3135..3252
/label=UAS
/bound_moiety="Gal4"
/note="upstream activating sequence mediating
Gal4-dependent induction"
primer_bind 3547..3570
/label=GAL1
/note="S. cerevisiae GAL1 promoter, forward primer"
primer_bind 3591..3607
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 3591..3607
/label=pBluescriptSK
/note="For pBluescript vector"
protein_bind 3615..3739
/gene="mutant version of attR"
/label=attR1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
promoter 3764..3794
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
CDS 3848..4507
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind complement(3914..3933)
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
CDS 4852..5157
/codon_start=1
/gene="ccdB"
/product="CcdB, a bacterial toxin that poisons DNA gyrase"
/label=ccdB
/note="Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains."
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
primer_bind 5036..5055
/label=ccdB-fwd
/note="ccdB gene, forward primer"
protein_bind complement(5198..5322)
/gene="mutant version of attR"
/label=attR2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
CDS 5381..5458
/codon_start=1
/product="calmodulin-binding peptide"
/label=CBP
/note="derived from skeletal muscle myosin light chain
kinase; binds calmodulin with nanomolar affinity in the
presence of calcium"
/translation="KRRWKKNFIAVSAANRFKKISSSGAL"
CDS 5486..5506
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQG"
terminator 6257..6504
/gene="S. cerevisiae CYC1"
/label=CYC1 terminator
/note="transcription terminator for CYC1"
primer_bind complement(6273..6291)
/label=CYC1
/note="CYC1 transcription termination signal, reverse
primer"
promoter complement(6527..6545)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(6566..6582)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6566..6582)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(6579..6601)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 6590..6606
/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(6614..6644)
/label=lac promoter
/note="promoter for the E. coli lac operon"
primer_bind complement(6797..6814)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(6968..7556)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(7727..8587)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 8350..8369
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(8588..8692)
/gene="bla"
/label=AmpR promoter
primer_bind 8760..8778
/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 accatatcga ctacgtcgta aggccgtttc tgacagagta aaattcttga gggaactttc
241 accattatgg gaaatggttc aagaaggtat tgacttaaac tccatcaaat ggtcaggtca
301 ttgagtgttt tttatttgtt gtattttttt ttttttagag aaaatcctcc aatatcaaat
361 taggaatcgt agtttcatga ttttctgtta cacctaactt tttgtgtggt gccctcctcc
421 ttgtcaatat taatgttaaa gtgcaattct ttttccttat cacgttgagc cattagtatc
481 aatttgctta cctgtattcc tttactatcc tcctttttct ccttcttgat aaatgtatgt
541 agattgcgta tatagtttcg tctaccctat gaacatattc cattttgtaa tttcgtgtcg
601 tttctattat gaatttcatt tataaagttt atgtacaaat atcataaaaa aagagaatct
661 ttttaagcaa ggattttctt aacttcttcg gcgacagcat caccgacttc ggtggtactg
721 ttggaaccac ctaaatcacc agttctgata cctgcatcca aaaccttttt aactgcatct
781 tcaatggcct taccttcttc aggcaagttc aatgacaatt tcaacatcat tgcagcagac
841 aagatagtgg cgatagggtc aaccttattc tttggcaaat ctggagcaga accgtggcat
901 ggttcgtaca aaccaaatgc ggtgttcttg tctggcaaag aggccaagga cgcagatggc
961 aacaaaccca aggaacctgg gataacggag gcttcatcgg agatgatatc accaaacatg
1021 ttgctggtga ttataatacc atttaggtgg gttgggttct taactaggat catggcggca
1081 gaatcaatca attgatgttg aaccttcaat gtagggaatt cgttcttgat ggtttcctcc
1141 acagtttttc tccataatct tgaagaggcc aaaacattag ctttatccaa ggaccaaata
1201 ggcaatggtg gctcatgttg tagggccatg aaagcggcca ttcttgtgat tctttgcact
1261 tctggaacgg tgtattgttc actatcccaa gcgacaccat caccatcgtc ttcctttctc
1321 ttaccaaagt aaatacctcc cactaattct ctgacaacaa cgaagtcagt acctttagca
1381 aattgtggct tgattggaga taagtctaaa agagagtcgg atgcaaagtt acatggtctt
1441 aagttggcgt acaattgaag ttctttacgg atttttagta aaccttgttc aggtctaaca
1501 ctaccggtac cccatttagg accacccaca gcacctaaca aaacggcatc aaccttcttg
1561 gaggcttcca gcgcctcatc tggaagtggg acacctgtag catcgatagc agcaccacca
1621 attaaatgat tttcgaaatc gaacttgaca ttggaacgaa catcagaaat agctttaaga
1681 accttaatgg cttcggctgt gatttcttga ccaacgtggt cacctggcaa aacgacgatc
1741 ttcttagggg cagacatagg ggcagacatt agaatggtat atccttgaaa tatatatata
1801 tattgctgaa atgtaaaagg taagaaaagt tagaaagtaa gacgattgct aaccacctat
1861 tggaaaaaac aataggtcct taaataatat tgtcaacttc aagtattgtg atgcaagcat
1921 ttagtcatga acgcttctct attctatatg aaaagccggt tccggcctct cacctttcct
1981 ttttctccca atttttcagt tgaaaaaggt atatgcgtca ggcgacctct gaaattaaca
2041 aaaaatttcc agtcatcgaa tttgattctg tgcgatagcg cccctgtgtg ttctcgttat
2101 gttgaggaaa aaaataatgg ttgctaagag attcgaactc ttgcatctta cgatacctga
2161 gtattcccac agttaactgc ggtcaagata tttcttgaat caggcgcctt agaccgctcg
2221 gccaaacaac caattacttg ttgagaaata gagtataatt atcctataaa tataacgttt
2281 ttgaacacac atgaacaagg aagtacagga caattgattt tgaagagaat gtggattttg
2341 atgtaattgt tgggattcca tttttaataa ggcaataata ttaggtatgt ggatatacta
2401 gaagttctcc tcgagggtcg atatgcggtg tgaaataccg cacagatgcg taaggagaaa
2461 ataccgcatc aggaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt
2521 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
2581 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
2641 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
2701 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
2761 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
2821 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
2881 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc gcgccattcg
2941 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
3001 cagctggcga aggggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
3061 cagtcacgac gttgtaaaac gacggccagt gaattgtaat acgactcact atagggcgaa
3121 ttggagctct agtacggatt agaagccgcc gagcgggcga cagccctccg acggaagact
3181 ctcctccgtg cgtcctcgtc ttcaccggtc gcgttcctga aacgcagatg tgcctcgcgc
3241 cgcactgctc cgaacaataa agattctaca atactagctt ttatggttat gaagaggaaa
3301 aattggcagt aacctggccc cacaaacctt caaattaacg aatcaaatta acaaccatag
3361 gatgataatg cgattagttt tttagcctta tttctggggt aattaatcag cgaagcgatg
3421 atttttgatc tattaacaga tatataaatg gaaaagctgc ataaccactt taactaatac
3481 tttcaacatt ttcagtttgt attacttctt attcaaatgt cataaaagta tcaacaaaaa
3541 attgttaata tacctctata ctttaacgtc aaggagaaaa aaccccggat tctagaacta
3601 gtggatcccc catcacaagt ttgtacaaaa aagctgaacg agaaacgtaa aatgatataa
3661 atatcaatat attaaattag attttgcata aaaaacagac tacataatac tgtaaaacac
3721 aacatatcca gtcactatgg cggccgcatt aggcacccca ggctttacac tttatgcttc
3781 cggctcgtat aatgtgtgga ttttgagtta ggatccgtcg agattttcag gagctaagga
3841 agctaaaatg gagaaaaaaa tcactggata taccaccgtt gatatatccc aatggcatcg
3901 taaagaacat tttgaggcat ttcagtcagt tgctcaatgt acctataacc agaccgttca
3961 gctggatatt acggcctttt taaagaccgt aaagaaaaat aagcacaagt tttatccggc
4021 ctttattcac attcttgccc gcctgatgaa tgctcatccg gaattccgta tggcaatgaa
4081 agacggtgag ctggtgatat gggatagtgt tcacccttgt tacaccgttt tccatgagca
4141 aactgaaacg ttttcatcgc tctggagtga ataccacgac gatttccggc agtttctaca
4201 catatattcg caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt
4261 tattgagaat atgtttttcg tctcagccaa tccctgggtg agtttcacca gttttgattt
4321 aaacgtggcc aatatggaca acttcttcgc ccccgttttc accatgggca aatattatac
4381 gcaaggcgac aaggtgctga tgccgctggc gattcaggtt catcatgccg tctgtgatgg
4441 cttccatgtc ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg
4501 ggcgtaaacg ccgcgtggat ccggcttact aaaagccaga taacagtatg cgtatttgcg
4561 cgctgatttt tgcggtataa gaatatatac tgatatgtat acccgaagta tgtcaaaaag
4621 aggtatgcta tgaagcagcg tattacagtg acagttgaca gcgacagcta tcagttgctc
4681 aaggcatata tgatgtcaat atctccggtc tggtaagcac aaccatgcag aatgaagccc
4741 gtcgtctgcg tgccgaacgc tggaaagcgg aaaatcagga agggatggct gaggtcgccc
4801 ggtttattga aatgaacggc tcttttgctg acgagaacag gggctggtga aatgcagttt
4861 aaggtttaca cctataaaag agagagccgt tatcgtctgt ttgtggatgt acagagtgat
4921 attattgaca cgcccgggcg acggatggtg atccccctgg ccagtgcacg tctgctgtca
4981 gataaagtct cccgtgaact ttacccggtg gtgcatatcg gggatgaaag ctggcgcatg
5041 atgaccaccg atatggccag tgtgccggtc tccgttatcg gggaagaagt ggctgatctc
5101 agccaccgcg aaaatgacat caaaaacgcc attaacctga tgttctgggg aatataaatg
5161 tcaggctccc ttatacacag ccagtctgca ggtcgaccat agtgactgga tatgttgtgt
5221 tttacagtat tatgtagtct gttttttatg caaaatctaa tttaatatat tgatatttat
5281 atcattttac gtttctcgtt cagctttctt gtacaaagtg gtgatgggct gcaggaattc
5341 gatatcaagc ttatcgatag gtggaccagg tggtggaatg aagcgacgat ggaaaaagaa
5401 tttcatagcc gtctcagcag ccaaccgctt taagaaaatc tcatcctccg gggcacttga
5461 ttatggtacc ccaactactg cttctgaaaa tctatatttt caaggtgaac taaaaactgc
5521 tgctttggct caacatgcga ttaagggtga agctcaaaaa cttaatgact ctcaagctcc
5581 aaaagctgat gcgcaacaaa ataacttcaa caaagatcaa caaagcgcct tctatgaaat
5641 cttgaacatg cctaacttaa acgaagcgca acgtaacggc ttcattcaaa gtcttaaaga
5701 cgacccaagc caaagcacta acgttttagg tgaagctaaa aaattaaacg aatctcaagc
5761 accgaaagct gataacaatt tcaacaaaga acaacaaaat gctttctatg aaatcttgaa
5821 tatgcctaac ttaaacgaag aacaacgcaa tggtttcatc caaagcttaa aagatgaccc
5881 aagccaaagt gctaacctat tgtcagaagc taaaaagtta aatgaatctc aagcaccgaa
5941 agcggataac aaattcaaca aagaacaaca aaatgctttc tatgaaatct tacatttacc
6001 taacttaaac gaagaacaac gcaatggttt catccaaagc ctaaaagatg acccaagcca
6061 aagcgctaac cttttagcag aagctaaaaa gctaaatgat gctcaagcac caaaagctga
6121 caacaaattc aacaaagaac aacaaaatgc tttctatgaa attttacatt tacctaactt
6181 aactgaagaa caacgtaacg gcttcatcca aagccttaaa gacgatcctt cggtgagcaa
6241 agaacagtca ctcgagtcat gtaattagtt atgtcacgct tacattcacg ccctcccccc
6301 acatccgctc taaccgaaaa ggaaggagtt agacaacctg aagtctaggt ccctatttat
6361 ttttttatag ttatgttagt attaagaacg ttatttatat ttcaaatttt tctttttttt
6421 ctgtacagac gcgtgtacgc atgtaacatt atactgaaaa ccttgcttga gaaggttttg
6481 ggacgctcga aggctttaat ttgcggccgg tacccagctt ttgttccctt tagtgagggt
6541 taattccgag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
6601 tcacaattcc acacaacata ggagccggaa gcataaagtg taaagcctgg ggtgcctaat
6661 gagtgaggta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
6721 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
6781 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
6841 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
6901 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
6961 tggcgttttt ccataggctc ggcccccctg acgagcatca caaaaatcga cgctcaagtc
7021 agaggtggcg aaacccgaca ggactataaa gataccaggc gttcccccct ggaagctccc
7081 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
7141 cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg
7201 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
7261 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
7321 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
7381 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
7441 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
7501 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
7561 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
7621 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
7681 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
7741 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactgc
7801 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
7861 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
7921 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
7981 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
8041 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
8101 aacgatcaag gcgagttaca tgatccccca tgttgtgaaa aaaagcggtt agctccttcg
8161 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
8221 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
8281 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
8341 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
8401 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
8461 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
8521 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
8581 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
8641 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
8701 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
8761 ataggcgtat cacgaggccc tttcgtc
//
| {
"Vector backbone": "pRS305",
"Backbone size w/o insert (bp)": "",
"Vector type": "Yeast Expression ; Gateway Destination",
"Selectable markers": "LEU2"
} | {
"Bacterial Resistance(s)": "Chloramphenicol and Ampicillin, 25 & 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "ccdB Survival",
"Copy number": "Unknown"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "A suite of Gateway cloning vectors for high-throughput genetic analysis in Saccharomyces cerevisiae",
"Publication": "Yeast. 2007 Oct . 24(10):913-9.",
"DOI": "10.1002/yea.1502",
"Authors": "Alberti S, Gitler AD, Lindquist S",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/17583893",
"PubMed ID": "PubMed 17583893"
} | {
"Gene/Insert name": "None",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": "TAP (C terminal on backbone)"
} | {
"Cloning method": "Gateway Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "GAL1",
"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": ""
} |
|
pUPD2-N-L-mCherry2 (B5) | 211849 | CDS for mCherry in domestication vector | Marcela Rojas-Pierce | Huang et al Plant Cell. 2024 Mar 6:koae072. doi: 10.1093/plcell/koae072. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/417749/90c0c546-78fe-4ad2-aec0-b9ae00f99603/addgene-plasmid-211849-sequence-417749.gbk | full | LOCUS sequence-417749- 2846 bp DNA circular SYN 27-AUG-2024
DEFINITION CDS for mCherry in domestication vector.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2846)
AUTHORS Huang L, Rojas-Pierce M
TITLE Rapid depletion of target proteins in plants by an inducible protein
degradation system.
JOURNAL Plant Cell. 2024 Mar 6:koae072. doi: 10.1093/plcell/koae072.
PUBMED 38446628
REFERENCE 2 (bases 1 to 2846)
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: "Plant Cell.
2024 Mar 6:koae072. doi: 10.1093/plcell/koae072."
FEATURES Location/Qualifiers
source 1..2846
/mol_type="other DNA"
/organism="synthetic DNA construct"
terminator 803..860
/label=his operon terminator
/note="This putative transcriptin terminator from the E.
coli his operon has a 2-bp deletion introduced during
synthesis. Its efficiency has not been determined."
primer_bind complement(884..901)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(1055..1643)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1135..1154)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
terminator complement(1826..1920)
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
CDS complement(1941..2600)
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind 2515..2534
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
primer_bind 2762..2780
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
terminator complement(2784..2827)
/label=bacterial terminator
/note="putative bacterial transcription terminator"
misc_feature join(2830..2846,1..5)
/label=BioBrick prefix
/note="BioBrick prefix for parts that do not start with
""ATG"""
ORIGIN
1 tagagcgatg agtggtctcg ttcggaggag gtggctcatc tggcggaggt atggtgagta
61 agggtgaaga agataatatg gcaataatta aagaatttat gcgttttaaa gtccatatgg
121 aaggatcagt caatggtcat gaattcgaaa ttgaggggga aggtgaagga aggccgtatg
181 aggggactca gaccgctaaa ctcaaggtga ccaagggtgg gcctttgccg tttgcatggg
241 atatcctctc tccacaattt atgtatggat caaaggctta cgtcaagcac ccggctgaca
301 tacctgacta tcttaaactc tctttcccgg aaggttttaa gtgggaaaga gtgatgaatt
361 tcgaagatgg tggggttgtg accgttacac aagattcaag tctgcaggat ggagagttta
421 tttacaaggt taaactcaga ggtaccaatt tcccgtctga cgggcctgtg atgcaaaaaa
481 agactatggg gtgggaggcc tcttcagaaa gaatgtatcc tgaggatgga gcactcaagg
541 gtgaaattaa acaaagattg aagcttaaag atggaggtca ttatgacgcc gaagtgaaaa
601 ctacatataa ggcaaagaag ccagtccaat tgccgggagc atataatgtg aatattaaac
661 tcgatatcac ctctcacaat gaagattata ctatcgtcga acagtatgaa agggccgaag
721 gtcgtcattc aaccggaggt atggatgagc tctataagtg agctttgaga ccactcatcg
781 cgactagtag cggccgctgc agtccggcaa aaaagggcaa ggtgtcacca ccctgccctt
841 tttctttaaa accgaaaaga ttacttcgcg ttatgcaggc ttcctcgctc actgactcgc
901 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
961 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
1021 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca caggctccgc ccccctgacg
1081 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
1141 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
1201 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
1261 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
1321 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
1381 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
1441 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
1501 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
1561 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
1621 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
1681 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
1741 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
1801 cttggtctga cagctcgagg cttggattct caccaataaa aaacgcccgg cggcaaccga
1861 gcgttctgaa caaatccaga tggagttctg aggtcattac tggatctatc aacaggagtc
1921 caagcgagct cgatatcaaa ttacgccccg ccctgccact cgtcgcagta ctgttgtaat
1981 tcattaagca ttctgccgac atggaagcca tcacaaacgg catgatgaac ctgaatcgcc
2041 agcggcatca gcaccttgtc gccttgcgta taatatttgc ccatggtgaa aacgggggcg
2101 aagaagttgt ccatattggc cacgtttaaa tcaaaactgg tgaaactcac ccagggattg
2161 gctgatacga aaaacatatt ctcaataaac cctttaggga aataggccag gttttcaccg
2221 taacacgcca catcttgcga atatatgtgt agaaactgcc ggaaatcgtc gtggtattca
2281 ctccagagtg atgaaaacgt ttcagtttgc tcatggaaaa cggtgtaaca agggtgaaca
2341 ctatcccata tcaccagctc accgtctttc attgccatac gaaattccgg atgagcattc
2401 atcaggcggg caagaatgtg aataaaggcc ggataaaact tgtgcttatt tttctttacg
2461 gtctttaaaa aggccgtaat atccagctga acggtctggt tataggtaca ttgagcaact
2521 gactgaaatg cctcaaaatg ttctttacga tgccattggg atatatcaac ggtggtatat
2581 ccagtgattt ttttctccat tttagcttcc ttagctcctg aaaatctcga taactcaaaa
2641 aatacgcccg gtagtgatct tatttcatta tggtgaaagt tggaacctct tacgtgcccg
2701 atcaactcga gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
2761 aaataggcgt atcacgaggc agaatttcag ataaaaaaaa tccttagctt tcgctaagga
2821 tgatttctgg aattcgcggc cgcttc
//
| {
"Vector backbone": "pUPD2",
"Backbone size w/o insert (bp)": "2105",
"Vector type": "Synthetic Biology",
"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 ; Takara Bio Limited Use Label License (formerly Clontech)",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Rapid depletion of target proteins in plants by an inducible protein degradation system",
"Publication": "Plant Cell. 2024 Mar 6:koae072. doi: 10.1093/plcell/koae072.",
"DOI": "10.1093/plcell/koae072.",
"Authors": "Huang L, Rojas-Pierce M /plcell/koae",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/38446628",
"PubMed ID": "PubMed 38446628"
} | {
"Gene/Insert name": "mCherry",
"Species": "Synthetic",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": "mCherry (C terminal on insert)"
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "BsmBI (destroyed during cloning)",
"3′ cloning site": "BsmBI (destroyed during cloning)",
"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": ""
} |
|
pH2B-miRFP718nano | 195750 | Histone 2B labelled with near-infrared fluorescent protein miRFP718nano | Vladislav Verkhusha | Oliinyk et al Nat Methods. 2022 Dec 1. doi: 10.1038/s41592-022-01683-0. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/384898/81d7cafb-da0e-4086-b0af-7e74578b039d/addgene-plasmid-195750-sequence-384898.gbk | full | LOCUS sequence-384898- 4852 bp DNA circular SYN 27-AUG-2024
DEFINITION Histone 2B labelled with near-infrared fluorescent protein
miRFP718nano.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4852)
AUTHORS Oliinyk OS, Ma C, Pletnev S, Baloban M, Toboada C, Sheng H, Yao J,
Verkhusha VV
TITLE Deep-tissue SWIR imaging using rationally designed small red-shifted
near-infrared fluorescent protein.
JOURNAL Nat Methods. 2022 Dec 1. doi: 10.1038/s41592-022-01683-0.
PUBMED 36456785
REFERENCE 2 (bases 1 to 4852)
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
Methods. 2022 Dec 1. doi: 10.1038/s41592-022-01683-0."
FEATURES Location/Qualifiers
source 1..4852
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 95..398
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 399..602
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 553..573
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
CDS 691..1068
/codon_start=1
/gene="HIST1H2BJ"
/product="human histone H2B"
/label=H2B
/translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
GELAKHAVSEGTKAITKYTSAK"
polyA_signal 1674..1795
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1711..1730)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 1765..1784
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(1802..2257)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1939..1960)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2151..2170
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2284..2388
/gene="bla"
/label=AmpR promoter
promoter 2390..2747
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
enhancer 2390..2581
/label=SV40 enhancer
/note="enhancer for the SV40 early promoter (Herr, 1993)"
primer_bind complement(2413..2433)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 2598..2733
/label=SV40 ori
/note="SV40 origin of replication"
CDS 2782..3576
/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 2782..3576
/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(2836..2855)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3446..3465
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(3764..3783)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 3808..3855
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4184..4772
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4673..4692
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 tgattctgtg gataaccgta ttaccgccat gcattagtta ttaatagtaa tcaattacgg
61 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
121 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
181 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
241 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
301 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
361 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
421 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
481 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
541 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
601 ctggtttagt gaaccgtcag atccgctagc gctaccggac tcagatctcg agctcaagct
661 tcgaattctg cagtcgacgg taccgccacc atgccagagc cagcgaagtc tgctcccgcc
721 ccgaaaaagg gctccaagaa ggcggtgact aaggcgcaga agaaaggcgg caagaagcgc
781 aagcgcagcc gcaaggagag ctattccatc tatgtgtaca aggttctgaa gcaggtccac
841 cctgacaccg gcatttcgtc caaggccatg ggcatcatga attcgtttgt gaacgacatt
901 ttcgagcgca tcgcaggtga ggcttcccgc ctggcgcatt acaacaagcg ctcgaccatc
961 acctccaggg agatccagac ggccgtgcgc ctgctgctgc ctggggagtt ggccaagcac
1021 gccgtgtccg agggtactaa ggccatcacc aagtacacca gcgctaagga tccaccggta
1081 atggcaaacc tggacaagat gctgaacacc atcgtgaccg aggtgcgcca gttcctgcaa
1141 gtggacagac tctgcgtgtt caagttcgag gaagattact ccggcaacat catctacgaa
1201 gccgtggacg acggatggtt gagcatcctg aagacccacg tgcgggactg ctacttcatg
1261 gaaaccagag gcgaggaata cctgcacggc agataccagg ccatcgccga catccaccag
1321 gccaatctgg cggagagcta tagagacttc ctgacccagt accaggtgcg ggccattgtg
1381 gctgtcccca tcctgaaggg caagaagctg tggggcttgt tcagcgccca ccaactggcc
1441 gcccctcgga gctggcaggc ctgggaaatc gagttcctga aacagcaagc cgtggtgatg
1501 ggcatcgcca tccagcagag ctagctgaat tctgcagata tccagcacag tggcggccgc
1561 gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
1621 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
1681 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
1741 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg
1801 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
1861 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
1921 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
1981 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
2041 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
2101 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
2161 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
2221 acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat
2281 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
2341 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa
2401 agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
2461 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
2521 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
2581 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
2641 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
2701 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag
2761 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
2821 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
2881 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
2941 tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg
3001 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
3061 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
3121 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
3181 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
3241 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
3301 ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
3361 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
3421 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
3481 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
3541 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
3601 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
3661 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
3721 atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga
3781 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg
3841 ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc
3901 caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc
3961 caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag
4021 cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
4081 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
4141 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
4201 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
4261 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
4321 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
4381 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
4441 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
4501 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
4561 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
4621 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
4681 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
4741 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
4801 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cc
//
| {
"Vector backbone": "pN1",
"Backbone size w/o insert (bp)": "",
"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": "Deep-tissue SWIR imaging using rationally designed small red-shifted near-infrared fluorescent protein",
"Publication": "Nat Methods. 2022 Dec 1. doi: 10.1038/s41592-022-01683-0.",
"DOI": "10.1038/s41592-022-01683-0.",
"Authors": "Oliinyk OS, Ma C, Pletnev S, Baloban M, Toboada C, Sheng H, Yao J, Verkhusha VV /s---",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/36456785",
"PubMed ID": "PubMed 36456785"
} | {
"Gene/Insert name": "H2B-miRFP718nano",
"Species": "H. sapiens (human), Synthetic",
"Insert Size (bp)": "831",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "CMV",
"Tag / Fusion Protein": "miRFP718nano (C terminal on insert)"
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "KpnI (not destroyed)",
"3′ cloning site": "NotI (not destroyed)",
"5′ sequencing primer": "CMV Forward",
"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": ""
} |
|
R777-E043 Hs.DUSP6 | 70327 | Gateway ORF clone of human DUSP6 [NM_001946.3] with stop codon (for native or N-terminal fusions) | Dominic Esposito | Esposito lab (unpublished) | https://media.addgene.org/snapgene-media/v2.0.0/sequences/125776/8cfda9fd-6a9e-45c9-b144-8b8420741dcb/addgene-plasmid-70327-sequence-125776.gbk | full | LOCUS sequence-125776- 3839 bp DNA circular SYN 26-AUG-2024
DEFINITION Gateway ORF clone of human DUSP6 [NM_001946.3] with stop codon (for
native or N-terminal fusions).
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3839)
TITLE Esposito lab
REFERENCE 2 (bases 1 to 3839)
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..3839
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 23..45
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 37..54
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 38..54
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 84..183
/gene="mutant version of attL"
/label=attL1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
protein_bind complement(1330..1429)
/gene="mutant version of attL"
/label=attL2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
CDS 1806..2597
/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 2690..3278
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3179..3198
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 3432..3449
/label=L4440
/note="L4440 vector, forward primer"
terminator complement(3608..3635)
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator complement(3727..3813)
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
ORIGIN
1 ttaacgctag catggatgtt ttcccagtca cgacgttgta aaacgacggc cagtcttaaa
61 attcgagctc ggtccgagga tccgaaataa tgattttatt ttgactgata gtgacctgtt
121 cgttgcaaca aattgataag caatgctttt ttataatgcc aactttgtac aaaaaagttg
181 gcaccatgat agatacgctc agacccgtgc ccttcgcgtc ggaaatggcg atcagcaaga
241 cggtggcgtg gctcaacgag cagctggagc tgggcaacga gcggctgctg ctgatggact
301 gccggccgca ggagctatac gagtcgtcgc acatcgagtc ggccatcaac gtggccatcc
361 cgggcatcat gctgcggcgc ctgcagaagg gtaacctgcc ggtgcgcgcg ctcttcacgc
421 gcggcgagga ccgggaccgc ttcacccggc gctgtggcac cgacacagtg gtgctctacg
481 acgagagcag cagcgactgg aacgagaata cgggcggcga gtcggtgctc gggctgctgc
541 tcaagaagct caaggacgag ggctgccggg cgttctacct ggaaggtggc ttcagtaagt
601 tccaagccga gttctccctg cattgcgaga ccaatctaga cggctcgtgt agcagcagct
661 cgccgccgtt gccagtgctg gggctcgggg gcctgcggat cagctctgac tcttcctcgg
721 acatcgagtc tgaccttgac cgagacccca atagtgcaac agactcggat ggtagtccgc
781 tgtccaacag ccagccttcc ttcccagtgg agatcttgcc cttcctctac ttgggctgtg
841 ccaaagactc caccaacttg gacgtgttgg aggaattcgg catcaagtac atcttgaacg
901 tcacccccaa tttgccgaat ctctttgaga acgcaggaga gtttaaatac aagcaaatcc
961 ccatctcgga tcactggagc caaaacctgt cccagttttt ccctgaggcc atttctttca
1021 tagatgaagc ccggggcaag aactgtggtg tcttggtaca ttgcttggct ggcattagcc
1081 gctcagtcac tgtgactgtg gcttacctta tgcagaagct caatctgtcg atgaacgatg
1141 cctatgacat tgtcaaaatg aaaaaatcca acatatcccc taacttcaac ttcatgggtc
1201 agctgctgga cttcgagagg acgctgggac tcagcagccc atgtgacaac agggttccag
1261 cacagcagct gtattttacc accccttcca accagaatgt ataccaggtg gactctctgc
1321 aatctacgta atcaactttc ttgtacaaag ttggcattat aggaaagcat tgcttatcaa
1381 tttgttgcaa cgaacaggtc actatcagtc aaaataaaat cattatttgc atgcaagctt
1441 agatcctcta gaccagccag gacagaaatg cctcgacttc gctgctaccc aaggttgccg
1501 ggtgacgcac accgtggaaa cggatgaagg cacgaaccca gtggacataa gcctgttcgg
1561 ttcgtaagct gtaatgcaag tagcgtatgc gctcacgcaa ctggtccaga accttgaccg
1621 aacgcagcgg tggtaacggc gcagtggcgg ttttcatggc ttgttatgac tgtttttttg
1681 gggtacagtc tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg ggtcgatgtt
1741 tgatgttatg gagcagcaac gatgttacgc agcagggcag tcgccctaaa acaaagttaa
1801 acattatgag ggaagcggtg atcgccgaag tatcgactca actatcagag gtagttggcg
1861 tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca tttgtacggc tccgcagtgg
1921 atggcggcct gaagccacac agtgatattg atttgctggt tacggtgacc gtaaggcttg
1981 atgaaacaac gcggcgagct ttgatcaacg accttttgga aacttcggct tcccctggag
2041 agagcgagat tctccgcgct gtagaagtca ccattgttgt gcacgacgac atcattccgt
2101 ggcgttatcc agctaagcgc gaactgcaat ttggagaatg gcagcgcaat gacattcttg
2161 caggtatctt cgagccagcc acgatcgaca ttgatctggc tatcttgctg acaaaagcaa
2221 gagaacatag cgttgccttg gtaggtccag cggcggagga actctttgat ccggttcctg
2281 aacaggatct atttgaggcg ctaaatgaaa ccttaacgct atggaactcg ccgcccgact
2341 gggctggcga tgagcgaaat gtagtgctta cgttgtcccg catttggtac agcgcagtaa
2401 ccggcaaaat cgcgccgaag gatgtcgctg ccgactgggc aatggagcgc ctgccggccc
2461 agtatcagcc cgtcatactt gaagctagac aggcttatct tggacaagaa gaagatcgct
2521 tggcctcgcg cgcagatcag ttggaagaat ttgtccacta cgtgaaaggc gagatcacca
2581 aggtagtcgg caaataaccc tcgagccacc catgaccaaa atcccttaac gtgagttacg
2641 cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
2701 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
2761 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
2821 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
2881 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
2941 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
3001 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
3061 tgagatacct acagcgtgag cattgagaaa gcgccacgct tcccgaaggg agaaaggcgg
3121 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
3181 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
3241 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
3301 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
3361 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
3421 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
3481 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
3541 aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc tagccaggaa gagtttgtag
3601 aaacgcaaaa aggccatccg tcaggatggc cttctgctta gtttgatgcc tggcagttta
3661 tggcgggcgt cctgcccgcc accctccggg ccgttgcttc acaacgttca aatccgctcc
3721 cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa aacgaaaggc
3781 ccagtcttcc gactgagcct ttcgttttat ttgatgcctg gcagttccct actctcgcg
//
| {
"Vector backbone": "pDonor-255",
"Backbone size w/o insert (bp)": "2693",
"Vector type": "Gateway Entry Clone",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Spectinomycin, 50 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": "",
"Publication": "",
"DOI": "",
"Authors": "",
"PubMed Link": "",
"PubMed ID": ""
} | {
"Gene/Insert name": "DUSP6",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "1146",
"Mutation": "",
"Entrez Gene": "DUSP6",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Gateway Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "M13F 23-mer (5'-CCCAGTCACGACGTTGTAAAACG)",
"3′ sequencing primer": "attLR 23-mer (5'-TAACATCAGAGATTTTGAGACAC)"
} | {
"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": ""
} |
|
Anti-Lgi1 [N283/7R-2b] | 222172 | Mammalian Expression Plasmid of anti-Lgi1 (Mouse). Derived from hybridoma N283/7. | James Trimmer | Mitchell et al Sci Rep. 2023 Sep 27;13(1):16200. doi: 10.1038/s41598-023-43233-4. | partial | {
"Vector backbone": "P1316-IgG2b",
"Backbone size w/o insert (bp)": "5925",
"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": "High-volume hybridoma sequencing on the NeuroMabSeq platform enables efficient generation of recombinant monoclonal antibodies and scFvs for neuroscience research",
"Publication": "Sci Rep. 2023 Sep 27;13(1):16200. doi: 10.1038/s41598-023-43233-4.",
"DOI": "10.1038/s41598-023-43233-4.",
"Authors": "Mitchell KG, Gong B, Hunter SS, Burkart-Waco D, Gavira-O'Neill CE, Templeton KM, Goethel ME, Bzymek M, MacNiven LM, Murray KD, Settles ML, Froenicke L, Trimmer JS /s---",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/37758930",
"PubMed ID": "PubMed 37758930"
} | {
"Gene/Insert name": "Anti-Lgi1 (Mus musculus) recombinant mouse monoclonal antibody.",
"Species": "M. musculus (mouse)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "Lgi1",
"Promoter": "Dual CMV",
"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": ""
} |
|||
pVITRO1-iRFP702-NES | 184457 | Cytosolic iRFP702 expression in mammalian cells | Chuan-Hsiang Huang | Yang et al Cell. 2021 Dec 9;184(25):6193-6206.e14. doi: 10.1016/j.cell.2021.11.005. Epub 2021 Nov 26. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/362894/ab13e9d3-e8f7-4261-9493-74ad662757f9/addgene-plasmid-184457-sequence-362894.gbk | full | LOCUS sequence-362894- 7466 bp DNA circular SYN 26-AUG-2024
DEFINITION Cytosolic iRFP702 expression in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7466)
AUTHORS Yang JM, Chi WY, Liang J, Takayanagi S, Iglesias PA, Huang CH
TITLE Deciphering cell signaling networks with massively multiplexed
biosensor barcoding.
JOURNAL Cell. 2021 Dec 9;184(25):6193-6206.e14. doi:
10.1016/j.cell.2021.11.005. Epub 2021 Nov 26.
PUBMED 34838160
REFERENCE 2 (bases 1 to 7466)
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.2021.11.005"; journalName: "Cell"; date: "2021-12-9-
9"; volume: "184"; issue: "25"; pages: "6193-6206.e14"
FEATURES Location/Qualifiers
source 1..7466
/mol_type="other DNA"
/organism="synthetic DNA construct"
polyA_signal 302..423
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(313..332)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 367..386
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
rep_origin 611..1199
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 1100..1119
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
enhancer 1279..1582
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 1697..3002
/label=rEF-1-alpha promoter
/note="strong constitutive promoter for rat elongation
factor EF-1-alpha"
misc_feature 2039..2970
/label=rEF-1-alpha intron
/note="intron upstream of the start codon of rat
EF-1-alpha"
regulatory 3009..3018
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 3015..3947
/codon_start=1
/product="near-infrared fluorescent protein with an
emission peak at 702 nm, engineered from a bacterial
phytochrome (Shcherbakova and Verkhusha, 2013)"
/label=iRFP702
/note="derived from Rhodopseudomonas palustris RpBphP6"
/translation="MARKVDLTSCDREPIHIPGSIQPCGCLLACDAQAVRITRITENAG
AFFGRETPRVGELLADYFGETEAHALRNALAQSSDPKRPALIFGWRDGLTGRTFDISLH
RHDGTSIIEFEPAAAEQADNPLRLTRQIIARTKELKSLEEMAARVPRYLQAMLGYHRVM
LYRFADDGSGKVIGEAKRSDLESFLGQHFPASLVPQQARLLYLKNAIRVVSDSRGISSR
IVPEHDASGAALDLSFAHLRSISPIHLEFLRNMGVSASMSLSIIIDGTLWGLIICHHYE
PRAVPMAQRVAAEMFADFLSLHFTAAHHQR"
misc_feature 4008..4452
/label=FMDV IRES
/note="internal ribosome entry site (IRES) of the
foot-and-mouth disease virus"
promoter 4485..4531
/label=EM7 promoter
/note="synthetic bacterial promoter "
promoter 4523..4541
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 4551..5576
/codon_start=1
/product="hygromycin B phosphotransferase"
/label=HygR
/note="confers resistance to hygromycin"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
E"
polyA_signal 5588..6160
/label=EF-1-alpha poly(A) signal
/note="human EF-1-alpha polyadenylation signal"
enhancer 6212..6403
/label=SV40 enhancer
/note="enhancer for the SV40 early promoter (Herr, 1993)"
primer_bind complement(6235..6255)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter join(6417..7466,1..257)
/label=mEF-1-alpha promoter
/note="strong constitutive promoter for mouse elongation
factor EF-1-alpha"
intron join(6744..7466,1..225)
/label=mEF-1-alpha intron
/note="intron upstream of the start codon of mouse
EF-1-alpha"
ORIGIN
1 ggtcctatcg gccgcaatag tcacctcggg cttttgagca cggctagtcg cggcgggggg
61 aggggatgta atggcgttgg agtttgttca catttggtgg gtggagacta gtcaggccag
121 cctggcgctg gaagtcattt ttggaatttg tccccttgag ttttgagcgg agctaattct
181 cgggcttctt agcggttcaa aggtatcttt taaacccttt tttaggtgtt gtgaaaacca
241 ccgctaattc aaagcaaccg gtgatatcaa agatcttgta cagctagctg gccagacatg
301 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
361 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
421 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
481 ttttaaagca agtaaaacct ctacaaatgt ggtatggaaa tgttaattaa ctagccatga
541 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
601 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
661 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
721 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
781 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
841 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
901 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
961 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
1021 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
1081 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
1141 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
1201 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
1261 tcttaattaa cctgcaggcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
1321 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
1381 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
1441 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
1501 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
1561 attagtcatc gctattacca tgatgatgcg gttttggcag tacatcaatg ggcgtggata
1621 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
1681 ttgactagtg gagccgagag taattcatac aaaaggaggg atcgccttcg caaggggaga
1741 gcccagggac cgtccctaaa ttctcacaga cccaaatccc tgtagccgcc ccacgacagc
1801 gcgaggagca tgcgcccagg gctgagcgcg ggtagatcag agcacacaag ctcacagtcc
1861 ccggcggtgg ggggaggggc gcgctgagcg ggggccaggg agctggcgcg gggcaaactg
1921 ggaaagtggt gtcgtgtgct ggctccgccc tcttcccgag ggtgggggag aacggtatat
1981 aagtgcggta gtcgccttgg acgttctttt tcgcaacggg tttgccgtca gaacgcaggt
2041 gagtggcggg tgtggcttcc gcgggccccg gagctggagc cctgctctga gcgggccggg
2101 ctgatatgcg agtgtcgtcc gcagggttta gctgtgagca ttcccacttc gagtggcggg
2161 cggtgcgggg gtgagagtgc gaggcctagc ggcaaccccg tagcctcgcc tcgtgtccgg
2221 cttgaggcct agcgtggtgt ccgccgccgc gtgccactcc ggccgcacta tgcgtttttt
2281 gtccttgctg ccctcgattg ccttccagca gcatgggcta acaaagggag ggtgtggggc
2341 tcactcttaa ggagcccatg aagcttacgt tggataggaa tggaagggca ggaggggcga
2401 ctggggcccg cccgccttcg gagcacatgt ccgacgccac ctggatgggg cgaggcctgt
2461 ggctttccga agcaatcggg cgtgagttta gcctacctgg gccatgtggc cctagcactg
2521 ggcacggtct ggcctggcgg tgccgcgttc ccttgcctcc caacaagggt gaggccgtcc
2581 cgcccggcac cagttgcttg cgcggaaaga tggccgctcc cggggccctg ttgcaaggag
2641 ctcaaaatgg aggacgcggc agcccggtgg agcgggcggg tgagtcaccc acacaaagga
2701 agagggcctt gcccctcgcc ggccgctgct tcctgtgacc ccgtggtcta tcggccgcat
2761 agtcacctcg ggcttctctt gagcaccgct cgtcgcggcg gggggagggg atctaatggc
2821 gttggagttt gttcacattt ggtgggtgga gactagtcag gccagcctgg cgctggaagt
2881 cattcttgga atttgcccct ttgagtttgg agcgaggcta attctcaagc ctcttagcgg
2941 ttcaaaggta ttttctaaac ccgtttccag gtgttgtgaa agccaccgct aattcaaagc
3001 aatccggagc caccatggcg cgtaaggtcg atctcacctc ctgcgatcgc gagccgatcc
3061 acatccccgg cagcattcag ccgtgcggct gcctgctagc ctgcgacgcg caggcggtgc
3121 ggatcacgcg cattacggaa aatgccggcg cgttctttgg acgcgaaact ccgcgggtcg
3181 gtgagctact cgccgattac ttcggcgaga ccgaagccca tgcgctgcgc aacgcactgg
3241 cgcagtcctc cgatccaaag cgaccggcgc tgatcttcgg ttggcgcgac ggcctgaccg
3301 gccgcacctt cgacatctcg ctgcatcgcc atgacggtac atcgatcatc gagttcgagc
3361 ctgcggcggc cgaacaggcc gacaatccgc tgcggctgac gcggcagatc atcgcgcgca
3421 ccaaagaact gaagtcgctc gaagagatgg ccgcacgggt gccgcgctat ctgcaggcga
3481 tgctcggcta tcaccgcgtg atgttgtacc gcttcgcgga cgacggctcc ggcaaagtga
3541 tcggcgaggc gaagcgcagc gacctcgaga gctttctcgg tcagcacttt ccggcgtcgc
3601 tggtcccgca gcaggcgcgg ctactgtact tgaagaacgc gatccgcgtg gtctcggatt
3661 cgcgcggcat cagcagccgg atcgtgcccg agcacgacgc ctccggcgcc gcgcttgatc
3721 tgtcgttcgc gcacctgcgc agcatctcgc ctatccatct cgaatttctg cggaacatgg
3781 gcgtcagcgc ctcgatgtcg ctgtcgatca tcattgacgg cacgctatgg ggattgatca
3841 tctgtcatca ttacgagccg cgtgccgtgc cgatggcgca gcgcgtcgcg gccgaaatgt
3901 tcgccgactt cttatcgctg cacttcaccg ccgcccacca ccaacgcgga agctccctcc
3961 agaagaaact ggaggaactg gaactgtgag gatcccgtac gcctaggagc aggtttcccc
4021 aatgacacaa aacgtgcaac ttgaaactcc gcctggtctt tccaggtcta gaggggtaac
4081 actttgtact gcgtttggct ccacgctcga tccactggcg agtgttagta acagcactgt
4141 tgcttcgtag cggagcatga cggccgtggg aactcctcct tggtaacaag gacccacggg
4201 gccaaaagcc acgcccacac gggcccgtca tgtgtgcaac cccagcacgg cgactttact
4261 gcgaaaccca ctttaaagtg acattgaaac tggtacccac acactggtga caggctaagg
4321 atgcccttca ggtaccccga ggtaacacgc gacactcggg atctgagaag gggactgggg
4381 cttctataaa agcgctcggt ttaaaaagct tctatgcctg aataggtgac cggaggtcgg
4441 cacctttcct ttgcaattac tgaccctatg aatacactga ctgtttgaca attaatcatc
4501 ggcatagtat atcggcatag tataatacga ctcactatag gagggccacc atgaagaaac
4561 ctgaactgac agcaacttct gttgagaagt ttctcattga aaaatttgat tctgtttctg
4621 atctcatgca gctgtctgaa ggtgaagaaa gcagagcctt ttcttttgat gttggaggaa
4681 gaggttatgt tctgagggtc aattcttgtg ctgatggttt ttacaaagac agatatgttt
4741 acagacactt tgcctctgct gctctgccaa ttccagaagt tctggacatt ggagaatttt
4801 ctgaatctct cacctactgc atcagcagaa gagcacaagg agtcactctc caggatctcc
4861 ctgaaactga gctgccagct gttctgcaac ctgttgctga agcaatggat gccattgcag
4921 cagctgatct gagccaaacc tctggatttg gtccttttgg tccccaaggc attggtcagt
4981 acaccacttg gagggatttc atttgtgcca ttgctgatcc tcatgtctat cactggcaga
5041 ctgtgatgga tgacacagtt tctgcttctg ttgctcaggc actggatgaa ctcatgctgt
5101 gggcagaaga ttgtcctgaa gtcagacacc tggtccatgc tgattttgga agcaacaatg
5161 ttctgacaga caatggcaga atcactgcag tcattgactg gtctgaagcc atgtttggag
5221 attctcaata tgaggttgcc aacatttttt tttggagacc ttggctggct tgcatggaac
5281 aacaaacaag atattttgaa agaagacacc cagaactggc tggttccccc agactgagag
5341 cctacatgct cagaattggc ctggaccaac tgtatcaatc tctggttgat ggaaactttg
5401 atgatgctgc ttgggcacaa ggaagatgtg atgccattgt gaggtctggt gctggaactg
5461 ttggaagaac tcaaattgca agaaggtctg ctgctgtttg gactgatgga tgtgttgaag
5521 ttctggctga ctctggaaac aggagaccct ccacaagacc cagagccaag gaatgaatat
5581 tagctagatt atccctaata cctgccaccc cactcttaat cagtggtgga agaacggtct
5641 cagaactgtt tgtttcaatt ggccatttaa gtttagtagt aaaagactgg ttaatgataa
5701 caatgcatcg taaaaccttc agaaggaaag gagaatgttt tgtggaccac tttggttttc
5761 ttttttgcgt gtggcagttt taagttatta gtttttaaaa tcagtacttt ttaatggaaa
5821 caacttgacc aaaaatttgt cacagaattt tgagacccat taaaaaagtt aaatgagaaa
5881 cctgtgtgtt cctttggtca acaccgagac atttaggtga aagacatcta attctggttt
5941 tacgaatctg gaaacttctt gaaaatgtaa ttcttgagtt aacacttctg ggtggagaat
6001 agggttgttt tccccccaca taattggaag gggaaggaat atcatttaaa gctatgggag
6061 ggttgctttg attacaacac tggagagaaa tgcagcatgt tgctgattgc ctgtcactaa
6121 aacaggccaa aaactgagtc cttgggttgc atagaaagct gcctgcaggg cctgaaataa
6181 cctctgaaag aggaacttgg ttaggtacct tctgaggcgg aaagaaccag ctgtggaatg
6241 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
6301 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
6361 gtatgcaaag catgcatctc aattagtcag caaccatagt cccactagtg gagccgagag
6421 taattcatac aaaaggaggg atcgccttcg caaggggaga gcccagggac cgtccctaaa
6481 ttctcacaga cccaaatccc tgtagccgcc ccacgacagc gcgaggagca tgcgctcagg
6541 gctgagcgcg gggagagcag agcacacaag ctcatagacc ctggtcgtgg gggggaggac
6601 cggggagctg gcgcggggca aactgggaaa gcggtgtcgt gtgctggctc cgccctcttc
6661 ccgagggtgg gggagaacgg tatataagtg cggcagtcgc cttggacgtt ctttttcgca
6721 acgggtttgc cgtcagaacg caggtgaggg gcgggtgtgg cttccgcggg ccgccgagct
6781 ggaggtcctg ctccgagcgg gccgggcccc gctgtcgtcg gcggggatta gctgcgagca
6841 ttcccgcttc gagttgcggg cggcgcggga ggcagagtgc gaggcctagc ggcaaccccg
6901 tagcctcgcc tcgtgtccgg cttgaggcct agcgtggtgt ccgcgccgcc gccgcgtgct
6961 actccggccg cactctggtc tttttttttt ttgttgttgt tgccctgctg ccttcgattg
7021 ccgttcagca ataggggcta acaaagggag ggtgcggggc ttgctcgccc ggagcccgga
7081 gaggtcatgg ttggggagga atggagggac aggagtggcg gctggggccc gcccgccttc
7141 ggagcacatg tccgacgcca cctggatggg gcgaggcctg gggtttttcc cgaagcaacc
7201 aggctggggt tagcgtgccg aggccatgtg gccccagcac ccggcacgat ctggcttggc
7261 ggcgccgcgt tgccctgcct ccctaactag ggtgaggcca tcccgtccgg caccagttgc
7321 gtgcgtggaa agatggccgc tcccgggccc tgttgcaagg agctcaaaat ggaggacgcg
7381 gcagcccggt ggagcgggcg ggtgagtcac ccacacaaag gaagagggcc tggtccctca
7441 ccggctgctg cttcctgtga ccccgt
//
| {
"Vector backbone": "pVITRO1-hygro-mcs",
"Backbone size w/o insert (bp)": "6491",
"Vector type": "Mammalian Expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Hygromycin, 200 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": "Deciphering cell signaling networks with massively multiplexed biosensor barcoding",
"Publication": "Cell. 2021 Dec 9;184(25):6193-6206.e14. doi: 10.1016/j.cell.2021.11.005. Epub 2021 Nov 26.",
"DOI": "10.1016/j.cell.2021.11.005.",
"Authors": "Yang JM, Chi WY, Liang J, Takayanagi S, Iglesias PA, Huang CH /jcell",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/34838160",
"PubMed ID": "PubMed 34838160"
} | {
"Gene/Insert name": "iRFP702-NES",
"Species": "Synthetic",
"Insert Size (bp)": "324",
"Mutation": "WT",
"Entrez Gene": "",
"Promoter": "Rat EF1α",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "BspEI (not destroyed)",
"3′ cloning site": "BamHI (not destroyed)",
"5′ sequencing primer": "N/A",
"3′ sequencing primer": "GGAAAGACCAGGCGGAGTTTC"
} | {
"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": ""
} |
|
pMCSGC-GB1 | 186786 | (Empty Backbone)E. coli expression vector containing N-terminal 6His-GB1-TEV fusion. | Alexei Savchenko | Venkatesan et al iScience. 2022 Sep 3;25(10):105054. doi: 10.1016/j.isci.2022.105054. eCollection 2022 Oct 21. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/369594/79dbd8cb-81f1-4e57-b44b-6b8bb1ab2142/addgene-plasmid-186786-sequence-369594.gbk | full | LOCUS sequence-369594- 5955 bp DNA circular SYN 26-AUG-2024
DEFINITION E. coli expression vector containing N-terminal 6His-GB1-TEV
fusion..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5955)
AUTHORS Venkatesan M, Semper C, Skrivergaard S, Di Leo R, Mesa N, Rasmussen
MK, Young JF, Therkildsen M, Stogios PJ, Savchenko A
TITLE Recombinant production of growth factors for application in cell
culture.
JOURNAL iScience. 2022 Sep 3;25(10):105054. doi: 10.1016/j.isci.2022.105054.
eCollection 2022 Oct 21.
PUBMED 36157583
REFERENCE 2 (bases 1 to 5955)
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: "iScience.";
date: "2022-09-3"; volume: "25(10):105054. doi"; pages: "
10.1016/j.isci.2022.105054. eCollection 2022 Oct 21"
FEATURES Location/Qualifiers
source 1..5955
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(99..118)
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 285..388
/gene="bla"
/label=AmpR promoter
CDS 389..1249
/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(607..626)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 1420..2008
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 1909..1928
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 2162..2179
/label=L4440
/note="L4440 vector, forward primer"
primer_bind complement(2259..2278)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
CDS complement(2434..3516)
/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 3478..3497
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(3517..3594)
/gene="lacI"
/label=lacI promoter
tRNA 3734..3809
/gene="ileX"
/product="E. coli tRNA-Ile recognizing AUA"
/label=ileX
primer_bind 3958..3977
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
tRNA 4079..4155
/gene="argU"
/product="E. coli tRNA-Arg recognizing AGA and AGG"
/label=argU
primer_bind 4430..4449
/label=T7
/note="T7 promoter, forward primer"
promoter 4430..4448
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 4449..4473
/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)."
RBS 4488..4510
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 4521..4538
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 4563..4721
/codon_start=1
/product="B1 domain of Streptococcal protein G "
/label=GB1
/note="effective as a solubilizing fusion partner (Cheng
and Patel, 2004)"
/translation="KLALNGKTLKGETTTEAVDAATAEKVFKQYANDNGVDGEWTYDDA
TKTFTVTE"
CDS 4728..4748
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQS"
CDS complement(4994..5299)
/codon_start=1
/gene="ccdB"
/product="CcdB, a bacterial toxin that poisons DNA gyrase"
/label=ccdB
/note="Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains."
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
primer_bind complement(5096..5115)
/label=ccdB-fwd
/note="ccdB gene, forward primer"
CDS 5799..5816
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(5869..5887)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 5883..5930
/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 acattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
301 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
361 tgcttcaata atttgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
421 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
481 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
541 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
601 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
661 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
721 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
781 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
841 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
901 accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact
961 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
1021 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
1081 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
1141 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
1201 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
1261 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
1321 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
1381 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
1441 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
1501 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
1561 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
1621 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
1681 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
1741 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
1801 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
1861 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
1921 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
1981 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
2041 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
2101 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
2161 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
2221 tctgtgcggt atttcacacc gcaatggtgc actctcagta caatctgctc tgatgccgca
2281 tagttaagcc agtatacact ccgctatcgc tacgtgactg tagtcatgcc ccgcgcccac
2341 cggaaggagc tgactgggtt gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa
2401 tgagtgagct aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
2461 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
2521 gggcgccagg gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac
2581 cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa
2641 atcctgtttg atggtggtta acggcgggat ataacatgag ctgtcttcgg tatcgtcgta
2701 tcccactacc gagatatccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc
2761 gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag
2821 catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat
2881 cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga
2941 gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg
3001 ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg
3061 gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc
3121 atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt
3181 gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct
3241 ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag
3301 ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc
3361 cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt
3421 cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc
3481 atactctgcg acatcgtata acgttactgg tttcacattc accaccctga attgactctc
3541 ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat
3601 ctcgacgctc tcccttatgc gactcctgca ttaggaagca gcccagtagt aggttgaggc
3661 cgttgagcac cgccgccgca aggaatggtg catgcggatt gcgacacgga gttactttat
3721 aatccgctac catggcccct tagctcagtg gttagagcag gcgactcata atcgcttggt
3781 cgctggttca agtccagcag gggccaccag atatagcaaa ggctgacgag aaatcgtcag
3841 cctttttgca tgcaaggaga tggcgcccaa cagtcccccg gccacggggc ctgccaccat
3901 acccacgccg aaacaagcgc tcatgagccc gaagtggcga gcccgatctt ccccatcggt
3961 gatgtcggcg atataggcgc cagcaaccgc acctgtggcg ccggcgagat caaaagccat
4021 tgactcagca agggttgacc gtataattca cgcgattaca ccgcattgcg gtatcaacgc
4081 gcccttagct cagttggata gagcaacgac cttctaagtc gtgggccgca ggttcgaatc
4141 ctgcagggcg cgccattaca attcaatcag ttacgccttc tttatatcct ccataattcc
4201 agagtgggac atatttggga cattatcacc aaaaatgtcg tctattttcc tcgcatgctc
4261 tgtcaaatga ttaggcgcaa ggtgagcata cctacgaacc atttctatgg actcccatcc
4321 gcccatttcc tgaagcactg ataatgggac gcctgactga atccaccggt gatgccggcc
4381 acgatgcgtc cggcgtagag gatcgagatc gatctcgatc ccgcgaaatt aatacgactc
4441 actatagggg aattgtgagc ggataacaat tcccctctag aaataatttt gtttaacttt
4501 aagaaggaga tatacatatg caccatcatc atcatcattc ttctggtgta gatctgggta
4561 ccaagcttgc tctgaacggt aaaaccctga aaggtgaaac caccaccgaa gctgttgacg
4621 ctgctaccgc ggaaaaagtt ttcaaacagt acgctaacga caacggtgtt gacggtgaat
4681 ggacctacga cgacgctacc aaaaccttca cggtaaccga aggtaccgag aacctgtact
4741 tccaatccaa tattgcgtgc gagcctcttt tttgtgtgac aaaataaaaa catctaccta
4801 ttcatatacg ctagtgtcat agtcctgaaa atcatctgca tcaagaacaa tttcacaact
4861 cttatacttt tctcttacaa gtcgttcggc ttcatctgga ttttcagcct ctatacttac
4921 taaacgtgat aaagtttctg taatttctac tgtatcgacc tgcagactgg ctgtgtataa
4981 gggagcctga catttatatt ccccagaaca tcaggttaat ggcgtttttg atgtcatttt
5041 cgcggtggct gagatcagcc acttcttccc cgataacgga gaccggcaca ctggccatat
5101 cggtggtcat catgcgccag ctttcatccc cgatatgcac caccgggtaa agttcacggg
5161 agactttatc tgacagcaga cgtgcactgg ccagggggat caccatccgt cgcccgggcg
5221 tgtcaataat atcactctgt acatccacaa acagacgata acggctctct cttttatagg
5281 tgtaaacctt aaactgcatt tcaccagccc ctgttctcgt cagcaaaaga gccgttcatt
5341 tcaataaacc gggcgacctc agccatccct tcctgatttt ccgctttcca gcgttcggca
5401 cgcagacgac gggcttcatt ctgcatggtt gtgcttacca gaccggagat attgacatca
5461 tatatgcctt gagcaactga tagctgtcgc tgtcaactgt cactgtaata cgctgcttca
5521 tagcatacct ctttttgaca tacttcgggt atacatatca gtatatattc ttataccgca
5581 aaaatcagcg cgcaaatacg catactgtta tctggctttt agtaagccgg atccacgcgg
5641 cgtttacgcc ccccctgcca ctcatcgcag tactgttgta attcattaag cattctgccg
5701 acatggaagc catcacagac ggcatgatga accaatattg gaagtggata acggatccga
5761 attcgagctc cgtcgacaag cttgcggccg cactcgagca ccaccaccac caccactgag
5821 atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat
5881 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag
5941 gaactatatc cggat
//
| {
"Vector backbone": "pMCSG53",
"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)": "ccdB Survival",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Recombinant production of growth factors for application in cell culture",
"Publication": "iScience. 2022 Sep 3;25(10):105054. doi: 10.1016/j.isci.2022.105054. eCollection 2022 Oct 21.",
"DOI": "10.1016/j.isci.2022.105054.",
"Authors": "Venkatesan M, Semper C, Skrivergaard S, Di Leo R, Mesa N, Rasmussen MK, Young JF, Therkildsen M, Stogios PJ, Savchenko A /jisci",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/36157583",
"PubMed ID": "PubMed 36157583"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Gibson Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "TAATACGACTCACTATAGGG",
"3′ sequencing primer": "GCTAGTTATTGCTCAGCGG"
} | {
"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-m4134324 | 152613 | Mammalian expression plasmid for SARS-CoV-2 nsp9 | 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/295625/e39eddeb-b3f0-479b-8f01-f5a1e5fa2e55/addgene-plasmid-152613-sequence-295625.gbk | full | LOCUS sequence-295625- 7451 bp DNA circular SYN 26-AUG-2024
DEFINITION Mammalian expression plasmid for SARS-CoV-2 nsp9.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7451)
TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for
protein expression and/or purification
REFERENCE 2 (bases 1 to 7451)
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..7451
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 347..367
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQS"
CDS 401..478
/codon_start=1
/product="calmodulin-binding peptide"
/label=CBP
/note="derived from skeletal muscle myosin light chain
kinase; binds calmodulin with nanomolar affinity in the
presence of calcium"
/translation="KRRWKKNFIAVSAANRFKKISSSGAL"
CDS 506..526
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQG"
polyA_signal 991..1112
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1002..1021)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 1056..1075
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
terminator 1227..1273
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
terminator 1303..1439
/label=rrnG terminator
/note="transcription terminator from the E. coli ribosomal
RNA rrnG operon (Albrechtsen et al., 1991)"
misc_feature 1484..1555
/label=3' beta-globin insulator
/note="insulator downstream of the human beta-globin locus
(Farrell et al., 2002)"
rep_origin complement(1781..2369)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1861..1880)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
promoter 2869..4078
/label=UbC promoter
/note="human ubiquitin C promoter"
primer_bind 3915..3934
/label=hUBCpro-F
/note="Human Ubiquitin C (UbC) promoter, forward primer"
CDS 4082..5110
/codon_start=1
/product="hygromycin B phosphotransferase"
/label=HygR
/note="confers resistance to hygromycin"
/translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
KE"
polyA_signal 5137..5185
/note="synthetic polyadenylation signal"
CDS 5451..6311
/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(5669..5688)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
terminator 6324..6460
/label=rrnG terminator
/note="transcription terminator from the E. coli ribosomal
RNA rrnG operon (Albrechtsen et al., 1991)"
misc_feature 6605..6676
/label=5' beta-globin insulator
/note="insulator upstream of the human beta-globin locus
(Farrell et al., 2002)"
terminator complement(6681..6727)
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
polyA_signal 6745..6793
/note="synthetic polyadenylation signal"
enhancer 6874..7177
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 7179..7381
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 7332..7352
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
ORIGIN
1 caccatgaat aacgagttaa gcccagtagc gcttcgccag atgagttgcg ccgcaggtac
61 aacccaaact gcttgtacgg atgacaacgc cctcgcttac tataatacca caaagggagg
121 ccgattcgtt ctggccctgt tgtcggatct gcaagacttg aaatgggcac gttttcccaa
181 gtccgacggg actggcacca tttacacaga actagagccg ccatgcaggt tcgtgactga
241 tacccctaaa ggacccaagg tcaaatatct gtattttatc aaggggctta acaatctcaa
301 tagaggcatg gtgttaggat ctctagccgc aacggtgcgg ctgcaagaaa acctttactt
361 tcagtcggga cggcgaatcc cggggctgat aaatccctgg aaaaggagat ggaagaagaa
421 ctttattgca gtgtctgcgg ctaaccggtt caaaaagatc agttcaagcg gcgccctaga
481 ttacgacatt cctacgaccg cctccgagaa tctctatttc cagggtgaat ttggcttggc
541 acaacatgac gaggccgtcg ataacaagtt taataaagaa cagcagaacg ctttctatga
601 gatccttcac ttaccaaatc tgaacgaaga gcaacgcaac gctttcattc agagcctgaa
661 agacgatccc tcgcagtctg caaatctcct ggccgaagcc aagaagctga atgacgcaca
721 ggctccaaaa gttgataaca agtttaacaa ggagcagcag aatgctttct acgagatatt
781 gcaccttcct aatctgaatg aggaacaaag aaatgcattt atccagtccc taaaagacga
841 tccatcacag agtgccaacc tgcttgcgga agccaaaaag ctcaatgatg cccaggcccc
901 gaaagtagac gcgaaccatc aataaggatc cgcaggcctc tgctagcttg actgactgag
961 atacagcgta ccttcagctc acagacatga taagatacat tgatgagttt ggacaaacca
1021 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
1081 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
1141 ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg
1201 gtattggccc atctctatcg gtatcgtagc ataacccctt ggggcctcta aacgggtctt
1261 gaggggtttt ttgtgcccct cgggccggat tgctatctac cggcattggc gcagaaaaaa
1321 atgcctgatg cgacgctgcg cgtcttatac tcccacatat gccagattca gcaacggata
1381 cggcttcccc aacttgccca cttccatacg tgtcctcctt accagaaatt tatccttaag
1441 gtcgtcagct atcctgcagg cgatctctcg atttcgatca agacattcct ttaatggtct
1501 tttctggaca ccactagggg tcagaagtag ttcatcaaac tttcttccct ccctaatctc
1561 attggttacc ttgggctatc gaaacttaat taaccagtca agtcagctac ttggcgagat
1621 cgacttgtct gggtttcgac tacgctcaga attgcgtcag tcaagttcga tctggtcctt
1681 gctattgcac ccgttctccg attacgagtt tcatttaaat catgtgagca aaaggccagc
1741 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
1801 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
1861 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
1921 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
1981 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
2041 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
2101 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
2161 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
2221 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
2281 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
2341 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
2401 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
2461 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
2521 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
2581 gtctatttcg ttcatccata gttgcattta aatttccgaa ctctccaagg ccctcgtcgg
2641 aaaatcttca aacctttcgt ccgatccatc ttgcaggcta cctctcgaac gaactatcgc
2701 aagtctcttg gccggccttg cgccttggct attgcttggc agcgcctatc gccaggtatt
2761 actccaatcc cgaatatccg agatcgggat cacccgagag aagttcaacc tacatcctca
2821 atcccgatct atccgagatc cgaggaatat cgaaatcggg gcgcgcctgg cctccgcgcc
2881 gggttttggc gcctcccgcg ggcgcccccc tcgtcacggc gagcgctgcc acgtcagacg
2941 aagggcgcag gagcgtcctg atccttccgc ccggacgctc aggacagcgg cccgctgctc
3001 ataagactcg gccttagaac cccagtatca gcagaaggac attttaggac gggacttggg
3061 tgactctagg gcactggttt tctttccaga gagcggaaca ggcgaggaaa agtagtccct
3121 tctcggcgat tctgcggagg gatctccgtg gggcggtgaa cgccgatgat tatataagga
3181 cgcgccgggt gtggcacagc tagttccgtc gcagccggga tttgggtcgc ggttcttgtt
3241 tgtggatcgc tgtgatcgtc acttggtgag tagcgggctg ctgggctggc cggggctttc
3301 gtggccgccg ggccgctcgg tgggacggaa gcgtgtggag agaccgccaa gggctgtagt
3361 ctgggtccgc gagcaaggtt gccctgaact gggggttggg gggagcgcag caaaatggcg
3421 gctgttcccg agtcttgaat ggaagacgct tgtgaggcgg gctgtgaggt cgttgaaaca
3481 aggtgggggg catggtgggc ggcaagaacc caaggtcttg agcccttcgc taatgcggga
3541 aagctcttat tcgggtgaga tgggctgggc accatctggg gaccctgacg tgaagtttgt
3601 cactgactgg agaactcggt ttgtcgtctg ttgcgggggc ggcagttatg gcggtgccgt
3661 tgggcagtgc acccgtacct ttgggagcgc gcgccctcgt cgtgtcgtga cgtcacccgt
3721 tctgttggct tataatgcag ggtggggcca cctgccggta ggtgtgcggt aggcttttct
3781 ccgtcgcagg acgcagggtt cgggcctagg gtaggctctc ctgaatcgac aggcgccgga
3841 cctctggtga ggggagggat aagtgaggcg tcagtttctt tggtcggttt tatgtaccta
3901 tcttcttaag tagctgaagc tccggttttg aactatgcgc tcggggttgg cgagtgtgtt
3961 ttgtgaagtt ttttaggcac cttttgaaat gtaatcattt gggtcaatat gtaattttca
4021 gtgttagact tgtaaattgt ccgctaaatt ctggccgttt ttggcttttt tgttagacaa
4081 catgggtaaa aagcctgaac tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt
4141 cgacagcgtc tccgacctga tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt
4201 cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa
4261 agatcgttat gtttatcggc actttgcatc cgccgcgctc ccgattccgg aagtgcttga
4321 cattggggag ttcagcgaga gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac
4381 gttgcaagac ctgcctgaaa ccgaactgcc cgctgttctg cagccggtcg cggaggcaat
4441 ggatgccatc gctgccgccg atcttagcca gacgagcggg ttcggcccat tcggaccgca
4501 aggaatcggt caatacacta catggcgtga tttcatatgc gcgattgctg atccccatgt
4561 gtatcactgg caaactgtga tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga
4621 tgagctgatg ctttgggccg aggactgccc cgaagtccgg cacctcgtgc acgcggattt
4681 cggctccaac aatgtcctga cggacaatgg ccgcataaca gcggtcattg actggagcga
4741 ggcgatgttc ggggattccc aatacgaggt cgccaacatc ttcttctgga ggccgtggtt
4801 ggcttgtatg gagcagcaga cgcgctactt cgagcggagg catccggagc ttgcaggatc
4861 gccccggctc cgggcgtata tgctccgcat tggtcttgac caactctatc agagcttggt
4921 tgacggcaat ttcgatgatg cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc
4981 cggagccggg actgtcgggc gtacacaaat cgcccgcaga agcgccgccg tctggaccga
5041 tggctgtgta gaagtactcg ccgatagtgg aaaccgacgc cccagcactc gtccgagggc
5101 aaaggaataa gctagtatgt aagcctagtc ttagataata aaatcgctat ccatcgaaga
5161 tggatgtgtg ttggtttttt gtgtgtgtaa cgctaggcgc gcctggtgta ccgagaacga
5221 tcctctcagt gcgagtctcg acgatccata tcgttgcttg gcagtcagcc agtcggaatc
5281 cagcttggga cccaggaagt ccaatcgtca gatattgtac tcaagcctgg tcacggcagc
5341 gtaccgatct gtttaaacct agatattgat agtctgatcg gtcaacgtat aatcgagtcc
5401 tagcttttgc aaacatctat caagagacag gatcagcagg aggctttcgc atgagtattc
5461 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
5521 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgcg cgagtgggtt
5581 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgct
5641 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
5701 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtatt
5761 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
5821 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatt ggaggaccga
5881 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
5941 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
6001 tggcaacaac cttgcgtaaa ctattaactg gcgaactact tactctagct tcccggcaac
6061 agttgataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
6121 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
6181 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
6241 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
6301 agcattggta accgattcta ggtgcattgg cgcagaaaaa aatgcctgat gcgacgctgc
6361 gcgtcttata ctcccacata tgccagattc agcaacggat acggcttccc caacttgccc
6421 acttccatac gtgtcctcct taccagaaat ttatccttaa gatcgtttaa actcgactct
6481 ggctctatcg aatctccgtc gtttcgagct tacgcgaaca gccgtggcgc tcatttgctc
6541 gtcgggcatc gaatctcgtc agctatcgtc agcttacctt tttggcagcg atcgcggctc
6601 ccgacatctt ggaccattag ctccacaggt atcttcttcc ctctagtggt cataacagca
6661 gcttcagcta cctctcaatt caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt
6721 tatgctatca atcgttgcgt tacacacaca aaaaaccaac acacatccat cttcgatgga
6781 tagcgatttt attatctaac tgctgatcga gtgtagccag atctagtaat caattacggg
6841 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
6901 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
6961 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
7021 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
7081 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
7141 gcagtacatc tacgtattag tcatcgctat taccatgctg atgcggtttt ggcagtacat
7201 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
7261 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
7321 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
7381 tggtttagtg aaccgtcaga tcagatcttt gtcgatccta ccatccactc gacacacccg
7441 ccagcggccg c
//
| {
"Vector backbone": "pTwist_CMV_Hygro_6",
"Backbone size w/o insert (bp)": "6525",
"Vector type": "Mammalian Expression",
"Selectable markers": "Hygromycin"
} | {
"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 nsp9",
"Species": "Severe acute respiratory syndrome coronavirus 2",
"Insert Size (bp)": "338",
"Mutation": "wt",
"Entrez Gene": "ORF1ab",
"Promoter": "CMV",
"Tag / Fusion Protein": "Cleavable TEV;TapTag (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": ""
} |
||
Smp_105450-Cd4-bio-His | 120653 | Expression of recombinant S. mansoni protein | Gavin Wright | Cécile et al bioRXiv | https://media.addgene.org/snapgene-media/v2.0.0/sequences/234473/06a8243f-ce25-4504-93e0-dbbb26bc16ee/addgene-plasmid-120653-sequence-234473.gbk | full | LOCUS sequence-234473- 7011 bp DNA circular SYN 26-AUG-2024
DEFINITION Expression of recombinant S. mansoni protein.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7011)
AUTHORS Cecile Crosnier, Anna V. Protasio, Gabriel Rinaldi, Shona Wilson,
Matthew Berriman, Gavin J. Wright
TITLE A library of cell-surface and secreted proteins from Schistosoma
mansoni identifies early serological markers of infection
JOURNAL bioRXiv
REFERENCE 2 (bases 1 to 7011)
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: "bioRXiv"
FEATURES Location/Qualifiers
source 1..7011
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 1..204
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 155..175
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 201..225
/label=LNCX
/note="Human CMV promoter, forward primer"
primer_bind 706..725
/label=pMT2-F
/note="Synthetic intron, forward primer"
regulatory 748..757
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 1789..1806
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
polyA_signal 1858..1913
/label=beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(1912..1931)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
primer_bind complement(4149..4167)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 4235..4339
/gene="bla"
/label=AmpR promoter
CDS 4340..5200
/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(4558..4577)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 5371..5959
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5860..5879
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 6113..6130
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 6247..6268
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 6283..6313
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6321..6337
/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 6326..6348
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 6345..6361
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 6345..6361
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
enhancer 6632..7011
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
ORIGIN
1 gtgatgcggt tttggcagta caccaatggg cgtggatagc ggtttgactc acggggattt
61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
121 tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa tgggcggtag gcgtgtacgg
181 tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcctcac tctcttccgc
241 atcgctgtct gcgagggcca gctgttgggc tcgcggttga ggacaaactc ttcgcggtct
301 ttccagtact cttggatcgg aaacccgtcg gcctccgaac ggtactccgc caccgaggga
361 cctgagcgag tccgcatcga ccggatcgga aaacctctcg agaaaggcgt ctaaccagtc
421 acagtcgcaa ggtaggctga gcaccgtggc gggcggcagc gggtggcggt cggggttgtt
481 tctggcggag gtgctgctga tgatgtaatt aaagtaggcg gtcttgagac ggcggatggt
541 cgaggtgagg tgtggcaggc ttgagatcca gctgttgggg tgagtactcc ctctcaaaag
601 cgggcattac ttctgcgcta agattgtcag tttccaaaaa cgaggaggat ttgatattca
661 cctggcccga tctggccata cacttgagtg acaatgacat ccactttgcc tttctctcca
721 caggtgtcca ctcccaggtc caagtttgcc accatggagt ttcagaccca ggtactcatg
781 tccctgctgc tctgcatgtc tggtgcggcc gcctacagcg tgaagaacgt ggacatgagc
841 aacaccaaag aaaccgtgga ctaccagctg gacaacaaca gcacctgtag catgtgcatc
901 cagttcatca ccaagtggca gacctacctg aacagcacca gcgtggacga gaagatcgag
961 cagttcatcc ggtctacctg cagcttctac ttcttcttcc ggtatcgctg cgaggaattc
1021 atggaacgct acgtgaacaa gaccgtgtac atcatccagc acaacaacgc caccaccctg
1081 tgcaagttca gcatcatcga gtgcatgctg atcatcctga agctgacccc cctgtgtggc
1141 gcgccgtcga cctccatcac ggcctataag agtgaggggg agtcagcgga gttctccttc
1201 ccactcaacc ttggagagga aagcctgcag ggagagttga gatggaaggc agagaaggct
1261 ccttcttccc agtcctggat caccttctcc ctaaagaacc aaaaggtgtc tgtgcagaag
1321 tctactagca accccaagtt ccagctgtcc gaaacgctcc cactcaccct tcagataccc
1381 caggtctccc ttcagtttgc tggttctggc aacctgaccc tgactctgga cagagggata
1441 ctgtatcagg aagtgaacct ggtggtgatg aaagtgactc agcccgacag caacactttg
1501 acctgtgagg tgatgggacc cacctcaccc aagatgagac tgatcttgaa gcaggagaat
1561 caggaggcca gggtctccag gcaggagaaa gtgattcaag tgcaggcccc tgaagcaggg
1621 gtgtggcaat gtctactgag tgaaggtgaa gaggtcaaga tggactccaa gatccaggtt
1681 ttatccaaag ggttgaattc cggcagcctg caccacatcc tggacgccca gaagatgctg
1741 tggaaccaca gggacaggaa cctgcctcct ctggcccctc tgggccccca ccaccatcac
1801 caccattgag tctagacacg tgattaatta aggatccccc gacctcgacc tctggctaat
1861 aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa
1921 ggacatatgg gagggcaaat catttggtcg agatccctcg gagatctcta gctagaggat
1981 cgatccccgc cccggacgaa ctaaacctga ctacgacatc tctgcccctt cttcgcgggg
2041 cagtgcatgt aatcccttca gttggttggt acaacttgcc aactgggccc tgttccacat
2101 gtgacacggg gggggaccaa acacaaaggg gttctctgac tgtagttgac atccttataa
2161 atggatgtgc acatttgcca acactgagtg gctttcatcc tggagcagac tttgcagtct
2221 gtggactgca acacaacatt gcctttatgt gtaactcttg gctgaagctc ttacaccaat
2281 gctgggggac atgtacctcc caggggccca ggaagactac gggaggctac accaacgtca
2341 atcagagggg cctgtgtagc taccgataag cggaccctca agagggcatt agcaatagtg
2401 tttataaggc ccccttgtta accctaaacg ggtagcatat gcttcccggg tagtagtata
2461 tactatccag actaacccta attcaatagc atatgttacc caacgggaag catatgctat
2521 cgaattaggg ttagtaaaag ggtcctaagg aacagcgata tctcccaccc catgagctgt
2581 cacggtttta tttacatggg gtcaggattc cacgagggta gtgaaccatt ttagtcacaa
2641 gggcagtggc tgaagatcaa ggagcgggca gtgaactctc ctgaatcttc gcctgcttct
2701 tcattctcct tcgtttagct aatagaataa ctgctgagtt gtgaacagta aggtgtatgt
2761 gaggtgctcg aaaacaaggt ttcaggtgac gcccccagaa taaaatttgg acggggggtt
2821 cagtggtggc attgtgctat gacaccaata taaccctcac aaaccccttg ggcaataaat
2881 actagtgtag gaatgaaaca ttctgaatat ctttaacaat agaaatccat ggggtgggga
2941 caagccgtaa agactggatg tccatctcac acgaatttat ggctatgggc aacacataat
3001 cctagtgcaa tatgatactg gggttattaa gatgtgtccc aggcagggac caagacaggt
3061 gaaccatgtt gttacactct atttgtaaca aggggaaaga gagtggacgc cgacagcagc
3121 ggactccact ggttgtctct aacacccccg aaaattaaac ggggctccac gccaatgggg
3181 cccataaaca aagacaagtg gccactcttt tttttgaaat tgtggagtgg gggcacgcgt
3241 cagcccccac acgccgccct gcggttttgg actgtaaaat aagggtgtaa taacttggct
3301 gattgtaacc ccgctaacca ctgcggtcaa accacttgcc cacaaaacca ctaatggcac
3361 cccggggaat acctgcataa gtaggtgggc gggccaagat aggggcgcga ttgctgcgat
3421 ctggaggaca aattacacac acttgcgcct gagcgccaag cacagggttg ttggtcctca
3481 tattcacgag gtcgctgaga gcacggtggg ctaatgttgc catgggtagc atatactacc
3541 caaatatctg gatagcatat gctatcctaa tctatatctg ggtagcatag gctatcctaa
3601 tctatatctg ggtagcatat gctatcctaa tctatatctg ggtagtatat gctatcctaa
3661 tttatatctg ggtagcatag gctatcctaa tctatatctg ggtagcatat gctatcctaa
3721 tctatatctg ggtagtatat gctatcctaa tctgtatccg ggtagcatat gctatcctaa
3781 tagagattag ggtagtatat gctatcctaa tttatatctg ggtagcatat actacccaaa
3841 tatctggata gcatatgcta tcctaatcta tatctgggta gcatatgcta tcctaatcta
3901 tatctgggta gcataggcta tcctaatcta tatctgggta gcatatgcta tcctaatcta
3961 tatctgggta gtatatgcta tcctaattta tatctgggta gcataggcta tcctaatcta
4021 tatctgggta gcatatgcta tcctaatcta tatctgggta gtatatgcta tcctaatctg
4081 tatccgggta gcatatgcta tcctcatgat aagctgtcaa acatgagaat tttcttgaag
4141 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
4201 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
4261 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
4321 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
4381 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
4441 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
4501 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
4561 atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca
4621 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
4681 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
4741 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
4801 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
4861 cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg
4921 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
4981 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
5041 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
5101 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
5161 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
5221 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
5281 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
5341 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
5401 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
5461 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
5521 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
5581 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
5641 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
5701 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
5761 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
5821 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
5881 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
5941 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
6001 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
6061 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
6121 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
6181 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
6241 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
6301 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
6361 ccatgattac gccaagctct agctagaggt cgaccaattc tcatgtttga cagcttatca
6421 tcgcagatcc gggcaacgtt gttgccattg ctgcaggcgc agaactggta ggtatggaag
6481 atctatacat tgaatcaata ttggcaatta gccatattag tcattggtta tatagcataa
6541 atcaatattg gctattggcc attgcatacg ttgtatctat atcataatat gtacatttat
6601 attggctcat gtccaatatg accgccatgt tgacattgat tattgactag ttattaatag
6661 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
6721 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
6781 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
6841 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct
6901 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg
6961 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat g
//
| {
"Vector backbone": "pTT3",
"Backbone size w/o insert (bp)": "6686",
"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 library of cell-surface and secreted proteins from Schistosoma mansoni identifies early serological markers of infection",
"Publication": "bioRXiv",
"DOI": "10.1101/593616",
"Authors": "Cécile Crosnier, Anna V Protasio, Gabriel Rinaldi, Shona Wilson, Matthew Berriman, Gavin J Wright",
"PubMed Link": "",
"PubMed ID": ""
} | {
"Gene/Insert name": "Smp_105450",
"Species": "Schistosoma mansoni",
"Insert Size (bp)": "325",
"Mutation": "",
"Entrez Gene": "Smp_105450.1",
"Promoter": "CMV",
"Tag / Fusion Protein": "ratCd4(d3+4)-bio-6His (C terminal on backbone)"
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "NotI (not destroyed)",
"3′ cloning site": "AscI (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": ""
} |
|
pAAV-S5E2-C1V1-eYFP | 135633 | AAV vector to drive the expression of C1V1-eYFP in PV cortical interneuronsunder the control of the E2 regulatory element | Jordane Dimidschstein | Vormstein-Schneider et al Nat Neurosci. 2020 Dec;23(12):1629-1636. doi: 10.1038/s41593-020-0692-9. Epub 2020 Aug 17. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/271298/0ccd6746-53fb-414c-a430-9d948c7e462a/addgene-plasmid-135633-sequence-271298.gbk | full | LOCUS sequence-271298- 6591 bp DNA circular SYN 26-AUG-2024
DEFINITION AAV vector to drive the expression of C1V1-eYFP in PV cortical
interneuronsunder the control of the E2 regulatory element.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6591)
AUTHORS Vormstein-Schneider D, Lin JD, Pelkey KA, Chittajallu R, Guo B,
Arias-Garcia MA, Allaway K, Sakopoulos S, Schneider G, Stevenson O,
Vergara J, Sharma J, Zhang Q, Franken TP, Smith J, Ibrahim LA, M
Astro KJ, Sabri E, Huang S, Favuzzi E, Burbridge T, Xu Q, Guo L,
Vogel I, Sanchez V, Saldi GA, Gorissen BL, Yuan X, Zaghloul KA,
Devinsky O, Sabatini BL, Batista-Brito R, Reynolds J, Feng G, Fu Z,
McBain CJ, Fishell G, Dimidschstein J
TITLE Viral manipulation of functionally distinct interneurons in mice,
non-human primates and humans.
JOURNAL Nat Neurosci. 2020 Dec;23(12):1629-1636. doi:
10.1038/s41593-020-0692-9. Epub 2020 Aug 17.
PUBMED 32807948
REFERENCE 2 (bases 1 to 6591)
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/s41593-020-0692-9"; journalName: "Nat Neurosci"; date:
"2020-12"; volume: "23"; issue: "12"; pages: "1629-1636"
FEATURES Location/Qualifiers
source 1..6591
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin complement(62..650)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(142..161)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(821..1681)
/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 1444..1463
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(1682..1786)
/gene="bla"
/label=AmpR promoter
primer_bind 1854..1872
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(1910..1932)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 2032..2051
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
rep_origin complement(2068..2523)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2205..2226)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2417..2436
/label=F1ori-R
/note="F1 origin, reverse primer"
repeat_region 2598..2738
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 2609..2738
/label=AAV2 ITR
primer_bind complement(2788..2807)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
polyA_signal 2817..2898
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind 2842..2861
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
misc_feature 3067..3655
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
CDS 3173..3184
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind 3582..3602
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(3680..4396)
/codon_start=1
/product="enhanced YFP"
/label=EYFP
/note="mammalian codon-optimized"
/translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
FVTAAGITLGMDELYK"
primer_bind complement(3725..3746)
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind 4074..4093
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 4333..4354
/label=EGFP-N
/note="EGFP, reverse primer"
intron 5526..5658
/label=chimeric intron
/note="chimera between introns from human beta-globin and
immunoglobulin heavy chain genes"
repeat_region 6462..6591
/label=AAV2 ITR (alternate)
/note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
1 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
61 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
121 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
181 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
241 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
301 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
361 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
421 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
481 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
541 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
601 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
661 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
721 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
781 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
841 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
901 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
961 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
1021 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
1081 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
1141 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
1201 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
1261 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
1321 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
1381 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
1441 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
1501 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
1561 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
1621 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
1681 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
1741 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
1801 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
1861 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
1921 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
1981 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
2041 agcagattgt actgagagtg caccataaaa ttgtaaacgt taatattttg ttaaaattcg
2101 cgttaaattt ttgttaaatc agctcatttt ttaaccaata gaccgaaatc ggcaaaatcc
2161 cttataaatc aaaagaatag cccgagatag agttgagtgt tgttccagtt tggaacaaga
2221 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
2281 atggcccact acgtgaacca tcacccaaat caagtttttt ggggtcgagg tgccgtaaag
2341 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
2401 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctaaggcg ctggcaagtg
2461 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
2521 cgtactatgg ttgctttgac gtatgcggtg tgaaataccg cacagatgcg taaggagaaa
2581 ataccgcatc aggcgcccct gcaggcagct gcgcgctcgc tcgctcactg aggccgcccg
2641 ggcaaagccc gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg
2701 cagagaggga gtggccaact ccatcactag gggttcctgc ggccgctcgg tccgcacgtg
2761 catgcggtat cgataagctg atacattgat gagtttggac aaaccacaac tagaatgcag
2821 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
2881 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
2941 gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat ggctgattat
3001 gatctagagt cgcggccagc ttgatggccg ctatcacaga tctctcgagc agcgctcggt
3061 atcgatgcgg ggaggcggcc caaagggaga tccgactcgt ctgagggcga aggcgaagac
3121 gcggaagagg ccgcagagcc ggcagcaggc cgcgggaagg aaggtccgct ggattgaggg
3181 ccgaagggac gtagcagaag gacgtcccgc gcagaatcca ggtggcaaca taggcgagca
3241 gccaaggaaa ggacgatgat ttccccgaca acaccacgga attgtcagtg cccaacagcc
3301 gagcccctgt ccagcagcgg gcaaggcagg cggcgatgag ttccgccgtg gcaataggga
3361 gggggaaagc gaaagtcccg gaaaggagct gacaggtggt ggcaatgccc caaccagtgg
3421 gggttgcgtc agcaaacaca gtgcacacca cgccacgttg cctgacaacg ggccacaact
3481 cctcataaag agacagcaac caggatttat acaaggagga gaaaatgaaa gccatacggg
3541 aagcaatagc atgatacaaa ggcattaaag cagcgtatcc acatagcgta aaaggagcaa
3601 catagttaag aataccagtc aatctttcac aaattttgta atccagaggt tgattatcga
3661 taagcttgat atcgaattct tacttgtaca gctcgtccat gccgagagtg atcccggcgg
3721 cggtcacgaa ctccagcagg accatgtgat cgcgcttctc gttggggtct ttgctcaggg
3781 cggactggta gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg
3841 tgttctgctg gtagtggtcg gcgagctgca cgctgccgtc ctcgatgttg tggcggatct
3901 tgaagttcac cttgatgccg ttcttctgct tgtcggccat gatatagacg ttgtggctgt
3961 tgtagttgta ctccagcttg tgccccagga tgttgccgtc ctccttgaag tcgatgccct
4021 tcagctcgat gcggttcacc agggtgtcgc cctcgaactt cacctcggcg cgggtcttgt
4081 agttgccgtc gtccttgaag aagatggtgc gctcctggac gtagccttcg ggcatggcgg
4141 acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg ggcgaagcac tgcaggccgt
4201 agccgaaggt ggtcacgagg gtgggccagg gcacgggcag cttgccggtg gtgcagatga
4261 acttcagggt cagcttgccg taggtggcat cgccctcgcc ctcgccggac acgctgaact
4321 tgtggccgtt tacgtcgccg tccagctcga ccaggatggg caccaccccg gtgaacagct
4381 cctcgccctt gctcaccacg ttgatgtcga tctggtccag ggggatgtac tcgccctcgc
4441 tggtgatcct gctcttggcg gccgcgtcct cctcttcagc caccagtgtc tcaacctcca
4501 tctcttggcc ggcgatcgta attttctgct tctttctgat gtcgccatac aacaggatgt
4561 gctcgtggat tttgacgcgc aggtaattcc ccagcacacc ccacatgttc tttgcaatca
4621 ggtcgaggat ggagtgccca atggcagatc cataagggct gatatgaccg aatccttccg
4681 tgcccagcag aaacaggact ggaaacattc cccaagccac aaagaatgtc caggccatca
4741 ctcggaccag ttctcgacaa atcccttttg gaactgtgtg gaatgcttcg atataaactt
4801 tagcagcatg aaaataggta tacataccat agctcaatga gatgaggaag aacaggatct
4861 tcgtccaccc cgtacacatg gcgctggttg cgccccacac gatgcatccc acatcagaaa
4921 ccaacaggcc catagtcctt ttgctataat cgtccttcag gcctgtgagg ttgctcagat
4981 ggatcagaag gaccgggcag gtgagcagcc aggtcgcgta acgaagccac acggtcttgt
5041 tgccgttgga tgagtagatc accgcaggtt cgtcaaactc atggaaatac tcgatgatga
5101 acttgatcat ctcgatcgtg gccacgtaaa tggtctccca gccgcaagta gacttccagg
5161 tctggtagcc gtagaacatc aggcagagcg ctgaaagcgc aaaagtaatc cactgcagaa
5221 tgttggcagc caacttctcg gcatttgttc cgttggattt aagccaagcc aagcagaagc
5281 actggccgtt gttcgggatg caaataacag aaccattgtt ctcaagagtg tatgaggttt
5341 ggaagagcat gcgctcgtgg gcacggtgga aaacgtagtc ggggccatcc tgagtcgcga
5401 ccggcaccgt cgcgtcactg ccagtcgagg ctcctgcgct gccggccgcc agcgccactg
5461 ccagcgctag ggcaagaagc catggcctcc gcgacatggt ggcctaccgg tacccagcga
5521 gctcgctgtg gagagaaagg caaagtggat gtcagtaaga ccaataggtg cctatcagaa
5581 acgcaagagt cttctctgtc tcgacaagcc cagtttctat tggtctcctt aaacctgtct
5641 tgtaaccttg atacttacgt gcccagtgcc tcacgaccaa cttctgcagc ttaaggcgcc
5701 gcgctctgct tctggaaggc tgcgctcctc gacttgcagg ctagaagcaa atgtaagcaa
5761 tagatggctc tgccctgact tttatgccca gcccgggctg caggaagttt aaaataactc
5821 tgagcaattg agactaaatg cctctgtata gagaaaagac tgccacagac aggaatgttc
5881 ttggtgtctc ttgggtgcac tgttccatca taccatgtgt cccatatagt gctttcatat
5941 agattgtaaa tactgcatta ttttaaggaa atcatagtta attaaggtaa gtatttccaa
6001 aatggggtta taacaattta ctccaacatc acatgttaat atgatctgag ggaattatgc
6061 atagtgtttg ccttttgttt ttgtttattt ttccattact gtttcatagc aatgaagaga
6121 caaatgctga ctctagcaga atttcctcag tctgctaagt atagctacat aattgccacc
6181 ttcaccagca ccattgggaa aaaaataacc tacattactt gactccatat agctatattg
6241 ccataacttt gtagggctta cagtaatgtg gtttctgtat aaaactgaat tgaaatctca
6301 gaattctaca gagagaggaa tggggacaag aaaatgagga gggaactgaa gtggcttgtg
6361 tcaaagcctt cttttaggaa gtgtgcactt aacttctagt cagtaagcta tagcagccat
6421 gttagattaa accactagtt taattaaacg cgtgcggccg caggaacccc tagtgatgga
6481 gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc
6541 ccgacgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag g
//
| {
"Vector backbone": "AAV-Dlx-GFP",
"Backbone size w/o insert (bp)": "",
"Vector type": "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": "Viral manipulation of functionally distinct interneurons in mice, non-human primates and humans",
"Publication": "Nat Neurosci. 2020 Dec;23(12):1629-1636. doi: 10.1038/s41593-020-0692-9. Epub 2020 Aug 17.",
"DOI": "10.1038/s41593-020-0692-9.",
"Authors": "Vormstein-Schneider D, Lin JD, Pelkey KA, Chittajallu R, Guo B, Arias-Garcia MA, Allaway K, Sakopoulos S, Schneider G, Stevenson O, Vergara J, Sharma J, Zhang Q, Franken TP, Smith J, Ibrahim LA, M Astro KJ, Sabri E, Huang S, Favuzzi E, Burbridge T, Xu Q, Guo L, Vogel I, Sanchez V, Saldi GA, Gorissen BL, Yuan X, Zaghloul KA, Devinsky O, Sabatini BL, Batista-Brito R, Reynolds J, Feng G, Fu Z, McBain CJ, Fishell G, Dimidschstein J /s---",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/32807948",
"PubMed ID": "PubMed 32807948"
} | {
"Gene/Insert name": "C1V1-eYFP",
"Species": "M. musculus (mouse)",
"Insert Size (bp)": "",
"Mutation": "None",
"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": ""
} |
|
AiP1965 - pAAV-AiE2475m-minBG-SYFP2-WPRE-BGHpA | 214595 | AiE2475m is an enhancer sequence, designed to drive AAV-mediated transgene expression in specific populations of brain cells | Ben-Simon et al bioRxiv, 2024.06.10.597244 | https://media.addgene.org/snapgene-media/v2.0.0/sequences/432597/923b3ca8-b79e-40df-9003-f2b803d30b8d/addgene-plasmid-214595-sequence-432597.gbk | full | LOCUS sequence-432597- 4888 bp DNA circular SYN 27-AUG-2024
DEFINITION AiE2475m is an enhancer sequence, designed to drive AAV-mediated
transgene expression in specific populations of brain cells.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4888)
AUTHORS Ben-Simon Y, Hooper M, Narayan S, Daigle T, Dwivedi D, Way SW, Oster
A, Stafford DA, Mich JK, Taormina MJ, Martinez RA, Opitz-Araya X,
Roth JR, Allen S, Ayala A, Bakken TE, Barcelli T, Barta S, Bendrick
J, Bertagnolli D, Bowlus J, Boyer G, Brouner K, Casian B, Casper T,
Chakka AB, Chakrabarty R, Chance RK, Chavan S, Departee M, Donadio
N, Dotson N, Egdorf T, Gabitto M, Gary A, Gasperini M, Goldy J, Gore
BB, Graybuck L, Greisman N, Haeseleer F, Halterman C, Helback O,
Hockemeyer D, Huang C, Huff S, Hunker A, Johansen N, Juneau Z,
Kalmbach B, Khem S, Kutsal R, Larsen R, Lee C, Lee AY, Leibly M,
Lenz GH, Liang E, Lusk N, Malone J, Mollenkopf T, Morin E, Newman D,
Ng L, Ngo K, Omstead V, Oyama A, Pham T, Pom CA, Potekhina L,
Ransford S, Rette D, Rimorin C, Rocha D, Ruiz A, Sanchez REA,
Sedeno-Cortes A, Sevigny JP, Shapovalova N, Shulga L, Sigler AR,
Siverts LA, Somasundaram S, Stewart K, Tieu M, Trader C, van
Velthoven CTJ, Walker M, Weed N, Wirthlin M, Wood T, Wynalda B, Yao
Z, Zhou T, Ariza J, Dee N, Reding M, Ronellenfitch K, Mufti S,
Sunkin SM, Smith KA, Esposito L, Waters J, Thyagarajan B, Yao S,
Lein ES, Zeng H, Levi BP, Ngai J, Ting J, Tasic B
TITLE A suite of enhancer AAVs and transgenic mouse lines for genetic
access to cortical cell types
JOURNAL bioRxiv, 2024.06.10.597244
REFERENCE 2 (bases 1 to 4888)
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,
2024.06.10.597244"
FEATURES Location/Qualifiers
source 1..4888
/mol_type="other DNA"
/organism="synthetic DNA construct"
repeat_region 503..632
/label=AAV2 ITR (alternate)
/note="Functional equivalent of wild-type AAV2 ITR"
rep_origin complement(694..1282)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(774..793)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(1453..2313)
/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 2076..2095
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(2314..2418)
/gene="bla"
/label=AmpR promoter
primer_bind 2486..2504
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(2542..2564)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 2664..2683
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
rep_origin complement(2700..3155)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2837..2858)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 3049..3068
/label=F1ori-R
/note="F1 origin, reverse primer"
repeat_region 3230..3370
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 3241..3370
/label=AAV2 ITR
polyA_signal 3488..3599
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind 3786..3806
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(3899..4615)
/codon_start=1
/product="monomeric optimized variant of EYFP, identical to
mVenus but with L68V (Kremers et al., 2006)"
/label=SYFP2
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KLICTTGKLPVPWPTLVTTLGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(3941..3962)
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind 4290..4309
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 4549..4570
/label=EGFP-N
/note="EGFP, reverse primer"
regulatory 4612..4621
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
ORIGIN
1 caaggcatgt agccatctct tccctctttc tcttcttcct tctagatctt gtcttcctgc
61 gccctcattg ctcgcgtttc tgattcatgt atctgaccac catcagcaga tctgaagagg
121 acaggaagca cccagttcga actcttccaa gtcggggcct gcacacccca cccccacccc
181 cttcctggag tctgtctggg aggggatccc caatgacgca ctcgggaagg cagtacagag
241 ccccacgcca ggcccgggga tcatgaatga gggcagagag aaggccaggc gcttcctcct
301 caccagccgg agctttggcc cccagcaggg tgagcagctg ccctcctgag gcagcccagg
361 caggctctat gcccagccct tgttggccag aagacaagct ggctgggtcc ccgcgaaaga
421 ggtcgtttca atagctttca ccctgcactc ccgcacacca ccccagcccc ggccccgcat
481 gctgtaacct ccttgcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg
541 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggcggcctc
601 agtgagcgag cgagcgcgca gctgcctgca ggacatgtga gcaaaaggcc agcaaaaggc
661 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
721 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
781 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
841 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
901 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
961 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
1021 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
1081 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
1141 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
1201 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
1261 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
1321 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
1381 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
1441 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
1501 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
1561 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
1621 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
1681 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
1741 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
1801 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
1861 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
1921 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
1981 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
2041 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
2101 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
2161 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
2221 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
2281 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
2341 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
2401 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
2461 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
2521 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
2581 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
2641 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccataa
2701 aattgtaaac gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
2761 ttttaaccaa tagaccgaaa tcggcaaaat cccttataaa tcaaaagaat agcccgagat
2821 agagttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
2881 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccaa
2941 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
3001 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
3061 gaaaggagcg ggcgctaagg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
3121 acccgccgcg cttaatgcgc cgctacaggg cgcgtactat ggttgctttg acgtatgcgg
3181 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgccc ctgcaggcag
3241 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
3301 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
3361 aggggttcct gcggccgctc ggtccgcacg tgagatctca tgcctgctat tgtcttccca
3421 atcctccccc ttgctgtcct gccccacccc accccccaga atagaatgac acctactcag
3481 acaatgcgat gcaatttcct cattttatta ggaaaggaca gtgggagtgg caccttccag
3541 ggtcaaggaa ggcacggggg aggggcaaac aacagatggc tggcaactag aaggcacagt
3601 cgaagatctc tcgagccacg gaattgtcag tgcccaacag ccgagcccct gtccagcagc
3661 gggcaaggca ggcggcgatg agttccgccg tggcaagaac taaccaggat ttatacaagg
3721 aggagaaaat gaaagccata cgggaagcaa tagcatgata caaaggcatt aaagcagcgt
3781 atccacatag cgtaaaagga gcaacatagt taagaatacc agtcaatctt tcacaaattt
3841 tgtaatccag aggttgatta tgatatcgaa ttcgcggccg cggcgcgccg tcgacttact
3901 tgtacagctc gtccatgccg agagtgatcc cggcggcggt cacgaactcc agcaggacca
3961 tgtgatcgcg cttctcgttg gggtctttgc tcagcttgga ctggtagctc aggtagtggt
4021 tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag tggtcggcga
4081 gctgcacgcc gccgtcctcg atgttgtggc ggatcttgaa gttggccttg atgccgttct
4141 tctgcttgtc ggcggtgata tagacgttgt ggctgttgta gttgtactcc agcttgtgcc
4201 ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg
4261 tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt gccgtcgtcc ttgaagaaga
4321 tggtgcgctc ctggacgtag ccttcgggca tggcggactt gaagaagtcg tgctgcttca
4381 tgtggtcggg gtagcgggcg aagcactgca cgccgtagcc cagggtggtc acgagggtgg
4441 gccagggcac gggcagcttg ccggtggtgc agatcagctt cagggtcagc ttgccgtagg
4501 tggcatcgcc ctcgccctcg ccggacacgc tgaacttgtg gccgtttacg tcgccgtcca
4561 gctcgaccag gatgggcacc accccggtga acagctcctc gcccttgctc accatggtgg
4621 cgaccggtag cgctagcttc gaaagatctg gatcccagaa gcaaatgtaa gcaatagatg
4681 gctctgccct gacttttatg cccagcccga gctccggcta cgtgacagtt ccctagagaa
4741 tggtacagct gcaagcctca acacccagca tgcaccgcag ctggggagtg ctgggagcct
4801 ttgaagagcc cgggcaggat gccacaggca tctgcgggaa gcaggggata cacaaggcag
4861 ggtcagcact gactgatttc aaccacta
//
| {
"Vector backbone": "pAAV2",
"Backbone size w/o insert (bp)": "",
"Vector type": "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": "A suite of enhancer AAVs and transgenic mouse lines for genetic access to cortical cell types",
"Publication": "bioRxiv, 2024.06.10.597244",
"DOI": "10.1101/2024.06.10.597244",
"Authors": "Ben-Simon Y, Hooper M, Narayan S, Daigle T, Dwivedi D, Way SW, Oster A, Stafford DA, Mich JK, Taormina MJ, Martinez RA, Opitz-Araya X, Roth JR, Allen S, Ayala A, Bakken TE, Barcelli T, Barta S, Bendrick J, Bertagnolli D, Bowlus J, Boyer G, Brouner K, Casian B, Casper T, Chakka AB, Chakrabarty R, Chance RK, Chavan S, Departee M, Donadio N, Dotson N, Egdorf T, Gabitto M, Gary A, Gasperini M, Goldy J, Gore BB, Graybuck L, Greisman N, Haeseleer F, Halterman C, Helback O, Hockemeyer D, Huang C, Huff S, Hunker A, Johansen N, Juneau Z, Kalmbach B, Khem S, Kutsal R, Larsen R, Lee C, Lee AY, Leibly M, Lenz GH, Liang E, Lusk N, Malone J, Mollenkopf T, Morin E, Newman D, Ng L, Ngo K, Omstead V, Oyama A, Pham T, Pom CA, Potekhina L, Ransford S, Rette D, Rimorin C, Rocha D, Ruiz A, Sanchez REA, Sedeno-Cortes A, Sevigny JP, Shapovalova N, Shulga L, Sigler AR, Siverts LA, Somasundaram S, Stewart K, Tieu M, Trader C, van Velthoven CTJ, Walker M, Weed N, Wirthlin M, Wood T, Wynalda B, Yao Z, Zhou T, Ariza J, Dee N, Reding M, Ronellenfitch K, Mufti S, Sunkin SM, Smith KA, Esposito L, Waters J, Thyagarajan B, Yao S, Lein ES, Zeng H, Levi BP, Ngai J, Ting J, Tasic B",
"PubMed Link": "",
"PubMed ID": ""
} | {
"Gene/Insert name": "SYFP2",
"Species": "Synthetic",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "minBG",
"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": ""
} |
||
pMOD_A4210 | 91057 | Module A, Promoter: Zmubi, Gene: Csy4-P2A-dTaCas9-VP64, Terminator: HSP | Daniel Voytas | Cermak et al Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/181850/7a0b55c2-fccb-4cdc-9b6c-a8055c7a7185/addgene-plasmid-91057-sequence-181850.gbk | full | LOCUS sequence-181850- 9276 bp DNA circular SYN 26-AUG-2024
DEFINITION Module A, Promoter: Zmubi, Gene: Csy4-P2A-dTaCas9-VP64, Terminator:
HSP.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9276)
AUTHORS Cermak T, Curtin SJ, Gil-Humanes J, Cegan R, Kono TJY, Konecna E,
Belanto JJ, Starker CG, Mathre JW, Greenstein RL, Voytas DF
TITLE A multi-purpose toolkit to enable advanced genome engineering in
plants.
JOURNAL Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi:
10.1105/tpc.16.00922.
PUBMED 28522548
REFERENCE 2 (bases 1 to 9276)
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: "Plant Cell.
2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922."
FEATURES Location/Qualifiers
source 1..9276
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin 80..668
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 569..588
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 822..839
/label=L4440
/note="L4440 vector, forward primer"
CDS 3453..3509
/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 3510..7613
/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 7614..7661
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
CDS 7677..7826
/codon_start=1
/product="tetrameric repeat of the minimal activation
domain of herpes simplex virus VP16 (Beerli et al., 1998)"
/label=VP64
/translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
DLDML"
terminator 7856..8104
/label=HSP terminator
/note="efficient transcription terminator from the
Arabidopsis thaliana heat shock protein 18.2 gene (Nagaya
et al., 2010)"
promoter 8184..8288
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gccttgacgg caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
61 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
121 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ctaactcttt
181 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
241 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
301 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
361 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgctcacagc
421 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
481 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
541 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
601 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
661 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
721 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
781 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
841 aagcggaaga gcgcccacct gctcatgatc tgcaggcgcg cctgcagtgc agcgtgaccc
901 ggtcgtgccc ctctctagag ataatgagca ttgcatgtct aagttataaa aaattaccac
961 atattttttt tgtcacactt gtttgaagtg cagtttatct atctttatac atatatttaa
1021 actttactct acgaataata taatctatag tactacaata atatcagtgt tttagagaat
1081 catataaatg aacagttaga catggtctaa aggacaattg agtattttga caacaggact
1141 ctacagtttt atctttttag tgtgcatgtg ttctcctttt tttttgcaaa tagcttcacc
1201 tatataatac ttcatccatt ttattagtac atccatttag ggtttagggt taatggtttt
1261 tatagactaa tttttttagt acatctattt tattctattt tagcctctaa attaagaaaa
1321 ctaaaactct attttagttt ttttatttaa taatttagat ataaaataga ataaaataaa
1381 gtgactaaaa attaaacaaa taccctttaa gaaattaaaa aaactaagga aacatttttc
1441 ttgtttcgag tagataatgc cagcctgtta aacgccgtcg acgagtctaa cggacaccaa
1501 ccagcgaacc agcagcgtcg cgtcgggcca agcgaagcag acggcacggc atctctgtcg
1561 ctgcctctgg acccctctcg agagttccgc tccaccgttg gacttgctcc gctgtcggca
1621 tccagaaatt gcgtggcgga gcggcagacg tgagccggca cggcaggcgg cctcctcctc
1681 ctctcacggc accggcagct acgggggatt cctttcccac cgctccttcg ctttcccttc
1741 ctcgcccgcc gtaataaata gacaccccct ccacaccctc tttccccaac ctcgtgttgt
1801 tcggagcgca cacacacaca accagatctc ccccaaatcc acccgtcggc acctccgctt
1861 caaggtacgc cgctcgtcct cccccccccc ctctctacct tctctagatc ggcgttccgg
1921 tccatggtta gggcccggta gttctacttc tgttcatgtt tgtgttagat ccgtgtttgt
1981 gttagatccg tgctgctagc gttcgtacac ggatgcgacc tgtacgtcag acacgttctg
2041 attgctaact tgccagtgtt tctctttggg gaatcctggg atggctctag ccgttccgca
2101 gacgggatcg atttcatgat tttttttgtt tcgttgcata gggtttggtt tgcccttttc
2161 ctttatttca atatatgccg tgcacttgtt tgtcgggtca tcttttcatg cttttttttg
2221 tcttggttgt gatgatgtgg tctggttggg cggtcgttct agatcggagt agaattaatt
2281 ctgtttcaaa ctacctggtg gatttattaa ttttggatct gtatgtgtgt gccatacata
2341 ttcatagtta cgaattgaag atgatggatg gaaatatcga tctaggatag gtatacatgt
2401 tgatgcgggt tttactgatg catatacaga gatgcttttt gttcgcttgg ttgtgatgat
2461 gtggtgtggt tgggcggtcg ttcattcgtt ctagatcgga gtagaatact gtttcaaact
2521 acctggtgta tttattaatt ttggaactgt atgtgtgtgt catacatctt catagttacg
2581 agtttaagat ggatggaaat atcgatctag gataggtata catgttgatg tgggttttac
2641 tgatgcatat acatgatggc atatgcagca tctattcata tgctctaacc ttgagtacct
2701 atctattata ataaacaagt atgttttata attattttga tcttgatata cttggatgat
2761 ggcatatgca gcagctatat gtggattttt ttagccctgc cttcatacgc tatttatttg
2821 cttggtactg tttcttttgt cgatgctcac cctgttgttt ggtgttactt cctgcaggaa
2881 gcatggacca ctacctcgac atcaggctca ggccagaccc agagttccca ccagcccagc
2941 tcatgtccgt cctcttcggc aagctccacc aggccctcgt ggcccagggc ggcgacagga
3001 tcggcgtgtc cttcccagac ctcgacgagt ccaggtccag gctcggcgag aggctccgca
3061 tccacgcctc cgccgacgac ctcagggccc tcctcgccag gccgtggctg gagggcctca
3121 gggaccacct ccagttcggc gagccagccg tggtgccaca cccaacccca tacaggcaag
3181 tgtccagggt gcaagccaag tccaacccag agaggctcag gaggaggctc atgaggaggc
3241 acgacctctc cgaggaagag gccaggaagc gcatcccaga caccgtggcc agggccctcg
3301 acctcccatt cgtgaccctc aggtcccagt ccaccggcca gcacttccgc ctcttcatca
3361 ggcacggccc actccaggtg accgccgagg agggcggctt tacctgctac ggcctctcca
3421 agggcggctt cgtgccgtgg ttcggctccg gcgccaccaa cttctccctc ctcaagcaag
3481 ccggcgacgt ggaggagaac ccaggcccaa tggacaagaa gtactcgatc ggcctcgcca
3541 tcgggacgaa ctcagttggc tgggccgtga tcaccgacga gtacaaggtg ccctctaaga
3601 agttcaaggt cctggggaac accgaccgcc attccatcaa gaagaacctc atcggcgctc
3661 tcctgttcga cagcggggag accgctgagg ctacgaggct caagagaacc gctaggcgcc
3721 ggtacacgag aaggaagaac aggatctgct acctccaaga gattttctcc aacgagatgg
3781 ccaaggttga cgattcattc ttccaccgcc tggaggagtc tttcctcgtg gaggaggata
3841 agaagcacga gcggcatccc atcttcggca acatcgtgga cgaggttgcc taccacgaga
3901 agtaccctac gatctaccat ctgcggaaga agctcgtgga ctccaccgat aaggcggacc
3961 tcagactgat ctacctcgct ctggcccaca tgatcaagtt ccgcggccat ttcctgatcg
4021 agggggatct caacccagac aacagcgatg ttgacaagct gttcatccaa ctcgtgcaga
4081 cctacaacca actcttcgag gagaacccga tcaacgcctc tggcgtggac gcgaaggcta
4141 tcctgtccgc gaggctctcg aagtccagga ggctggagaa cctgatcgct cagctcccag
4201 gcgagaagaa gaacggcctg ttcgggaacc tcatcgctct cagcctgggg ctcaccccga
4261 acttcaagtc gaacttcgat ctcgctgagg acgccaagct gcaactctcc aaggacacct
4321 acgacgatga cctcgataac ctcctggccc agatcggcga tcaatacgcg gacctgttcc
4381 tcgctgccaa gaacctgtcg gacgccatcc tcctgtcaga tatcctccgc gtgaacaccg
4441 agatcacgaa ggctccactc tctgcctcca tgatcaagcg ctacgacgag caccatcagg
4501 atctgaccct cctgaaggcg ctggtccgcc aacagctccc ggagaagtac aaggagattt
4561 tcttcgatca gtcgaagaac ggctacgctg ggtacatcga cggcggggcc tcacaagagg
4621 agttctacaa gttcatcaag ccaatcctgg agaagatgga cggcacggag gagctcctgg
4681 tgaagctcaa cagggaggac ctcctgcgga agcagagaac cttcgataac ggcagcatcc
4741 cccaccaaat ccatctcggg gagctgcacg ccatcctgag aaggcaagag gacttctacc
4801 ctttcctcaa ggataaccgg gagaagatcg agaagatcct gaccttcaga atcccatact
4861 acgtcggccc tctcgcgcgg gggaactcaa gattcgcttg gatgacccgc aagtctgagg
4921 agaccatcac gccgtggaac ttcgaggagg tggtggacaa gggcgctagc gctcagtcgt
4981 tcatcgagag gatgaccaac ttcgacaaga acctgcccaa cgagaaggtg ctccctaagc
5041 actcgctcct gtacgagtac ttcaccgtct acaacgagct cacgaaggtg aagtacgtca
5101 ccgagggcat gcgcaagcca gcgttcctgt ccggggagca gaagaaggct atcgtggacc
5161 tcctgttcaa gaccaaccgg aaggtcacgg ttaagcaact caaggaggac tacttcaaga
5221 agatcgagtg cttcgattcg gtcgagatca gcggcgttga ggaccgcttc aacgccagcc
5281 tcgggaccta ccacgatctc ctgaagatca tcaaggataa ggacttcctg gacaacgagg
5341 agaacgagga tatcctggag gacatcgtgc tgaccctcac gctgttcgag gacagggaga
5401 tgatcgagga gcgcctgaag acgtacgccc atctcttcga tgacaaggtc atgaagcaac
5461 tcaagcgccg gagatacacc ggctggggga ggctgtcccg caagctcatc aacggcatcc
5521 gggacaagca gtccgggaag accatcctcg acttcctcaa gagcgatggc ttcgccaaca
5581 ggaacttcat gcaactgatc cacgatgaca gcctcacctt caaggaggat atccaaaagg
5641 ctcaagtgag cggccagggg gactcgctgc acgagcatat cgcgaacctc gctggctccc
5701 ccgcgatcaa gaagggcatc ctccagaccg tgaaggttgt ggacgagctc gtgaaggtca
5761 tgggccggca caagcctgag aacatcgtca tcgagatggc cagagagaac caaaccacgc
5821 agaaggggca aaagaactct agggagcgca tgaagcgcat cgaggagggc atcaaggagc
5881 tggggtccca aatcctcaag gagcacccag tggagaacac ccaactgcag aacgagaagc
5941 tctacctgta ctacctccag aacggcaggg atatgtacgt ggaccaagag ctggatatca
6001 accgcctcag cgattacgac gtcgatgcta tcgttcccca gtctttcctg aaggatgact
6061 ccatcgacaa caaggtcctc accaggtcgg acaagaaccg cggcaagtca gataacgttc
6121 catctgagga ggtcgttaag aagatgaaga actactggag gcagctcctg aacgccaagc
6181 tgatcacgca aaggaagttc gacaacctca ccaaggctga gagaggcggg ctctcagagc
6241 tggacaaggc cggcttcatc aagcggcagc tggtcgagac cagacaaatc acgaagcacg
6301 ttgcgcaaat cctcgactct cggatgaaca cgaagtacga tgagaacgac aagctgatca
6361 gggaggttaa ggtgatcacc ctgaagtcta agctcgtctc cgacttcagg aaggatttcc
6421 agttctacaa ggttcgcgag atcaacaact accaccatgc ccatgacgct tacctcaacg
6481 ctgtggtcgg caccgctctg atcaagaagt acccaaagct ggagtccgag ttcgtgtacg
6541 gggactacaa ggtttacgat gtgcgcaaga tgatcgccaa gtcggagcaa gagatcggca
6601 aggctaccgc caagtacttc ttctactcaa acatcatgaa cttcttcaag accgagatca
6661 cgctggccaa cggcgagatc cggaagagac cgctcatcga gaccaacggc gagacggggg
6721 agatcgtgtg ggacaagggc agggatttcg cgaccgtccg caaggttctc tccatgcccc
6781 aggtgaacat cgtcaagaag accgaggtcc aaacgggcgg gttctcaaag gagtctatcc
6841 tgcctaagcg gaacagcgac aagctcatcg ccagaaagaa ggactgggac ccaaagaagt
6901 acggcgggtt cgacagccct accgtggcct actcggtcct ggttgtggcg aaggttgaga
6961 agggcaagtc caagaagctc aagagcgtga aggagctcct ggggatcacc atcatggaga
7021 ggtccagctt cgagaagaac ccaatcgact tcctggaggc caagggctac aaggaggtga
7081 agaaggacct gatcatcaag ctcccgaagt actctctctt cgagctggag aacggcagga
7141 agagaatgct ggcttccgct ggcgagctcc agaaggggaa cgagctcgcg ctgccaagca
7201 agtacgtgaa cttcctctac ctggcttccc actacgagaa gctcaagggc agcccggagg
7261 acaacgagca aaagcagctg ttcgtcgagc agcacaagca ttacctcgac gagatcatcg
7321 agcaaatctc cgagttcagc aagcgcgtga tcctcgccga cgcgaacctg gataaggtcc
7381 tctccgccta caacaagcac cgggacaagc ccatcagaga gcaagcggag aacatcatcc
7441 atctcttcac cctgacgaac ctcggcgctc ctgctgcttt caagtacttc gacaccacga
7501 tcgatcggaa gagatacacc tccacgaagg aggtcctgga cgcgaccctc atccaccagt
7561 cgatcaccgg cctgtacgag acgaggatcg acctctcaca actcggcggg gataagagac
7621 ccgcagcaac caagaaggca gggcaagcaa agaagaagaa gggatccgga cgggctgacg
7681 cattggacga ttttgatctg gatatgctgg gaagtgacgc cctcgatgat tttgaccttg
7741 acatgcttgg ttcggatgcc cttgatgact ttgacctcga catgctcggc agtgacgccc
7801 ttgatgattt cgacctggac atgctgatta actctagatg acgcgtactc gagtatatga
7861 agatgaagat gaaatatttg gtgtgtcaaa taaaaagctt gtgtgcttaa gtttgtgttt
7921 ttttcttggc ttgttgtgtt atgaatttgt ggctttttct aatattaaat gaatgtaaga
7981 tcacattata atgaataaac aaatgtttct ataatccatt gtgaatgttt tgttggatct
8041 cttctgcagc atataactac tgtatgtgct atggtatgga ctatggaata tgattaaaga
8101 taaggagctc gttaaccggt gacggacgtg ggcaggtgtg atgaaaagcc ttagggtcag
8161 gtggcacttt tcggggaaat gtgcgatgaa cccctatttg tttatttttc taaatacatt
8221 caaatatgta tccgctaatg agacaataac cctgataaat gcttcaatta tattgaaaaa
8281 ggaagagtat gagtatccaa catttccgtg tcgcccttat tccctttttt gcggccttct
8341 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
8401 tgggtgcccg agtgggttac attgaactgg atctcaacag cggcaaaatc ttagagagtt
8461 ttcgccccga agaacgcttc ccaatgatga gcactttcaa agttctgcta tgtggcgcgg
8521 tgttatcccg tattgatgct gggcaagagc aactcggtcg ccgcatacac tattctcaga
8581 atgacttggt tgagtattca ccagtcacag aaaagcatct tacggatggc atgacagtaa
8641 gagaattatg tagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
8701 caacgattgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
8761 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
8821 ccacgatgcc tgtagcaatg gcaacaactc tccgcaaact attaactggc gaactactta
8881 ctcttgcttc acgccagcaa ctcattgact ggatggaggc ggacaaagtt gcaggaccac
8941 ttctgcgctc ggcacttccg gctggctggt ttattgctga taaatctgga gccggtgagc
9001 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
9061 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
9121 taggtgcctc actgattaag cattggtaac tgtcagacga agggaaataa atagtagccc
9181 gcctgatgtg cgggcttttt tttggtggtg gttaccagtg tcgcttagct accgtaggtc
9241 ctcttaggcg gtccgattag cgacatagtc atcaat
//
| {
"Vector backbone": "pMOD_A1001",
"Backbone size w/o insert (bp)": "",
"Vector type": "CRISPR",
"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 multi-purpose toolkit to enable advanced genome engineering in plants",
"Publication": "Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922.",
"DOI": "10.1105/tpc.16.00922.",
"Authors": "Cermak T, Curtin SJ, Gil-Humanes J, Cegan R, Kono TJY, Konecna E, Belanto JJ, Starker CG, Mathre JW, Greenstein RL, Voytas DF /tpc",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/28522548",
"PubMed ID": "PubMed 28522548"
} | {
"Gene/Insert name": "Csy4-P2A-dTaCas9-VP64",
"Species": "Synthetic",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "Zmubi",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Gibson Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "unknown",
"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": ""
} |
|
T4 Lysozyme T26H WT* | 18251 | Brian Matthews | T4 Lysozyme Plasmids - Tray 1 (unpublished) | https://media.addgene.org/snapgene-media/v2.0.0/sequences/24038/bf420d2b-cf7e-433a-87fc-a2811e23904d/addgene-plasmid-18251-sequence-24038.gbk | partial | LOCUS sequence-24038-e 853 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 853)
TITLE T4 Lysozyme Plasmids - Tray 1
REFERENCE 2 (bases 1 to 853)
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..853
/mol_type="genomic DNA"
/organism="unspecified"
ORIGIN
1 gaaaagcccg ctcattaggc gggctcagat ctgctcatgt ttgacagctt atcatcgata
61 agcttcggca ataaattctt gatatgtttt catatgtttc ctttaaatat tttaattagt
121 aattgtctat tcaagtaatt gtgaatatac tatcacaatt ccaagagaaa gtaaacagct
181 ttatagattt ttatacgcgt cccaagtgcc agttctaaac gttgtaatga ctcgttttgc
241 gcgattaggt gtttgattat accatctact tttagctaag ttaactgctg cttcatccca
301 gcgtttttgt tgaagcatac gtaaagagtt agtaaatcct gccacaccgg tttctcccat
361 ttggaaaacc atattaatca atgcagcgcg acgaaccgca tcaagagaat cataaaccgg
421 ttttaattta gcatttctca gaattccgcg aacagcagca tcaacatcct gattaaagag
481 tttttcagcc tcatcttttg taattacacc attagtatta cgcccaatag ctttatctaa
541 ttcagattta gcagcattaa gtgatggact ttttgtaagc aaatgaccga tgccaatatg
601 ataatagcct tctgtgtctt tatagatttt aagtctaaga ccttcatcta tacgtaacat
661 ttcaaatata ttcataatac ctcctaagca tcgatggatc ctgtttcctg tgtgaattgt
721 tatccgctca cattccacac attatacgag ccgatgatan tgtcacaggg gatgggagta
781 gctganctgt tctggngaat gtancgncca tccccataac ngcgagatat gcacggggag
841 gnnanccngn cnc
//
| {
"Vector backbone": "pHS1403",
"Backbone size w/o insert (bp)": "5000",
"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": "T4 Lysozyme",
"Species": "Bacteriophage T4",
"Insert Size (bp)": "600",
"Mutation": "T26H WT*",
"Entrez Gene": "e",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "BamHI (not destroyed)",
"3′ cloning site": "HindIII (not destroyed)",
"5′ sequencing primer": "na",
"3′ sequencing primer": "GGCGTATCACGAGGCCC"
} | {
"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": ""
} |
||
pH6HTC_FAM98A-Halo | 175329 | Production of HaloTag fusion protein | Claudia Kutter | Atanasoai et al bioRxiv 2021.11.08.467743 | https://media.addgene.org/snapgene-media/v2.0.0/sequences/345583/433d6ca6-d94c-459a-9f02-717af06e5444/addgene-plasmid-175329-sequence-345583.gbk | full | LOCUS sequence-345583- 4992 bp DNA circular SYN 25-AUG-2024
DEFINITION Production of HaloTag fusion protein.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4992)
AUTHORS Atanasoai I, Papavasileiou S, Preiß N, Kutter C
TITLE Large-scale identification of RBP-RNA interactions by RAPseq refines
essentials of post-transcriptional gene regulation
JOURNAL bioRxiv 2021.11.08.467743
REFERENCE 2 (bases 1 to 4992)
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: "bioRxiv
2021.11.08.467743"
FEATURES Location/Qualifiers
source 1..4992
/mol_type="other DNA"
/organism="synthetic DNA construct"
terminator 37..64
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
primer_bind 146..165
/label=T7
/note="T7 promoter, forward primer"
promoter 146..164
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory 1317..1326
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 1783..1803
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="EDLYFQS"
CDS 1819..2703
/codon_start=1
/product="modified bacterial dehalogenase that forms
covalent bonds with chloroalkane derivatives"
/label=HaloTag(R)
/translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
SEIARWLSTLEISG"
CDS 2704..2721
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(2732..2750)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 2746..2793
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
promoter 3022..3126
/gene="bla"
/label=AmpR promoter
CDS 3127..3987
/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(3345..3364)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin complement(4145..4733)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4225..4244)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
terminator 4851..4937
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
ORIGIN
1 gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg acggatggcc
61 tttttgcgtt tctacaaact cttttgttta tttttctaaa tacattcaaa tatgtatccg
121 ctcatggatc tcgatcccgc gaaattaata cgactcacta tagggtattt taactttact
181 aaggagcgat cgcttccgaa ttcctaccgc ggatatatgg agtgtgacct catggagact
241 gacatcttgg agtcgttgga agatctaggt tacaagggcc cattgttgga agatggagcg
301 ctctctcagg cagtctctgc tggagccagt tcccccgagt ttaccaaact ctgtgcttgg
361 ctggtgtctg aattaagagt gctctgtaaa ctagaggaaa acgtgcaagc aactaacagt
421 ccgagtgaag ctgaagaatt ccagcttgag gtgagtgggc tactagggga gatgaactgc
481 ccgtatcttt cactgacatc tggggatgtg accaagcgcc ttctcattca gaagaactgc
541 ctcctcttgc tcacatacct catctcagaa ctagaagctg ccagaatgct ctgtgtgaat
601 gctcctccaa aaaaagctca agaaggaggc ggtagtgagg tctttcaaga gttgaaaggc
661 atatgtattg ctctaggaat gtccaaacct ccagccaata taactatgtt ccaattcttc
721 agcgggattg aaaaaaaatt aaaggaaaca ttagcaaaag ttccacctaa tcatgtggga
781 aagcctttac tgaagaagcc aatgggacca gcccactggg aaaagataga agcaattaac
841 caagccatag ccaatgaata tgaagtccgg agaaagctgc taataaaacg tttggatgtc
901 actgtacaat cctttggctg gtctgacaga gctaagagcc agacagaaaa attagccaag
961 gtttaccagc cgaaacgttc agtcttatcc cctaaaacta ctatttctgt tgcccatctt
1021 ttggctgcaa ggcaggactt gtcaaagatt ttaaggacaa gcagcggctc tataagagaa
1081 aagactgcct gtgccatcaa taaggtgttg atgggcaggg tgcctgacag aggtggtaga
1141 cccaatgaaa tcgaacctcc acccccagag atgccaccgt ggcagaagag gcaagatggc
1201 ccccagcagc aaacaggagg ccgaggagga gggagaggtg gctatgaaca ttcctcatac
1261 ggaggacgag gaggtcatga acaaggaggc gggagaggtg gacgtggtgg ctatgaccat
1321 ggtggccgag ggggaggaag aggaaataag catcaaggag gctggacaga tggagggagt
1381 ggtggaggag gtggctacca agatggtggt tatcgagatt caggtttcca gccaggtggc
1441 tatcatggtg gccacagcag tggtggctat caaggcggag gttatggtgg cttccaaaca
1501 tcttcttcat atacaggaag tggataccag ggtggtggct accagcagga caatagatac
1561 caagatggcg ggcaccatgg tgatcgtggt ggtggtcgtg gtgggcgagg tggtcgtgga
1621 ggccgaggtg gtcgtgcagg ccagggagga ggctggggag gaagagggag ccagaattat
1681 caccaagggg gtcaatttga acagcatttc cagcatggag gttatcagta taatcattct
1741 ggatttggac agggaagaca ttacactagt gggccaacca ctgaggatct gtactttcag
1801 agcgataacg atggatccga aatcggtact ggctttccat tcgaccccca ttatgtggaa
1861 gtcctgggcg agcgcatgca ctacgtcgat gttggtccgc gcgatggcac ccctgtgctg
1921 ttcctgcacg gtaacccgac ctcctcctac gtgtggcgca acatcatccc gcatgttgca
1981 ccgacccatc gctgcattgc tccagacctg atcggtatgg gcaaatccga caaaccagac
2041 ctgggttatt tcttcgacga ccacgtccgc ttcatggatg ccttcatcga agccctgggt
2101 ctggaagagg tcgtcctggt cattcacgac tggggctccg ctctgggttt ccactgggcc
2161 aagcgcaatc cagagcgcgt caaaggtatt gcatttatgg agttcatccg ccctatcccg
2221 acctgggacg aatggccaga atttgcccgc gagaccttcc aggccttccg caccaccgac
2281 gtcggccgca agctgatcat cgatcagaac gtttttatcg agggtacgct gccgatgggt
2341 gtcgtccgcc cgctgactga agtcgagatg gaccattacc gcgagccgtt cctgaatcct
2401 gttgaccgcg agccactgtg gcgcttccca aacgagctgc caatcgccgg tgagccagcg
2461 aacatcgtcg cgctggtcga agaatacatg gactggctgc accagtcccc tgtcccgaag
2521 ctgctgttct ggggcacccc aggcgttctg atcccaccgg ccgaagccgc tcgcctggcc
2581 aaaagcctgc ctaactgcaa ggctgtggac atcggcccgg gtctgaatct gctgcaagaa
2641 gacaacccgg acctgatcgg cagcgagatc gcgcgctggc tgtctactct ggagatttcc
2701 ggtcatcatc accatcacca ctaatagtca accgctgagc aataactagc ataacccctt
2761 ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggttcg
2821 cttgctgtcc ataaaaccgc ccagtctagc tatcgccatg taagcccact gcaagctacc
2881 tgctttctct ttgcgcttgc gttttccctt gtccagatag cccagtagct gacattcatc
2941 cggggtcagc accgtttctg cggactggct ttctacgtaa tggtttctta gacgtcaggt
3001 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
3061 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
3121 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
3181 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
3241 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
3301 cgccccgaag aacgttttcc aatgatgagc actttcaaag ttctgctatg tggcgcggta
3361 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
3421 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
3481 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
3541 actatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
3601 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
3661 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
3721 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
3781 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
3841 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
3901 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
3961 ggtgcctcac tgattaagca ttggtaattc gaaatgaccg accaagcgac gcccaaccgg
4021 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
4081 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
4141 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
4201 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
4261 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
4321 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
4381 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
4441 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
4501 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
4561 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
4621 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
4681 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggattt caagaagatc
4741 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggcata
4801 tgatggtagt gtggggtctc cccatgcgag agtagggaac tgccaggcat caaataaaac
4861 gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc
4921 tcctgagtag gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag
4981 ggtggcgggc ag
//
| {
"Vector backbone": "pH6HTC_minus_cer-region",
"Backbone size w/o insert (bp)": "",
"Vector type": "Bacterial Expression ; Cell Free 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 ; HaloTag Vector Limited Use Label License",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Large-scale identification of RBP-RNA interactions by RAPseq refines essentials of post-transcriptional gene regulation",
"Publication": "bioRxiv 2021.11.08.467743",
"DOI": "10.1101/2021.11.08.467743",
"Authors": "Atanasoai I, Papavasileiou S, Preiß N, Kutter C",
"PubMed Link": "",
"PubMed ID": ""
} | {
"Gene/Insert name": "FAM98A",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "FAM98A",
"Promoter": "",
"Tag / Fusion Protein": "HaloTag-6xHis (C terminal on backbone)"
} | {
"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": ""
} |
|
pPS22 (pET23a-Cas10/Csm/csm2H6N) | 70038 | Low | Bacterial expression of the S. epidermidis Cas10·Csm complex | Luciano Marraffini | Hatoum-Aslan et al J Biol Chem. 2013 Sep 27;288(39):27888-97. doi: 10.1074/jbc.M113.499244. Epub 2013 Aug 9. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/220331/064f8877-989f-4531-a7cd-5fe5e1c74477/addgene-plasmid-70038-sequence-220331.gbk | full | LOCUS sequence-220331- 12393 bp DNA circular SYN 25-AUG-2024
DEFINITION Bacterial expression of the S. epidermidis Cas10·Csm complex.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 12393)
AUTHORS Hatoum-Aslan A, Samai P, Maniv I, Jiang W, Marraffini LA
TITLE A ruler protein in a complex for antiviral defense determines the
length of small interfering CRISPR RNAs.
JOURNAL J Biol Chem. 2013 Sep 27;288(39):27888-97. doi:
10.1074/jbc.M113.499244. Epub 2013 Aug 9.
PUBMED 23935102
REFERENCE 2 (bases 1 to 12393)
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:
"10.1074/jbc.M113.499244"; journalName: "J Biol Chem"; date:
"2013-09-27- 27"; volume: "288"; issue: "39"; pages: "27888-97"
FEATURES Location/Qualifiers
source 1..12393
/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 493..597
/gene="bla"
/label=AmpR promoter
CDS 598..1458
/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(816..835)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 1629..2217
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2118..2137
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 2371..2388
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 2403..2542
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(2468..2487)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 2628..2650
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(2644..2835)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
primer_bind 3344..3363
/label=T7
/note="T7 promoter, forward primer"
promoter 3344..3362
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 7238..7255
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(12307..12325)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 12321..12368
/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 attaacgctt acaatttagg tggcactttt
481 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
541 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
601 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
661 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
721 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
781 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
841 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
901 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
961 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
1021 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
1081 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
1141 gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
1201 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
1261 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
1321 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
1381 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
1441 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
1501 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
1561 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
1621 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
1681 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
1741 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
1801 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
1861 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
1921 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
1981 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
2041 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
2101 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
2161 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
2221 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
2281 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
2341 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
2401 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg caatggtgca
2461 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct
2521 acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg
2581 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
2641 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc
2701 agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag
2761 tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt
2821 ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat
2881 accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt
2941 actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat
3001 cactcagggt caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca
3061 gcagcatcct gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc
3121 cagactttac gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt
3181 tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt
3241 aaggcaaccc cgccagccta gccgggtcct caacgacagg agcacgatca tgcgcacccg
3301 tggccaggac ccaacgctgc ccgagatctc gatcccgcga aattaatacg actcactata
3361 gggagaccac aacggtttcc ctctagaagt gatggcattt gttaaagtat cggatcgata
3421 cccaccccga agaaaagggg acgagaacac gtatgccgaa gtatataaat catcagtaca
3481 aaggatcgat acccaccccg aagaaaaggg gacgagaact agtaataatt gtcatttgca
3541 tacgttacat cgatgatcga tacccacccc gaagaaaagg ggacgagaac tagtacggtc
3601 gtgaacattt tttcttgatt ctctgatcga tagccacccc gaagaaaagg gggcagagtg
3661 taaatctaac aacactctaa aaaattgtag attttgaata aaatacgcat tgattaattt
3721 ctaggggatg gaaaaatgaa agatgttatt tatgtagaaa atcattactt tgttactgcg
3781 aaggaaaata gcattaaatt tagaaatgtc atagataaaa gtgaaaaatt ttatttgttt
3841 gaagaaattg aagctattat ttttgatcat tataaaagct atttttcaca taaattggta
3901 attaaatgta tagagaatga tattgcgatt atattttgtg acaaaaagca ttctccagtt
3961 acacagctca tttcttcata tggcatggtt aatcgtctta aaagaattca aagtcaattt
4021 cagttatcgg gaagaacaaa agatagaatt tggaaaaaga ttgttataaa taaaattttt
4081 aaccaaacac gatgtcttga aaataattta cataatgaaa atgttaagct gatgttaggt
4141 tttgcaaaag aagtgagttc aggagacaaa agtaataaag aagcacatgc tactcgtatt
4201 tattttaaag atttatttgg caaacaattt aaacgtggac gctataatga tgttatcaat
4261 tcaggattga attatggtta ttcaattctc agatctttta tcaacaaaga gttagctatt
4321 catggatttg aaatgagcct aggtattaaa catcagtcaa aagaaaatcc atttaattta
4381 gcagatgata tcattgaagt ttttcgtcct tttgtagata atatagtgta cgagatagtt
4441 tttaagaaaa atattgatac atttgatata aacgaaaaga aattattatt aaatgttttg
4501 tatgaaaggt gcattataga taaaaaagta gtgcgattac ttgatagtgt gaagatagtt
4561 attcaatcgc taattagatg ttatgaagaa aatacaccta cttatttatt actacctaag
4621 atgatagagg tgggaaacta atgtatttgt tagttagttt tgatttacct cgagatacta
4681 aactagaacg tcgaattgct agtaaatatc gtcttagatt aatagaactt ggatttagta
4741 tgaagcaatt tagtttatac gagagatatg tcagtaatgt tcaaaagaaa gataaaatct
4801 tagaaatatt aaagcaggaa atacctgata ctggaagtat taccttatac atattgcctg
4861 atgaagtgaa tagtaatcag attactatat taggaaaaga agtaaagctt gctgtcaaca
4921 aagaaccaaa actaatcttt atataaatgg aggttaaaat gaataaaaaa aatatattaa
4981 tgtatggctc tttattacat gatataggga aaattatata tcgaagtggt gatcatacat
5041 tttcaagagg tacgcattca aaattaggtc atcaattttt gtcccaattt tcagaattta
5101 aagacaacga agtgcttgat aacgttgctt atcatcatta caaagaactc gcaaaagcta
5161 atttagataa tgataataca gcttatatta cctatattgc ggataatatt gcgagtggta
5221 ttgatagaag agatattata gaagaaggcg atgaagaata cgaaaaacaa ctatttaatt
5281 ttgataaata tacaccgcta tatagtgtgt ttaatattgt gaattctgaa aaattgaaac
5341 aaacaaacgg gaagtttaaa ttttctaatg aaagtaatat tgaatatcct aaaactgaaa
5401 acattcaata ttcaagtgga aattatacaa cattaatgaa agatatgagt catgatttag
5461 agcacaaatt aagtattaaa gaaggtacat ttccttcatt attacaatgg acggaaagtc
5521 tatggcaata tgtacctagt tcgacaaata aaaaccaatt aattgatatt tctctttatg
5581 atcatagtcg tattacatgt gccatcgcca gttgtatatt tgattattta aatgaaaata
5641 acatacataa ttacaaagat gaattgttct caaagtatga aaataccaaa tcattttatc
5701 aaaaagaagc ttttttacta cttagtatgg atatgagtgg tattcaagat tttatttaca
5761 atataagcgg ttctaaagca ttaaagagtc taagatctcg tagtttttat ttagaactca
5821 tgcttgaagt aatcgttgat caattattag aaagattaga attagcacga gcaaatcttt
5881 tgtatacagg tggtggccat gcttatttat tagtgtctaa tactgataaa gtgaagaaaa
5941 aaataactca atttaataat gaattaaaaa aatggtttat gtcagaattt actacagatc
6001 tttcattatc aatggctttt gaaaaatgta gtggcgatga tttaatgaat acaagtggta
6061 attatagaac tatttggcgt aatgttagca gcaaactttc tgatattaaa gcgcataaat
6121 attccgcgga agatatatta aaattaaatc attttcattc gtatggagat cgggaatgta
6181 aagaatgttt aagaagtgac atagatatta atgatgatgg actatgtagt atatgtgaag
6241 gaattattaa tatatcaaat gatttaagag ataaatcatt ctttgtactg tcagaaactg
6301 gaaaattaaa aatgccattc aataaattta tatcggttat tgattatgaa gaggcagaaa
6361 tgttagtaca aaataataat caagttcgta tttacagtaa aaataaacca tatataggca
6421 taggaatatc aacaaattta tggatgtgtg attacgacta tgctagtcaa aatcaagata
6481 tgagagaaaa aggtattgga agttatgtag atagagaaga aggggttaag cgtttaggcg
6541 tggtacgtgc cgatatagat aatctcggtg ctacatttat atctggaatt ccagaaaaat
6601 ataattcaat ttcaagaaca gctacattgt ctcgtcaatt atcattattt tttaaatacg
6661 aattaaatca tttattagaa aattatcaaa ttactgctat atattcaggc ggtgacgatt
6721 tatttttaat cggtgcatgg gatgacatta tagaagcaag catttatata aatgacaaat
6781 ttaaagagtt tactcttgat aaactaacat tgtctgccgg ggttggaatg tttagtggta
6841 agtacccagt ttctaaaatg gcttttgaga caggacgact tgaagaagcg gctaagactg
6901 gtgaaaaaaa tcagatatct ctttggttac aagaaaaagt atataactgg gatgagttta
6961 aaaagaatat cttagaagaa aaacttctcg ttttacaaca ggggttttct caaacagatg
7021 aacacgggaa agccttcatt tataaaatgc tcgctttact gagaaataat gaagctatta
7081 atattgctcg tttagcttac ttattagcaa gaagcaagat gaatgaggat tttacgtcta
7141 aaatttttaa ttgggctcaa aacgacaaag ataaaaatca attaattaca gcgttagagt
7201 attatattta tcaaataagg gaggctgatt gagtatgcat caccatcacc atcacgctag
7261 catattagct aaaactaaaa gtggtaaaac gatagatttg acttttgcac atgaggtcgt
7321 aaaaagtaat gtaaaaaatg ttaaagatag aaaaggcaaa gaaaaacaag ttttatttaa
7381 cgggcttaca acaagcaagt taagaaattt aatggagcag gtaaatagac tttatactat
7441 tgcatttaat tcgaatgagg atcaattgaa tgaagaattc attgatgaat tagaatattt
7501 aaaaattaaa ttttattatg aagcaggacg agaaaaaagc gttgatgaat ttttaaaaaa
7561 aacattgatg tttccaatta ttgatagagt gataaaaaaa gaatcaaaaa aatttttctt
7621 agattattgt aaatactttg aagctttagt tgcatacgct aaatattatc aaaaggagga
7681 ttaatatgta ttcaaaaatt aaaatttcag gaacaattga agtagttact ggtttacaca
7741 tcggcggagg cggtgaatct agtatgattg gagcaattga ttctcctgta gttagagatt
7801 tgcaaaccaa attacctatc atacctggca gttcaatcaa aggaaaaatg agaaatttat
7861 tagcaaaaca ttttggcttg aaaatgaaac aagagagtca taaccaagat gatgaacgtg
7921 tattaagatt atttggctca agtgaaaaag gaaatatcca aagagctcgt ctacaaattt
7981 ctgatgcatt cttttctgaa aagacaaaag agcattttgc gcaaaatgat attgcctata
8041 cagaaacgaa atttgagaat acaattaatc gtttaactgc agttgcaaac ccaagacaaa
8101 ttgaaagagt aacaagagga tctgagtttg actttgtatt tatttacaat gtcgatgaag
8161 agtcgcaagt tgaggatgat tttgagaata ttgaaaaagc gattcactta ttagagaatg
8221 actatcttgg tggcggcgga actagaggta acggacgtat tcaatttaaa gatacaaata
8281 tcgagacagt tgttggagaa tacgatagta caaatcttaa aattaagtag gtgaatgaca
8341 ttggcaacaa aagtatttaa actttctttt aagactcctg ttcattttgg aaaaaaacgg
8401 ttgtcagatg gtgaaatgac aatcactgct gatactttgt ttagtgcttt atttattgaa
8461 acacttcaat tgggtaaaga taccgattgg ttattaaatg atttaatcat tagtgataca
8521 tttccttatg agaatgagct ttattatctt cctaaacctt tgataaaaat tgactctaaa
8581 gaagaagata accataaagc atttaaaaag ttaaaatatg ttccggttca tcactataat
8641 caatatttaa atggagagtt aagcgctgaa gatgcgacag atttaaatga tatttttaat
8701 attgggtatt tttctctaca aacaaaggtt tcattaatag cacaagaaac tgattcaagt
8761 gctgatagtg aaccttattc agtgggaaca tttacttttg aacctgaagc gggtttatat
8821 tttattgcaa aaggatcaga agaaacgctt gaccatttaa ataatattat gactgcatta
8881 cagtattcag gtttaggtgg taaacgtaat gcaggatacg gacaatttga atatgaaata
8941 ataaataatc aacaactatc taagttactg aatcaaaatg gaaaacattc tattctttta
9001 tcaacggcaa tggctaaaaa agaagagata gagagtgctt taaaagaggc gagatacatt
9061 ttaactaaac gttctggttt cgtacaatca acgaattatt ctgaaatgct agttaaaaaa
9121 agtgatttct atagcttttc ttcgggttca gtttttaaaa atatctttaa tggtgatatt
9181 tttaatgttg gccataatgg taaacaccca gtctatcgct atgcaaaacc tttatggttg
9241 gaggtataag tatgacaata aaaaattatg aagtcgttat taaaacttta ggtccaattc
9301 atattggtag tggtcaagtt atgaagaagc aagattacat ttatgacttt tataattcta
9361 aagtttatat gattaatgga aataaactag ttaaattttt aaaaagaaaa aatttacttt
9421 atacatatca aaactttttg aggtacccac caaaaaatcc aagagaaaat ggacttaaag
9481 actatttaga cgctcaaaat gttaagcaaa gtgaatggga agcatttgtg agttattctg
9541 aaaaggtcaa tcaaggtaag aaatatggta acacacgtcc taaaccgcta aatgatttac
9601 acttaatggt aagagacggt caaaataaag tgtatcttcc aggtagttca atcaaaggtg
9661 ctatcaaaac aactctcgtg tcaaaatata ataatgaaaa aaacaaagac atttatagca
9721 aaattaaagt cagcgattca aaacctattg atgaaagtaa tttagcgatt tatcaaaaaa
9781 tagacattaa taaaagtgaa aaatcaatgc ctttatatag agagtgcata gatgtaaata
9841 ccgaaataaa atttaagtta acaattgaag atgaaattta ttctattaat gaaattgaac
9901 aaagtatcca agatttttac aaaaactatt atgataaatg gttagtcgga ttcaaagaaa
9961 caaaaggtgg aagacgattt gcattagagg gcggtatacc agatgtccta aatcagaata
10021 ttttgttctt aggtgctggg acaggatttg ttagtaaaac aacacactat caattaaaaa
10081 atcgaaaaca agctaaacaa gattcttttg agattttaac taaaaaattc cgaggaactt
10141 atgggaaaat gaaggaaata ccttctaacg taccagttgc tttaaaagga acaactaatc
10201 aaagtcgtca tacttcatat cagcaaggaa tgtgtaaagt gagttttcaa gagttaaata
10261 atgaggtgct ataatgaaaa tattatttag tccaataggt aattcagatc catggagaaa
10321 tgatagagat ggtgcgatgc ttcatatcgt gcgtcattat aatttagata aggttgtatt
10381 atattttact agaactattt gggaaggaaa tgaaaataga aaaggccata aaatttacga
10441 atgggaaaaa attatccaaa ctgtttctcc aaatactgaa gtagaaatca ttattgaaaa
10501 tgtagacaat gctcaagatt atgatgtctt caaggagaaa tttcataagt atttaaaaat
10561 aattgaagat agctatgaag attgtgaaat aattttgaat gtcactagtg gtacacccca
10621 aatggaatca acattatgtt tagaatatat cgtctatcct gaaaataaga agtgtgtaca
10681 agtgagcacg ccaactaagg atagtaatgc aggtatcgaa tattcaaatc ctaaagataa
10741 agtagaagaa tttgaaatag tcaatgaagt agaaaagaaa tctgaaaaac gttgtaaaga
10801 aataaacatt ttaagcttta gagaagcaat gattagatct caaattctcg gtttaataga
10861 taattatgat tatgaaggtg ctcttaattt agtaagtaat caaaaatctt ttcgcaatgg
10921 gaaattatta agaaaaaaac tactatcatt aacaaaacaa attaaaacac atgaagtttt
10981 tccagaaatt aatgagaagt acagagatga tgctttaaaa aaatcactat ttcattattt
11041 actgttaaat atgagatata atcgtcttga tgtagctgaa acgttaatta gagtaaaatc
11101 tattgctgag tttatactta aaacatacat tgagattcat tggcctactt taataattga
11161 aaaagatggt aaaccttatc taaatgatga agataattta tcttttgttt ataaatataa
11221 tctgttatta gaaaaaagaa aacaaaattt tgatgtttca agaattttgg gacttcctgc
11281 attcattgat atactcacaa ttttagaacc taattctcaa ctattaaaag aagtcaatgc
11341 agtaaacgat ataaatggtt taagaaattc catagcccat aatttagata cattaaattt
11401 ggataaaaat aaaaattata agaaaataat gttatctgtt gaagcgataa agaatatgtt
11461 acacatctca tttcctgaga tagaggaaga agactataat tattttgaag aaaaaaataa
11521 ggaatttaaa gagctattat gataaataaa attacagtag agttagactt gccagaaagt
11581 attcggtttc aatatttagg aagtgtttta catggtgtgt taatggatta tctatctgat
11641 gatattgctg accaattaca tcatgaattt gcttatagcc cattgaaaca aagaatatat
11701 cataaaaata aaaaaatcat ttgggaaatt gtatgtatgt cagataattt atttaaagag
11761 gttgttaaac tatttagttc taaaaatagt ttgcttttga aatattatca aacaaatatt
11821 gacattcaat catttcaaat tgagaagata aatgttcaga acatgatgaa ccaactgtta
11881 caagtagaag atctaagtcg ttatgtacgt cttaatatac aaacacctat gtcttttaaa
11941 tatcagaaca gttacatgat ttttcctgat gttaaacgtt tttttagaag tattatgata
12001 caatttgacg cgttttttga agaatataga atgtacgaca aagaaacatt aaattttcta
12061 gaaaagaatg ttaatattgt tgactacaaa ttgaaaagta cacgttttaa cttggaaaaa
12121 gttaaaattc cttcatttac aggagaaata gtatttaaaa ttaaaggacc cttacctttt
12181 ctacagttaa ctcatttttt attaaagttt ggcgaatttt caggttcagg tataaaaaca
12241 agcttaggta tgggaaaata tagtataatt taattaagac aaaggaagct gagttggctg
12301 ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg
12361 gttttttgct gaaaggagga actatatccg gat
//
| {
"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": "Low Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "A ruler protein in a complex for antiviral defense determines the length of small interfering CRISPR RNAs",
"Publication": "J Biol Chem. 2013 Sep 27;288(39):27888-97. doi: 10.1074/jbc.M113.499244. Epub 2013 Aug 9.",
"DOI": "10.1074/jbc.M113.499244.",
"Authors": "Hatoum-Aslan A, Samai P, Maniv I, Jiang W, Marraffini LA /jbcM",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/23935102",
"PubMed ID": "PubMed 23935102"
} | {
"Gene/Insert name": "Cas10/Csm complex",
"Species": "S. epidermidis",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "T7",
"Tag / Fusion Protein": "6xHIS on N terminus of Csm2"
} | {
"Cloning method": "Gibson Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "T7",
"3′ sequencing primer": "T7 terminal"
} | {
"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": ""
} |
pHBS1383 TDP43 RRM-GFP FYW-S | 118791 | To test the effect of sequence on TDP43 RRM-GFP droplet properties | Rajat Rohatgi | Schmidt et al Nat Commun. 2019 Oct 25;10(1):4890. doi: 10.1038/s41467-019-12740-2. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/229668/b3040788-1666-4716-bfc5-6d4c75edd2b3/addgene-plasmid-118791-sequence-229668.gbk | full | LOCUS sequence-229668- 5677 bp DNA circular SYN 26-AUG-2024
DEFINITION To test the effect of sequence on TDP43 RRM-GFP droplet properties.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5677)
AUTHORS Schmidt HB, Barreau A, Rohatgi R
TITLE Phase separation-deficient TDP43 remains functional in splicing.
JOURNAL Nat Commun. 2019 Oct 25;10(1):4890. doi: 10.1038/s41467-019-12740-2.
PUBMED 31653829
REFERENCE 2 (bases 1 to 5677)
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
Commun."; date: "2019-10-25"; volume: "10(1):4890. doi"; pages: "
10.1038/s41467-019-12740-2"
FEATURES Location/Qualifiers
source 1..5677
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 42..1019
/label=CMV IE94 promoter
/note="enhancer/promoter region of simian cytomegalovirus
major immediate early transcription unit IE94"
promoter 1054..1072
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 1054..1071
/label=SP6
/note="SP6 promoter, forward primer"
protein_bind 1127..1151
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
regulatory 1170..1179
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
protein_bind complement(2679..2703)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
polyA_signal 2748..2882
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2772..2791)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 2826..2845
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind complement(2995..3015)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(2995..3013)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(3034..3050)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3034..3050)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3047..3069)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3058..3074
/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(3082..3112)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3127..3148
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(3265..3282)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3436..4024)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3516..3535)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4195..5055)
/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 4818..4837
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5056..5160)
/gene="bla"
/label=AmpR promoter
rep_origin complement(5186..5641)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5323..5344)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 5535..5554
/label=F1ori-R
/note="F1 origin, reverse primer"
ORIGIN
1 ggcgatcggt gcgggcctct tcgctattac gccagtcgac cgccaattca atatggcgta
61 tatggactca tgccaattca atatggtgga tctggacctg tgccaattca atatggcgta
121 tatggactcg tgccaattca atatggtgga tctggacccc agccaattca atatggcgga
181 cttggcacca tgccaattca atatggcgga cctggcactg tgccaactgg ggaggggtct
241 acttggcacg gtgccaagtt tgaggagggg tcttggccct gtgccaagtc cgccatattg
301 aattggcatg gtgccaataa tggcggccat attggctata tgccaggatc aatatatagg
361 caatatccaa tatggcccta tgccaatatg gctattggcc aggttcaata ctatgtattg
421 gccctatgcc atatagtatt ccatatatgg gttttcctat tgacgtagat agcccctccc
481 aatgggcggt cccatatacc atatatgggg cttcctaata ccgcccatag ccactccccc
541 attgacgtca atggtctcta tatatggtct ttcctattga cgtcatatgg gcggtcctat
601 tgacgtatat ggcgcctccc ccattgacgt caattacggt aaatggcccg cctggctcaa
661 tgcccattga cgtcaatagg accacccacc attgacgtca atgggatggc tcattgccca
721 ttcatatccg ttctcacgcc ccctattgac gtcaatgacg gtaaatggcc cacttggcag
781 tacatcaata tctattaata gtaacttggc aagtacatta ctattggaag tacgccaggg
841 tacattggca gtactcccat tgacgtcaat ggcggtaaat ggcccgcgat ggctgccaag
901 tacatcccca ttgacgtcaa tggggagggg caatgacgca aatgggcgtt ccattgacgt
961 aaatgggcgg taggcgtgcc taatgggagg tctatataag caatgctcgt ttagggaacc
1021 gccattctgc ctggggacgt cggagcaagc ttgatttagg tgacactata gaatacaagc
1081 tacttgttct ttttgcagga tcccatcgat tcgaattcaa ggatcaacaa gtttgtacaa
1141 aaaagcaggc tggcgccggt accaccggtg ccaccatggg atccatgtct gaatatattc
1201 gggtaaccga agatgagaac gatgagccca ttgaaatacc atcggaagac gatgggacgg
1261 tgctgctctc cacggttaca gcccagtttc caggggcgtg tgggcttcgc tacaggaatc
1321 cagtgtctca gtgtatgaga ggtgtccggc tggtagaagg aattctgcat gccccagatg
1381 ctggctgggg aaatctggtg tatgttgtca actatccaaa agataacaaa agaaaaatgg
1441 atgagacaga tgcttcatca gcagtgaaag tgaaaagagc agtccagaaa acatccagta
1501 aaggagagga actatttaca ggtactgtgc ctatcaaagt cgagctggac ggcgacgtaa
1561 acggccacaa gttcagcgtg cgtggcgagg gcgagggcga tgccaccgag ggcaagctga
1621 ccctgaagtt catctgcacc accggcaagc tgccggtgcc gtggccgacc ctcgtgacca
1681 ccctgaccta cggcgtgcag tgcttcagcc gctacccgga ccacatgaag cgtcacgact
1741 tcttcaagtc cgccatgccg gaaggctacg tccaggagcg caccatcgag ttcaaggacg
1801 acggcaccta caagacccgc gcggaggtga agttcgaggg cgacaccctg gtgaaccgca
1861 tcgagctgaa gggcaacgac ttcaaggagg acggcaacat cctgggtcac aagctggagt
1921 acaaccacaa cagccacaac gtccgtatcg aagccgacaa gcagaagaac ggcatcaagg
1981 ctaacttcaa gatccgccac aacgttgagg acggcagcca gcaggaagcc gaccacaaac
2041 agcagaacac cccgatcggc gacggcccgg tgcgcctgcc ggacaaccac tatctgagca
2101 cccagaccac gctgagcaaa gacccgaacg agaagcgcga tcacatggtc ctgaaggagt
2161 tcgtgacggc agccggtatc actaaaggtg aagatgaaag tggatccaat gccgaaccta
2221 agcataatag caacagacaa ctcgaacgaa gcgggcggag tgggggcaac cccggcggat
2281 ctggtaatca aggcggaagc ggcaatagcc ggggtggggg agccggtctc ggcaataatc
2341 aaggatctaa catgggcggc ggtatgaact ctggcgcatc cagtatcaac ccggccatga
2401 tggcagcagc acaggccgcc ctgcaaagtt catggggaat gatgggtatg ttggcaagtc
2461 agcagaacca gtcaggacct tccgggaata atcaaaacca agggaacatg caacgcgaac
2521 ccaatcaagc ctcaggatca ggcaataata gcagttccgg ctcaaattcc ggcgcggcga
2581 tagggtctgg ctccgcctca aatgcgggaa gcggatcagg gagtaacgga ggatcaggct
2641 catctatgga ttcaaaatcc agcggcagtg gtatgtagac ccagctttct tgtacaaagt
2701 ggttgatcct ctcgagcctc tagaactata gtgagtcgta ttacgtagat ccagacatga
2761 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
2821 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
2881 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
2941 tttaattcgc ggccgcggcg ccaatgcatt gggcccggta cccagctttt gttcccttta
3001 gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
3061 ttatccgctc acaattccac acaacatacg agccgggagc ataaagtgta aagcctgggg
3121 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
3181 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
3241 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
3301 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
3361 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
3421 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
3481 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
3541 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
3601 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
3661 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
3721 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
3781 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
3841 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
3901 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
3961 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
4021 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
4081 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
4141 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
4201 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
4261 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
4321 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
4381 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
4441 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
4501 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
4561 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
4621 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
4681 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
4741 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
4801 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
4861 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
4921 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
4981 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
5041 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
5101 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
5161 cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg
5221 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
5281 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
5341 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
5401 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
5461 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
5521 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
5581 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
5641 tcccattcgc cattcaggct gcgcaactgt tgggaag
//
| {
"Vector backbone": "pCS2-Dest",
"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": "Phase separation-deficient TDP43 remains functional in splicing",
"Publication": "Nat Commun. 2019 Oct 25;10(1):4890. doi: 10.1038/s41467-019-12740-2.",
"DOI": "10.1038/s41467-019-12740-2.",
"Authors": "Schmidt HB, Barreau A, Rohatgi R /s---",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/31653829",
"PubMed ID": "PubMed 31653829"
} | {
"Gene/Insert name": "TARDBP mutant",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "All FYW to S in CTD of TDP43 RRM-GFP reporter",
"Entrez Gene": "TARDBP",
"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": ""
} |
|
pJ2040_N2only | 140683 | Control (“C”) lacking sg-4 | Yolanda Schaerli | Santos-Moreno et al Nat Commun. 2020 Jun 2;11(1):2746. doi: 10.1038/s41467-020-16574-1. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/305196/d9d41b45-ed66-498b-9b2f-a301ab4f4662/addgene-plasmid-140683-sequence-305196.gbk | full | LOCUS sequence-305196- 7295 bp DNA circular SYN 26-AUG-2024
DEFINITION Control ("C") lacking sg-4.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7295)
AUTHORS Santos-Moreno J, Tasiudi E, Stelling J, Schaerli Y
TITLE Multistable and dynamic CRISPRi-based synthetic circuits.
JOURNAL Nat Commun. 2020 Jun 2;11(1):2746. doi: 10.1038/s41467-020-16574-1.
PUBMED 32488086
REFERENCE 2 (bases 1 to 7295)
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
Commun."; date: "2020-06-2"; volume: "11(1):2746. doi"; pages: "
10.1038/s41467-020-16574-1"
FEATURES Location/Qualifiers
source 1..7295
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS complement(20..898)
/codon_start=1
/gene="araC"
/product="L-arabinose regulatory protein"
/label=araC
/translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
EKVNDVAVKLS"
promoter 925..1209
/gene="araBAD"
/label=araBAD promoter
/note="promoter of the L-arabinose operon of E. coli; the
araC regulatory gene is transcribed in the opposite
direction (Guzman et al., 1995)"
primer_bind 1132..1151
/label=pBAD Forward
/note="For vectors with E. coli araBAD promoter, forward
primer"
protein_bind 1318..1337
/label=Csy4 Site
/bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
/note="cleavage site for the Csy4 endoribonuclease (Cermak
et al. 2017)"
RBS 1342..1364
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 1372..2025
/codon_start=1
/product="monomeric Kusabira-Orange 2 fluorescent protein"
/label=mKO2
/translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
LKLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAHS"
promoter 2745..2779
/label=BBa_J23102
/note="Anderson constitutive promoter
(https://parts.igem.org/Promoters/Catalog/Anderson).
Measured strength relative to BBa_J223100: 0.86"
protein_bind 2929..2948
/label=Csy4 Site
/bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
/note="cleavage site for the Csy4 endoribonuclease (Cermak
et al. 2017)"
RBS 2953..2975
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 2983..3696
/codon_start=1
/product="GFP variant that folds robustly even when fused
to poorly folded proteins (Pedelacq et al., 2006)"
/label=superfolder GFP
/translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
FVTAAGITHGMDELYK"
protein_bind 4116..4135
/label=Csy4 Site
/bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
/note="cleavage site for the Csy4 endoribonuclease (Cermak
et al. 2017)"
RBS 4140..4162
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
terminator 5389..5416
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
CDS complement(5604..6419)
/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(5604..6419)
/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 6327..6346
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
promoter complement(6420..6511)
/gene="bla"
/label=AmpR promoter
rep_origin complement(6529..7164)
/direction=LEFT
/label=ColA ori
/note="Plasmids containing the ColA origin of replication
can be propagated in E. coli cells that contain additional
plasmids with compatible origins."
primer_bind complement(6674..6693)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 gggtctgatt cgttaccaat tatgacaact tgacggctac atcattcact ttttcttcac
61 aaccggcacg gaactcgctc gggctggccc cggtgcattt tttaaatacc cgcgagaaat
121 agagttgatc gtcaaaacca acattgcgac cgacggtggc gataggcatc cgggtggtgc
181 tcaaaagcag cttcgcctgg ctgatacgtt ggtcctcgcg ccagcttaag acgctaatcc
241 ctaactgctg gcggaaaaga tgtgacagac gcgacggcga caagcaaaca tgctgtgcga
301 cgctggcgat atcaaaattg ctgtctgcca ggtgatcgct gatgtactga caagcctcgc
361 gtacccgatt atccatcggt ggatggagcg actcgttaat cgcttccatg cgccgcagta
421 acaattgctc aagcagattt atcgccagca gctccgaata gcgcccttcc ccttgcccgg
481 cgttaatgat ttgcccaaac aggtcgctga aatgcggctg gtgcgcttca tccgggcgaa
541 agaaccccgt attggcaaat attgacggcc agttaagcca ttcatgccag taggcgcgcg
601 gacgaaagta aacccactgg tgataccatt cgcgagcctc cggatgacga ccgtagtgat
661 gaatctctcc tggcgggaac agcaaaatat cacccggtcg gcaaacaaat tctcgtccct
721 gatttttcac caccccctga ccgcgaatgg tgagattgag aatataacct ttcattccca
781 gcggtcggtc gataaaaaaa tcgagataac cgttggcctc aatcggcgtt aaacccgcca
841 ccagatgggc attaaacgag tatcccggca gcaggggatc attttgcgct tcagccatac
901 ttttcatact cccgccattc agagaagaaa ccaattgtcc atattgcatc agacattgcc
961 gtcactgcgt cttttactgg ctcttctcgc taaccaaacc ggtaaccccg cttattaaaa
1021 gcattctgta acaaagcggg accaaagcca tgacaaaaac gcgtaacaaa agtgtctata
1081 atcacggcag aaaagtccac attgattatt tgcacggcgt cacactttgc tatgccatag
1141 catttttatc cataagatta gcggttccta cctgacgctt tttatcgcaa ctctctactg
1201 tttctccata ccgaattcat aggatagatt ctggaaactt taccgtccga gctcaggctt
1261 accttactcg agcaataaac agttgatagg gcttctccgt tacagcctgc ggtccgggtt
1321 cactgccgta taggcagtaa ttttgtttaa ctttaagaag gagatataca tatggtttcg
1381 gttatcaaac cagagatgaa aatgcgttac tatatggatg gttcagtaaa tggtcacgaa
1441 tttactattg agggcgaggg tacgggacgc ccatacgagg ggcaccagga aatgacttta
1501 cgcgtcacaa tggctgaagg cgggcctatg ccgtttgcgt tcgatcttgt tagtcatgtc
1561 ttttgttacg gtcaccgtgt atttactaaa taccccgagg aaattccaga ctatttcaaa
1621 caagccttcc cggaaggttt gtcttgggag cgcagtttag agtttgaaga cggtggctcg
1681 gccagcgtgt cagctcatat tagtcttcgc ggcaatacat tttatcacaa gtcaaagttc
1741 accggcgtga acttccccgc agacggccca atcatgcaga atcaaagtgt tgattgggaa
1801 ccgtccacag agaagattac agcttccgat ggagtcttaa agggcgatgt aaccatgtac
1861 ttaaaattag aagggggagg gaaccataaa tgtcagatga agactaccta taaggccgca
1921 aaagagattc ttgaaatgcc cggagaccac tacattgggc atcgtttggt ccgtaagaca
1981 gaaggaaata ttactgaaca ggtcgaagac gctgtggcac acagcatgtc ccgccgtaat
2041 actgacgcca tcacaatcca cagcatcctg gattggattg aagacaactt ggagtcgccg
2101 ttgagtttag aaaaagttag tgaacgtagt ggttactcaa agtggcacct tcagcgcatg
2161 tttaagaagg aaacgggtca ttcattgggt caatatattc gttctcgcaa gatgactgaa
2221 attgcccaga aattgaaaga gtctaatgaa cctattttgt acctggcgga gcgttacggc
2281 tttgaaagtc agcaaaccct tacacgtacc ttcaagaatt actttgacgt tccaccacac
2341 aaatatcgta tgaccaacat gcagggtgag tcacgttttt tgcatccgtt gaatcattac
2401 aattcctaat aatcgctggg acgcccgcca tggttcagcc aaaaaactta agaccgccgg
2461 tcttgtccac taccttgcag taatgcggtg gacaggatcg gcggttttct tttctcttct
2521 caattcttct gacctgtaac gaataataga tagtaaagta gtctccgatt gagttttctc
2581 tgccgagtcc cacccagttc tgtgatttca gtaagttggt aattgataca ctgttgcgag
2641 aactgctgcc tggtagtaga taggttgtta ttgagtaaga aggtaaagtg aacgaaatcc
2701 ctgaaactga gactgtagaa aataagcttc agcctgcggt ccggttgaca gctagctcag
2761 tcctaggtac tgtgctagct cgctgggacg cccggggact acacttacga aactattgat
2821 tgctcagcct gcggtccggc catacatact ctaagatgtg tctcgctggg acgcccggga
2881 tccaagagat ttctacacga ttgagcactg tctcagcctg cggtccgggt tcactgccgt
2941 ataggcagta attttgttta actttaagaa ggagatatac atatgcgtaa aggcgaagaa
3001 ctgtttaccg gtgtggttcc gattctggtg gaactggacg gcgatgttaa tggtcataaa
3061 ttcagtgttc gcggcgaagg tgaaggcgat gcgacgaacg gcaaactgac cctgaaattt
3121 atctgcacca cgggtaaact gccggtcccg tggccgacgc tggtgaccac gctgacctat
3181 ggcgttcaat gttttgcgcg ttacccggat cacatgaaac agcacgactt tttcaaatcg
3241 gccatgccgg aaggctatgt gcaggaacgt acgattagct ttaaagacga tggtacgtat
3301 aaaacccgcg cggaagtgaa attcgaaggc gataccctgg ttaaccgtat cgaactgaaa
3361 ggtatcgatt tcaaagaaga cggcaatatt ctgggtcata aactggaata taacttcaat
3421 tcccacaacg tgtacatcac cgcggataaa cagaaaaacg gcattaaagc caatttcaaa
3481 atccgccata atgtggaaga tggtagcgtt cagctggccg accactatca gcaaaacacg
3541 ccgattggtg atggcccggt cctgctgccg gacaatcact acctgagtac ccagtccgtg
3601 ctgtcaaaag atccgaacga aaaacgtgac cacatggtcc tgctggaatt tgtgacggct
3661 gcgggtatca cccacggcat ggacgaactg tataaaatgt cccgccgtaa tactgacgcc
3721 atcacaatcc acagcatcct ggattggatt gaagactaat aatcgctggg acgcccgcct
3781 gcaggctcgg taccaaattc cagaaaagag gcctcccgaa aggggggcct tttttcgttt
3841 tggtcctaat agataaagga taggtctggt agtgttgttc gttctcgcag gtaaatcaat
3901 aatactcagc agttccgtag acttttcagt gggacagggt agcgataaca gatagattgt
3961 aataagacac agtaggtgct cgtagttgcg tgaagagaac cgctcaggaa atccagtcag
4021 aagtattggt aatcgttgaa aactcagtcg acgcacttac tgaagacgtc ctattacact
4081 cgtcgttgga aactgaagat cagcctgcgg tccgggttca ctgccgtata ggcagtaatt
4141 ttgtttaact ttaagaagga gatatacata tgaatcagtc attcatctcg gacatcttat
4201 atgccgacat cgaatcgaag gctaaggaac ttacagtcaa ttccaacaat actgtccagc
4261 cggtcgcgct tatgcgctta ggagttttcg ttcccaaacc ttccaagagc aaaggagaaa
4321 gtaaggaaat tgacgccacc aaagccttct ctcaactgga gattgctaaa gcagagggca
4381 tggttagtaa aggagaagaa aataacatgg cactgattaa ggagaacatg cacatgaagc
4441 tgtacatgga gggcaccgtg aacaaccacc acttcaagtg cacatccgag ggcgaaggca
4501 agccctacga gggcacccag accatgagaa tcaaggccgt cgagggcggc cctctcccct
4561 tcgccttcga catcctggct accagcttca tgtacggcag caaaaccttc atcaaccaca
4621 cccagggcat ccccgacttc tttaagcagt ccttccctga gggcttcaca tgggagagag
4681 tcaccacata cgaagacggg ggcgtgctga ccgctaccca ggacaccagc ctccaggacg
4741 gctgcctcat ctacaacgtc aagatcagag gggtgaactt cccatccaac ggccctgtga
4801 tgcagaagaa aacactcggc tgggaggcct ccaccgagac cctgtacccc gctgacggcg
4861 gcctggaagg cagagccgac atggccctga agctcgtggg cgggggccac ctgatctgca
4921 acttgaagac cacatacaga tccaagaaac ccgctaagaa cctcaagatg cccggcgtct
4981 actatgtgga cagaagactg gaaagaatca aggaggccga caaagagacc tacgtcgagc
5041 agcacgaggt ggctgtggcc agatactgcg acctccctag caaactgggg cacagagtct
5101 aataatcgct gggacgcccg gcggccgcgg aaacacagaa aaaagcccgc acctgacagt
5161 gcgggctttt tttttcgacc aaaggtagcg aacgacgagt cactgttgag gataaatact
5221 ttctctacta ggcgcctgtt acacaggtcc tcagcggcgc gcctttgtcg gtgaacgctc
5281 tcctgagtag gacaaatccg ccgggagcgg atttgaacgt tgtgaagcaa cggcccggag
5341 ggtggcgggc aggacgcccg ccataaactg ccaggcatca aactaagcag aaggccatcc
5401 tgacggatgg cctttttgcg tttcagatct accggtaaac cagcaataga cataagcggc
5461 tatttaacga ccctgccctg aaccgacgac aagctgacga ccgggtctcc gcaagtggca
5521 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
5581 tgtatccgct catgaattaa ttcttagaaa aactcatcga gcatcaaatg aaactgcaat
5641 ttattcatat caggattatc aataccatat ttttgaaaaa gccgtttctg taatgaagga
5701 gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc tgcgattccg
5761 actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag gttatcaagt
5821 gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagttt atgcatttct
5881 ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact cgcatcaacc
5941 aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa atacgcggtc gctgttaaaa
6001 ggacaattac aaacaggaat cgaatgcaac cggcgcagga acactgccag cgcatcaaca
6061 atattttcac ctgaatcagg atattcttct aatacctgga atgctgtttt cccggggatc
6121 gcagtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaaga
6181 ggcataaatt ccgtcagcca gtttagtctg accatctcat ctgtaacatc attggcaacg
6241 ctacctttgc catgtttcag aaacaactct ggcgcatcgg gcttcccata caatcgatag
6301 attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca
6361 tccatgttgg aatttaatcg cggcctagag caagacgttt cccgttgaat atggctcata
6421 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
6481 atatttgaat gtatttagaa aaataaacaa ataggcatgc tagcgcagaa acgtcctaga
6541 agatgccagg aggatactta gcagagagac aataaggccg gagcgaagcc gtttttccat
6601 aggctccgcc cccctgacga acatcacgaa atctgacgct caaatcagtg gtggcgaaac
6661 ccgacaggac tataaagata ccaggcgttt ccccctgatg gctccctctt gcgctctcct
6721 gttcccgtcc tgcggcgtcc gtgttgtggt ggaggcttta cccaaatcac cacgtcccgt
6781 tccgtgtaga cagttcgctc caagctgggc tgtgtgcaag aaccccccgt tcagcccgac
6841 tgctgcgcct tatccggtaa ctatcatctt gagtccaacc cggaaagaca cgacaaaacg
6901 ccactggcag cagccattgg taactgagaa ttagtggatt tagatatcga gagtcttgaa
6961 gtggtggcct aacagaggct acactgaaag gacagtattt ggtatctgcg ctccactaaa
7021 gccagttacc aggttaagca gttccccaac tgacttaacc ttcgatcaaa ccgcctcccc
7081 aggcggtttt ttcgtttaca gagcaggaga ttacgacgat cgtaaaagga tctcaagaag
7141 atcctttacg gattcccgac accatcactc tagatttcag tgcaatttat ctcttcaaat
7201 gtagcacctg aagtcagccc catacgatat aagttgtaat tctcatgtta gtcatgcccc
7261 gcgcccaccg gaaggagctg actgggttgc tccta
//
| {
"Vector backbone": "pCOLA",
"Backbone size w/o insert (bp)": "",
"Vector type": "Bacterial Expression, Synthetic Biology",
"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": "Multistable and dynamic CRISPRi-based synthetic circuits",
"Publication": "Nat Commun. 2020 Jun 2;11(1):2746. doi: 10.1038/s41467-020-16574-1.",
"DOI": "10.1038/s41467-020-16574-1.",
"Authors": "Santos-Moreno J, Tasiudi E, Stelling J, Schaerli Y /s---",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/32488086",
"PubMed ID": "PubMed 32488086"
} | {
"Gene/Insert name": "Control (“C”) lacking sg-4",
"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": ""
} |
|
TFORF0123 | 141528 | Lentiviral vector for overexpressing the OLIG2 transcription factor ORF with a unique 24-bp barcode. 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/276720/9f1c8d60-5ad4-452b-b6da-e69b4c529e3b/addgene-plasmid-141528-sequence-276720.gbk | full | LOCUS sequence-276720- 9337 bp DNA circular SYN 25-AUG-2024
DEFINITION Lentiviral vector for overexpressing the OLIG2 transcription factor
ORF with a unique 24-bp barcode. 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 9337)
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 9337)
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..9337
/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"
regulatory 4109..4118
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
misc_feature 5194..5782
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(5247..5267)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(5665..5676)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 5854..6087
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 6165..6286
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(6202..6221)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 6256..6275
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 6326..6461
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(6481..6500)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(6482..6500)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(6510..6527)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(6510..6526)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6519..6541)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 6668..7123
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(6755..6774)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 6965..6986
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 7149..7253
/gene="bla"
/label=AmpR promoter
CDS 7254..8114
/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(7472..7491)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 8285..8873
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 8774..8793
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 9027..9044
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 9161..9182
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 9197..9227
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 9235..9251
/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 9240..9262
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 9259..9275
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 9259..9275
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 9294..9314
/label=T3
/note="T3 promoter, forward primer"
promoter 9296..9314
/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 caccatggac tcggacgcca gcctggtgtc
4141 cagccgcccg tcgtcgccag agcccgatga cctttttctg ccggcccgga gtaagggcag
4201 cagcggcagc gccttcactg ggggcaccgt gtcctcgtcc accccgagtg actgcccgcc
4261 ggagctgagc gccgagctgc gcggcgctat gggctctgcg ggcgcgcatc ctggggacaa
4321 gctaggaggc agtggcttca agtcatcctc gtccagcacc tcgtcgtcta cgtcgtcggc
4381 ggctgcgtcg tccaccaaga aggacaagaa gcaaatgaca gagccggagc tgcagcagct
4441 gcgtctcaag atcaacagcc gcgagcgcaa gcgcatgcac gacctcaaca tcgccatgga
4501 tggcctccgc gaggtcatgc cgtacgcaca cggcccttcg gtgcgcaagc tttccaagat
4561 cgccacgctg ctgctggcgc gcaactacat cctcatgctc accaactcgc tggaggagat
4621 gaagcgactg gtgagcgaga tctacggggg ccaccacgct ggcttccacc cgtcggcctg
4681 cggcggcctg gcgcactccg cgcccctgcc cgccgccacc gcgcacccgg cagcagcagc
4741 gcacgccgca catcaccccg cggtgcacca ccccatcctg ccgcccgccg ccgcagcggc
4801 tgctgccgcc gctgcagccg cggctgtgtc cagcgcctct ctgcccggat ccgggctgcc
4861 gtcggtcggc tccatccgtc caccgcacgg cctactcaag tctccgtctg ctgccgcggc
4921 cgccccgctg gggggcgggg gcggcggcag tggggcgagc gggggcttcc agcactgggg
4981 cggcatgccc tgcccctgca gcatgtgcca ggtgccgccg ccgcaccacc acgtgtcggc
5041 tatgggcgcc ggcagcctgc cgcgcctcac ctccgacgcc aagtagtaat gaactagtcc
5101 gtaacttgaa agtatttcga tttcttggct ttatatatct tgtggaaagg acgacttgcc
5161 tgtatgcggg ccctggccac gcgttaagtc gacaatcaac ctctggatta caaaatttgt
5221 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
5281 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
5341 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
5401 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
5461 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
5521 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
5581 tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc
5641 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
5701 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
5761 tccctttggg ccgcctcccc gcgtcgactt taagaccaat gacttacaag gcagctgtag
5821 atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa
5881 gacaagatct gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg
5941 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
6001 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
6061 tttagtcagt gtggaaaatc tctagcagta cgtatagtag ttcatgtcat cttattattc
6121 agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg tttattgcag
6181 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
6241 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggctct
6301 agctatcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
6361 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
6421 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagggacgt acccaattcg
6481 ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg
6541 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
6601 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
6661 gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
6721 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
6781 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
6841 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
6901 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
6961 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
7021 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
7081 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg
7141 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
7201 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
7261 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
7321 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
7381 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
7441 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
7501 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
7561 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
7621 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
7681 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
7741 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
7801 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
7861 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
7921 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
7981 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
8041 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
8101 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
8161 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
8221 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
8281 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
8341 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
8401 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
8461 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
8521 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
8581 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
8641 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
8701 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
8761 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
8821 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
8881 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
8941 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
9001 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
9061 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
9121 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
9181 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
9241 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag cgcgcaatta
9301 accctcacta aagggaacaa aagctggagc tgcaagc
//
| {
"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": "OLIG2",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "1073",
"Mutation": "",
"Entrez Gene": "OLIG2",
"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": ""
} |
|
pAAV-EF1a-NCre-MBP6-WPRE | 193916 | Expresses one of the components of Cre-DOR_N6C1 (RFP-dependent Cre) | Tatsushi Onaka | Inutsuka et al Commun Biol. 2022 Sep 16;5(1):979. doi: 10.1038/s42003-022-03944-2. | partial | {
"Vector backbone": "pAAV vector",
"Backbone size w/o insert (bp)": "5338",
"Vector type": "Mammalian Expression, AAV, Cre/Lox, Synthetic Biology, Affinity Reagent/ Antibody",
"Selectable markers": ""
} | {
"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": "Nanobody-based RFP-dependent Cre recombinase for selective anterograde tracing in RFP-expressing transgenic animals",
"Publication": "Commun Biol. 2022 Sep 16;5(1):979. doi: 10.1038/s42003-022-03944-2.",
"DOI": "10.1038/s42003-022-03944-2.",
"Authors": "Inutsuka A, Maejima S, Mizoguchi H, Kaneko R, Nomura R, Takanami K, Sakamoto H, Onaka T /s---",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/36114373",
"PubMed ID": "PubMed 36114373"
} | {
"Gene/Insert name": "NCre-MBP6",
"Species": "Synthetic",
"Insert Size (bp)": "723",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "EF1a",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "BamHI (not destroyed)",
"3′ cloning site": "EcoRI (not destroyed)",
"5′ sequencing primer": "TCAAGCCTCAGACAGTGGTTC",
"3′ sequencing primer": "gcgtaaaaggagcaacatag"
} | {
"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": ""
} |
|||
pET-32a(+)-TGFB3 | 120288 | High | Produce human recombinant TGFB3 in DE3 cells | Paul Burridge | Kuo et al Stem Cell Reports. 2020 Jan 9. pii: S2213-6711(19)30446-1. doi: 10.1016/j.stemcr.2019.12.007. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/234994/119cabe1-016d-48e5-a1d0-81a23b850a96/addgene-plasmid-120288-sequence-234994.gbk | full | LOCUS sequence-234994- 6237 bp DNA circular SYN 25-AUG-2024
DEFINITION Produce human recombinant TGFB3 in DE3 cells.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6237)
AUTHORS Kuo HH, Gao X, DeKeyser JM, Fetterman KA, Pinheiro EA, Weddle CJ,
Fonoudi H, Orman MV, Romero-Tejeda M, Jouni M, Blancard M, Magdy T,
Epting CL, George AL Jr, Burridge PW
TITLE Negligible-Cost and Weekend-Free Chemically Defined Human iPSC
Culture.
JOURNAL Stem Cell Reports. 2020 Jan 9. pii: S2213-6711(19)30446-1. doi:
10.1016/j.stemcr.2019.12.007.
PUBMED 31928950
REFERENCE 2 (bases 1 to 6237)
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: "Stem Cell
Reports. 2020 Jan 9. pii: S2213-6711(19)30446-1. doi:
10.1016/j.stemcr.2019.12.007."
FEATURES Location/Qualifiers
source 1..6237
/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 493..597
/gene="bla"
/label=AmpR promoter
CDS 598..1458
/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(816..835)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 1629..2217
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2118..2137
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 2371..2388
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 2403..2542
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(2468..2487)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 2628..2650
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(2644..2835)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(3644..4726)
/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 4688..4707
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(4727..4804)
/gene="lacI"
/label=lacI promoter
primer_bind 5010..5029
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind 5117..5136
/label=T7
/note="T7 promoter, forward primer"
promoter 5117..5135
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 5136..5160
/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)."
RBS 5175..5197
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 5205..5531
/codon_start=1
/gene="trxA"
/product="E. coli thioredoxin"
/label=TrxA
/translation="MSDKIIHLTDDSFDTDVLKADGAILVDFWAEWCGPCKMIAPILDE
IADEYQGKLTVAKLNIDQNPGTAPKYGIRGIPTLLLFKNGEVAATKVGALSKGQLKEFL
DANLA"
CDS 5553..5570
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 5580..5597
/codon_start=1
/product="thrombin recognition and cleavage site"
/label=thrombin site
/translation="LVPRGS"
CDS 5604..5648
/codon_start=1
/product="affinity and epitope tag derived from pancreatic
ribonuclease A"
/label=S-Tag
/translation="KETAAAKFERQHMDS"
CDS 5664..5678
/codon_start=1
/product="enterokinase recognition and cleavage site"
/label=enterokinase site
/translation="DDDDK"
CDS 6081..6098
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(6151..6169)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 6165..6212
/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 attaacgctt acaatttagg tggcactttt
481 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
541 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
601 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
661 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
721 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
781 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
841 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
901 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
961 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
1021 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
1081 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
1141 gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
1201 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
1261 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
1321 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
1381 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
1441 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
1501 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
1561 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
1621 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
1681 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
1741 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
1801 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
1861 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
1921 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
1981 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
2041 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
2101 acgagggagt ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
2161 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
2221 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
2281 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
2341 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
2401 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg caatggtgca
2461 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct
2521 acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg
2581 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
2641 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc
2701 agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag
2761 tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt
2821 ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat
2881 accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt
2941 actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat
3001 cactcagggt caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca
3061 gcagcatcct gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc
3121 cagactttac gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt
3181 tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt
3241 aaggcaaccc cgccagccta gccgggtcct caacgacagg agcacgatca tgcgcacccg
3301 tggggccgcc atgccggcga taatggcctg cttctcgccg aaacgtttgg tggcgggacc
3361 agtgacgaag gcttgagcga gggcgtgcaa gattccgaat accgcaagcg acaggccgat
3421 catcgtcgcg ctccagcgaa agcggtcctc gccgaaaatg acccagagcg ctgccggcac
3481 ctgtcctacg agttgcatga taaagaagac agtcataagt gcggcgacga tagtcatgcc
3541 ccgcgcccac cggaaggagc tgactgggtt gaaggctctc aagggcatcg gtcgagatcc
3601 cggtgcctaa tgagtgagct aacttacatt aattgcgttg cgctcactgc ccgctttcca
3661 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
3721 tttgcgtatt gggcgccagg gtggtttttc ttttcaccag tgagacgggc aacagctgat
3781 tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca
3841 gcaggcgaaa atcctgtttg atggtggtta acggcgggat ataacatgag ctgtcttcgg
3901 tatcgtcgta tcccactacc gagatgtccg caccaacgcg cagcccggac tcggtaatgg
3961 cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc
4021 cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc
4081 gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca
4141 gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg
4201 cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga
4261 tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca
4321 cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg
4381 cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca
4441 ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg
4501 gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca
4561 gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt
4621 cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag
4681 agacaccggc atactctgcg acatcgtata acgttactgg tttcacattc accaccctga
4741 attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg
4801 tgtccgggat ctcgacgctc tcccttatgc gactcctgca ttaggaagca gcccagtagt
4861 aggttgaggc cgttgagcac cgccgccgca aggaatggtg catgcaagga gatggcgccc
4921 aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc
4981 ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc
5041 gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat cgagatcgat
5101 ctcgatcccg cgaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc
5161 cctctagaaa taattttgtt taactttaag aaggagatat acatatgagc gataaaatta
5221 ttcacctgac tgacgacagt tttgacacgg atgtactcaa agcggacggg gcgatcctcg
5281 tcgatttctg ggcagagtgg tgcggtccgt gcaaaatgat cgccccgatt ctggatgaaa
5341 tcgctgacga atatcagggc aaactgaccg ttgcaaaact gaacatcgat caaaaccctg
5401 gcactgcgcc gaaatatggc atccgtggta tcccgactct gctgctgttc aaaaacggtg
5461 aagtggcggc aaccaaagtg ggtgcactgt ctaaaggtca gttgaaagag ttcctcgacg
5521 ctaacctggc cggttctggt tctggccata tgcaccatca tcatcatcat tcttctggtc
5581 tggtgccacg cggttctggt atgaaagaaa ccgctgctgc taaattcgaa cgccagcaca
5641 tggacagccc agatctgggt accgacgacg acgacaaggc catggctgat atcggatccg
5701 cgctggatac caactattgc tttcgcaacc tggaagaaaa ctgctgcgtg cgcccgctgt
5761 atattgattt tcgccaggat ctgggctgga aatgggtgca tgaaccgaaa ggctattatg
5821 cgaacttttg cagcggcccg tgcccgtatc tgcgcagcgc ggataccacc catagcaccg
5881 tgctgggcct gtataacacc ctgaacccgg aagcgagcgc gagcccgtgc tgcgtgccgc
5941 aggatctgga accgctgacc attctgtatt atgtgggccg caccccgaaa gtggaacagc
6001 tgagcaacat ggtggtgaaa agctgcaaat gcagctgagg gaattcgagc tccgtcgaca
6061 agcttgcggc cgcactcgag caccaccacc accaccactg agatccggct gctaacaaag
6121 cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg
6181 gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata tccggat
//
| {
"Vector backbone": "pET-32a (+)",
"Backbone size w/o insert (bp)": "5900",
"Vector type": "Bacterial Expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "BL21 Star/DE3",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Negligible-Cost and Weekend-Free Chemically Defined Human iPSC Culture",
"Publication": "Stem Cell Reports. 2020 Jan 9. pii: S2213-6711(19)30446-1. doi: 10.1016/j.stemcr.2019.12.007.",
"DOI": "10.1016/j.stemcr.2019.12.007.",
"Authors": "Kuo HH, Gao X, DeKeyser JM, Fetterman KA, Pinheiro EA, Weddle CJ, Fonoudi H, Orman MV, Romero-Tejeda M, Jouni M, Blancard M, Magdy T, Epting CL, George AL Jr, Burridge PW /jstemcr",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/31928950",
"PubMed ID": "PubMed 31928950"
} | {
"Gene/Insert name": "Transforming Growth Factor-β3",
"Species": "H. sapiens (human), Synthetic",
"Insert Size (bp)": "339",
"Mutation": "Codon optimized for E. Coli production",
"Entrez Gene": "TGFB3",
"Promoter": "T7 promoter",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "BamHI (not destroyed)",
"3′ cloning site": "EcoRI (not destroyed)",
"5′ sequencing primer": "taatacgactcactatagg",
"3′ sequencing primer": "ctagcataaccccttggggcctctaaacgggtcttgaggggttttttg"
} | {
"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": ""
} |
pHR: pPGK PYL1-sfGFP-HP1A | 174132 | Expression of PYL1-sfGFP-HP1A | Stanley Qi | Gao et al Mol Cell. 2021 Aug 13. pii: S1097-2765(21)00612-2. doi: 10.1016/j.molcel.2021.07.034. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/343507/90fc716f-cc49-482e-b063-953624ae49f0/addgene-plasmid-174132-sequence-343507.gbk | full | LOCUS sequence-343507- 10843 bp DNA circular SYN 26-AUG-2024
DEFINITION Expression of PYL1-sfGFP-HP1A.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10843)
AUTHORS Gao Y, Han M, Shang S, Wang H, Qi LS
TITLE Interrogation of the dynamic properties of higher-order
heterochromatin using CRISPR-dCas9.
JOURNAL Mol Cell. 2021 Aug 13. pii: S1097-2765(21)00612-2. doi:
10.1016/j.molcel.2021.07.034.
PUBMED 34428454
REFERENCE 2 (bases 1 to 10843)
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.
2021 Aug 13. pii: S1097-2765(21)00612-2. doi:
10.1016/j.molcel.2021.07.034."
FEATURES Location/Qualifiers
source 1..10843
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(4..20)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5..21)
/label=pBluescriptKS
/note="For pBluescript vector"
LTR 111..344
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
promoter complement(369..387)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(370..387)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(677..696)
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind complement(910..929)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 1029..1051
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(1089..1107)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 1175..1279
/gene="bla"
/label=AmpR promoter
CDS 1280..2140
/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(1498..1517)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 2311..2899
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2800..2819
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 3053..3070
/label=L4440
/note="L4440 vector, forward primer"
primer_bind complement(3140..3160)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 3145..3474
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3325..3460
/label=SV40 ori
/note="SV40 origin of replication"
intron 4608..4673
/label=small t intron
/note="SV40 (simian virus 40) small t antigen intron"
CDS 4803..4823
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
polyA_signal 5248..5382
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5285..5304)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 5339..5358
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
LTR 5550..6183
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 6230..6355
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 6852..7085
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 7270..7314
/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 7599..7716
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 7781..8280
/label=PGK promoter
/note="mouse phosphoglycerate kinase 1 promoter"
primer_bind complement(7816..7839)
/label=MSCV-rev
/note="Murine stem cell virus, reverse primer"
primer_bind 8213..8232
/label=mPGK-F
/note="Mouse PGK promoter, forward primer"
CDS 8914..9624
/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="SKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGT
YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF
VTAAGITHGMDELYK"
CDS 8914..9624
/codon_start=1
/product="GFP variant that folds robustly even when fused
to poorly folded proteins (Nager et al., 2011)"
/label=superfolder GFP
/translation="SKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGT
YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF
VTAAGITHGMDELYK"
primer_bind complement(8941..8969)
/label=GFP-R
/note="GFP, reverse primer. Does NOT anneal to EGFP"
primer_bind 9564..9584
/label=GFP-F
/note="GFP, forward primer. Does NOT anneal to EGFP"
misc_feature join(10256..10843,1)
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(10309..10329)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(10727..10738)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
ORIGIN
1 catcgatacc gtcgacctcg agggaattaa ttcgagctcg gtacctttaa gaccaatgac
61 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct
121 aattcactcc caacgaagac aagatctgct ttttgcttgt actgggtctc tctggttaga
181 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
241 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
301 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagcatct agaattaatt
361 ccgtgtattc tatagtgtca cctaaatcgt atgtgtatga tacataaggt tatgtattaa
421 ttgtagccgc gttctaacga caatatgtac aagcctaatt gtgtagcatc tggcttactg
481 aagcagaccc tatcatctct ctcgtaaact gccgtcagag tcggtttggt tggacgaacc
541 ttctgagttt ctggtaacgc cgtcccgcac ccggaaatgg tcagcgaacc aatcagcagg
601 gtcatcgcta gccagatcct ctacgccgga cgcatcgtgg ccggcatcac cggcgccaca
661 ggtgcggttg ctggcgccta tatcgccgac atcaccgatg gggaagatcg ggctcgccac
721 ttcgggctca tgagcgcttg tttcggcgtg ggtatggtgg caggccccgt ggccggggga
781 ctgttgggcg ccatctcctt gcatgcacca ttccttgcgg cggcggtgct caacggcctc
841 aacctactac tgggctgctt cctaatgcag gagtcgcata agggagagcg tcgaatggtg
901 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
961 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
1021 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
1081 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
1141 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
1201 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
1261 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
1321 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
1381 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
1441 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
1501 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
1561 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
1621 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
1681 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
1741 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
1801 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
1861 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
1921 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
1981 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
2041 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
2101 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
2161 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
2221 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
2281 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
2341 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
2401 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
2461 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
2521 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
2581 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
2641 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
2701 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
2761 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
2821 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
2881 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
2941 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
3001 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
3061 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgtcggcc
3121 gattcattaa tgcagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
3181 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
3241 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
3301 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
3361 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
3421 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctt
3481 ggacacaaga caggcttgcg agatatgttt gagaatacca ctttatcccg cgtcagggag
3541 aggcagtgcg taaaaagacg cggactcatg tgaaatactg gtttttagtg cgccagatct
3601 ctataatctc gcgcaaccta ttttcccctc gaacactttt taagccgtag ataaacaggc
3661 tgggacactt cacatgagcg aaaaatacat cgtcacctgg gacatgttgc agatccatgc
3721 acgtaaactc gcaagccgac tgatgccttc tgaacaatgg aaaggcatta ttgccgtaag
3781 ccgtggcggt ctgtaccggg tgcgttactg gcgcgtgaac tgggtattcg tcatgtcgat
3841 accgtttgta tttccagcta cgatcacgac aaccagcgcg agcttaaagt gctgaaacgc
3901 gcagaaggcg atggcgaagg cttcatcgtt attgatgacc tggtggatac cggtggtact
3961 gcggttgcga ttcgtgaaat gtatccaaaa gcgcactttg tcaccatctt cgcaaaaccg
4021 gctggtcgtc cgctggttga tgactatgtt gttgatatcc cgcaagatac ctggattgaa
4081 cagccgtggg atatgggcgt cgtattcgtc ccgccaatct ccggtcgcta atcttttcaa
4141 cgcctggcac tgccgggcgt tgttcttttt aacttcaggc gggttacaat agtttccagt
4201 aagtattctg gaggctgcat ccatgacaca ggcaaacctg agcgaaaccc tgttcaaacc
4261 ccgctttaaa catcctgaaa cctcgacgct agtccgccgc tttaatcacg gcgcacaacc
4321 gcctgtgcag tcggcccttg atggtaaaac catccctcac tggtatcgca tgattaaccg
4381 tctgatgtgg atctggcgcg gcattgaccc acgcgaaatc ctcgacgtcc aggcacgtat
4441 tgtgatgagc gatgccgaac gtaccgacga tgatttatac gatacggtga ttggctaccg
4501 tggcggcaac tggatttatg agtgggcccc ggatctttgt gaaggaacct tacttctgtg
4561 gtgtgacata attggacaaa ctacctacag agatttaaag ctctaaggta aatataaaat
4621 ttttaagtgt ataatgtgtt aaactactga ttctaattgt ttgtgtattt tagattccaa
4681 cctatggaac tgatgaatgg gagcagtggt ggaatgcctt taatgaggaa aacctgtttt
4741 gctcagaaga aatgccatct agtgatgatg aggctactgc tgactctcaa cattctactc
4801 ctccaaaaaa gaagagaaag gtagaagacc ccaaggactt tccttcagaa ttgctaagtt
4861 ttttgagtca tgctgtgttt agtaatagaa ctcttgcttg ctttgctatt tacaccacaa
4921 aggaaaaagc tgcactgcta tacaagaaaa ttatggaaaa atattctgta acctttataa
4981 gtaggcataa cagttataat cataacatac tgttttttct tactccacac aggcatagag
5041 tgtctgctat taataactat gctcaaaaat tgtgtacctt tagcttttta atttgtaaag
5101 gggttaataa ggaatatttg atgtatagtg ccttgactag agatcataat cagccatacc
5161 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
5221 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
5281 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
5341 ggtttgtcca aactcatcaa tgtatcttat catgtctgga tcaactggat aactcaagct
5401 aaccaaaatc atcccaaact tcccacccca taccctatta ccactgccaa ttacctgtgg
5461 tttcatttac tctaaacctg tgattcctct gaattatttt cattttaaag aaattgtatt
5521 tgttaaatat gtactacaaa cttagtagtt ggaagggcta attcactccc aaagaagaca
5581 agatatcctt gatctgtgga tctaccacac acaaggctac ttccctgatt agcagaacta
5641 cacaccaggg ccaggggtca gatatccact gacctttgga tggtgctaca agctagtacc
5701 agttgagcca gataaggtag aagaggccaa taaaggagag aacaccagct tgttacaccc
5761 tgtgagcctg catgggatgg atgacccgga gagagaagtg ttagagtgga ggtttgacag
5821 ccgcctagca tttcatcacg tggcccgaga gctgcatccg gagtacttca agaactgctg
5881 atatcgagct tgctacaagg gactttccgc tggggacttt ccagggaggc gtggcctggg
5941 cgggactggg gagtggcgag ccctcagatc ctgcatataa gcagctgctt tttgcctgta
6001 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
6061 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
6121 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
6181 gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc tctctcgacg
6241 caggactcgg cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac
6301 gccaaaaatt ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat
6361 taagcggggg agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa
6421 aaaatataaa ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa
6481 tcctggcctg ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc
6541 ccttcagaca ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg
6601 tgtgcatcaa aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga
6661 gcaaaacaaa agtaagacca ccgcacagca agcggccggc cgctgatctt cagacctgga
6721 ggaggagata tgagggacaa ttggagaagt gaattatata aatataaagt agtaaaaatt
6781 gaaccattag gagtagcacc caccaaggca aagagaagag tggtgcagag agaaaaaaga
6841 gcagtgggaa taggagcttt gttccttggg ttcttgggag cagcaggaag cactatgggc
6901 gcagcgtcaa tgacgctgac ggtacaggcc agacaattat tgtctggtat agtgcagcag
6961 cagaacaatt tgctgagggc tattgaggcg caacagcatc tgttgcaact cacagtctgg
7021 ggcatcaagc agctccaggc aagaatcctg gctgtggaaa gatacctaaa ggatcaacag
7081 ctcctgggga tttggggttg ctctggaaaa ctcatttgca ccactgctgt gccttggaat
7141 gctagttgga gtaataaatc tctggaacag atttggaatc acacgacctg gatggagtgg
7201 gacagagaaa ttaacaatta cacaagctta atacactcct taattgaaga atcgcaaaac
7261 cagcaagaaa agaatgaaca agaattattg gaattagata aatgggcaag tttgtggaat
7321 tggtttaaca taacaaattg gctgtggtat ataaaattat tcataatgat agtaggaggc
7381 ttggtaggtt taagaatagt ttttgctgta ctttctatag tgaatagagt taggcaggga
7441 tattcaccat tatcgtttca gacccacctc ccaaccccga ggggacccga caggcccgaa
7501 ggaatagaag aagaaggtgg agagagagac agagacagat ccattcgatt agtgaacgga
7561 tctcgacggt atcgccaaat ggcagtattc atccacaatt ttaaaagaaa aggggggatt
7621 ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat acaaactaaa
7681 gaattacaaa aacaaattac aaaaattcaa aattttcggg tttattacag ggacagcaga
7741 gatccagttt ggatcgataa gcttgatatc gaattctacc gggtagggga ggcgcttttc
7801 ccaaggcagt ctggagcatg cgctttagca gccccgctgg gcacttggcg ctacacaagt
7861 ggcctctggc ctcgcacaca ttccacatcc accggtaggc gccaaccggc tccgttcttt
7921 ggtggcccct tcgcgccacc ttctactcct cccctagtca ggaagttccc ccccgccccg
7981 cagctcgcgt cgtgcaggac gtgacaaatg gaagtagcac gtctcactag tctcgtgcag
8041 atggacagca ccgctgagca atggaagcgg gtaggccttt ggggcagcgg ccaatagcag
8101 ctttgctcct tcgctttctg ggctcagagg ctgggaaggg gtgggtccgg gggcgggctc
8161 aggggcgggc tcaggggcgg ggcgggcgcc cgaaggtcct ccggaggccc ggcattctgc
8221 acgcttcaaa agcgcacgtc tgccgcgctg ttctcctctt cctcatctcc gggcctttcg
8281 acctgcagcc caagcttacg cgtctgcagg atatcaagct tgcggtacga ctgagtcatg
8341 ggtgggggcg cgccaactca agacgaattc acccaactct cccaatcaat cgccgagttc
8401 cacacgtacc aactcggtaa cggccgttgc tcatctctcc tagctcagcg aatccacgcg
8461 ccgccggaaa cagtatggtc cgtggtgaga cgtttcgata ggccacagat ttacaaacac
8521 ttcatcaaaa gctgtaacgt gagtgaagat ttcgagatgc gagtgggatg cacgcgcgac
8581 gtgaacgtga taagtggatt accggcgaat acgtctcgag agagattaga tctgttggac
8641 gatgatcgga gagtgactgg gtttagtata accggtggtg aacataggct gaggaattat
8701 aaatcggtta cgacggttca tagatttgag aaagaagaag aagaagaaag gatctggacc
8761 gttgttttgg aatcttatgt tgttgatgta ccggaaggta attcggagga agatacgaga
8821 ttgtttgctg atacggttat tagattgaat cttcagaaac ttgcttcgat cactgaagct
8881 atgaacgtgg atccaggtgg aggtggaagc ggtagcaaag gagaagaact tttcactgga
8941 gttgtcccaa ttcttgttga attagatggt gatgttaatg ggcacaaatt ttctgtccgt
9001 ggagagggtg aaggtgatgc tacaaacgga aaactcaccc ttaaatttat ttgcactact
9061 ggaaaactac ctgttccgtg gccaacactt gtcactactc tgacctatgg tgttcaatgc
9121 ttttcccgtt atccggatca catgaaacgg catgactttt tcaagagtgc catgcccgaa
9181 ggttatgtac aggaacgcac tatatctttc aaagatgacg ggacctacaa gacgcgtgct
9241 gaagtcaagt ttgaaggtga tacccttgtt aatcgtatcg agttaaaggg tattgatttt
9301 aaagaagatg gaaacattct tggacacaaa ctcgagtaca actttaactc acacaatgta
9361 tacatcacgg cagacaaaca aaagaatgga atcaaagcta acttcaaaat tcgccacaac
9421 gttgaagatg gttccgttca actagcagac cattatcaac aaaatactcc aattggcgat
9481 ggccctgtcc ttttaccaga caaccattac ctgtcgacac aatctgtcct ttcgaaagat
9541 cccaacgaaa agcgtgacca catggtcctt cttgagtttg taactgctgc tgggattaca
9601 catggcatgg atgagctcta caaaggtgga ggtcggaccg gatccggaaa gaaaaccaag
9661 cggacagctg acagttcttc ttcagaggat gaggaggagt atgtcgtgga gaaggtgcta
9721 gacaggcgcg tggttaaggg acaagtggaa tatctactga agtggaaagg cttttctgag
9781 gagcacaata cttgggaacc tgagaaaaac ttggattgcc ctgagctaat ttctgaattt
9841 atgaaaaagt ataagaagat gaaggagggt gaaaataata aacccaggga gaagtcagaa
9901 agtaacaaga ggaaatccaa tttctcaaac agtgccgatg acatcaaatc taaaaaaaag
9961 agagagcaga gcaatgatat cgctcggggc tttgagagag gactggaacc agaaaagatc
10021 attggggcaa cagattcctg tggtgattta atgttcctaa tgaaatggaa agacacagat
10081 gaagctgacc tggttcttgc aaaagaagct aatgtgaaat gtccacaaat tgtgatagca
10141 ttttatgaag agagactgac atggcatgca tatcctgagg atgcggaaaa caaagagaaa
10201 gaaacagcaa agagctaacc tgcaggcatg caagcttgat atcaagctta tcgataatca
10261 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
10321 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
10381 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
10441 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
10501 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
10561 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
10621 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg
10681 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
10741 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
10801 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccg
//
| {
"Vector backbone": "pHR",
"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": "Unknown"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Interrogation of the dynamic properties of higher-order heterochromatin using CRISPR-dCas9",
"Publication": "Mol Cell. 2021 Aug 13. pii: S1097-2765(21)00612-2. doi: 10.1016/j.molcel.2021.07.034.",
"DOI": "10.1016/j.molcel.2021.07.034.",
"Authors": "Gao Y, Han M, Shang S, Wang H, Qi LS /jmolcel",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/34428454",
"PubMed ID": "PubMed 34428454"
} | {
"Gene/Insert name": "PYL1-sfGFP-HP1A",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "PGK",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Unknown",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "CATTCTGCACGCTTCAAAAG",
"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": ""
} |
|
pNIC28-BSA4-G-S18 | 127825 | Production of recombinant human ribosomal protein S18 | Ivanhoe Leung | Correddu et al Sci Rep. 2019 Jun 20;9(1):8884. doi: 10.1038/s41598-019-45323-8. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/250135/cdf117d9-0db0-4d26-aa7d-321940aadbea/addgene-plasmid-127825-sequence-250135.gbk | full | LOCUS sequence-250135- 5820 bp DNA circular SYN 26-AUG-2024
DEFINITION Production of recombinant human ribosomal protein S18.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5820)
AUTHORS Correddu D, Montano Lopez JJ, Vadakkedath PG, Lai A, Pernes JI,
Watson PR, Leung IKH
TITLE An improved method for the heterologous production of soluble human
ribosomal proteins in Escherichia coli.
JOURNAL Sci Rep. 2019 Jun 20;9(1):8884. doi: 10.1038/s41598-019-45323-8.
PUBMED 31222068
REFERENCE 2 (bases 1 to 5820)
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 Rep.";
date: "2019-06-20"; volume: "9(1):8884. doi"; pages: "
10.1038/s41598-019-45323-8"
FEATURES Location/Qualifiers
source 1..5820
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 1..20
/label=T7
/note="T7 promoter, forward primer"
promoter 1..19
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 20..44
/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)."
RBS 59..81
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 92..109
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 134..154
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQS"
CDS 685..702
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(755..773)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 769..816
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
rep_origin 853..1308
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(940..959)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 1150..1171
/label=F1ori-F
/note="F1 origin, forward primer"
CDS complement(1400..2215)
/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(1400..2215)
/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 2123..2142
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
primer_bind 2207..2226
/label=pENTR-R
/note="pENTR vectors, reverse primer"
rep_origin 2337..2925
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2826..2845
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 3079..3096
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 3111..3250
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(3176..3195)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 3336..3358
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(3352..3543)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(4352..5434)
/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 5396..5415
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(5435..5512)
/gene="lacI"
/label=lacI promoter
primer_bind 5718..5737
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
ORIGIN
1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
61 tgtttaactt taagaaggag atatacatat gcaccatcat catcatcatt cttctggtgt
121 agatctgggt accgagaacc tgtacttcca atccggcggc ggcatgtctc tagtgatccc
181 tgaaaagttc cagcatattt tgcgagtact caacaccaac atcgatgggc ggcggaaaat
241 agcctttgcc atcactgcca ttaagggtgt gggccgaaga tatgctcatg tggtgttgag
301 gaaagcagac attgacctca ccaagagggc gggagaactc actgaggatg aggtggaacg
361 tgtgatcacc attatgcaga atccacgcca gtacaagatc ccagactggt tcttgaacag
421 acagaaggat gtaaaggatg gaaaatacag ccaggtccta gccaatggtc tggacaacaa
481 gctccgtgaa gacctggagc gactgaagaa gattcgggcc catagagggc tgcgtcactt
541 ctggggcctt cgtgtccgag gccagcacac caagaccact ggccgccgtg gccgcaccgt
601 gggtgtgtcc aagaagaaat aacagtaaag gtggatacgg atccgaattc gagctccgtc
661 gacaagcttg cggccgcact cgagcaccac caccaccacc actgagatcc ggctgctaac
721 aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc
781 cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga
841 ttggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
901 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
961 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
1021 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
1081 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
1141 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
1201 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
1261 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt
1321 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
1381 tccgctcatg aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat
1441 tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa
1501 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc
1561 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga
1621 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc
1681 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac
1741 cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac
1801 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat
1861 tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag
1921 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca
1981 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac
2041 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg
2101 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca
2161 tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcataacac
2221 cccttgtatt actgtttatg taagcagaca gttttattgt tcatgaccaa aatcccttaa
2281 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
2341 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
2401 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
2461 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
2521 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
2581 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
2641 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
2701 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
2761 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
2821 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
2881 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
2941 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
3001 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
3061 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
3121 tattttctcc ttacgcatct gtgcggtatt tcacaccgca atggtgcact ctcagtacaa
3181 tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt
3241 catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
3301 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
3361 ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg
3421 aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag
3481 cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt
3541 cactgatgcc tccgtgtaag ggggatttct gttcatgggg gtaatgatac cgatgaaacg
3601 agagaggatg ctcacgatac gggttactga tgatgaacat gcccggttac tggaacgttg
3661 tgagggtaaa caactggcgg tatggatgcg gcgggaccag agaaaaatca ctcagggtca
3721 atgccagcgc ttcgttaata cagatgtagg tgttccacag ggtagccagc agcatcctgc
3781 gatgcagatc cggaacataa tggtgcaggg cgctgacttc cgcgtttcca gactttacga
3841 aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca
3901 gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc taaccagtaa ggcaaccccg
3961 ccagcctagc cgggtcctca acgacaggag cacgatcatg cgcacccgtg gggccgccat
4021 gccggcgata atggcctgct tctcgccgaa acgtttggtg gcgggaccag tgacgaaggc
4081 ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca tcgtcgcgct
4141 ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct gccggcacct gtcctacgag
4201 ttgcatgata aagaagacag tcataagtgc ggcgacgata gtcatgcccc gcgcccaccg
4261 gaaggagctg actgggttga aggctctcaa gggcatcggt cgagatcccg gtgcctaatg
4321 agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
4381 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
4441 gcgccagggt ggtttttctt ttcaccagtg agacgggcaa cagctgattg cccttcaccg
4501 cctggccctg agagagttgc agcaagcggt ccacgctggt ttgccccagc aggcgaaaat
4561 cctgtttgat ggtggttaac ggcgggatat aacatgagct gtcttcggta tcgtcgtatc
4621 ccactaccga gatatccgca ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc
4681 ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc tcattcagca
4741 tttgcatggt ttgttgaaaa ccggacatgg cactccagtc gccttcccgt tccgctatcg
4801 gctgaatttg attgcgagtg agatatttat gccagccagc cagacgcaga cgcgccgaga
4861 cagaacttaa tgggcccgct aacagcgcga tttgctggtg acccaatgcg accagatgct
4921 ccacgcccag tcgcgtaccg tcttcatggg agaaaataat actgttgatg ggtgtctggt
4981 cagagacatc aagaaataac gccggaacat tagtgcaggc agcttccaca gcaatggcat
5041 cctggtcatc cagcggatag ttaatgatca gcccactgac gcgttgcgcg agaagattgt
5101 gcaccgccgc tttacaggct tcgacgccgc ttcgttctac catcgacacc accacgctgg
5161 cacccagttg atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg
5221 ccagactgga ggtggcaacg ccaatcagca acgactgttt gcccgccagt tgttgtgcca
5281 cgcggttggg aatgtaattc agctccgcca tcgccgcttc cactttttcc cgcgttttcg
5341 cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt ctgataagag acaccggcat
5401 actctgcgac atcgtataac gttactggtt tcacattcac caccctgaat tgactctctt
5461 ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg tccgggatct
5521 cgacgctctc ccttatgcga ctcctgcatt aggaagcagc ccagtagtag gttgaggccg
5581 ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg
5641 gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc gaagtggcga
5701 gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg
5761 ccggtgatgc cggccacgat gcgtccggcg tagaggatcg agatctcgat cccgcgaaat
//
| {
"Vector backbone": "pNIC28-Bsa4",
"Backbone size w/o insert (bp)": "",
"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": "An improved method for the heterologous production of soluble human ribosomal proteins in Escherichia coli",
"Publication": "Sci Rep. 2019 Jun 20;9(1):8884. doi: 10.1038/s41598-019-45323-8.",
"DOI": "10.1038/s41598-019-45323-8.",
"Authors": "Correddu D, Montano Lopez JJ, Vadakkedath PG, Lai A, Pernes JI, Watson PR, Leung IKH /s---",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/31222068",
"PubMed ID": "PubMed 31222068"
} | {
"Gene/Insert name": "RPS18",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "RPS18",
"Promoter": "T7",
"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": ""
} |
|
Gma-pCAMBIA-1301-STTM171 | 84080 | Disturb the expression of miR171 in soybean | Guiliang Tang | Yan et al Plant Cell. 2012 Feb;24(2):415-27. doi: 10.1105/tpc.111.094144. Epub 2012 Feb 17. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/174468/9f51d69c-267a-4188-8b7b-ecc7cd360214/addgene-plasmid-84080-sequence-174468.gbk | full | LOCUS sequence-174468- 14716 bp DNA circular SYN 26-AUG-2024
DEFINITION Disturb the expression of miR171 in soybean.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 14716)
AUTHORS Yan J, Gu Y, Jia X, Kang W, Pan S, Tang X, Chen X, Tang G
TITLE Effective small RNA destruction by the expression of a short tandem
target mimic in Arabidopsis.
JOURNAL Plant Cell. 2012 Feb;24(2):415-27. doi: 10.1105/tpc.111.094144. Epub
2012 Feb 17.
PUBMED 22345490
REFERENCE 2 (bases 1 to 14716)
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:
"10.1105/tpc.111.094144"; journalName: "Plant Cell"; date:
"2012-02"; volume: "24"; issue: "2"; pages: "415-27"
FEATURES Location/Qualifiers
source 1..14716
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 1..346
/label=CaMV 35S promoter
/note="strong constitutive promoter from cauliflower mosaic
virus"
polyA_signal 507..683
/label=CaMV poly(A) signal
/note="cauliflower mosaic virus polyadenylation signal"
promoter complement(1245..1263)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(1246..1263)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind 1902..1921
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
promoter complement(2024..2052)
/label=tet promoter
/note="E. coli promoter for tetracycline efflux protein
gene"
primer_bind complement(2311..2328)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(2311..2327)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2320..2342)
/label=M13/pUC Forward
/note="In lacZ gene"
promoter 2704..3049
/label=CaMV 35S promoter
/note="strong constitutive promoter from cauliflower mosaic
virus"
CDS join(3066..3080,3271..5088)
/codon_start=1
/product="beta-glucuronidase"
/label=GUS
/note="This version of the gusA gene has a 5' extension
with a catalase intron to ensure expression in plants but
not bacteria."
/translation="MVDLRNRRLVRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWES
ALQESRAIAVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGK
VWVNNQEVMEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGK
KKQSYFHDFFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSV
ELRDADQQVVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRS
VAVKGEQFLINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYP
YAEEMLDWADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQA
IKELIARDKNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCD
AHTDTISDLFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDT
LAGLHSMYTDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKK
GIFTRDRKPKSAAFLLQKRWTGMNFGEKPQQGGKQ"
intron 3081..3270
/label=cat1 intron
/note="castor bean catalase intron, modified"
CDS 5095..5112
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
terminator 5147..5399
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
misc_feature 5421..5445
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
CDS 6746..7375
/codon_start=1
/product="stability protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/label=pVS1 StaA
/translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
CDS 7804..8877
/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 8943..9137
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
primer_bind complement(9373..9395)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
misc_feature 9481..9621
/label=bom
/note="basis of mobility region from pBR322"
primer_bind 9536..9555
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(9636..9653)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(9807..10395)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(10482..11276)
/codon_start=1
/gene="aphA-3"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin"
/translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
misc_feature 11701..11725
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
polyA_signal 11803..11977
/label=CaMV poly(A) signal
/note="cauliflower mosaic virus polyadenylation signal"
CDS complement(12018..13043)
/codon_start=1
/product="hygromycin B phosphotransferase"
/label=HygR
/note="confers resistance to hygromycin"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP
ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
K"
promoter complement(13110..13786)
/label=CaMV 35S promoter (enhanced)
/note="cauliflower mosaic virus 35S promoter with a
duplicated enhancer region"
protein_bind 13977..13998
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 14013..14043
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 14051..14067
/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 14056..14078
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 14075..14091
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 14075..14091
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 14291..14310
/label=T7
/note="T7 promoter, forward primer"
promoter 14291..14309
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
1 tgagactttt caacaaaggg taatttcggg aaacctcctc ggattccatt gcccagctat
61 ctgtcacctc atcgaaagga cagtagaaaa ggaagatggc tcctacaaat gccatcattg
121 cgataaagga aaggctatca ttcaagatgc ctctgccgac agtggtccca aagatggacc
181 cccacccacg aggagcatcg tggaaaaaga agacgtccca accacgtctt caaagcaagt
241 ggattgatgt gacatctcca ctgacataag ggatgacgca cagccccact atccttcgca
301 agacccttcc tctatataag gaagttcatt tcatttggag aggacagccc aagcttgata
361 ttggcgccta ggctcaatca gttgttgttg ttatggtcta atttaaatat ggtctaaaga
421 agaagaattg agatattgac tacgcggctc aagaattcgg tacgctgaaa tcaccagtct
481 ctctctacaa atctatctct ctctattttc tccataaata atgtgtgagt agtttcccga
541 taagggaaat tagggttctt atagggtttc gctcatgtgt tgagcatata agaaaccctt
601 agtatgtatt tgtatttgta aaatacttct atcaataaaa tttctaattc ctaaaaccaa
661 aatccagtac taaaatccag atctcctaaa gtccctatag atctttgtcg tgaatataaa
721 ccagacacga gacgactaga cctggagccc agacgccgtt cgaagctaga agtaccgctt
781 aggcaggagg ccgttaggga aaagatgcta aggcagggtt ggttacgttg actcccccgt
841 aggtttggtt taaatatgat gaagtggacg gaaggaagga ggaagacaag gaaggataag
901 gttgcaggcc ctgtgcaagg taagaagatg gaaatttgat agaggtacgc tactatactt
961 atactatacg ctaagggaat gattgtattt ataccctata ccccctaata accccttatc
1021 aatttaagaa ataatccgca taagcccccg cttaaaaatt ggtatcagag ccatgaatag
1081 gtctatgacc aaaactcaag aggataaaac ctcaccaaaa tacgaaagag tccttaactc
1141 taaagataaa agatctttca agatcaaaac tagttccctc acaccggtga cggggatcgc
1201 atgcgacctc tagaccatca cattggagct tcagctgctc gagttctata gtgtcaccta
1261 aatcgtatgt gtatgataca taaggttatg tattaattgt agccgcgttc ttacgtcgac
1321 gtaccaatta cgccccgccc tgtcactcat cgcagtactg ttgtaattca ttaagcattc
1381 tgccgacatg gaagccatca caaacggcat gatgaacctg aatcgccagc ggcatcagca
1441 ccttgtcgcc ttgcgtataa tatttgccca tggtgaaaac gggggcgaag aagttgtcca
1501 tattggccac gtttaaatca aaactggtga aactcaccca gggattggct gagacgaaaa
1561 acatattctc aataaaccct ttagggaaat aggccaggtt ctcaccgtaa cacgtcacat
1621 cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg gtattcactc cagagtgatg
1681 aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg gtgaacacta tcccatatta
1741 ccagctcacc gtctttcatt gccatacgga actccggatg agcattcatc aggcgggcaa
1801 gaatgtgaat aaaggccgga taaaacttgt gcttattttt ctttacggtc tttaaaaagg
1861 ccgtaatatc cagctgaacg gtctggttat aggtacattg agcaactgac tgaaatgcct
1921 caaaatgttc tttacgatgc cattgggata tatcaacggt ggtatatcca gtgatttctt
1981 tctccatttt agcttcctta gctcctgaaa atctcgataa ctcattaaag cttatcgatg
2041 ataagctgtc aaacatgaga atttagcccg cctaatgggc gggctttttg ttgtgatccg
2101 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
2161 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt cccccctgga
2221 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
2281 cttaattaac tgcaggcatg caagcttggc actggccgtc gttttacaac gtcgtgactg
2341 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
2401 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
2461 cgaatgctag agcagcttga gcttggatca gattgtcgtt tcccgccttc agtttagctt
2521 catggagtca aagattcaaa tagaggacct aacagaactc gccgtaaaga ctggcgaaca
2581 gttcatacag agtctcttac gactcaatga caagaagaaa atcttcgtca acatggtgga
2641 gcacgacaca cttgtctact ccaaaaatat caaagataca gtctcagaag accaaagggc
2701 aattgagact tttcaacaaa gggtaatatc cggaaacctc ctcggattcc attgcccagc
2761 tatctgtcac tttattgtga agatagtgga aaaggaaggt ggctcctaca aatgccatca
2821 ttgcgataaa ggaaaggcca tcgttgaaga tgcctctgcc gacagtggtc ccaaagatgg
2881 acccccaccc acgaggagca tcgtggaaaa agaagacgtt ccaaccacgt cttcaaagca
2941 agtggattga tgtgatatct ccactgacgt aagggatgac gcacaatccc actatccttc
3001 gcaagaccct tcctctatat aaggaagttc atttcatttg gagagaacac gggggactct
3061 tgaccatggt agatctgagg gtaaatttct agtttttctc cttcattttc ttggttagga
3121 cccttttctc tttttatttt tttgagcttt gatctttctt taaactgatc tattttttaa
3181 ttgattggtt atggtgtaaa tattacatag ctttaactga taatctgatt actttatttc
3241 gtgtgtctat gatgatgatg atagttacag aaccgacgac tcgtccgtcc tgtagaaacc
3301 ccaacccgtg aaatcaaaaa actcgacggc ctgtgggcat tcagtctgga tcgcgaaaac
3361 tgtggaattg atcagcgttg gtgggaaagc gcgttacaag aaagccgggc aattgctgtg
3421 ccaggcagtt ttaacgatca gttcgccgat gcagatattc gtaattatgc gggcaacgtc
3481 tggtatcagc gcgaagtctt tataccgaaa ggttgggcag gccagcgtat cgtgctgcgt
3541 ttcgatgcgg tcactcatta cggcaaagtg tgggtcaata atcaggaagt gatggagcat
3601 cagggcggct atacgccatt tgaagccgat gtcacgccgt atgttattgc cgggaaaagt
3661 gtacgtatca ccgtttgtgt gaacaacgaa ctgaactggc agactatccc gccgggaatg
3721 gtgattaccg acgaaaacgg caagaaaaag cagtcttact tccatgattt ctttaactat
3781 gccggaatcc atcgcagcgt aatgctctac accacgccga acacctgggt ggacgatatc
3841 accgtggtga cgcatgtcgc gcaagactgt aaccacgcgt ctgttgactg gcaggtggtg
3901 gccaatggtg atgtcagcgt tgaactgcgt gatgcggatc aacaggtggt tgcaactgga
3961 caaggcacta gcgggacttt gcaagtggtg aatccgcacc tctggcaacc gggtgaaggt
4021 tatctctatg aactgtgcgt cacagccaaa agccagacag agtgtgatat ctacccgctt
4081 cgcgtcggca tccggtcagt ggcagtgaag ggcgaacagt tcctgattaa ccacaaaccg
4141 ttctacttta ctggctttgg tcgtcatgaa gatgcggact tgcgtggcaa aggattcgat
4201 aacgtgctga tggtgcacga ccacgcatta atggactgga ttggggccaa ctcctaccgt
4261 acctcgcatt acccttacgc tgaagagatg ctcgactggg cagatgaaca tggcatcgtg
4321 gtgattgatg aaactgctgc tgtcggcttt aacctctctt taggcattgg tttcgaagcg
4381 ggcaacaagc cgaaagaact gtacagcgaa gaggcagtca acggggaaac tcagcaagcg
4441 cacttacagg cgattaaaga gctgatagcg cgtgacaaaa accacccaag cgtggtgatg
4501 tggagtattg ccaacgaacc ggatacccgt ccgcaaggtg cacgggaata tttcgcgcca
4561 ctggcggaag caacgcgtaa actcgacccg acgcgtccga tcacctgcgt caatgtaatg
4621 ttctgcgacg ctcacaccga taccatcagc gatctctttg atgtgctgtg cctgaaccgt
4681 tattacggat ggtatgtcca aagcggcgat ttggaaacgg cagagaaggt actggaaaaa
4741 gaacttctgg cctggcagga gaaactgcat cagccgatta tcatcaccga atacggcgtg
4801 gatacgttag ccgggctgca ctcaatgtac accgacatgt ggagtgaaga gtatcagtgt
4861 gcatggctgg atatgtatca ccgcgtcttt gatcgcgtca gcgccgtcgt cggtgaacag
4921 gtatggaatt tcgccgattt tgcgacctcg caaggcatat tgcgcgttgg cggtaacaag
4981 aaagggatct tcactcgcga ccgcaaaccg aagtcggcgg cttttctgct gcaaaaacgc
5041 tggactggca tgaacttcgg tgaaaaaccg cagcagggag gcaaacaagc tagccaccac
5101 caccaccacc acgtgtgaat tacaggtgac cagctcgaat ttccccgatc gttcaaacat
5161 ttggcaataa agtttcttaa gattgaatcc tgttgccggt cttgcgatga ttatcatata
5221 atttctgttg aattacgtta agcatgtaat aattaacatg taatgcatga cgttatttat
5281 gagatgggtt tttatgatta gagtcccgca attatacatt taatacgcga tagaaaacaa
5341 aatatagcgc gcaaactagg ataaattatc gcgcgcggtg tcatctatgt tactagatcg
5401 ggaattaaac tatcagtgtt tgacaggata tattggcggg taaacctaag agaaaagagc
5461 gtttattaga ataatcggat atttaaaagg gcgtgaaaag gtttatccgt tcgtccattt
5521 gtatgtgcat gccaaccaca gggttcccct cgggatcaaa gtactttgat ccaacccctc
5581 cgctgctata gtgcagtcgg cttctgacgt tcagtgcagc cgtcttctga aaacgacatg
5641 tcgcacaagt cctaagttac gcgacaggct gccgccctgc ccttttcctg gcgttttctt
5701 gtcgcgtgtt ttagtcgcat aaagtagaat acttgcgact agaaccggag acattacgcc
5761 atgaacaaga gcgccgccgc tggcctgctg ggctatgccc gcgtcagcac cgacgaccag
5821 gacttgacca accaacgggc cgaactgcac gcggccggct gcaccaagct gttttccgag
5881 aagatcaccg gcaccaggcg cgaccgcccg gagctggcca ggatgcttga ccacctacgc
5941 cctggcgacg ttgtgacagt gaccaggcta gaccgcctgg cccgcagcac ccgcgaccta
6001 ctggacattg ccgagcgcat ccaggaggcc ggcgcgggcc tgcgtagcct ggcagagccg
6061 tgggccgaca ccaccacgcc ggccggccgc atggtgttga ccgtgttcgc cggcattgcc
6121 gagttcgagc gttccctaat catcgaccgc acccggagcg ggcgcgaggc cgccaaggcc
6181 cgaggcgtga agtttggccc ccgccctacc ctcaccccgg cacagatcgc gcacgcccgc
6241 gagctgatcg accaggaagg ccgcaccgtg aaagaggcgg ctgcactgct tggcgtgcat
6301 cgctcgaccc tgtaccgcgc acttgagcgc agcgaggaag tgacgcccac cgaggccagg
6361 cggcgcggtg ccttccgtga ggacgcattg accgaggccg acgccctggc ggccgccgag
6421 aatgaacgcc aagaggaaca agcatgaaac cgcaccagga cggccaggac gaaccgtttt
6481 tcattaccga agagatcgag gcggagatga tcgcggccgg gtacgtgttc gagccgcccg
6541 cgcacgtctc aaccgtgcgg ctgcatgaaa tcctggccgg tttgtctgat gccaagctgg
6601 cggcctggcc ggccagcttg gccgctgaag aaaccgagcg ccgccgtcta aaaaggtgat
6661 gtgtatttga gtaaaacagc ttgcgtcatg cggtcgctgc gtatatgatg cgatgagtaa
6721 ataaacaaat acgcaagggg aacgcatgaa ggttatcgct gtacttaacc agaaaggcgg
6781 gtcaggcaag acgaccatcg caacccatct agcccgcgcc ctgcaactcg ccggggccga
6841 tgttctgtta gtcgattccg atccccaggg cagtgcccgc gattgggcgg ccgtgcggga
6901 agatcaaccg ctaaccgttg tcggcatcga ccgcccgacg attgaccgcg acgtgaaggc
6961 catcggccgg cgcgacttcg tagtgatcga cggagcgccc caggcggcgg acttggctgt
7021 gtccgcgatc aaggcagccg acttcgtgct gattccggtg cagccaagcc cttacgacat
7081 atgggccacc gccgacctgg tggagctggt taagcagcgc attgaggtca cggatggaag
7141 gctacaagcg gcctttgtcg tgtcgcgggc gatcaaaggc acgcgcatcg gcggtgaggt
7201 tgccgaggcg ctggccgggt acgagctgcc cattcttgag tcccgtatca cgcagcgcgt
7261 gagctaccca ggcactgccg ccgccggcac aaccgttctt gaatcagaac ccgagggcga
7321 cgctgcccgc gaggtccagg cgctggccgc tgaaattaaa tcaaaactca tttgagttaa
7381 tgaggtaaag agaaaatgag caaaagcaca aacacgctaa gtgccggccg tccgagcgca
7441 cgcagcagca aggctgcaac gttggccagc ctggcagaca cgccagccat gaagcgggtc
7501 aactttcagt tgccggcgga ggatcacacc aagctgaaga tgtacgcggt acgccaaggc
7561 aagaccatta ccgagctgct atctgaatac atcgcgcagc taccagagta aatgagcaaa
7621 tgaataaatg agtagatgaa ttttagcggc taaaggaggc ggcatggaaa atcaagaaca
7681 accaggcacc gacgccgtgg aatgccccat gtgtggagga acgggcggtt ggccaggcgt
7741 aagcggctgg gttgcctgcc ggccctgcaa tggcactgga acccccaagc ccgaggaatc
7801 ggcgtgagcg gtcgcaaacc atccggcccg gtacaaatcg gcgcggcgct gggtgatgac
7861 ctggtggaga agttgaaggc cgcgcaggcc gcccagcggc aacgcatcga ggcagaagca
7921 cgccccggtg aatcgtggca agcggccgct gatcgaatcc gcaaagaatc ccggcaaccg
7981 ccggcagccg gtgcgccgtc gattaggaag ccgcccaagg gcgacgagca accagatttt
8041 ttcgttccga tgctctatga cgtgggcacc cgcgatagtc gcagcatcat ggacgtggcc
8101 gttttccgtc tgtcgaagcg tgaccgacga gctggcgagg tgatccgcta cgagcttcca
8161 gacgggcacg tagaggtttc cgcagggccg gccggcatgg ccagtgtgtg ggattacgac
8221 ctggtactga tggcggtttc ccatctaacc gaatccatga accgataccg ggaagggaag
8281 ggagacaagc ccggccgcgt gttccgtcca cacgttgcgg acgtactcaa gttctgccgg
8341 cgagccgatg gcggaaagca gaaagacgac ctggtagaaa cctgcattcg gttaaacacc
8401 acgcacgttg ccatgcagcg tacgaagaag gccaagaacg gccgcctggt gacggtatcc
8461 gagggtgaag ccttgattag ccgctacaag atcgtaaaga gcgaaaccgg gcggccggag
8521 tacatcgaga tcgagctagc tgattggatg taccgcgaga tcacagaagg caagaacccg
8581 gacgtgctga cggttcaccc cgattacttt ttgatcgatc ccggcatcgg ccgttttctc
8641 taccgcctgg cacgccgcgc cgcaggcaag gcagaagcca gatggttgtt caagacgatc
8701 tacgaacgca gtggcagcgc cggagagttc aagaagttct gtttcaccgt gcgcaagctg
8761 atcgggtcaa atgacctgcc ggagtacgat ttgaaggagg aggcggggca ggctggcccg
8821 atcctagtca tgcgctaccg caacctgatc gagggcgaag catccgccgg ttcctaatgt
8881 acggagcaga tgctagggca aattgcccta gcaggggaaa aaggtcgaaa aggtctcttt
8941 cctgtggata gcacgtacat tgggaaccca aagccgtaca ttgggaaccg gaacccgtac
9001 attgggaacc caaagccgta cattgggaac cggtcacaca tgtaagtgac tgatataaaa
9061 gagaaaaaag gcgatttttc cgcctaaaac tctttaaaac ttattaaaac tcttaaaacc
9121 cgcctggcct gtgcataact gtctggccag cgcacagccg aagagctgca aaaagcgcct
9181 acccttcggt cgctgcgctc cctacgcccc gccgcttcgc gtcggcctat cgcggccgct
9241 ggccgctcaa aaatggctgg cctacggcca ggcaatctac cagggcgcgg acaagccgcg
9301 ccgtcgccac tcgaccgccg gcgcccacat caaggcaccc tgcctcgcgc gtttcggtga
9361 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
9421 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
9481 cgcagccatg acccagtcac gtagcgatag cggagtgtat actggcttaa ctatgcggca
9541 tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta
9601 aggagaaaat accgcatcag gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
9661 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
9721 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
9781 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
9841 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
9901 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
9961 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
10021 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
10081 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
10141 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
10201 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
10261 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
10321 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
10381 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
10441 gaaaactcac gttaagggat tttggtcatg cattctaggt actaaaacaa ttcatccagt
10501 aaaatataat attttatttt ctcccaatca ggcttgatcc ccagtaagtc aaaaaatagc
10561 tcgacatact gttcttcccc gatatcctcc ctgatcgacc ggacgcagaa ggcaatgtca
10621 taccacttgt ccgccctgcc gcttctccca agatcaataa agccacttac tttgccatct
10681 ttcacaaaga tgttgctgtc tcccaggtcg ccgtgggaaa agacaagttc ctcttcgggc
10741 ttttccgtct ttaaaaaatc atacagctcg cgcggatctt taaatggagt gtcttcttcc
10801 cagttttcgc aatccacatc ggccagatcg ttattcagta agtaatccaa ttcggctaag
10861 cggctgtcta agctattcgt atagggacaa tccgatatgt cgatggagtg aaagagcctg
10921 atgcactccg catacagctc gataatcttt tcagggcttt gttcatcttc atactcttcc
10981 gagcaaagga cgccatcggc ctcactcatg agcagattgc tccagccatc atgccgttca
11041 aagtgcagga cctttggaac aggcagcttt ccttccagcc atagcatcat gtccttttcc
11101 cgttccacat cataggtggt ccctttatac cggctgtccg tcatttttaa atataggttt
11161 tcattttctc ccaccagctt atatacctta gcaggagaca ttccttccgt atcttttacg
11221 cagcggtatt tttcgatcag ttttttcaat tccggtgata ttctcatttt agccatttat
11281 tatttccttc ctcttttcta cagtatttaa agatacccca agaagctaat tataacaaga
11341 cgaactccaa ttcactgttc cttgcattct aaaaccttaa ataccagaaa acagcttttt
11401 caaagttgtt ttcaaagttg gcgtataaca tagtatcgac ggagccgatt ttgaaaccgc
11461 ggtgatcaca ggcagcaacg ctctgtcatc gttacaatca acatgctacc ctccgcgaga
11521 tcatccgtgt ttcaaacccg gcagcttagt tgccgttctt ccgaatagca tcggtaacat
11581 gagcaaagtc tgccgcctta caacggctct cccgctgacg ccgtcccgga ctgatgggct
11641 gcctgtatcg agtggtgatt ttgtgccgag ctgccggtcg gggagctgtt ggctggctgg
11701 tggcaggata tattgtggtg taaacaaatt gacgcttaga caacttaata acacattgcg
11761 gacgttttta atgtactgaa ttaacgccga attaattcgg gggatctgga ttttagtact
11821 ggattttggt tttaggaatt agaaatttta ttgatagaag tattttacaa atacaaatac
11881 atactaaggg tttcttatat gctcaacaca tgagcgaaac cctataggaa ccctaattcc
11941 cttatctggg aactactcac acattattat ggagaaactc gagcttgtcg atcgacagat
12001 cccggtcggc atctactcta tttctttgcc ctcggacgag tgctggggcg tcggtttcca
12061 ctatcggcga gtacttctac acagccatcg gtccagacgg ccgcgcttct gcgggcgatt
12121 tgtgtacgcc cgacagtccc ggctccggat cggacgattg cgtcgcatcg accctgcgcc
12181 caagctgcat catcgaaatt gccgtcaacc aagctctgat agagttggtc aagaccaatg
12241 cggagcatat acgcccggag tcgtggcgat cctgcaagct ccggatgcct ccgctcgaag
12301 tagcgcgtct gctgctccat acaagccaac cacggcctcc agaagaagat gttggcgacc
12361 tcgtattggg aatccccgaa catcgcctcg ctccagtcaa tgaccgctgt tatgcggcca
12421 ttgtccgtca ggacattgtt ggagccgaaa tccgcgtgca cgaggtgccg gacttcgggg
12481 cagtcctcgg cccaaagcat cagctcatcg agagcctgcg cgacggacgc actgacggtg
12541 tcgtccatca cagtttgcca gtgatacaca tggggatcag caatcgcgca tatgaaatca
12601 cgccatgtag tgtattgacc gattccttgc ggtccgaatg ggccgaaccc gctcgtctgg
12661 ctaagatcgg ccgcagcgat cgcatccata gcctccgcga ccggttgtag aacagcgggc
12721 agttcggttt caggcaggtc ttgcaacgtg acaccctgtg aacggcggga gatgcaatag
12781 gtcaggctct cgctaaactc cccaatgtca agcacttccg gaatcgggag cgcggccgat
12841 gcaaagtgcc gataaacata acgatctttg tagaaaccat cggcgcagct atttacccgc
12901 aggacatatc cacgccctcc tacatcgaag ctgaaagcac gagattcttc gccctccgag
12961 agctgcatca ggtcggagac gctgtcgaac ttttcgatca gaaacttctc gacagacgtc
13021 gcggtgagtt caggcttttt catatctcat tgccccccgg atctgcgaaa gctcgagaga
13081 gatagatttg tagagagaga ctggtgattt cagcgtgtcc tctccaaatg aaatgaactt
13141 ccttatatag aggaagggtc ttgcgaagga tagtgggatt gtgcgtcatc ccttacgtca
13201 gtggagatat cacatcaatc cacttgcttt gaagacgtgg ttggaacgtc ttctttttcc
13261 acgatgctcc tcgtgggtgg gggtccatct ttgggaccac tgtcggcaga ggcatcttga
13321 acgatagcct ttcctttatc gcaatgatgg catttgtagg tgccaccttc cttttctact
13381 gtccttttga tgaagtgaca gatagctggg caatggaatc cgaggaggtt tcccgatatt
13441 accctttgtt gaaaagtctc aatagccctt tggtcttctg agactgtatc tttgatattc
13501 ttggagtaga cgagagtgtc gtgctccacc atgttcacat caatccactt gctttgaaga
13561 cgtggttgga acgtcttctt tttccacgat gctcctcgtg ggtgggggtc catctttggg
13621 accactgtcg gcagaggcat cttgaacgat agcctttcct ttatcgcaat gatggcattt
13681 gtaggtgcca ccttcctttt ctactgtcct tttgatgaag tgacagatag ctgggcaatg
13741 gaatccgagg aggtttcccg atattaccct ttgttgaaaa gtctcaatag ccctttggtc
13801 ttctgagact gtatctttga tattcttgga gtagacgaga gtgtcgtgct ccaccatgtt
13861 ggcaagctgc tctagccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
13921 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
13981 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
14041 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga catgattacg
14101 aattcgagct cggtacccgg ggatcctcta gctagattaa ttaagtttgc aagcagcaga
14161 ttacgcgcag aaaaaaagga tctcaaggag atcctttgga tcacaaaaaa aaagcccgct
14221 cattaggcgg gttaaattcc cgggtactgc agccgactca ttaatgcagg ttaacctggc
14281 ttatcgaaat taatacgact cactataggg agaccggcag atctgatccc tactccaaaa
14341 atgtcaaaga tacagtctca gaagaccaaa gggctattga gacttttcaa caaagggtaa
14401 tttcgggaaa cctcctcgga ttccattgcc cagctatctg tcacttcatc gaaaggacag
14461 tggaaaagga aggtggctcc tacaaatgcc atcattgcga taaaggaaag gctatcattc
14521 aagatgcctc tgccgacagt ggtcccaaag atggaccccc acccacgagg agcatcgtgg
14581 gaaaagaaga cgttccaacc acgtcttcga agcaagtgga ttgatgtgac atctccactg
14641 acgtaaggga tgacgcacaa tcccacccct actccaaaaa tgtcaaagat actgtctcag
14701 aagaccaaag ggctat
//
| {
"Vector backbone": "pCAMBIA-1301",
"Backbone size w/o insert (bp)": "11827",
"Vector type": "Bacterial Expression, Yeast Expression, Plant Expression",
"Selectable markers": "Hygromycin"
} | {
"Bacterial Resistance(s)": "Chloramphenicol and Kanamycin, 25 & 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": "Effective small RNA destruction by the expression of a short tandem target mimic in Arabidopsis",
"Publication": "Plant Cell. 2012 Feb;24(2):415-27. doi: 10.1105/tpc.111.094144. Epub 2012 Feb 17.",
"DOI": "10.1105/tpc.111.094144.",
"Authors": "Yan J, Gu Y, Jia X, Kang W, Pan S, Tang X, Chen X, Tang G /tpc",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/22345490",
"PubMed ID": "PubMed 22345490"
} | {
"Gene/Insert name": "miR171",
"Species": "Soybean",
"Insert Size (bp)": "2846",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "2x35S",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "PacI (unknown if destroyed)",
"3′ cloning site": "PacI (unknown if destroyed)",
"5′ sequencing primer": "35S promoter",
"3′ sequencing primer": "AGGTTTAGTCGTCTCGTGTCTG"
} | {
"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": ""
} |
|
pEft-3::Cerulean-Venus::Tomm-7 | 68046 | Ubiquitously expressed mitochondrial Tomm-7 tagged wtih oxCerulean-oxVenus (dFP, or double fluorescent protein) for monitoring mitophagy. | Dana Miller | Chapin et al Aging (Albany NY). 2015 Jun 30. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/122817/6c5d9563-0818-4c09-9ba5-755bac16cf54/addgene-plasmid-68046-sequence-122817.gbk | full | LOCUS sequence-122817- 10278 bp DNA circular SYN 25-AUG-2024
DEFINITION Ubiquitously expressed mitochondrial Tomm-7 tagged wtih
oxCerulean-oxVenus (dFP, or double fluorescent protein) for
monitoring mitophagy..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10278)
AUTHORS Chapin HA, Okada M, Merz AJ, Miller DL
TITLE Tissue-specific autophagy responses to aging and stress in C.
elegans.
JOURNAL Aging (Albany NY). 2015 Jun 30.
PUBMED 26142908
REFERENCE 2 (bases 1 to 10278)
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: "Aging
(Albany NY). 2015 Jun 30."
FEATURES Location/Qualifiers
source 1..10278
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(214..233)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 333..355
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(393..411)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 479..583
/gene="bla"
/label=AmpR promoter
CDS 584..1444
/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(802..821)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 1615..2203
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2104..2123
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3839..3850)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
protein_bind 5916..5936
/gene="mutant version of attB"
/label=attB4
/bound_moiety="BP Clonase(TM)"
/note="core recombination site for the Gateway(R) BP
reaction"
promoter 5937..6536
/gene="eft-3"
/label=eft-3 promoter
/note="promoter for the C. elegans elongation factor
1-alpha gene (Mitrovich and Anderson, 2000)"
protein_bind 6545..6569
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
CDS 7400..7420
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQG"
protein_bind complement(8172..8196)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
protein_bind complement(8746..8766)
/gene="mutant version of attB"
/label=attB3
/bound_moiety="BP Clonase(TM)"
/note="core recombination site for the Gateway(R) BP
reaction"
primer_bind complement(8774..8791)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(8774..8790)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
ORIGIN
1 cacgtgacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
61 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
121 gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat tttctcctta cgcatctgtg
181 cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
241 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
301 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
361 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
421 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
481 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
541 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
601 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
661 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
721 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
781 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
841 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
901 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
961 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
1021 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
1081 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
1141 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
1201 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
1261 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
1321 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
1381 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
1441 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
1501 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
1561 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
1621 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
1681 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
1741 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
1801 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
1861 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
1921 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
1981 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
2041 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
2101 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
2161 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
2221 ctttttacgg ttcctggtct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
2281 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgatacca gcggataaca
2341 atttcacagg gccctgtcgg attatgggct cttcttttat cctctacttt ctttctagaa
2401 aatgcaaatg ttgagtcaga aattgctgaa tggagtacaa ctgcaatggg tttagcttat
2461 gtttctggaa caattactgg tggaattgtg attgaaaagt aaggttatat ttcatttcaa
2521 taaatgttac cacagtataa caaaaacaag ttacattgtt cagagtatgt ctgaatgtat
2581 tattattatt gtatttgtgc atcagtactg tagttgtact gaagaagggt ttaaaataat
2641 ctcaacgttt ttaagacgag ttagtgagaa tacggtcgca gttaggatgt ttggacatag
2701 ttttattaga taatcgtgaa ctaactcact ataaaaaccc gctaactcat tgaataagat
2761 agttttaaac gttcaaaatt tgaagacaaa attgtatgaa taagtttaca gagttggtcg
2821 acgcaagctt ctccttctct tcacatttct caataattta gctctaattg cattcgtatt
2881 cttcgcaaaa atacgtattc tgattgaccc gatgaaatac ggatgcctag ttgcattgat
2941 catctacggt tatacttatg ggtgaggttt ttcaaatata ttttgattta atcgaataat
3001 gaaatttcag aactggagtt ggtccaattt cgtggtttat atcctctgaa ctggtacctc
3061 aaaaacatcg tagtattgct caatctgtgg catattcaat gaatactgta atggtggtta
3121 tcagtacatt tacagtatta cctctatatt ccgttattgg tagttatgct ttccttattc
3181 tatattcaat tccttcattt ataagtatgc ttatattatt tcgctactta ccggaaacca
3241 aaggacgaga gattcatgat attgtgaatg aacttaaaaa aaaatgaaaa cctttgaaaa
3301 cgttcatacg gccgaaattt tacttgatcg agccaattga acttttgtta agacttacag
3361 aaaagtgaga gattttgtta caaaaaagtg agaaaaaagc gaaaatgaac aggaaaaaga
3421 agaaaaatac acccgaagta tcatataaaa attaaaaaga ttgtttgacc tggcggaact
3481 tttttgcaaa aactctgaat atccatggca catgcttcgt gcaaaacagg atccaaatga
3541 atatcaacaa gagattcttg aagacgacga gccacttgct cagagcattg cttatctcga
3601 atgagaccct tgttaaaatc atttttcaaa cattatatga cattgtcgaa atgtcctcct
3661 gattccatga tggtagcaaa ctcacttcgt ctttgagcca atttatccaa gtccttgtaa
3721 aaagtattcg aaaattgtta acggataaaa tgtttattat aatatcaaaa acaatttgtc
3781 agttgaccac tttttgatat atttgacaga aaccgggatg aattggctca aagttagggc
3841 gcccttctat tacagggttt ctgataacaa accggttatt aactcccaac aagggatgat
3901 ttcaattcat catgctcaaa ttgacccaaa ttaagttaca tgacaaattc atcgcccttt
3961 tcaactcttc ttggctcatc atctgttatt ctgttcctat tctgtctgca ccctataccc
4021 ctttgcatac tctctcgtca ttcctttctt ggatagtgtc ttcttctccc cagctctgct
4081 acttctatga cttgcgcccg ctgcttttcc gctcgttcct ctcctctctg acgtcttctt
4141 ctgcttcttt tctccttcct ttttcccttc tcaatcttcc tctttttccc atttctgtca
4201 atcattcgaa gaagaagaag aagaagaccc tcattcattt catttttttt tctgtcggtg
4261 tgtgctgctc ggttaagagt gagctctcta ttccacgtct tcttcttttt cttcttcgat
4321 tcgaatcaat cactccacca aaacgcattc gtttttggga ttcaccccgc ggttcgcaac
4381 aggtttcttt ttcaaattat tagcgttata aatagaaaaa tggtggagtt tcaataaaaa
4441 tgataattta caaagtgtat tttgattaca tgtactcaaa aaggttgaaa tttttcatac
4501 cagttttccg gaaatccatc tgatatcatt atcgtatttt tctttttaaa aatgttttca
4561 aaaaaaaaca aaatatagct ggttatttgg caccctctaa ttaccatttt cctgtcacac
4621 cacactcttt tccttcttcc acttctttcc gttttcagcc gcttccaacc aaaccgatat
4681 gaaagccgag caacaacaat cgattccacc cggctcggcg accttcccgt cgcaggtgag
4741 actcagaaaa ctagagaaac cgctcaacta actcttgata tccgatttca ttcttttctc
4801 ttttcttttt gttgaacttt tccatatttc cagatgccac ggccaccacc aagcaccgaa
4861 caaggaatca caacggaatc ggagcttgcg aagaaagctc aaatcactcc gaacgacgtt
4921 ttagcacttc cgggaatcac tcaaggtatc ttgctccctt tggatttcag tttaaaacat
4981 aaatttagga ttcttatgct ccccatctgc gaacatctat aacatcgagt tcaccaagtt
5041 ccaaatccgt gatctggaca ctgagcaagt gcttttcgag atcgccaaac cggagaacga
5101 tcaggagaat gatgagtcgc cacaggagtc ggcaagatac gtgcgttata gatttgctcc
5161 aaactttttg aaactcaaaa cggtcggagc aactgtggaa ttcaaagtag gagacatccc
5221 aatccatcat ttccgaatga tcgaacgtca cttcttcaaa gatcgccttc tgaagtgttt
5281 tgactttgaa ttcggattct gtattccgaa ttcacgaaac aactgtgaac atatctatga
5341 gttccctcaa ctctctcaac aactcagtga gtcattattc taaaaagtac aattcaaaag
5401 actaatctct ttttcagtgg acgacatgat caacaatcca aacgagactc gttctgacag
5461 cttctatttc gtcgataaca aactcgtcat gcacaacaaa gccgactact catatgatgc
5521 ataaatattt aatacaaaaa tgttctggat aattattctg tcgaatagaa aaaaaactcc
5581 aaatgtgatt aaattccaat aattcctgtc tagtttgttc cttccccttc cccttctcat
5641 gttcaatgca ttcctaagct tttcagttcc ccccttgttt ctatattttt tcgctgtcct
5701 gtcacactcg ctaaaaacac taatcacacg gaaatctgtt ttcaataaaa actccaactt
5761 taactcattt tcaatttcaa ctgaaagatt ttttcattag agaatgtcta gaactaggcc
5821 cgggctacgt aatacgactc acttaaggcc ttgactagag ggtaccagag ctcacctagg
5881 caggaacagc tatgaccatg attacgccaa gctatcaact ttgtatagaa aagttggcac
5941 ctttggtctt ttattgtcaa cttccattgg ttcttccatt gtttctgtta aattaatgaa
6001 tttttcataa aataaagaca ttatacaata taaaaatgaa gaatttattg aaaataaact
6061 gccagagaga aaaagtatgc aacactcccg ccgagagtgt ttgaaatggt gtacggtaca
6121 ttttcgtgct aggagttaga tgtgcaggca gcaacgagag ggggagagat ttttttgggc
6181 cttgtgaaat taacgtgagt tttctggtca tctgactaat catgttggtt ttttgttggt
6241 ttattttgtt tttatctttg tttttatcca gattaggaaa tttaaatttt atgaatttat
6301 aatgaggtca aacattcagt cccagcgttt ttcctgttct cactgtttag tcgaattttt
6361 attttaggct ttcaacaaat gttctaactg tcttatttgt gacctcactt tttatatttt
6421 tttaattttt aaaaatatta gaagtttcta ggataatttt ttcgactttt attctctcta
6481 ccgtccgcac tcttcttact tttaaattaa attgtttttt tttcagttgg gaaacacttt
6541 gctcacaagt ttgtacaaaa aagcaggctc aatggtgtca aagggcgaag agctgtttac
6601 tggagtcgtc ccaattctgg tggagctgga tggagatgtc aacgggcata agttttcagt
6661 gcgaggagag ggagaaggag acgctaccaa cggcaagctg acactgaaat tcatcagcac
6721 cacagggaag ctgcccgtgc cttgggtaag tttaaacata tatatactaa ctaaccctga
6781 ttatttaaat tttcagccaa ccctggtcac taccctgagt tggggcgtcc agtcatttgc
6841 ccgataccca gaccacatga agcagcatga tttctttaaa tccgctatgc ccgagggcta
6901 cgtgcaggaa cggacaattt tctttaagga cgatggaaca tacaaaacta gtaagtttaa
6961 acagttcggt actaactaac catacatatt taaattttca ggagcagagg tgaagttcga
7021 aggagacact ctggtcaaca ggatcgagct gaagggcatt gactttaaag aagatggaaa
7081 tatcctgggc cacaaactgg agtacaacgc catccatggc aacgtgtaca ttacagctga
7141 taagcagaaa aacgggatta aggcaaattt cggactgaac tgcaatgtgg aagacgggag
7201 tgtccagctg gccgatcact accagcagaa cactcctatc ggcgacgggc ccgtgctgct
7261 gcctgataat cattatctga gcacccagtc cgtcctgtct aaggacccca atgagaaaag
7321 agatcacatg gtgctgctgg agtttgtgac cgctgctggc attaccctgg ggatggacga
7381 gctgtacaag ggaccaggcg agaatttgta ttttcagggt cgtcttgtca agttccttgt
7441 tggaggacca ggcatggtgt ctaaaggcga ggaactgttc actggggtgg tgcccatcct
7501 ggtggagctg gacggcgatg tgaatgggca caagtttagc gtgcgcggag agggagaagg
7561 ggacgctacc aacggaaagc tgacactgaa actgatttcc accacaggca agctgcccgt
7621 gccttggcca acactggtga ctaccctggg atacggcctc cagagcttcg cccggtatcc
7681 agaccacatg aagcagcatg atttctttaa atcagctatg cccgagggct acgtgcagga
7741 acggacaatc ttctttaagg acgatgggac atataaaact agagccgagg tgaagtttga
7801 aggggacact ctggtgaaca ggatcgagct gaagggaatt gacttcaaag aagatggaaa
7861 tatcctgggc cacaagctgg agtacaactt caacagccat aacgtgtata tcaccgccga
7921 taagcagaaa aacggcatca aggctaattt caaaattagg cacaatgtgg aagacggcgg
7981 ggtgcagctg gctgatcatt accagcagaa cacacctatc ggggacggac ccgtgctgct
8041 gcctgataat cactacctga gctatcagtc cgtgctgtct aaggacccaa acgagaaacg
8101 ggatcatatg gtgctgaaag aatttgtgac tgccgctggg attaccctgg gaatggatga
8161 gctgtataag cacccagctt tcttgtacaa agtgggaatg aagctctccc cagctaccaa
8221 gtctttcatc ggtaagacag tcgatatttc cacatttgca atccaatggg gattcgtgcc
8281 gtttgtggtc taccttggat ttaagaaagg cgctgagcca atgccaaatg gacaggtaaa
8341 aactagaaaa agaaacattt agttttaaaa tcacgtctcc gagagagcgt aataaataaa
8401 aaatgcgtcc cataatacac aagattcaaa catttatata gataccgcat tttctttaca
8461 agtttctcgt tgtgtaaata gaacaattgc atgcgagacg ctgccaatta ttttttaatt
8521 tatttgtcaa aagtttaagt tttaccaaaa acacctatgc tttcaattca aagatttaag
8581 aattcatatt tcagatcctc cctctttccg ccatgagtct tctctgggga tagtttatct
8641 acgaaagcag ccgggtggcc gttggatcgt tctgaatctc atagttttgt tagcctccac
8701 tcattgtttt cttcttttat taataaacag ttaaagtttc cttgccaact ttattataca
8761 tagttgataa ttcactggcc gtcgttttac actcgagacg tacggtgcgc gcgatgcatt
8821 cgaagatctg cccactagtg agtcgtatta taagtgcaag taagatcagt gtttgtttcg
8881 atatcagtct gtttcgtaac ggtcttctgt ataactacaa aaaagaataa aaaaccgtat
8941 cacaactgag aaaaaagaca aaacttattg aacagtgttc ctttcaaaaa attattttgt
9001 aaaaaatcga gtttttctcc aaaataacgc acgcaatcaa aaacaagaaa actaacgtgt
9061 tctcctgggc gtggaactcc ttatcaagat taccattatt gtctgtcaga attgatattt
9121 caattatttc atatctagac aaacccgggt gtcttgcatt tattagtact gtgagtttac
9181 atacaaacga ggcttactca gataacatag agtcacctct aattagcaaa aacccatttc
9241 taattagcta tttcgaccag ttctaaagtc accgtaatta gaaatggttt ttgtagcttt
9301 tctaattaga aatgaccccg agataagtga aatatggcat tttctgggtt ttctagtaag
9361 aaatgatccc gcacgtagtg attccgcgtt acaagccaaa ttcaaattaa aaaagtatat
9421 tacacaattc aagttacatt attttcagga aacaatggcg aagagcaaag cagagattca
9481 aaaggaggaa agagacaaca gatatgaacg aagacttctt gaaaagagaa agagacatga
9541 ggtgagtaac catgataata aataatcaag taagggctat agatttataa attctggagt
9601 taacagtttc gaaaaaaagt agttttcata tgtgcgaagg aatatcaaaa ataggggaat
9661 agcctaaacg agtgagaaaa ttaaatgttt gttatttttt caaaaatgtc tgcagggccg
9721 ttatcctttc aaatgcactc tatcaaacat agatcatatc catcatatct tatccagaat
9781 ttatatcata tccaagattg agagcgtttc aaggaaaatc tcaaacaacc atacaatttt
9841 gtagatcgac aacaaggcct ccacttcttc atcgatggcc tctgcagcac ctgcaccacc
9901 aagtttgtat ttttcatatt aatataaagt gagccacgca tagtcgtttt taggattttt
9961 gccagatcga tgtcgcgcaa acagactgtt tttttttctt tgaaagccgt gcgtgacatg
10021 ggccacggat tgtaacgccg cgctggatgg ataaaaattt attcagtttt ttagccttta
10081 caaaatatgg ttcgactact ttctttattt ttacattttt ataggtgctc attgagctga
10141 gatattttgg gtataatcgg aaaatcgctg attttggtca tttttctgca ctaaagatta
10201 cacaaaaagt tcagtattta gattcattgt tgaaaattgg ggatgatcta cagatacata
10261 tcgccgtgtt caaaaaca
//
| {
"Vector backbone": "pCFJ150",
"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": "Tissue-specific autophagy responses to aging and stress in C. elegans",
"Publication": "Aging (Albany NY). 2015 Jun 30.",
"DOI": "",
"Authors": "Chapin HA, Okada M, Merz AJ, Miller DL [pii]",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/26142908",
"PubMed ID": "PubMed 26142908"
} | {
"Gene/Insert name": "pEft-3::Cerulean-Venus::Tomm-7",
"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": ""
} |
|
pGEL670 | 196818 | (Empty Backbone)LrCas9 cytosine base editing plasmid in rice | Yong Zhang | Zhong et al Nat Commun. 2023 Sep 29;14(1):6102. doi: 10.1038/s41467-023-41802-9. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/388278/c719fdb0-7953-489d-95a8-4c25cea9b982/addgene-plasmid-196818-sequence-388278.gbk | full | LOCUS sequence-388278- 17379 bp DNA circular SYN 27-AUG-2024
DEFINITION LrCas9 cytosine base editing plasmid in rice.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 17379)
AUTHORS Zhong Z, Liu G, Tang Z, Xiang S, Yang L, Huang L, He Y, Fan T, Liu
S, Zheng X, Zhang T, Qi Y, Huang J, Zhang Y
TITLE Efficient plant genome engineering using a probiotic sourced
CRISPR-Cas9 system.
JOURNAL Nat Commun. 2023 Sep 29;14(1):6102. doi: 10.1038/s41467-023-41802-9.
PUBMED 37773156
REFERENCE 2 (bases 1 to 17379)
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
Commun."; date: "2023-09-29"; volume: "14(1):6102. doi"; pages: "
10.1038/s41467-023-41802-9"
FEATURES Location/Qualifiers
source 1..17379
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 8..2004
/label=Ubi promoter
/note="maize polyubiquitin gene promoter"
CDS 6130..6150
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 6358..6423
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
CDS 7027..7047
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 7054..7305
/codon_start=1
/product="uracil-DNA glycosylase inhibitor from a Bacillus
subtilis bacteriophage (Mol et al., 1995)"
/label=UGI
/translation="MTNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHT
AYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
CDS 7306..7359
/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 7546..7566
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 7579..7599
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 7612..7632
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
terminator 7643..7891
/label=HSP terminator
/note="efficient transcription terminator from the
Arabidopsis thaliana heat shock protein 18.2 gene (Nagaya
et al., 2010)"
CDS complement(8153..8458)
/codon_start=1
/gene="ccdB"
/product="CcdB, a bacterial toxin that poisons DNA gyrase"
/label=ccdB
/note="Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains."
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
primer_bind complement(8255..8274)
/label=ccdB-fwd
/note="ccdB gene, forward primer"
protein_bind complement(8941..8965)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
misc_feature 8995..9019
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
CDS 10320..10949
/codon_start=1
/product="stability protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/label=pVS1 StaA
/translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
CDS 11378..12451
/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 12517..12711
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
primer_bind complement(12947..12969)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
misc_feature 13045..13185
/label=bom
/note="basis of mobility region from pBR322"
primer_bind 13100..13119
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(13200..13217)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(13371..13959)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(13451..13470)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(14046..14840)
/codon_start=1
/gene="aphA-3"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin"
/translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
misc_feature 15265..15289
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
polyA_signal 15367..15541
/label=CaMV poly(A) signal
/note="cauliflower mosaic virus polyadenylation signal"
promoter complement(16631..17307)
/label=CaMV 35S promoter (enhanced)
/note="cauliflower mosaic virus 35S promoter with a
duplicated enhancer region"
primer_bind complement(16674..16694)
/label=35S promoter
/note="CaMV 35S promoter, forward primer"
ORIGIN
1 ggcgcgcctg cagtgcagcg tgacccggtc gtgcccctct cttgagataa ggtgagcatt
61 gcatgtctaa gttataaaaa attaccacat attttttttg tcacacttgt ttgaagtgca
121 gtttatctat ctttatacat atatttaaac tttactctac gaataatata atctatagta
181 ctacaataat atcagtgttt tagagaatca tataaatgaa cagttagaca tggtctaaag
241 gacaattgag tattttgaca acaggactct acagttttat ctttttagtg tgcatgtgtt
301 ctcctttttt tttgcaaata gcttcaccta tataatactt catccatttt attagtacat
361 ccatttaggg tttagggtta atggttttta tagactaatt tttttagtac atctatttta
421 ttctatttta gcctctaaat taagaaaact aaaactctat tttagttttt ttatttaata
481 atttagatat aaaatagaat aaaataaagt gactaaaaat taaacaaata ccctttaaga
541 aattaaaaaa actaaggaaa catttttctt gtttcgagta gataatgcca gcctgttaaa
601 cgccgtcgac gagtctaacg gacaccaacc agcgaaccag cagcgtcgcg tcgggccaag
661 cgaagcagac ggcacggcat ctctgtcgct gcctctggac ccctctcgag agttccgctc
721 caccgttgga cttgctccgc tgtcggcatc cagaaattgc gtggcggagc ggcagacgtg
781 agccggcacg gcaggcggcc tcctcctcct ctcacggcac cggcagctac gggggattcc
841 tttcccaccg ctccttcgct ttcccttcct cgcccgccgt aataaataga caccccctcc
901 acaccctctt tccccaacct cgtgttgttc ggagcgcaca cacacacaac cagaactccc
961 ccaaatccac ccgtcggcac ctccgcttca aggtacgccg ctcgtcctcc ccccccccct
1021 ctctaccttc tcaagatcgg cgttccggtc catggttagg gcccggtagt tctacttctg
1081 ttcatgtttg tgttagatcc gtgtttgtgt tagatccgtg ctactagcgt tcgtacacgg
1141 atgcgacctg tacgtcagac acgttctgat tgctaacttg ccagtgtttc tctttgggga
1201 atcctgggat ggctctagcc gttccgcaga cgggatcgat ttcatgattt tttttgtttc
1261 gttgcatagg gtttggtttg cccttttcct ttatttcaat atatgccgtg cacttgtttg
1321 tcgggtcatc ttttcatgct tttttttgtc ttggttgtga tgatgtggtc tggttgggcg
1381 gtcgttcaag atcggagtag aattaattct gtttcaaact acctggtgga tttattaatt
1441 ttggatctgt atgtgtgtgc catacatatt catagttacg aattgaagat gatggatgga
1501 aatatcgatc taggataggt atacatgttg atgcgggttt tactgatgca tatacagaga
1561 tgctttttgt tcgcttggtt gtgatgatgt ggtgtggttg ggcggtcgtt cattcgttca
1621 agatcggagt agaatactgt ttcaaactac ctggtgtatt tattaatttt ggaactgtat
1681 gtgtgtgtca tacatcttca tagttacgag tttaagatgg atggaaatat cgatctagga
1741 taggtataca tgttgatgtg ggttttactg atgcatatac atgatggcat atgcagcatc
1801 tattcatatg ctctaacctt gagtacctat ctattataat aaacaagtat gttttataat
1861 tattttgatc ttgatatact tggatgatgg catatgcagc agctatatgt ggattttttt
1921 agccctgcct tcatacgcta tttatttgct tggtactgtt tcttttgtcg atgctcaccc
1981 tgttgtttgg tgttacttct gcagcctgca ggtagatcgc tcgtcgacat gacaaagctg
2041 aaccagccat acggcatcgg cctcgctatt ggctcaaact ctatcggctt cgcggtggtc
2101 gacgccaatt cccacctcct gcgcctgaag ggcgagacag ccatcggcgc caggctcttc
2161 cgcgagggac agtcagccgc ggatcggagg ggctctagga ccacacggag gcgcctctcc
2221 cggacaaggt ggcgcctcag cttcctgcgc gacttcttcg cgccacatat caccaagatt
2281 gaccctgatt tcttcctccg ccagaagtac tccgagatca gccctaagga caaggatcgg
2341 ttcaagtacg agaagaggct gttcaacgac cgcaccgatg ccgagttcta cgaggattac
2401 ccatcaatgt accacctgag gctccatctg atgacccaca cacataaggc cgacccgagg
2461 gagattttcc tcgccatcca ccatattctc aagtctcggg gccacttcct cacacctgga
2521 gcggccaagg atttcaatac cgacaaggtg gatctggagg acatcttccc agccctcaca
2581 gaggcgtacg cccaggtcta cccggacctg gagctgacct tcgacctcgc gaaggccgac
2641 gatttcaagg cgaagctcct ggatgagcag gccaccccgt ccgacacaca gaaggcgctg
2701 gtgaacctcc tgctctccag cgatggcgag aaggagattg tcaagaagag gaagcaggtg
2761 ctgacagagt tcgcgaaggc catcaccggc ctcaagacaa agttcaacct cgccctgggc
2821 accgaggtgg atgaggccga tgcctcaaat tggcagttct ctatgggcca gctcgacgat
2881 aagtggtcca acatcgagac atccatgaca gaccagggca ccgagatttt cgagcagatt
2941 caggagctgt accgggcgag gctgctcaac ggcattgtgc cagccggcat gtccctgagc
3001 caggccaagg tcgccgatta cggccagcac aaggaggacc tggagctgtt caagacatac
3061 ctcaagaagc tgaatgatca tgagctggcg aagaccatcc ggggcctcta cgacaggtac
3121 attaacggcg acgatgccaa gccgttcctg cgcgaggact tcgtcaaggc gctcaccaag
3181 gaggtgacag cccatccaaa cgaggtgtcc gagcagctgc tcaacaggat gggccaggcc
3241 aatttcatgc tcaagcagag gaccaaggcc aacggcgcaa tcccaattca gctccagcag
3301 cgcgagctgg atcagatcat tgcgaaccag agcaagtact acgactggct cgccgcccca
3361 aatccagtgg aggcacacag gtggaagatg ccgtaccagc tcgatgagct gctcaatttc
3421 catatcccat actacgtcgg ccctctgatt acaccgaagc agcaggcgga gtcaggcgag
3481 aacgtgttcg cctggatggt caggaaggac ccgtctggca acatcacccc atacaatttc
3541 gatgagaagg tggaccggga ggcgtcagcc aatacattca tccagaggat gaagaccaca
3601 gatacctacc tgattggcga ggacgtgctc cctaagcagt ctctgctcta ccagaagtac
3661 gaggtgctca acgagctgaa caatgtccgc atcaacaatg agtgcctggg cacagatcag
3721 aagcagcggc tcatcaggga ggtgttcgag cggcactcat ctgtgaccat taagcaggtc
3781 gcggataatc tcgtggcaca tggcgacttc gcccggaggc cggagatcag gggcctggcg
3841 gacgagaaga ggttcctctc cagcctgtcc acataccacc agctcaagga gatcctgcat
3901 gaggccattg acgatccaac caagctgctc gatatcgaga acatcattac ctggagcaca
3961 gtgttcgagg accacaccat cttcgagaca aagctggccg agattgagtg gctcgacccg
4021 aagaagatca acgagctgtc aggcattcgc taccggggat ggggacagtt ctctcgcaag
4081 ctgctcgacg gcctcaagct gggcaacggc cacacagtca ttcaggagct gatgctgagc
4141 aaccataatc tcatgcagat cctggccgat gagacactca aggagacaat gacagagctg
4201 aaccaggaca agctcaagac agacgatatc gaggatgtga ttaacgacgc gtacacctca
4261 ccatctaata agaaggccct ccggcaggtc ctgagggtgg tcgaggatat taagcatgcc
4321 gcgaatggcc aggacccttc atggctcttc atcgagacag ccgacggcac cggcacagcc
4381 ggcaagcgca cacagtctcg gcagaagcag atccagaccg tgtacgcgaa cgccgcgcag
4441 gagctgattg attccgccgt gaggggcgag ctggaggata agatcgcgga caaggcctct
4501 ttcacagacc gcctcgtgct gtacttcatg cagggcggca gggatatcta cacaggcgcc
4561 cctctgaata ttgaccagct ctcccactac gacatcgatc atattctccc gcagtcactg
4621 attaaggacg attctctgga caacagggtg ctcgtcaacg cgacaatcaa tcgcgagaag
4681 aacaatgtgt tcgcctccac actcttcgcg ggcaagatga aggcaacctg gcggaagtgg
4741 catgaggcag gcctgatttc aggacgcaag ctccggaatc tcatgctgag gccggatgag
4801 atcgacaagt tcgccaaggg attcgtggca aggcagctgg tggagacaag gcagatcatt
4861 aagctcacag agcagatcgc tgccgcccag taccctaaca ccaagatcat tgccgtgaag
4921 gcgggcctca gccaccagct gagggaggag ctggatttcc cgaagaacag ggacgtgaat
4981 cattaccacc atgcgttcga tgccttcctc gccgcaagga ttggcacata cctgctcaag
5041 cgctacccga agctggcgcc tttcttcacc tacggcgagt tcgccaaggt ggacgtgaag
5101 aagttcaggg agttcaactt catcggcgcg ctcacacacg ccaagaagaa tatcattgcg
5161 aaggataccg gcgagattgt gtgggataag gagagggaca tccgcgagct ggacaggatc
5221 tacaatttca agcgcatgct cattacccat gaggtctact tcgagacagc cgatctcttc
5281 aagcagacca tctacgccgc aaaggactcc aaggagaggg gcggcagcaa gcagctgatt
5341 cctaagaagc agggctaccc gacacaggtg tacggcggat acacccagga gtcaggcagc
5401 tacaatgcgc tcgtgcgcgt ggccgaggca gataccacag cctaccaggt catcaagatt
5461 tccgcgcaga acgcctccaa gatcgcgagc gccaatctca agagcaggga gaagggcaag
5521 cagctgctca atgagattgt ggtcaagcag ctggcgaagc gccggaagaa ctggaagcct
5581 tcagccaatt ctttcaagat cgtgattccg aggttcggca tgggcacact gttccagaac
5641 gccaagtacg gcctcttcat ggtcaactca gacacctact accgcaatta ccaggagctg
5701 tggctgtccc gggagaatca gaagctgctc aagaagctct tcagcatcaa gtacgagaag
5761 acacagatga accacgatgc cctccaggtg tacaaggcca tcattgacca ggtcgagaag
5821 ttcttcaagc tgtacgatat caatcagttc agggcgaagc tctccgacgc cattgagcgc
5881 ttcgagaagc tgccgatcaa caccgatggc aataagattg gcaagaccga gacactccgg
5941 cagatcctca ttggcctcca ggcgaacggc acaaggtcca acgtgaagaa tctgggcatc
6001 aagaccgacc tcggcctgct ccaggtcggc agcggcatca agctcgacaa ggatacccag
6061 attgtctacc agtcaccgtc tggcctgttc aagaggcgca tcccactcgc cgacctgtca
6121 agggctgatc ctaagaagaa gaggaaggtt ggaggtggcg gtacaggagg gggtgggtcc
6181 gctgagtatg tcagggcgtt gttcgacttc aatggaaacg acgaggaaga tctgcctttt
6241 aaaaagggag atattctcag gatcagagat aagccggaag aacaatggtg gaacgctgaa
6301 gactctgaag gtaagagagg tatgattctt gtcccctacg tcgagaagta ttcgggtgac
6361 tataaagacc acgatggaga ttataaggac cacgatatag attataagga tgatgatgat
6421 aagagcggaa tgaccgatgc agagtacgtc aggattcatg agaaacttga catctacacg
6481 tttaagaaac agtttttcaa caacaaaaaa tctgttagtc accgctgtta cgtgctgttc
6541 gaattgaaac gcagaggtga gaggagagcc tgcttttggg gctatgccgt caacaagccg
6601 caaagcggca cagaaagggg cattcacgcg gagatattta gcattagaaa ggtcgaggaa
6661 taccttcggg ataatcccgg gcaattcact atcaattggt actcttcatg gtccccgtgt
6721 gcagattgcg ctgaaaagat actggagtgg tataatcaag aactcagagg aaacggtcac
6781 accctcaaga tttgggcttg caagctttac tacgagaaaa atgcaaggaa ccagatcggc
6841 ctctggaact tgcgcgacaa cggcgtgggg ttgaatgtga tggtgtcgga gcattaccag
6901 tgctgccgga agatattcat tcagtcgtca cataatcaat tgaacgagaa taggtggctc
6961 gaaaaaaccc tgaagcgggc cgagaagtgg aggagtgaac tctcgataat gatccaggtt
7021 aaaggaccga agaagaagcg caaggtgggg accatgacta atctctcaga tataatcgag
7081 aaggaaacag gaaagcaact ggtcatccaa gaatcgattt tgatgcttcc cgaagaagtc
7141 gaagaagtta taggaaataa gcccgagtct gacatactgg ttcacacagc gtacgatgaa
7201 agtacggacg agaatgtcat gttgctgaca tcggacgcac ctgaatacaa gccttgggct
7261 ctggtcatac aagatagtaa cggagaaaat aagattaaaa tgcttgaagg aaggggctcc
7321 ctgctgacat gtggagatgt ggaagagaat ccaggaccat ccaggatgat agagcaagat
7381 ggttgtaccc aagtgctgcg gcccctcgga tggcgggggt acagcgccat gactggccat
7441 aatagacagt ccgctgctct catgcctcca tgcagcggat gtcagaggag gggagctagg
7501 ttcttcctgt ctagacctac ctgccccgtg ccctcaaggg ctgatcctaa gaagaagagg
7561 aaggtttcta gggcagatcc aaagaagaag cggaaggtgt ctagagctga tccgaagaag
7621 aagcgcaagg tgtgactcga gatatgaaga tgaagatgaa atatttggtg tgtcaaataa
7681 aaagcttgtg tgcttaagtt tgtgtttttt tcttggcttg ttgtgttatg aatttgtggc
7741 tttttctaat attaaatgaa tgtaagatca cattataatg aataaacaaa tgtttctata
7801 atccattgtg aatgttttgt tggatctctt ctgcagcata taactactgt atgtgctatg
7861 gtatggacta tggaatatga ttaaagataa gaggatccgc ggatcatgaa ccaacggcct
7921 ggctgtattt ggtggttgtg tagggagatg gggagaagaa aagcccgatt ctcttcgctg
7981 tgatgggctg gatgcatgcg ggggagcggg aggcccaagt acgtgcacgg tgagcggccc
8041 acagggcgag tgtgagcgcg agaggcggga ggaacagttt agtaccacat tgcccagcta
8101 actcgaacgc gaccaactta taaacccgcg cgctgtcgct tgtgtggaga ccttatattc
8161 cccagaacat caggttaatg gcgtttttga tgtcattttc gcggtggctg agatcagcca
8221 cttcttcccc gataacggaa accggcacac tggccatatc ggtggtcatc atgcgccagc
8281 tttcatcccc gatatgcacc accgggtaaa gttcacggga gactttatct gacagcagac
8341 gtgcactggc cagggggatc accatccgtc gcccgggcgt gtcaataata tcactctgta
8401 catccacaaa cagacgataa cggctctctc ttttataggt gtaaacctta aactgcattt
8461 caccagcccc tgttctcgtc agcaaaagag ccgttcattt caataaaccg ggcgacctca
8521 gccatccctt cctgattttc cgctttccag cgttcggcac gcagacgacg ggcttcattc
8581 tgcatggttg tgcttaccag accggagata ttgacatcat atatgccttg agcaactgat
8641 agctgtcgct gtcaactgtc actgtaatac gctgcttcat agcatacctc tttttgacat
8701 acttcgggta tacatatcag tatatattct tataccgcaa aaatcagcgc gcaaatacgc
8761 atactgttat ctggcttggt accggtctca gtctcaggta gatgtcagat caatcagaaa
8821 tgattgatct gacatctacg agttgagatc aaacaaagct tcagctgagt ttcaatttct
8881 gagcccatgt tgggccatac atatgccacc cgagtgcaaa tcgggtggct ttttttctag
8941 acccagcttt cttgtacaaa gttggcatta ctgcgagctc aaactatcag tgtttgacag
9001 gatatattgg cgggtaaacc taagagaaaa gagcgtttat tagaataatc ggatatttaa
9061 aagggcgtga aaaggtttat ccgttcgtcc atttgtatgt gcatgccaac cacagggttc
9121 ccctcgggat caaagtactt tgatccaacc cctccgctgc tatagtgcag tcggcttctg
9181 acgttcagtg cagccgtctt ctgaaaacga catgtcgcac aagtcctaag ttacgcgaca
9241 ggctgccgcc ctgccctttt cctggcgttt tcttgtcgcg tgttttagtc gcataaagta
9301 gaatacttgc gactagaacc ggagacatta cgccatgaac aagagcgccg ccgctggcct
9361 gctgggctat gcccgcgtca gcaccgacga ccaggacttg accaaccaac gggccgaact
9421 gcacgcggcc ggctgcacca agctgttttc cgagaagatc accggcacca ggcgcgaccg
9481 cccggagctg gccaggatgc ttgaccacct acgccctggc gacgttgtga cagtgaccag
9541 gctagaccgc ctggcccgca gcacccgcga cctactggac attgccgagc gcatccagga
9601 ggccggcgcg ggcctgcgta gcctggcaga gccgtgggcc gacaccacca cgccggccgg
9661 ccgcatggtg ttgaccgtgt tcgccggcat tgccgagttc gagcgttccc taatcatcga
9721 ccgcacccgg agcgggcgcg aggccgccaa ggcccgaggc gtgaagtttg gcccccgccc
9781 taccctcacc ccggcacaga tcgcgcacgc ccgcgagctg atcgaccagg aaggccgcac
9841 cgtgaaagag gcggctgcac tgcttggcgt gcatcgctcg accctgtacc gcgcacttga
9901 gcgcagcgag gaagtgacgc ccaccgaggc caggcggcgc ggtgccttcc gtgaggacgc
9961 attgaccgag gccgacgccc tggcggccgc cgagaatgaa cgccaagagg aacaagcatg
10021 aaaccgcacc aggacggcca ggacgaaccg tttttcatta ccgaagagat cgaggcggag
10081 atgatcgcgg ccgggtacgt gttcgagccg cccgcgcacg gctcaaccgt gcggctgcat
10141 gaaatcctgg ccggtttgtc tgatgccaag ctggcggcct ggccggccag cttggccgct
10201 gaagaaaccg agcgccgccg tctaaaaagg tgatgtgtat ttgagtaaaa cagcttgcgt
10261 catgcggtcg ctgcgtatat gatgcgatga gtaaataaac aaatacgcaa ggggaacgca
10321 tgaaggttat cgctgtactt aaccagaaag gcgggtcagg caagacgacc atcgcaaccc
10381 atctagcccg cgccctgcaa ctcgccgggg ccgatgttct gttagtcgat tccgatcccc
10441 agggcagtgc ccgcgattgg gcggccgtgc gggaagatca accgctaacc gttgtcggca
10501 tcgaccgccc gacgattgac cgcgacgtga aggccatcgg ccggcgcgac ttcgtagtga
10561 tcgacggagc gccccaggcg gcggacttgg ctgtgtccgc gatcaaggca gccgacttcg
10621 tgctgattcc ggtgcagcca agcccttacg acatatgggc caccgccgac ctggtggagc
10681 tggttaagca gcgcattgag gtcacggatg gaaggctaca agcggccttt gtcgtgtcgc
10741 gggcgatcaa aggcacgcgc atcggcggtg aggttgccga ggcgctggcc gggtacgagc
10801 tgcccattct tgagtcccgt atcacgcagc gcgtgagcta cccaggcact gccgccgccg
10861 gcacaaccgt tcttgaatca gaacccgagg gcgacgctgc ccgcgaggtc caggcgctgg
10921 ccgctgaaat taaatcaaaa ctcatttgag ttaatgaggt aaagagaaaa tgagcaaaag
10981 cacaaacacg ctaagtgccg gccgtccgag cgcacgcagc agcaaggctg caacgttggc
11041 cagcctggca gacacgccag ccatgaagcg ggtcaacttt cagttgccgg cggaggatca
11101 caccaagctg aagatgtacg cggtacgcca aggcaagacc attaccgagc tgctatctga
11161 atacatcgcg cagctaccag agtaaatgag caaatgaata aatgagtaga tgaattttag
11221 cggctaaagg aggcggcatg gaaaatcaag aacaaccagg caccgacgcc gtggaatgcc
11281 ccatgtgtgg aggaacgggc ggttggccag gcgtaagcgg ctgggttgcc tgccggccct
11341 gcaatggcac tggaaccccc aagcccgagg aatcggcgtg agcggtcgca aaccatccgg
11401 cccggtacaa atcggcgcgg cgctgggtga tgacctggtg gagaagttga aggccgcgca
11461 ggccgcccag cggcaacgca tcgaggcaga agcacgcccc ggtgaatcgt ggcaagcggc
11521 cgctgatcga atccgcaaag aatcccggca accgccggca gccggtgcgc cgtcgattag
11581 gaagccgccc aagggcgacg agcaaccaga ttttttcgtt ccgatgctct atgacgtggg
11641 cacccgcgat agtcgcagca tcatggacgt ggccgttttc cgtctgtcga agcgtgaccg
11701 acgagctggc gaggtgatcc gctacgagct tccagacggg cacgtagagg tttccgcagg
11761 gccggccggc atggccagtg tgtgggatta cgacctggta ctgatggcgg tttcccatct
11821 aaccgaatcc atgaaccgat accgggaagg gaagggagac aagcccggcc gcgtgttccg
11881 tccacacgtt gcggacgtac tcaagttctg ccggcgagcc gatggcggaa agcagaaaga
11941 cgacctggta gaaacctgca ttcggttaaa caccacgcac gttgccatgc agcgtacgaa
12001 gaaggccaag aacggccgcc tggtgacggt atccgagggt gaagccttga ttagccgcta
12061 caagatcgta aagagcgaaa ccgggcggcc ggagtacatc gagatcgagc tagctgattg
12121 gatgtaccgc gagatcacag aaggcaagaa cccggacgtg ctgacggttc accccgatta
12181 ctttttgatc gatcccggca tcggccgttt tctctaccgc ctggcacgcc gcgccgcagg
12241 caaggcagaa gccagatggt tgttcaagac gatctacgaa cgcagtggca gcgccggaga
12301 gttcaagaag ttctgtttca ccgtgcgcaa gctgatcggg tcaaatgacc tgccggagta
12361 cgatttgaag gaggaggcgg ggcaggctgg cccgatccta gtcatgcgct accgcaacct
12421 gatcgagggc gaagcatccg ccggttccta atgtacggag cagatgctag ggcaaattgc
12481 cctagcaggg gaaaaaggtc gaaaaggcct ctttcctgtg gatagcacgt acattgggaa
12541 cccaaagccg tacattggga accggaaccc gtacattggg aacccaaagc cgtacattgg
12601 gaaccggtca cacatgtaag tgactgatat aaaagagaaa aaaggcgatt tttccgccta
12661 aaactcttta aaacttatta aaactcttaa aacccgcctg gcctgtgcat aactgtctgg
12721 ccagcgcaca gccgaagagc tgcaaaaagc gcctaccctt cggtcgctgc gctccctacg
12781 ccccgccgct tcgcgtcggc ctatcgcggc cgctggccgc tcaaaaatgg ctggcctacg
12841 gccaggcaat ctaccagggc gcggacaagc cgcgccgtcg ccactcgacc gccggcgccc
12901 acatcaaggc accctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
12961 agctcccgga aacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcgcgtc
13021 agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt
13081 gtatactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg
13141 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc
13201 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
13261 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
13321 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
13381 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
13441 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
13501 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
13561 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
13621 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
13681 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
13741 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
13801 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
13861 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
13921 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
13981 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgcattct
14041 aggtactaaa acaattcatc cagtaaaata taatatttta ttttctccca atcaggcttg
14101 atccccagta agtcaaaaaa tagctcgaca tactgttctt ccccgatatc ctccctgatc
14161 gaccggacgc agaaggcaat gtcataccac ttgtccgccc tgccgcttct cccaagatca
14221 ataaagccac ttactttgcc atctttcaca aagatgttgc tgtctcccag gtcgccgtgg
14281 gaaaagacaa gttcctcttc gggcttttcc gtctttaaaa aatcatacag ctcgcgcgga
14341 tctttaaatg gagtgtcttc ttcccagttt tcgcaatcca catcggccag atcgttattc
14401 agtaagtaat ccaattcggc taagcggctg tctaagctat tcgtataggg acaatccgat
14461 atgtcgatgg agtgaaagag cctgatgcac tccgcataca gctcgataat cttttcaggg
14521 ctttgttcat cttcatactc ttccgagcaa aggacgccat cggcctcact catgagcaga
14581 ttgctccagc catcatgccg ttcaaagtgc aggacctttg gaacaggcag ctttccttcc
14641 agccatagca tcatgtcctt ttcccgttcc acatcatagg tggtcccttt ataccggctg
14701 tccgtcattt ttaaatatag gttttcattt tctcccacca gcttatatac cttagcagga
14761 gacattcctt ccgtatcttt tacgcagcgg tatttttcga tcagtttttt caattccggt
14821 gatattctca ttttagccat ttattatttc cttcctcttt tctacagtat ttaaagatac
14881 cccaagaagc taattataac aagacgaact ccaattcact gttccttgca ttctaaaacc
14941 ttaaatacca gaaaacagct ttttcaaagt tgttttcaaa gttggcgtat aacatagtat
15001 cgacggagcc gattttgaaa ccgcggtgat cacaggcagc aacgctctgt catcgttaca
15061 atcaacatgc taccctccgc gagatcatcc gtgtttcaaa cccggcagct tagttgccgt
15121 tcttccgaat agcatcggta acatgagcaa agtctgccgc cttacaacgg ctctcccgct
15181 gacgccgtcc cggactgatg ggctgcctgt atcgagtggt gattttgtgc cgagctgccg
15241 gtcggggagc tgttggctgg ctggtggcag gatatattgt ggtgtaaaca aattgacgct
15301 tagacaactt aataacacat tgcggacgtt tttaatgtac tgaattaacg ccgaattaat
15361 tcgggggatc tggattttag tactggattt tggttttagg aattagaaat tttattgata
15421 gaagtatttt acaaatacaa atacatacta agggtttctt atatgctcaa cacatgagcg
15481 aaaccctata ggaaccctaa ttcccttatc tgggaactac tcacacatta ttatggagaa
15541 aatagagctt gtcgatcgtt aattaatcac ctccaagatc ccggacgagt gctggggcgt
15601 cggtttccac tatcggcgag tacttctaca cagccatcgg tccagacggc cgcgcttctg
15661 cgggcgattt gtgtacgccc gacagtcccg gctccggatc ggacgattgc gtcgcatcga
15721 ccctgcgccc aagctgcatc atcgaaattg ccgtcaacca agctctgata gagttggtca
15781 agaccaatgc ggagcatata cgcccggagc cgcggcgatc ctgcaagctc cggatgcctc
15841 cgctcgaagt agcgcgtctg ctgctccata caagccaacc acggcctcca gaagaagatg
15901 ttggcgacct cgtattggga atccccgaac atcgcctcgc tccagtcaat gaccgctgtt
15961 atgcggccat tgtccgtcag gacattgttg gagccgaaat ccgcgtgcac gaggtgccgg
16021 acttcggggc agtcctcggc ccaaagcatc agctcatcga gagcctgcgc gacggacgca
16081 ctgacggtgt cgtccatcac agtttgccag tgatacacat ggggatcagc aatcgcgcat
16141 atgaaatcac gccatgtagt gtattgaccg attccttgcg gtccgaatgg gccgaacccg
16201 ctcgtctggc taagatcggc cgcagcgatc gcatccatgg cctccgcgac cggctgcaga
16261 acagcgggca gttcggtttc aggcaggtct tgcaacgtga caccctgtgc acggcgggag
16321 atgcaatagg tcaggctctc gctgaattcc ccaatgtcaa gcacttccgg aatcgggagc
16381 gcggccgatg caaagtgccg ataaacataa cgatctttgt agaaaccatc ggcgcagcta
16441 tttacccgca ggacatatcc acgccctcct acatcgaagc tgaaagcacg agattcttcg
16501 ccctccgaga gctgcatcag gtcggacacg ctgtcgaact tttcgatcag aaacttctcg
16561 acagacgtcg cggtgagttc aggctttttc atcctaggag agatagattt gtagagagag
16621 actggtgatt tcagcgtgtc ctctccaaat gaaatgaact tccttatata gaggaagggt
16681 cttgcgaagg atagtgggat tgtgcgtcat cccttacgtc agtggagata tcacatcaat
16741 ccacttgctt tgaagacgtg gttggaacgt cttctttttc cacgatgctc ctcgtgggtg
16801 ggggtccatc tttgggacca ctgtcggcag aggcatcttg aacgatagcc tttcctttat
16861 cgcaatgatg gcatttgtag gtgccacctt ccttttctac tgtccttttg atgaagtgac
16921 agatagctgg gcaatggaat ccgaggaggt ttcccgatat taccctttgt tgaaaagtct
16981 caatagccct ttggtcttct gagactgtat ctttgatatt cttggagtag acgagagtgt
17041 cgtgctccac catgttcaca tcaatccact tgctttgaag acgtggttgg aacgtcttct
17101 ttttccacga tgctcctcgt gggtgggggt ccatctttgg gaccactgtc ggcagaggca
17161 tcttgaacga tagcctttcc tttatcgcaa tgatggcatt tgtaggtgcc accttccttt
17221 tctactgtcc ttttgatgaa gtgacagata gctgggcaat ggaatccgag gaggtttccc
17281 gatattaccc tttgttgaaa agtctcaata gccctttggt cttctgagac tgtatctttg
17341 atattcttgg agtagacgag agtgtcgtgc tccaccatg
//
| {
"Vector backbone": "pZHY988",
"Backbone size w/o insert (bp)": "",
"Vector type": "Plant Expression, CRISPR, Synthetic Biology",
"Selectable markers": "Hygromycin"
} | {
"Bacterial Resistance(s)": "Kanamycin, 50 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "ccdB Survival",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Efficient plant genome engineering using a probiotic sourced CRISPR-Cas9 system",
"Publication": "Nat Commun. 2023 Sep 29;14(1):6102. doi: 10.1038/s41467-023-41802-9.",
"DOI": "10.1038/s41467-023-41802-9.",
"Authors": "Zhong Z, Liu G, Tang Z, Xiang S, Yang L, Huang L, He Y, Fan T, Liu S, Zheng X, Zhang T, Qi Y, Huang J, Zhang Y /s---",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/37773156",
"PubMed ID": "PubMed 37773156"
} | {
"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": ""
} |
|
pMD19T-slr0230-PL31-sgRNANT1-KmR | 73221 | Contains sgRNA which targets GFPmut3b. Under an aTc inducible promoter. Suicide vector inserts into slr0230 site of Synechocystis. Carries kanamycin resistance. Propogates in E. coli | Paul Hudson | Yao et al ACS Synth Biol. 2015 Dec 28. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/135054/b75bdbfb-0ad2-4f85-85fd-3477be0a7f2d/addgene-plasmid-73221-sequence-135054.gbk | full | LOCUS sequence-135054- 6094 bp DNA circular SYN 26-AUG-2024
DEFINITION Contains sgRNA which targets GFPmut3b. Under an aTc inducible
promoter. Suicide vector inserts into slr0230 site of Synechocystis.
Carries kanamycin resistance. Propogates in E. coli.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6094)
AUTHORS Yao L, Cengic I, Anfelt J, Hudson EP
TITLE Multiple Gene Repression in Cyanobacteria Using CRISPRi.
JOURNAL ACS Synth Biol. 2015 Dec 28.
PUBMED 26689101
REFERENCE 2 (bases 1 to 6094)
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. 2015 Dec 28."
FEATURES Location/Qualifiers
source 1..6094
/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"
misc_feature 1476..1497
/label=BioBrick prefix
/note="BioBrick prefix for parts that do not start with
""ATG"""
protein_bind 1498..1516
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
misc_RNA 1575..1650
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
terminator 1674..1745
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 1761..1788
/label=T7Te terminator
/note="phage T7 early transcription terminator"
CDS complement(1839..2654)
/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(1839..2654)
/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 2562..2581
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
primer_bind 2646..2665
/label=pENTR-R
/note="pENTR vectors, reverse primer"
misc_feature 2769..2789
/label=BioBrick suffix
/note="universal suffix for all parts"
primer_bind complement(3873..3889)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3873..3889)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3886..3908)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3897..3913
/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(3921..3951)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3966..3987
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4104..4121)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4275..4863)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4355..4374)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5034..5894)
/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 5657..5676
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5895..5999)
/gene="bla"
/label=AmpR promoter
primer_bind 6067..6085
/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 agaactcggt acgcgcggat
421 cttccagaga tttgccacct gacgtctaag aaaccataac cgttgtccct tttgtttcat
481 cgaccaacaa ccgccgggga aacgggagag tttgtattac aaagatgatg actatcgcct
541 gagttttctc tacggcagtt atctcacttt gactaatttg agtgccaagg aatggcaacg
601 catagaacag ttaaggcttt ctcccctcta cgtttccatc catgccacgg aagcatcggt
661 gcgggaaaga ttgttaaaaa atcatcgggc cggtcagatt cttgaccagt tggcttggtt
721 ccaagctaga agattacaga tccacgccca ggtggtggtt tgtcctggtg ttaatgatgg
781 taagcactta gagcaaactt tgagggactt agcccagttt caccaagggg aaaccccagc
841 ggtgatttca gtggcggtgg tgccggtggg tctaacccgt tttcgtcccc aggaagatga
901 gttaagcccc gttggtcaag ctaaagccac ggaggtaatt gcccaggttc aagctttgca
961 aaaggaattt gcccagcaac taggtggtaa ctttgtttgg ttggccgatg aatggtttct
1021 cattgcccgt caacctttgc caccggaaag ccattacgaa gactatcccc aaattggcaa
1081 cggggtaggg tctatccgtc aattcattaa ggagtttcaa caacaagctg cggaatttct
1141 ccccccggcg atcgccgaag ccaagacgtt gacctgggta gtgggcaatg ctgtagaaca
1201 agcgtttgag ctactggtgg aacagttaaa tcaggtaaag ggtttaacag ttaatttagc
1261 gcccttaaac agtgactatt ggggtcagga aattacggtg acgggactat tgacggggca
1321 ggatttaatc gctaaattgg caggcagaga tttaggggat ggtattctat tgcctgctct
1381 gatgttgaaa catgatgata ctcgcttctt ggatgacctc cgggtggccg atgtggctca
1441 aaagttgggc acaaccattt accctgtggc tgatggaatt cgcggccgct tctagagtcc
1501 ctatcagtga tagagattga catccctatc agtgatagat ataatgtgtg gtaccatcta
1561 attcaacaag aattgtttta gagctagaaa tagcaagtta aaataaggct agtccgttat
1621 caacttgaaa aagtggcacc gagtcggtgc ttttttttac tagagccagg catcaaataa
1681 aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg
1741 ctctctacta gagtcacact ggctcacctt cgggtgggcc tttctgcgtt tatatactag
1801 aggctctgcc agtgttacaa ccaattaacc aattctgatt agaaaaactc atcgagcatc
1861 aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg aaaaagccgt
1921 ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat
1981 cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa
2041 ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaaa
2101 agcttatgca tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa
2161 tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagcgag acgaaatacg
2221 cgatcgctgt taaaaggaca attacaaaca ggaatcgaat gcaaccggcg caggaacact
2281 gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaatgct
2341 gttttcccgg ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc
2401 ttgatggtcg gaagaggcat aaattccgtc agccagttta gtctgaccat ctcatctgta
2461 acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc
2521 ccatacaatc gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac
2581 ccatataaat cagcatccat gttggaattt aatcgcggcc tggagcaaga cgtttcccgt
2641 tgaatatggc tcataacacc ccttgtatta ctgtttatgt aagcagacag ttttattgtt
2701 catgatgata tatttttatc ttgtgcaatg taacatcaga gattttgaga cacaacgtgg
2761 ctttgttgta ctagtagcgg ccgctgcagc ggagtttaca taatctgaat cagtttctgg
2821 aattgacccc tctgatggcc accagggtca ccgatgccga tggcagtgct ttagttctaa
2881 tgagagaggg ggaaatatct atctttgaac aaatccatgg ccacaaaaat agtcttaagg
2941 gcaccattaa aggcgctttg cagaaggccc gccaggttaa cctcacattg gattcttcca
3001 ctgtcctgag ttattttgac cgtcaattgc gccaagaatt accggcgatc gcctgttata
3061 acacccccat tttgagtcac caagaggaag tgggccgctt atatatcttt agtcaagatc
3121 gtaactatag ctggactccc acccgtcgta agttactaca gctaatttct gatcaaaccg
3181 ctgtggcgat cgccaatagt gatttaaacc aaaagttgag atccagagaa agtcaggatc
3241 gggaattgga aattgcttca gaaattcaaa atcaactatt gccccgttgt tgtccccaaa
3301 ttaatggttt ggacattgct gcccagtgta aaaccgccag tcgggtaggt ggggattact
3361 acgatttcat tcctgccaac tatgaccaac tgcgccaggg ggattggtta tgtcgtaaca
3421 caagccattt gggggtgccc tggagtattg tcatcggcga tgtcatgggt aaaggagtac
3481 cggcgggtct aattatgacc atgactaggg ggatgctgcg ggcagaagtg ctgaaccgcc
3541 atagtccagc ccaaattctt aaccacctca accgggtaat gtatgccgac ctggaaaatt
3601 cccatcgctt tgtcaccttg ttttattctg aatataaccc cgaaacgagc atactttcct
3661 atagcaacgc cgcccataac cctcccctat tgtggcgggc cggcagtgac agcccccaat
3721 gtttaattcc cctggatacg gaaggagctt taatcggctt ggagtcggat tccaattacc
3781 gggatgccca gattcagctt gttcccccag tcgggaaacc tgtcgtgcca gaaatcgtcg
3841 aacggcaggc gtgcaaactt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
3901 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
3961 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
4021 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
4081 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
4141 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
4201 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
4261 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
4321 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
4381 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
4441 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
4501 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
4561 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
4621 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
4681 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
4741 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
4801 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
4861 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
4921 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
4981 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
5041 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
5101 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
5161 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
5221 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
5281 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
5341 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
5401 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
5461 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
5521 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
5581 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
5641 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
5701 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
5761 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
5821 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
5881 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
5941 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatggg ggttccgcgc
6001 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
6061 tataaaaata ggcgtatcac gaggcccttt cgtc
//
| {
"Vector backbone": "pMD19T simple",
"Backbone size w/o insert (bp)": "",
"Vector type": "Bacterial 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": "Multiple Gene Repression in Cyanobacteria Using CRISPRi",
"Publication": "ACS Synth Biol. 2015 Dec 28.",
"DOI": "10.1021/acssynbio.5b00264",
"Authors": "Yao L, Cengic I, Anfelt J, Hudson EP",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/26689101",
"PubMed ID": "PubMed 26689101"
} | {
"Gene/Insert name": "PL31-sgRNA NT1",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "PL31",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "EcoRI (not destroyed)",
"3′ cloning site": "SpeI (destroyed during cloning)",
"5′ sequencing primer": "GCAACGGGGTAGGGTCTATC",
"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 LIC 6A CARD8-FLAG R394E + S297A | 169955 | Mammalian expression of CARD8-FLAG with a site III mutation (R394E) with an additional autoprocessing mutation (S297A) | Hao Wu | Sharif et al Immunity. 2021 Jul 13;54(7):1392-1404.e10. doi: 10.1016/j.immuni.2021.04.024. Epub 2021 May 20. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/332597/81453100-4e95-44e0-9ef2-6a6011410305/addgene-plasmid-169955-sequence-332597.gbk | full | LOCUS sequence-332597- 7087 bp DNA circular SYN 26-AUG-2024
DEFINITION Mammalian expression of CARD8-FLAG with a site III mutation (R394E)
with an additional autoprocessing mutation (S297A).
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7087)
AUTHORS Sharif H, Hollingsworth LR, Griswold AR, Hsiao JC, Wang Q,
Bachovchin DA, Wu H
TITLE Dipeptidyl peptidase 9 sets a threshold for CARD8 inflammasome
formation by sequestering its active C-terminal fragment.
JOURNAL Immunity. 2021 Jul 13;54(7):1392-1404.e10. doi:
10.1016/j.immuni.2021.04.024. Epub 2021 May 20.
PUBMED 34019797
REFERENCE 2 (bases 1 to 7087)
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.immuni.2021.04.024"; journalName: "Immunity"; date:
"2021-07-13- 13"; volume: "54"; issue: "7"; pages: "1392-1404.e10"
FEATURES Location/Qualifiers
source 1..7087
/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"
protein_bind complement(2310..2334)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
CDS 2342..2365
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
promoter complement(2406..2424)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(2407..2424)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(2444..2461)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2450..2674
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2720..3148
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2807..2826)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3017..3038
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(3157..3177)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 3162..3491
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3342..3477
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3404..3423
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3558..4352
/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(3612..3631)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4222..4241
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 4526..4647
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4563..4582)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4617..4636
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4696..4712)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4696..4712)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4709..4731)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4720..4736
/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(4744..4774)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4789..4810
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4927..4944)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5098..5686)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5178..5197)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5857..6717)
/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 6480..6499
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6718..6822)
/gene="bla"
/label=AmpR promoter
primer_bind complement(6897..6916)
/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 acccaagctt
661 ggtaccgagc tcggatccct acttccaatc caatgcaatg gaaaaaaagg agtgtccaga
721 aaagagtagc agcagtgagg aagagctgcc gagacgggac agtggatcca gtaggaacat
781 agatgcatcc aaactcatta gactacaagg atcacggaaa ctgttggttg acaatagcat
841 acgggaactg caatacacaa aaactggaat tttttttcag gctgaggcct gtgtgacaaa
901 tgatacggta tacagggagc taccctgtgt ttctgagacc ctttgtgaca tctcacattt
961 tttccaagaa gatgatgaga cagaggcaga gccattattg ttccgtgctg ttcctgagtg
1021 tcaactatct gggggggaca ttcccagtgt atcagaagag caggaatctt cagagggaca
1081 agattcagga gacatttgct cagaagagaa tcaaatagtt tcctcttatg cttctaaagt
1141 ctgttttgag atcgaagaag attataaaaa tcgtcagttt ctggggcctg aaggaaatgt
1201 ggatgttgag ttgattgata agagcacaaa cagatacagc gtttggttcc ccactgctgg
1261 ctggtatctg tggtcagcca caggcctcgg cttcctggta agggatgagg tcacagtgac
1321 gattgcgttt ggttcctgga gtcagcacct ggccctggac ctgcagcacc atgaacagtg
1381 gctggtgggc ggccccttgt ttgatgtcac tgcagagcca gaggaggctg tcgccgaaat
1441 ccacctcccc cacttcatct ccctccaagc aggtgaggtg gacgtctcct ggtttctcgt
1501 tgcccatttt aagaatgaag ggatggtcct ggagcatcca gcccgggtgg agcctttcta
1561 tgctgtcctg gaaagcccca gcttcgccct gatgggcatc ctgctgcgga tcgccagtgg
1621 gactcgcctc tccatcccca tcacttccaa cacattgatc tattatcacc cccaccccga
1681 agatattaag ttccacttgt accttgtccc cagcgacgcc ttgctaacaa aggcgataga
1741 tgatgaggaa gatcgcttcc atggtgtgcg cctgcagact tcgcccccaa tggaacccct
1801 gaactttggt tccagttata ttgtgtctaa ttctgctaac ctgaaagtaa tgcccaagga
1861 gttgaaattg tcctacgaga gccctggaga aattcagcac ttctcaaaat tctatgctgg
1921 gcagatgaag gaacccattc aacttgagat tactgaaaaa agacatggga ctttggtgtg
1981 ggatactgag gtgaagccag tggatctcca gcttgtagct gcatcagccc ctcctccttt
2041 ctcaggtgca gcctttgtga aggagaacca ccggcaactc caagccagga tgggggacct
2101 gaaaggggtg ctcgatgatc tccaagacaa tgaggttctt actgagaatg agaaggagct
2161 ggtggagcag gaaaagacac ggcagagcaa gaatgaggcc ttgctgagca tggtggagaa
2221 gaaaggggac ctggccctgg acgtgctctt cagaagcatt agtgaaaggg acccttacct
2281 cgtgtcctat cttagacagc agaatttgga cccagctttc ttgtacaaag tggttgatat
2341 cgactacaag gacgatgacg acaagtagta ataacattgg aagtggataa tctagagggc
2401 cctattctat agtgtcacct aaatgctaga gctcgctgat cagcctcgac tgtgccttct
2461 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
2521 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
2581 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
2641 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
2701 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
2761 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
2821 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
2881 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
2941 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
3001 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
3061 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
3121 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
3181 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
3241 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
3301 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
3361 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
3421 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
3481 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
3541 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
3601 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
3661 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
3721 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
3781 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
3841 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
3901 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
3961 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
4021 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
4081 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
4141 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
4201 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
4261 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
4321 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
4381 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
4441 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
4501 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
4561 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
4621 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
4681 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
4741 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
4801 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
4861 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
4921 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
4981 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
5041 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
5101 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
5161 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
5221 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
5281 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
5341 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
5401 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
5461 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
5521 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
5581 tggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
5641 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
5701 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
5761 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
5821 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
5881 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
5941 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
6001 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
6061 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
6121 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
6181 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
6241 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
6301 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
6361 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
6421 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
6481 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
6541 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
6601 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
6661 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
6721 cttccttttt cattattatt gaagcattta tcagggttat tgtctcatga gcggatacat
6781 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
6841 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca
6901 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc
6961 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
7021 atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat
7081 acgcgtt
//
| {
"Vector backbone": "pcDNA3.1 LIC 6A",
"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": "Dipeptidyl peptidase 9 sets a threshold for CARD8 inflammasome formation by sequestering its active C-terminal fragment",
"Publication": "Immunity. 2021 Jul 13;54(7):1392-1404.e10. doi: 10.1016/j.immuni.2021.04.024. Epub 2021 May 20.",
"DOI": "10.1016/j.immuni.2021.04.024.",
"Authors": "Sharif H, Hollingsworth LR, Griswold AR, Hsiao JC, Wang Q, Bachovchin DA, Wu H /jimmuni",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/34019797",
"PubMed ID": "PubMed 34019797"
} | {
"Gene/Insert name": "CARD8",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "S297A, R394E",
"Entrez Gene": "CARD8",
"Promoter": "CMV",
"Tag / Fusion Protein": "FLAG (C terminal on insert)"
} | {
"Cloning method": "Ligation Independent Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "CMV F",
"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": ""
} |
|
pAc5.1B-EGFP-Dm4E-T_1-58-del28-43_T | 147596 | Insect Expression of Dm4E-T_1-58-del28-43 | Elisa Izaurralde | Igreja et al Nat Commun. 2014 Sep 2;5:4790. doi: 10.1038/ncomms5790. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/290043/8a62870d-7d54-4aa8-b976-68accb300f02/addgene-plasmid-147596-sequence-290043.gbk | full | LOCUS sequence-290043- 6200 bp DNA circular SYN 26-AUG-2024
DEFINITION Insect Expression of Dm4E-T_1-58-del28-43.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6200)
AUTHORS Igreja C, Peter D, Weiler C, Izaurralde E
TITLE 4E-BPs require non-canonical 4E-binding motifs and a lateral surface
of eIF4E to repress translation.
JOURNAL Nat Commun. 2014 Sep 2;5:4790. doi: 10.1038/ncomms5790.
PUBMED 25179781
REFERENCE 2 (bases 1 to 6200)
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
Commun."; date: "2014-09-2"; volume: "5:4790. doi"; pages: "
10.1038/ncomms5790"
FEATURES Location/Qualifiers
source 1..6200
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 56..2568
/label=Ac5 promoter
/note="Drosophila melanogaster actin 5C promoter"
CDS 2588..3304
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 2588..3304
/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(2633..2654)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(2894..2913)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 3241..3262
/label=EGFP-C
/note="EGFP, forward primer"
CDS 3485..3526
/codon_start=1
/product="epitope tag from simian virus 5"
/label=V5 tag
/translation="GKPIPNPLLGLDST"
CDS 3536..3553
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(3576..3593)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 3626..3760
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3650..3669)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 3704..3723
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
rep_origin complement(4382..4970)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4462..4481)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5141..6001)
/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 5764..5783
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6002..6106)
/gene="bla"
/label=AmpR promoter
primer_bind 6174..6192
/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 taccaacatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
2641 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
2701 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
2761 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
2821 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
2881 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
2941 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
3001 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
3061 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
3121 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
3181 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
3241 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
3301 caagccaagc ttgaattcta tggatacatc aaagattagt gccaggtact cgaaagtaga
3361 cctgttagct ctaagatatg aaggtaaaag tcgacagcgc attaatctga acacagctgc
3421 gttaaccgtg tccagcgcct attcttaagg cggccgctcg agtctagagg gcccgcggtt
3481 cgaaggtaag cctatcccta accctctcct cggtctcgat tctacgcgta ccggtcatca
3541 tcaccatcac cattgagttt aaacccgctg atcagcctcg actgtgcctt ctaaggcctg
3601 agctcgctga tcagcctcga tcgaggatcc agacatgata agatacattg atgagtttgg
3661 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
3721 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
3781 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
3841 caaatgtggt atggctgatt atgatcagtc gaccgatgcc cttgagagcc ttcaacccag
3901 tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct
3961 ttatcatgca actcgtagga caggtgccgg cagcgctctg ggtcattttc ggcgaggacc
4021 gctttcgctg gagcgcgacg atgatcggcc tgtcgcttgc ggtattcgga atcttgcacg
4081 ccctcgctca agccttcgtc actggtcccg ccaccaaacg tttcggcgag aagcaggcca
4141 ttatcgccgg catggcggcc gacgcgctgg gctacgtctt gctggcgttc gcgacgcgag
4201 gctggatggc cttccccatt atgattcttc tcgcttccgg cggcatcggg atgcccgcgt
4261 tgcaggccat gctgtccagg caggtagatg acgaccatca gggacagctt caaggatcgc
4321 tcgcggctct taccagccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
4381 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
4441 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
4501 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
4561 gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
4621 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
4681 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
4741 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
4801 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
4861 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
4921 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
4981 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
5041 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
5101 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
5161 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
5221 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
5281 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
5341 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
5401 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctgcag
5461 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
5521 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
5581 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
5641 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
5701 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaacac
5761 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
5821 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
5881 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
5941 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
6001 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
6061 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
6121 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
6181 gtatcacgag gccctttcgt
//
| {
"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": "4E-BPs require non-canonical 4E-binding motifs and a lateral surface of eIF4E to repress translation",
"Publication": "Nat Commun. 2014 Sep 2;5:4790. doi: 10.1038/ncomms5790.",
"DOI": "10.1038/ncomms5790.",
"Authors": "Igreja C, Peter D, Weiler C, Izaurralde E /ncomms",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/25179781",
"PubMed ID": "PubMed 25179781"
} | {
"Gene/Insert name": "Dm4E-T_1-58-del28-43",
"Species": "D. melanogaster (fly)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "4E-T",
"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": ""
} |
|
pWT057f | 96869 | dHHR-bsgRNA-HEK3 | David Liu | Tang et al Nat Commun. 2017 Jun 28;8:15939. doi: 10.1038/ncomms15939. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/199245/b3a56540-b02d-4a68-b346-1f005588e100/addgene-plasmid-96869-sequence-199245.gbk | full | LOCUS sequence-199245- 3818 bp DNA circular SYN 26-AUG-2024
DEFINITION dHHR-bsgRNA-HEK3.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3818)
AUTHORS Tang W, Hu JH, Liu DR
TITLE Aptazyme-embedded guide RNAs enable ligand-responsive genome editing
and transcriptional activation.
JOURNAL Nat Commun. 2017 Jun 28;8:15939. doi: 10.1038/ncomms15939.
PUBMED 28656978
REFERENCE 2 (bases 1 to 3818)
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
Commun."; date: "2017-06-28"; volume: "8:15939. doi"; pages: "
10.1038/ncomms15939"
FEATURES Location/Qualifiers
source 1..3818
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 82..330
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA
(Domitrovich & Kunkel, 2003)"
primer_bind 82..102
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 253..272
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
misc_RNA 431..506
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
promoter 570..669
/label=CMVd1 promoter
/note="truncated human cytomegalovirus (CMV) immediate
early promoter that yields slightly reduced expression
relative to the full-length promoter"
primer_bind 620..640
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
misc_feature 692..772
/label=MCS
/note="multiple cloning site"
regulatory 774..783
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 780..1715
/codon_start=1
/product="near-infrared fluorescent protein with an
emission peak at 670 nm, engineered from a bacterial
phytochrome (Shcherbakova and Verkhusha, 2013)"
/label=iRFP670
/note="derived from Rhodopseudomonas palustris RpBphP6"
/translation="MARKVDLTSCDREPIHIPGSIQPCGCLLACDAQAVRITRITENAG
AFFGRETPRVGELLADYFGETEAHALRNALAQSSDPKRPALIFGWRDGLTGRTFDISLH
RHDGTSIIEFEPAAAEQADNPLRLTRQIIARTKELKSLEEMAARVPRYLQAMLGYHRVM
LYRFADDGSGMVIGEAKRSDLESFLGQHFPASLVPQQARLLYLKNAIRVVSDSRGISSR
IVPEHDASGAALDLSFAHLRSISPCHLEFLRNMGVSASMSLSIIIDGTLWGLIICHHYE
PRAVPMAQRVAAEMFADFLSLHFTAAHHQR"
polyA_signal 1837..1958
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1874..1893)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 1928..1947
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(2069..2657)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2149..2168)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(2828..3688)
/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 3451..3470
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(3689..3793)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gacgtcgcta gctgtacaaa aaagcaggct ttaaaggaac caattcagtc gactggatcc
61 ggtaccaagg tcgggcagga agagggccta tttcccatga ttccttcata tttgcatata
121 cgatacaagg ctgttagaga gataattaga attaatttga ctgtaaacac aaagatatta
181 gtacaaaata cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta
241 tgttttaaaa tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct
301 ttatatatct tgtggaaagg acgaaacacc cgtgctcagt ctgggccggt acatccagct
361 gatgagtccc aaataggacg agacgcgctt cggtgcgtcc tggattccac ggcccagact
421 gagcacgtga gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac
481 ttgaaaaagt ggcaccgagt cggtgctttt tttaagcttg ggccgctcga ggtacctctc
541 tacatatgac atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
601 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
661 agcagagctg gtttagtgaa ccgtcagatc cgctagcgct accggactca gatctcgagc
721 tcaagcttcg aattctgcag tcgacggtac cgcgggcccg ggatccaccg gtcgccacca
781 tggcgcgtaa ggtcgatctc acctcctgcg atcgcgagcc gatccacatc cccggcagca
841 ttcagccgtg cggctgcctg ctagcctgcg acgcgcaggc ggtgcggatc acgcgcatta
901 cggaaaatgc cggcgcgttc tttggacgcg aaactccgcg ggtcggtgag ctactcgccg
961 attacttcgg cgagaccgaa gcccatgcgc tgcgcaacgc actggcgcag tcctccgatc
1021 caaagcgacc ggcgctgatc ttcggttggc gcgacggcct gaccggccgc accttcgaca
1081 tctcactgca tcgccatgac ggtacatcga tcatcgagtt cgagcctgcg gcggccgaac
1141 aggccgacaa tccgctgcgg ctgacgcggc agatcatcgc gcgcaccaaa gaactgaagt
1201 cgctcgaaga gatggccgca cgggtgccgc gctatctgca ggcgatgctc ggctatcacc
1261 gcgtgatgtt gtaccgcttc gcggacgacg gctccgggat ggtgatcggc gaggcgaagc
1321 gcagcgacct cgagagcttt ctcggtcagc actttccggc gtcgctggtc ccgcagcagg
1381 cgcggctact gtacttgaag aacgcgatcc gcgtggtctc ggattcgcgc ggcatcagca
1441 gccggatcgt gcccgagcac gacgcctccg gcgccgcgct cgatctgtcg ttcgcgcacc
1501 tgcgcagcat ctcgccctgc catctcgaat ttctgcggaa catgggcgtc agcgcctcga
1561 tgtcgctgtc gatcatcatt gacggcacgc tatggggatt gatcatctgt catcattacg
1621 agccgcgtgc cgtgccgatg gcgcagcgcg tcgcggccga aatgttcgcc gacttcttat
1681 cgctgcactt caccgccgcc caccaccaac gctaagcggc cgcgactcta gatcataatc
1741 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
1801 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
1861 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
1921 tctagttgtg gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa
1981 tattttgtta aaattcgcgt taaattttta tgtgagcaaa aggccagcaa aaggccagga
2041 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
2101 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
2161 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
2221 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
2281 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
2341 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
2401 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
2461 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
2521 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
2581 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
2641 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
2701 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
2761 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
2821 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
2881 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
2941 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
3001 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
3061 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
3121 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
3181 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
3241 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
3301 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
3361 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
3421 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
3481 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
3541 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
3601 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
3661 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
3721 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
3781 taggggttcc gcgcacattt ccccgaaaag tgccacct
//
| {
"Vector backbone": "pFYF1320",
"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": "Aptazyme-embedded guide RNAs enable ligand-responsive genome editing and transcriptional activation",
"Publication": "Nat Commun. 2017 Jun 28;8:15939. doi: 10.1038/ncomms15939.",
"DOI": "10.1038/ncomms15939.",
"Authors": "Tang W, Hu JH, Liu DR /ncomms",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/28656978",
"PubMed ID": "PubMed 28656978"
} | {
"Gene/Insert name": "dHHR-bsgRNA-HEK3",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"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": ""
} |
|
Rtn2b-GFP | 186599 | Encodes human full-length Reticulon 2 isoform B fused to GFP | Gia Voeltz | Zamponi et al Dev Cell. 2022 Jun 6;57(11):1369-1382.e6. doi: 10.1016/j.devcel.2022.05.002. Epub 2022 May 23. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/369196/a7428670-e114-4660-bb0c-ea604bb60221/addgene-plasmid-186599-sequence-369196.gbk | full | LOCUS sequence-369196- 6118 bp DNA circular SYN 25-AUG-2024
DEFINITION Encodes human full-length Reticulon 2 isoform B fused to GFP.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6118)
AUTHORS Zamponi E, Meehl JB, Voeltz GK
TITLE The ER ladder is a unique morphological feature of developing
mammalian axons.
JOURNAL Dev Cell. 2022 Jun 6;57(11):1369-1382.e6. doi:
10.1016/j.devcel.2022.05.002. Epub 2022 May 23.
PUBMED 35609616
REFERENCE 2 (bases 1 to 6118)
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.2022.05.002"; journalName: "Dev Cell"; date:
"2022-06-6- 6"; volume: "57"; issue: "11"; pages: "1369-1382.e6"
FEATURES Location/Qualifiers
source 1..6118
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 1..304
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 305..508
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 459..479
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
CDS 2005..2724
/codon_start=1
/product="Aequorea coerulescens GFP"
/label=AcGFP1
/note="mammalian codon-optimized"
/translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
GFVTAAAITHGMDELYK"
polyA_signal 2846..2967
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2883..2902)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2937..2956
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(2974..3429)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3111..3132)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 3323..3342
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 3456..3560
/gene="bla"
/label=AmpR promoter
promoter 3562..3919
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
enhancer 3562..3753
/label=SV40 enhancer
/note="enhancer for the SV40 early promoter (Herr, 1993)"
primer_bind complement(3585..3605)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 3770..3905
/label=SV40 ori
/note="SV40 origin of replication"
CDS 3954..4748
/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 3954..4748
/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(4008..4027)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4618..4637
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(4936..4955)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 4980..5027
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 5356..5944
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5845..5864
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
541 ccggactcag atctcgagat ggggcaggtc ctgccggtct tcgcccactg caaagaagct
601 ccgtctacag cctcctcaac tcctgattcc acagaaggag ggaacgacga ctctgatttt
661 cgagagctgc acacagcccg ggaattctca gaggaggacg aggaggagac cacgtcgcag
721 gactggggca ccccccggga gctgaccttc tcctacatcg cctttgatgg tgtagtgggc
781 tccgggggcc gcagggattc aactgcccgc cgcccccgcc cccagggccg ctcagtctcg
841 gaaccacgag accagcaccc tcagcccagc ctgggcgaca gcttggagag catccccagc
901 ctgagccaat ccccggagcc tggacgacgg ggtgatcctg acaccgcgcc tccatccgag
961 cgccctctgg aagacctgag gcttcggttg gaccatctgg gctgggtggc ccggggaacg
1021 ggatccgggg aggactcttc caccagcagc tccaccccgc tggaagacga agaaccccaa
1081 gaacccaaca gattggagac aggagaagct ggggaagaac tggacctacg actccgactt
1141 gctcagccct catcgcccga ggtcttgact ccccagctca gtccgggctc tgggacaccc
1201 caggccggta ctccgtcccc atcccgatcg cgagattcga actctgggcc cgaagagcca
1261 ttgctggaag aggaagaaaa gcagtggggg ccactggagc gagagccagt aaggggacag
1321 tgcctcgata gcacggacca attagaattc acggtggagc cacgccttct agtggcggac
1381 ctgctgtact ggaaggacac gaggacgtca ggagtggtct tcacaggcct gatggtctcc
1441 ctcctctgcc tcctgcactt tagcatcgtg tccgtggccg cgcacttggc tctgttgctg
1501 ctctgcggca ccatctctct cagggtttac cgcaaagtgc tgcaggccgt gcaccggggg
1561 gatggagcca accctttcca ggcctacctg gatgtggacc tcaccctgac tcgggagcag
1621 acggaacgtt tgtcccacca gatcacctcc cgcgtggtct cggcggccac gcagctgcgg
1681 cacttcttcc tggtagaaga cctcgtggat tccctcaagc tggccctcct cttctacatc
1741 ttgaccttcg tgggtgccat cttcaatggt ttgactcttc tcattctggg agtgattggt
1801 ctattcacca tccccctgct gtaccggcag caccaggctc agatcgacca atatgtgggg
1861 ttggtgacca atcagttgag ccacatcaaa gctaagatcc gagctaaaat cccagggacc
1921 ggagccctgg cctctgcagc agccgcagtc tccggatcca aagccaaagc cgaacgggta
1981 ccgcgggccc gggatccacc ggtcatggtg agcaagggcg ccgagctgtt caccggcatc
2041 gtgcccatcc tgatcgagct gaatggcgat gtgaatggcc acaagttcag cgtgagcggc
2101 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
2161 aagctgcctg tgccctggcc caccctggtg accaccctga gctacggcgt gcagtgcttc
2221 tcacgctacc ccgatcacat gaagcagcac gacttcttca agagcgccat gcctgagggc
2281 tacatccagg agcgcaccat cttcttcgag gatgacggca actacaagtc gcgcgccgag
2341 gtgaagttcg agggcgatac cctggtgaat cgcatcgagc tgaccggcac cgatttcaag
2401 gaggatggca acatcctggg caataagatg gagtacaact acaacgccca caatgtgtac
2461 atcatgaccg acaaggccaa gaatggcatc aaggtgaact tcaagatccg ccacaacatc
2521 gaggatggca gcgtgcagct ggccgaccac taccagcaga atacccccat cggcgatggc
2581 cctgtgctgc tgcccgataa ccactacctg tccacccaga gcgccctgtc caaggacccc
2641 aacgagaagc gcgatcacat gatctacttc ggcttcgtga ccgccgccgc catcacccac
2701 ggcatggatg agctgtacaa gtgagcggcc gcgactctag atcataatca gccataccac
2761 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
2821 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
2881 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
2941 tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat attttgttaa
3001 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
3061 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
3121 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
3181 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
3241 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
3301 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
3361 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
3421 agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
3481 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
3541 aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt
3601 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
3661 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
3721 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
3781 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
3841 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
3901 aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg
3961 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
4021 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
4081 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg
4141 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
4201 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
4261 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
4321 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
4381 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
4441 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg
4501 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
4561 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
4621 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
4681 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
4741 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
4801 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
4861 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
4921 tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac
4981 ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt
5041 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
5101 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
5161 agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat
5221 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
5281 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
5341 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
5401 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
5461 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
5521 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
5581 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
5641 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
5701 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
5761 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
5821 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
5881 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
5941 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
6001 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta
6061 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccg
//
| {
"Vector backbone": "GFP-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": "Unknown"
} | {
"Academic/Nonprofit Terms": "UBMTA ; Takara Bio Limited Use Label License (formerly Clontech)",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "The ER ladder is a unique morphological feature of developing mammalian axons",
"Publication": "Dev Cell. 2022 Jun 6;57(11):1369-1382.e6. doi: 10.1016/j.devcel.2022.05.002. Epub 2022 May 23.",
"DOI": "10.1016/j.devcel.2022.05.002.",
"Authors": "Zamponi E, Meehl JB, Voeltz GK /jdevcel",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/35609616",
"PubMed ID": "PubMed 35609616"
} | {
"Gene/Insert name": "Human Reticulon 2 isoform B",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "1416",
"Mutation": "",
"Entrez Gene": "RTN2",
"Promoter": "CMV",
"Tag / Fusion Protein": "GFP (C terminal on backbone)"
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "XhoI (not destroyed)",
"3′ cloning site": "KpnI (not destroyed)",
"5′ sequencing primer": "cgccattcagctcgatcagg",
"3′ sequencing primer": "caaatgggcggtaggcgt"
} | {
"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": ""
} |
|
pDSP31 | 199373 | Encodes dual cistronic nuclear localized GCaMP6f and mScarlet-I optimized for C. elegans expression | Hang Lu | plasmids optimized for C. elegans expression (unpublished) | https://media.addgene.org/snapgene-media/v2.0.0/sequences/390590/9b546a75-f9d0-4f9e-98d2-e5ccea3e9efd/addgene-plasmid-199373-sequence-390590.gbk | full | LOCUS sequence-390590- 5387 bp DNA circular SYN 26-AUG-2024
DEFINITION Encodes dual cistronic nuclear localized GCaMP6f and mScarlet-I
optimized for C. elegans expression.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5387)
TITLE plasmids optimized for C. elegans expression
REFERENCE 2 (bases 1 to 5387)
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..5387
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(29..51)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 53..69
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
CDS 227..247
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 263..280
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 320..343
/codon_start=1
/product="Xpress(TM) epitope tag, including an enterokinase
recognition and cleavage site"
/label=Xpress(TM) tag
/translation="DLYDDDDK"
CDS 371..427
/codon_start=1
/product="calmodulin-binding peptide from avian smooth
muscle myosin light chain kinase"
/label=calmodulin-binding peptide
/translation="RRKWNKTGHAVRAIGRLSS"
CDS 2122..2142
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
terminator 3087..3410
/gene="tbb-2"
/label=tbb-2 terminator
/note="transcription terminator for the C. elegans tubulin
beta-2 gene"
primer_bind complement(3508..3524)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3508..3524)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
rep_origin complement(3615..4203)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3695..3714)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4374..5189)
/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="MSHIQRETSCLKPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNGDRVFRLAQAQSRMNNGLVGA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind 5097..5116
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
promoter complement(5190..5292)
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
primer_bind 5360..5378
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
ORIGIN
1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagtaaaacg
61 acggccagtt tatctagtca gcttgattct agctgatcgt ggaccggaag gtgagccagt
121 gagttgattg cagtccagtt acgctggagt ctgaggctcg tcctgaatga tatgcggccg
181 catggtttcc aagggagagg ctgttatcaa ggaattcatg acttacccaa agaagaagcg
241 taaggtccag gttatgggat cccaccacca ccaccaccac ggaatggcct ctatgaccgg
301 aggacaacaa atgggacgcg acctttacga tgacgacgac aaggaccttg ctaccatggt
361 tgattcttct cgccgcaagt ggaacaagac tggacatgct gtccgtgcca tcggacgtct
421 ttcctccctt gagaacgttt acatcaaggc tgataagcaa aagaacggaa ttaaggctaa
481 cttcaagatc cgtcacaaca tcgaagatgg aggagtccaa cttgcttacc attaccaaca
541 aaacactcca atcggagatg gaccagttct tcttccagat aaccactacc tttccgttca
601 atccaagctc tctaaggtaa gtttaaacat atatatacta actaaccctg attatttaaa
661 ttttcaggat ccaaacgaga agcgtgacca catggtcctt cttgagttcg tcaccgctgc
721 tggaatcacc cttggaatgg atgagcttta taagggagga actggaggat ctatggtttc
781 taagggagag gagcttttca ctggagttgt cccaatcctt gtcgagcttg atggagatgt
841 taacggacac aagttctctg tttctggaga gggagaggga gacgctacct acggaaagct
901 tactcttaag ttcatctgca ctactggaaa gcttccagtt ccatggccaa cccttgttac
961 cacccttact tacggagttc aatgcttctc tcgttaccca gatcatatga agcaacatga
1021 tttcttcaag tctgctatgc cagagggata catccaagag cgtactatct tcttcaaggt
1081 aagtttaaac agttcggtac taactaacca tacatattta aattttcagg atgatggaaa
1141 ctacaagacc cgtgctgagg ttaagttcga gggagatacc cttgttaacc gtatcgagct
1201 taagggaatc gatttcaagg aagatggaaa catccttgga cataagcttg agtacaacct
1261 tccagatcaa cttaccgagg aacaaatcgc tgagttcaag gaggaattct cccttttcga
1321 taaggatgga gacggaacta tcactaccaa ggtaagtttt ctagagacct ttactaacta
1381 actaggtctc atagattttc aggagcttgg aaccgttatg cgttcccttg gacaaaaccc
1441 aactgaggct gagcttcaag atatgatcaa cgaggttgac gctgatggag atggaactat
1501 cgatttccca gagttcctta ctatgatggc tcgcaagatg aagtaccgcg atactgagga
1561 ggagatccgc gaggctttcg gagtcttcga caaggacgga aacggataca tttccgctgc
1621 tgagctccgt cacgttatga ctaacctcgg agagaagctt accgacgagg aggtcgacga
1681 gatgatccgt gaggctgata tcgacggaga cggacaagtt aactacgagg agttcgttca
1741 aatgatgact gccaagtccc gtacgcgtaa ggccaaccca accaagctct ccgagaacgc
1801 caagaagctc gccaaggagg tcgagaacta agctgtctca tcctactttc acctagttaa
1861 ctgcttgtct taaaatctat gcttctcttt agtatctaaa attttcctag aagcttacaa
1921 gtatataaat ggtctcttct caataaaggt tgtatattta ttcatcttat tgaatctgcc
1981 atttcctcgt ttttgcgagt ttatatacct tccaattttc tttctattgt attttcaact
2041 tctaatttta attcagggaa actgcttcaa cgcatcatgg tttccaaggg agaggctgtt
2101 atcaaggaat tcatgactta cccaaagaag aagcgtaagg tcgaacttat caaggagttt
2161 atgcgcttta aggttcacat ggaaggatct atgaacggac acgaattcga aatcgaagga
2221 gaaggagaag gacgtccata cgagggaact caaactgcta agcttaaggt aagtttatct
2281 aaaaagtttt tcattcaaaa tgtgtaaaaa ttcatttaaa ataacccaaa aatcattaat
2341 cctcgatatt ttcaggttac taaaggagga ccacttccat tctcttggga tatcctttct
2401 ccacaattca tgtacggatc tcgcgctttc atcaagcacc cagctgatat cccagattac
2461 tacaagcaat ctttcccaga aggattcaaa tgggagcgtg ttatgaactt cgaagatgga
2521 ggagctgtta ccgttaccca agatacctcc cttgaggatg gaacccttat ctacaaggta
2581 ttttcctgca tttttcaact gggaaaatga aagaaaatcg ataatttcag gttaagcttc
2641 gtggaactaa tttcccacca gatggaccag ttatgcaaaa gaagactatg ggatgggagg
2701 cttctaccga gcgcctttac ccagaggatg gagtccttaa gggagatatc aagatggctc
2761 ttcgtcttaa ggatggagga cgttaccttg ctgatttcaa ggtaagtttt ctagagacct
2821 ttactaacta actaggtctc atagattttc agactactta caaggctaag aagccagttc
2881 aaatgccggg agcttacaac gttgatcgta agcttgatat cacctctcac aacgaggatt
2941 acaccgttgt tgagcaatac gagcgttctg agggacgtca ctctaccgga ggaatggatg
3001 agctttacaa gtcccgtacg cgtaaggcca acccaaccaa gctctccgag aacgccaaga
3061 agctcgccaa ggaggtcgag aactaaatgc aaaatccttt caagcattcc cttcttctct
3121 atcactcttc tttctttttg tcaaaaaatt ctctcgctaa tttatttgct tttttaatgt
3181 tattatttta tgacttttta tagtcactga aaagtttgca tctgagtgaa gtgaatgcta
3241 tcaaaatgtg attctgtctg atgtactttc acaatctctc ttcaattcca ttttgaagtg
3301 ctttaaaccc gaaaggttga gaaaaatgcg agcgctcaaa tatttgtatt gtgttcgttg
3361 agtgacccaa caaaaagagg aaactttatt gtgccgccaa gaaaaaagtc tcaatcagtt
3421 ctggaccagc gagctgtgct gcgatgatct tacggcatta tacgtatgat cggtccacga
3481 tcagctagat tatctagtca gcttgatgtc atagctgttt cctgacatgt gagcaaaagg
3541 ccagcaaaag gccaggaacc gtaaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
3601 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
3661 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
3721 agctccctcg tgcgctctcc tgttccgacc ctgtcgctta ccggatacct gtccgccttt
3781 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
3841 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
3901 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
3961 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
4021 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
4081 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
4141 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
4201 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
4261 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
4321 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttagaaa
4381 aactcatcga gcatcaagtg aaactgcaat ttattcatat caggattatc aataccatat
4441 ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg
4501 gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat
4561 ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc
4621 ggtgagaatg gcaagagttt atgcatttct ttccagactt gttcaacagg ccagccatta
4681 cgctcgtcat caaaatcact cgcaccaacc aaaccgttat tcattcgtga ttgcgcctga
4741 gcgagacgaa atacgcgatc gccgttaaaa ggacaattac aaacaggaat cgaatgcaac
4801 cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct
4861 aatacctgga atgctgtttt ccctgggatc gcagtggtga gtaaccatgc atcatcagga
4921 gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagcctg
4981 accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct
5041 ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg
5101 cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg cggcttcaag
5161 caagacgttt cccgttgaat atggctcatt ttagcttcct tagctcctga aaatctcgat
5221 aactcaaaaa atacgcccgg tagtgatctt atttcattat ggtgaaagtt ggaacctctt
5281 acgtgccgat cacacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
5341 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtc
//
| {
"Vector backbone": "pDSP1",
"Backbone size w/o insert (bp)": "",
"Vector type": "Bacterial 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": "Not Available to Industry"
} | {
"Title": "",
"Publication": "",
"DOI": "",
"Authors": "",
"PubMed Link": "",
"PubMed ID": ""
} | {
"Gene/Insert name": "NLS::GCaMP6f::egl-13NLS::SL2::NLS::mScarlet-I::egl-13NLS",
"Species": "C. elegans (nematode)",
"Insert Size (bp)": "",
"Mutation": "no mutations, except added subcellular localization sequences",
"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": ""
} |
|
pPN013 | 91587 | Express sgRNA targeting human CSMD1 | Lindy Barrett | Hazelbaker et al Stem Cell Reports. 2017 Oct 10;9(4):1315-1327. doi: 10.1016/j.stemcr.2017.09.006. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/180147/d0c7211c-c356-49a7-b504-57563b33bbff/addgene-plasmid-91587-sequence-180147.gbk | full | LOCUS sequence-180147- 3023 bp DNA circular SYN 26-AUG-2024
DEFINITION Express sgRNA targeting human CSMD1.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3023)
AUTHORS Hazelbaker DZ, Beccard A, Bara AM, Dabkowski N, Messana A, Mazzucato
P, Lam D, Manning D, Eggan K, Barrett LE
TITLE A Scaled Framework for CRISPR Editing of Human Pluripotent Stem
Cells to Study Psychiatric Disease.
JOURNAL Stem Cell Reports. 2017 Oct 10;9(4):1315-1327. doi:
10.1016/j.stemcr.2017.09.006.
PUBMED 29020615
REFERENCE 2 (bases 1 to 3023)
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.stemcr.2017.09"; journalName: "Stem Cell Reports"; date:
"2017-10-10- 10"; volume: "9"; issue: "4"; pages: "1315-1327"
FEATURES Location/Qualifiers
source 1..3023
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 1..249
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA
(Domitrovich & Kunkel, 2003)"
primer_bind 1..21
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 172..191
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
misc_RNA 271..346
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
primer_bind complement(371..387)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(371..387)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(384..406)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 395..411
/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(419..449)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 464..485
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(602..619)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(773..1361)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(853..872)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(1532..2392)
/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 2155..2174
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(2393..2497)
/gene="bla"
/label=AmpR promoter
primer_bind 2565..2583
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(2621..2643)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 2743..2762
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 2956..2978
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 2970..2987
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 2971..2987
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
ORIGIN
1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
61 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
241 cgaaacaccg tcatcacggg cgagcgcaat gttttagagc tagaaatagc aagttaaaat
301 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt ttaagcttgg
361 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
421 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
481 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
541 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
601 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
661 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
721 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
781 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
841 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
901 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
961 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
1021 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
1081 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
1141 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
1201 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
1261 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
1321 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
1381 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
1441 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
1501 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
1561 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
1621 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
1681 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
1741 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
1801 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
1861 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
1921 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
1981 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
2041 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
2101 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
2161 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
2221 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
2281 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
2341 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
2401 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
2461 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
2521 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
2581 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
2641 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
2701 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
2761 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
2821 gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
2881 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
2941 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa ttcgagctcg
3001 gtacccgggg atccgttaat taa
//
| {
"Vector backbone": "pUC19-U6 promoter",
"Backbone size w/o insert (bp)": "",
"Vector type": "CRISPR",
"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 Scaled Framework for CRISPR Editing of Human Pluripotent Stem Cells to Study Psychiatric Disease",
"Publication": "Stem Cell Reports. 2017 Oct 10;9(4):1315-1327. doi: 10.1016/j.stemcr.2017.09.006.",
"DOI": "10.1016/j.stemcr.2017.09.006.",
"Authors": "Hazelbaker DZ, Beccard A, Bara AM, Dabkowski N, Messana A, Mazzucato P, Lam D, Manning D, Eggan K, Barrett LE /jstemcr",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/29020615",
"PubMed ID": "PubMed 29020615"
} | {
"Gene/Insert name": "CSMD1",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "21",
"Mutation": "",
"Entrez Gene": "CSMD1",
"Promoter": "U6",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "BbsI (destroyed during cloning)",
"3′ cloning site": "BbsI (destroyed during cloning)",
"5′ sequencing primer": "M13F (-21)",
"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": ""
} |
|
pSGE_975_Nhe2_EcR-mut | 71179 | Luciferase reporter | Alexander Stark | Stampfel et al Nature. 2015 Nov 9. doi: 10.1038/nature15545. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/146510/4a12bf7e-7604-44ba-a95f-6b7d8bd5fbe9/addgene-plasmid-71179-sequence-146510.gbk | full | LOCUS sequence-146510- 5437 bp DNA circular SYN 26-AUG-2024
DEFINITION Luciferase reporter.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5437)
AUTHORS Stampfel G, Kazmar T, Frank O, Wienerroither S, Reiter F, Stark A
TITLE Transcriptional regulators form diverse groups with
context-dependent regulatory functions.
JOURNAL Nature. 2015 Nov 9. doi: 10.1038/nature15545.
PUBMED 26550828
REFERENCE 2 (bases 1 to 5437)
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.
2015 Nov 9. doi: 10.1038/nature15545."
FEATURES Location/Qualifiers
source 1..5437
/mol_type="other DNA"
/organism="synthetic DNA construct"
polyA_signal 39..160
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(50..69)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 104..123
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind complement(408..425)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(579..1167)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(659..678)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(1338..2198)
/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 1961..1980
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(2199..2303)
/gene="bla"
/label=AmpR promoter
rep_origin 2330..2785
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2417..2436)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2627..2648
/label=F1ori-F
/note="F1 origin, forward primer"
polyA_signal 2916..2964
/note="synthetic polyadenylation signal"
misc_feature 2978..3069
/label=pause site
/note="RNA polymerase II transcriptional pause signal from
the human alpha-2 globin gene"
primer_bind 3018..3037
/label=RVprimer3
/note="pGL3 vector, forward primer"
protein_bind 3083..3107
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
protein_bind complement(3562..3586)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
regulatory 3777..3786
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 3783..5435
/codon_start=1
/gene="luc+"
/product="firefly luciferase"
/label=luciferase
/note="enhanced luc+ version of the luciferase gene"
/translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
primer_bind complement(3849..3867)
/label=LucNrev
/note="Firefly luciferase, reverse primer"
ORIGIN
1 ctagagtcgg ggcggccggc cgcttcgagc agacatgata agatacattg atgagtttgg
61 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
121 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
181 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
241 caaatgtggt aaaatcgata aggatccgtc gaccgatgcc cttgagagcc ttcaacccag
301 tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct
361 ttatcatgca actcgtagga caggtgccgg cagcgctctt ccgcttcctc gctcactgac
421 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
481 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
541 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
601 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
661 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
721 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
781 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
841 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
901 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
961 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
1021 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
1081 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
1141 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
1201 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
1261 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
1321 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
1381 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
1441 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
1501 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
1561 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
1621 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
1681 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
1741 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
1801 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
1861 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
1921 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
1981 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
2041 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
2101 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
2161 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
2221 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
2281 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgcgccctgt
2341 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
2401 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
2461 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
2521 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
2581 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
2641 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
2701 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
2761 aacaaaatat taacgcttac aatttgccat tcgccattca ggctgcgcaa ctgttgggaa
2821 gggcgatcgg tgcgggcctc ttcgctatta cgccagccca agctaccatg ataagtaagt
2881 aatattaagg tacgggaggt acttggagcg gccgcaataa aatatcttta ttttcattac
2941 atctgtgtgt tggttttttg tgtgaatcga tagtactaac atacgctctc catcaaaaca
3001 aaacgaaaca aaacaaacta gcaaaatagg ctgtccccag tgcaagtgca ggtgccagaa
3061 catttctcta tcgataggta ccacaagttt gtacaaaaaa gcaggctccg aattcgccct
3121 tccagttggc ctgcgataag atctgatttg cattattggc gaaattttat tagcagcagt
3181 agcgaaagct tatctgtcga gtgcggaatg actaaagact cccaattgag caattggtgt
3241 gtacatcgag tgtaattttc gagatgctga taaatctctg agggatttgt acgagtcgga
3301 ttcagtaatt gtcccgactt gagcaattgc atccgttggt attcgctatc tgctggaatg
3361 agtgtcgccc atctcattat ataaaattgg tagtcaattt aggatgatgc ctcatgcatt
3421 tttccgtctc catattcaat tgttctaacc agtggcctag gccttccaca aattcaattt
3481 ggtgttttaa tttaagtatc ttgactgtga ttacagagtg ttgacagagt acagcaatat
3541 ttgaaaacaa gggcgaattc gacccagctt tcttgtacaa agtggtagat ctgcgatcga
3601 gctcgcccgg ggatcgagcg cagcggtata aaagggcgcg gggtggctga gagcatcagt
3661 tgtgaatgaa tgttcgagcc gagcagacgt gccgctgcct tcgttaatat cctttgaata
3721 agccaacttt gaatcacaag acgcatacca aacggcattc cggtactgtt ggtaaagcca
3781 ccatggaaga cgccaaaaac ataaagaaag gcccggcgcc attctatccg ctggaagatg
3841 gaaccgctgg agagcaactg cataaggcta tgaagagata cgccctggtt cctggaacaa
3901 ttgcttttac agatgcacat atcgaggtgg acatcactta cgctgagtac ttcgaaatgt
3961 ccgttcggtt ggcagaagct atgaaacgat atgggctgaa tacaaatcac agaatcgtcg
4021 tatgcagtga aaactctctt caattcttta tgccggtgtt gggcgcgtta tttatcggag
4081 ttgcagttgc gcccgcgaac gacatttata atgaacgtga attgctcaac agtatgggca
4141 tttcgcagcc taccgtggtg ttcgtttcca aaaaggggtt gcaaaaaatt ttgaacgtgc
4201 aaaaaaagct cccaatcatc caaaaaatta ttatcatgga ttctaaaacg gattaccagg
4261 gatttcagtc gatgtacacg ttcgtcacat ctcatctacc tcccggtttt aatgaatacg
4321 attttgtgcc agagtccttc gatagggaca agacaattgc actgatcatg aactcctctg
4381 gatctactgg tctgcctaaa ggtgtcgctc tgcctcatag aactgcctgc gtgagattct
4441 cgcatgccag agatcctatt tttggcaatc aaatcattcc ggatactgcg attttaagtg
4501 ttgttccatt ccatcacggt tttggaatgt ttactacact cggatatttg atatgtggat
4561 ttcgagtcgt cttaatgtat agatttgaag aagagctgtt tctgaggagc cttcaggatt
4621 acaagattca aagtgcgctg ctggtgccaa ccctattctc cttcttcgcc aaaagcactc
4681 tgattgacaa atacgattta tctaatttac acgaaattgc ttctggtggc gctcccctct
4741 ctaaggaagt cggggaagcg gttgccaaga ggttccatct gccaggtatc aggcaaggat
4801 atgggctcac tgagactaca tcagctattc tgattacacc cgagggggat gataaaccgg
4861 gcgcggtcgg taaagttgtt ccattttttg aagcgaaggt tgtggatctg gataccggga
4921 aaacgctggg cgttaatcaa agaggcgaac tgtgtgtgag aggtcctatg attatgtccg
4981 gttatgtaaa caatccggaa gcgaccaacg ccttgattga caaggatgga tggctacatt
5041 ctggagacat agcttactgg gacgaagacg aacacttctt catcgttgac cgcctgaagt
5101 ctctgattaa gtacaaaggc tatcaggtgg ctcccgctga attggaatcc atcttgctcc
5161 aacaccccaa catcttcgac gcaggtgtcg caggtcttcc cgacgatgac gccggtgaac
5221 ttcccgccgc cgttgttgtt ttggagcacg gaaagacgat gacggaaaaa gagatcgtgg
5281 attacgtcgc cagtcaagta acaaccgcga aaaagttgcg cggaggagtt gtgtttgtgg
5341 acgaagtacc gaaaggtctt accggaaaac tcgacgcaag aaaaatcaga gagatcctca
5401 taaaggccaa gaagggcgga aagatcgccg tgtaatt
//
| {
"Vector backbone": "unknown",
"Backbone size w/o insert (bp)": "",
"Vector type": "Insect Expression, Luciferase",
"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 ; Luciferase Limited Use Label License",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Transcriptional regulators form diverse groups with context-dependent regulatory functions",
"Publication": "Nature. 2015 Nov 9. doi: 10.1038/nature15545.",
"DOI": "10.1038/nature15545.",
"Authors": "Stampfel G, Kazmar T, Frank O, Wienerroither S, Reiter F, Stark A /nature",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/26550828",
"PubMed ID": "PubMed 26550828"
} | {
"Gene/Insert name": "975_Nhe2_EcR-mut",
"Species": "Synthetic",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "DSCP Drosophila synthetic core promoter",
"Tag / Fusion Protein": "luciferase (C terminal on backbone)"
} | {
"Cloning method": "Gateway Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "RVprimer3",
"3′ sequencing primer": "LucNRev"
} | {
"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": ""
} |
|
pEN_hUmiRc2 | 25747 | Low | (Empty Backbone)Entry vector for cloning miR30-based shRNA driven by human U6 promoter. | Iain Fraser | Shin et al Proc Natl Acad Sci U S A. 2006 Sep 12. 103(37):13759-64. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/12072/6dd7461f-bdef-4ddd-bb52-32ca98b4e760/addgene-plasmid-25747-sequence-12072.gbk | full | LOCUS sequence-12072-e 4608 bp DNA circular SYN 26-AUG-2024
DEFINITION Entry vector for cloning miR30-based shRNA driven by human U6
promoter..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4608)
AUTHORS Shin KJ, Wall EA, Zavzavadjian JR, Santat LA, Liu J, Hwang JI,
Rebres R, Roach T, Seaman W, Simon MI, Fraser ID
TITLE A single lentiviral vector platform for microRNA-based conditional
RNA interference and coordinated transgene expression.
JOURNAL Proc Natl Acad Sci U S A. 2006 Sep 12. 103(37):13759-64.
PUBMED 16945906
REFERENCE 2 (bases 1 to 4608)
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. 2006 Sep 12. 103(37):13759-64."
FEATURES Location/Qualifiers
source 1..4608
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 25..48
/label=pENTR-F
/note="pENTR vectors, forward primer"
protein_bind 67..166
/gene="mutant version of attL"
/label=attL1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
promoter 426..675
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA
(Domitrovich & Kunkel, 2003)"
primer_bind 426..446
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 598..617
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
ncRNA 730..851
/label=5' miR-30a
/note="sequence upstream of the 71-nt precursor of the
human miR-30a microRNA (Zeng et al., 2002)"
/ncRNA_class="miRNA"
CDS 887..1192
/codon_start=1
/gene="ccdB"
/product="CcdB, a bacterial toxin that poisons DNA gyrase"
/label=ccdB
/note="Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains."
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRIVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
primer_bind 1071..1090
/label=ccdB-fwd
/note="ccdB gene, forward primer"
ncRNA 1213..1339
/label=3' miR-30a
/note="sequence downstream of the 71-nt precursor of the
human miR-30a microRNA (Zeng et al., 2002)"
/ncRNA_class="miRNA"
protein_bind complement(1378..1477)
/gene="mutant version of attL"
/label=attL2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
primer_bind complement(1589..1608)
/label=pENTR-R
/note="pENTR vectors, reverse primer"
primer_bind complement(1667..1686)
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
CDS complement(2260..2793)
/codon_start=1
/gene="aacC1"
/product="gentamycin acetyltransferase"
/label=GmR
/note="confers resistance to gentamycin"
/translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
EEVMHFDIDPSTAT"
promoter complement(2982..3010)
/gene="intI1 (promoter lies within the coding sequence)"
/label=Pc promoter
/note="class 1 integron promoter"
rep_origin 3667..4255
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4156..4175
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
terminator 4420..4506
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator join(4598..4608,1..17)
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
ORIGIN
1 ctgacggatg gcctttttgc gtttctacaa actcttcctg ttagttagtt acttaagctc
61 gggccccaaa taatgatttt attttgactg atagtgacct gttcgttgca acaaattgat
121 aagcaatgct tttttataat gccaactttg tacaaaaaag caggctttaa aggaaccaat
181 tcagtcgact ggatcccccc gagtccaaca cccgtgggaa tcccatgggc accatggccc
241 ctcgctccaa aaatgctttc gcgtctcgca gacactgctc ggtagtttcg gggatcagcg
301 tttgagtaag agcccgcgtc tgaaccctcc gcgccgcccc ggcccagtgg aaagacgcgc
361 aggcaaaacg caccacgtga cggagcgtga ccgcgcgccg agcgcgcgcc aaggtcgggc
421 aggaagaggg cctatttccc atgattcctt catatttgca tatacgatac aaggctgtta
481 gagagataat tagaattaat ttgactgtaa acacaaagat attagtacaa aatacgtgac
541 gtagaaagta ataatttctt gggtagtttg cagtttttaa aattatgttt taaaatggac
601 tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata tatcttgtgg
661 aaaggacgaa acaccgtgct cgcttcggca gcacatatac tagtcgacta gggataacag
721 ggtaattgtt tgaatgaggc ttcagtactt tacagaatcg ttgcctgcac atcttggaaa
781 cacttgctgg gattacttct tcaggttaac ccaacagaag gctcgagaag gtatattgct
841 gttgacagtg agcgtgctgc aggtaagctt gaggtgtgct tccagaatgc agtttaaggt
901 ttacacctat aaaagagaga gccgttatcg tctgtttgtg gatgtacaga gtgatattat
961 tgacacgccc gggcgacgga tagtgatccc cctggccagt gcacgtctgc tgtcagataa
1021 agtctcccgt gaactttacc cggtggtgca tatcggggat gaaagctggc gcatgatgac
1081 caccgatatg gccagtgtgc cggtctccgt tatcggggaa gaagtggctg atctcagcca
1141 ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc tggggaatat agaattccaa
1201 cctgctgctt gcctactgcc tcggaattca aggggctact ttaggagcaa ttatcttgtt
1261 tactaaaact gaataccttg ctatctcttt gatacatttt tacaaagctg aattaaaatg
1321 gtataaatta aatcactttt ttcaattgga agactaatgc gttttcgaga tatctagacc
1381 cagctttctt gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg
1441 aacaggtcac tatcagtcaa aataaaatca ttatttgcca tccagctgca gctctggccc
1501 gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc atgaacaata
1561 aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa
1621 acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct
1681 cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg
1741 ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg
1801 gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt
1861 actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc attccaggta
1921 ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt gtccctgcgc
1981 cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc
2041 gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag
2101 cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca
2161 ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg
2221 aaattaggcc gggaagccga tctcggcttg aacgaattgt taggtggcgg tacttgggtc
2281 gatatcaaag tgcatcactt cttcccgtat gcccaacttt gtatagagag ccactgcggg
2341 atcgtcaccg taatctgctt gcacgtagat cacataagca ccaagcgcgt tggcctcatg
2401 cttgaggaga ttgatgagcg cggtggcaat gccctgcctc cggtgctcgc cggagactgc
2461 gagatcatag atatagatct cactacgcgg ctgctcaaac ctgggcagaa cgtaagccgc
2521 gagagcgcca acaaccgctt cttggtcgaa ggcagcaagc gcgatgaatg tcttactacg
2581 gagcaagttc ccgaggtaat cggagtccgg ctgatgttgg gagtaggtgg ctacgtctcc
2641 gaactcacga ccgaaaagat caagagcagc ccgcatggat ttgacttggt cagggccgag
2701 cctacatgtg cgaatgatgc ccatacttga gccacctaac tttgttttag ggcgactgcc
2761 ctgctgcgta acatcgttgc tgctgcgtaa catcgttgct gctccataac atcaaacatc
2821 gacccacggc gtaacgcgct tgctgcttgg atgcccgagg catagactgt acaaaaaaac
2881 agtcataaca agccatgaaa accgccactg cgccgttacc accgctgcgt tcggtcaagg
2941 ttctggacca gttgcgtgag cgcatacgct acttgcatta cagtttacga accgaacagg
3001 cttatgtcaa ctgggttcgt gccttcatcc gtttccacgg tgtgcgtcac ccggcaacct
3061 tgggcagcag cgaagtcgag gcatttctgt cctggctggc gaacgagcgc aaggtttcgg
3121 tctccacgca tcgtcaggca ttggcggcct tgctgttctt ctacggcaag gtgctgtgca
3181 cggatctgcc ctggcttcag gagatcggaa gacctcggcc gtcgcggcgc ttgccggtgg
3241 tgctgacccc ggatgaagtg gttcgcatcc tcggttttct ggaaggcgag catcgtttgt
3301 tcgcccagga ctctagctat agttctagtg gttggctact aataggttgt attgatgttg
3361 gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg
3421 agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata
3481 tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagaa ttggttaatt
3541 ggttgtaaca ctggcagagc attacgctga cttgacggga cggcgcaagc tcatgaccaa
3601 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
3661 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
3721 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
3781 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
3841 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
3901 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
3961 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
4021 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
4081 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
4141 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
4201 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
4261 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
4321 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gctagcatgg atctcgggga
4381 cgtctaacta ctaagcgaga gtagggaact gccaggcatc aaataaaacg aaaggctcag
4441 tcggaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg
4501 acaaatccgc cgggagcgga tttgaacgtt gtgaagcaac ggcccggagg gtggcgggca
4561 ggacgcccgc cataaactgc caggcatcaa actaagcaga aggccatc
//
| {
"Vector backbone": "pENTR1A-Gent",
"Backbone size w/o insert (bp)": "",
"Vector type": "Entry vector",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Gentamicin, 10 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DB3.1",
"Copy number": "Unknown"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "A single lentiviral vector platform for microRNA-based conditional RNA interference and coordinated transgene expression",
"Publication": "Proc Natl Acad Sci U S A. 2006 Sep 12. 103(37):13759-64.",
"DOI": "10.1073/pnas.0606179103",
"Authors": "Shin KJ, Wall EA, Zavzavadjian JR, Santat LA, Liu J, Hwang JI, Rebres R, Roach T, Seaman W, Simon MI, Fraser ID",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/16945906",
"PubMed ID": "PubMed 16945906"
} | {
"Gene/Insert name": "None",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "attL1 (not destroyed)",
"3′ cloning site": "attL2 (not destroyed)",
"5′ sequencing primer": "hU6-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": ""
} |
pcDNA3-sACE2v6 | 145153 | Mammalian expression plasmid for soluble ACE2 (protease domain) high affinity variant 6 | Erik Procko | Chan et al Science. 2020 Sep 4;369(6508):1261-1265. doi: 10.1126/science.abc0870. Epub 2020 Aug 4. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/292591/3b4e5e8b-48a6-46b0-9f02-7332fee67d2e/addgene-plasmid-145153-sequence-292591.gbk | full | LOCUS sequence-292591- 7210 bp DNA circular SYN 26-AUG-2024
DEFINITION Mammalian expression plasmid for soluble ACE2 (protease domain) high
affinity variant 6.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7210)
AUTHORS Chan KK, Dorosky D, Sharma P, Abbasi SA, Dye JM, Kranz DM, Herbert
AS, Procko E
TITLE Engineering human ACE2 to optimize binding to the spike protein of
SARS coronavirus 2.
JOURNAL Science. 2020 Sep 4;369(6508):1261-1265. doi:
10.1126/science.abc0870. Epub 2020 Aug 4.
PUBMED 32753553
REFERENCE 2 (bases 1 to 7210)
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:
"10.1126/science.abc0870"; journalName: "Science"; date: "2020-09-4-
4"; volume: "369"; issue: "6508"; pages: "1261-1265"
FEATURES Location/Qualifiers
source 1..7210
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 143..522
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 523..726
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 677..697
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 771..790
/label=T7
/note="T7 promoter, forward primer"
promoter 771..789
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(2709..2726)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2715..2939
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2985..3413
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3072..3091)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3282..3303
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(3422..3442)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 3427..3756
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3607..3742
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3669..3688
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3823..4617
/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(3877..3896)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4487..4506
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 4791..4912
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4828..4847)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4882..4901
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4961..4977)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4961..4977)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4974..4996)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4985..5001
/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(5009..5039)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5054..5075
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(5192..5209)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5363..5951)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5443..5462)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6122..6982)
/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 6745..6764
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6983..7087)
/gene="bla"
/label=AmpR promoter
primer_bind complement(7162..7181)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
61 ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat
121 gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt
181 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
241 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
301 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
361 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
421 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
481 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
541 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
601 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
661 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
721 agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat taatacgact
781 cactataggg agacccaagc tggctagcgc caccatgtca agctcttcct ggctccttct
841 cagccttgtt gctgtaactg ctgctcagtc caccattgag gaacaggcca agtacttttt
901 ggacaagttt aaccacgaag ccgaagacct gttctatctg agttcacttg cttcttggaa
961 ttataacacc aatattactg aagagaatgt ccaaaacatg aataatgctg gggacaaatg
1021 gtctgccttt ttaaaggaac agtccacaac tgcccaaatg tatccactac aagaaattca
1081 gaatctccaa gtcaagcttc agctgcaggc tcttcagcaa aatgggtctt cagtgctctc
1141 agaagacaag agcaaacggt tgaacacaat tctaaataca atgagcacca tctacagtac
1201 tggaaaagtt tgtaacccag ataatccaca agaatgctta ttacttgaac caggtttgaa
1261 tgaaataatg gcaaacagtt tagactacaa tgagaggctc tgggcttggg aaagctggag
1321 atctgaggtc ggcaagcagc tgaggccatt atatgaagag tatgtggtct tgaaaaatga
1381 gatggcaaga gcaaatcatt atgaggacta tggggattat tggagaggag actatgaagt
1441 aaatggggta gatggctatg actacagccg cggccagttg attgaagatg tggaacatac
1501 ctttgaagag attaaaccat tatatgaaca tcttcatgcc tatgtgaggg caaagttgat
1561 gaatgcctat ccttcctata tcagtccaat tggatgcctc cctgctcatt tgcttggtga
1621 tatgtggggt agattttgga caaatctgta ctctttgaca gttccctttg gacagaaacc
1681 aaacatagat gttactgatg caatggtgga ccaggcctgg gatgcacaga gaatattcaa
1741 ggaggccgag aagttctttg tatctgttgg tcttcctaat atgactccag gattctggga
1801 atattccatg ctaacggacc caggaaatgt tcagaaagca gtctgccatc ccacagcttg
1861 ggacctgggg aagggcgact tcaggattct tatgtgcaca aaggtgacaa tggacgactt
1921 cctgacagct catcatgaga tggggcatat ccagtatgat atggcatatc ttgcacaacc
1981 ttttctgcta agaaatggag ctaatgaagg attccatgaa gctgttgggg aaatcatgtc
2041 actttctgca gccacaccta agcatttaaa atccattggt cttctgtcac ccgattttca
2101 agaagacaat gaaacagaaa taaacttcct gctcaaacaa gcactcacga ttgttgggac
2161 tctgccattt acttacatgt tagagaagtg gaggtggatg gtctttaaag gggaaattcc
2221 caaagaccag tggatgaaaa agtggtggga gatgaagcga gagatagttg gggtggtgga
2281 acctgtgccc catgatgaaa catactgtga ccccgcatct ctgttccatg tttctaatga
2341 ttactcattc attcgatatt acacaaggac cctttaccaa ttccagtttc aagaagcact
2401 ttgtcaagca gctaaacatg aaggccctct gcacaaatgt gacatctcaa actctacaga
2461 agctggacag aaactgttca atatgctgag gcttggaaaa tcagaaccct ggaccctagc
2521 attggaaaat gttgtaggag caaagaacat gaatgtaagg ccactgctca actactttga
2581 gcccttattt acctggctga aagaccagaa caagaattct tttgtgggat ggagtaccga
2641 ctggagtcca tatgcagacg gatcctgata actcgagtct agagggcccg tttaaacccg
2701 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
2761 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
2821 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
2881 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
2941 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
3001 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
3061 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
3121 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
3181 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
3241 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
3301 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
3361 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
3421 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
3481 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
3541 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
3601 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
3661 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
3721 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
3781 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
3841 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
3901 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
3961 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
4021 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
4081 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
4141 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
4201 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
4261 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
4321 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
4381 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
4441 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
4501 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
4561 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
4621 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
4681 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
4741 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
4801 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
4861 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
4921 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
4981 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
5041 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
5101 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
5161 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
5221 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
5281 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
5341 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
5401 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
5461 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
5521 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
5581 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
5641 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
5701 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
5761 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
5821 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
5881 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
5941 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
6001 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
6061 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
6121 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
6181 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
6241 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
6301 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
6361 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
6421 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
6481 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
6541 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
6601 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
6661 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
6721 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
6781 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
6841 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
6901 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
6961 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
7021 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
7081 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga
7141 tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct
7201 gctccctgct
//
| {
"Vector backbone": "pcDNA3.1(+)",
"Backbone size w/o insert (bp)": "5332",
"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": "Engineering human ACE2 to optimize binding to the spike protein of SARS coronavirus 2",
"Publication": "Science. 2020 Sep 4;369(6508):1261-1265. doi: 10.1126/science.abc0870. Epub 2020 Aug 4.",
"DOI": "10.1126/science.abc0870.",
"Authors": "Chan KK, Dorosky D, Sharma P, Abbasi SA, Dye JM, Kranz DM, Herbert AS, Procko E /scienceabc",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/32753553",
"PubMed ID": "PubMed 32753553"
} | {
"Gene/Insert name": "Angiotensin-converting enzyme 2",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "1875",
"Mutation": "Extracellular, soluble protease domain (a.a. M1-D615) with mutations T27Y, Q42L, L79T, T92Q, Q325P, N330Y, A386L",
"Entrez Gene": "ACE2",
"Promoter": "CMV",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "NheI (not destroyed)",
"3′ cloning site": "XhoI (not destroyed)",
"5′ sequencing primer": "CMV-fwd",
"3′ sequencing primer": "BGH-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": ""
} |
|
LIMK1 gRNA (BRDN0001144894) | 76047 | 3rd generation lentiviral gRNA plasmid targeting human LIMK1 | Doench et al Nat Biotechnol. 2016 Jan 18. doi: 10.1038/nbt.3437. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/185078/d9f41d71-e211-490a-af78-ec1fba5fb6c5/addgene-plasmid-76047-sequence-185078.gbk | full | LOCUS sequence-185078- 8323 bp DNA circular SYN 26-AUG-2024
DEFINITION 3rd generation lentiviral gRNA plasmid targeting human LIMK1.
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 cgtccggctt atactcccag cggttttaga 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": "LIMK1",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "LIMK1",
"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": ""
} |
||
pGL0_78 [ORI_BC_3] | 198979 | Level 0 part | Gilbert et al bioRxiv 2023.05.19.541510 | https://media.addgene.org/snapgene-media/v2.0.0/sequences/401287/1c828073-6645-4b03-8bfd-ff2389a5268c/addgene-plasmid-198979-sequence-401287.gbk | full | LOCUS sequence-401287- 2313 bp DNA circular SYN 27-AUG-2024
DEFINITION Level 0 part.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2313)
AUTHORS Gilbert C, Brumwell SL, Crits-Christoph A, Kang SC, Martin-Moldes Z,
Alsharif W, Esmurria A, Nguyen M-A, Lee HH, Ostrov N
TITLE A scalable framework for high-throughput identification of
functional origins of replication in non-model bacteria
JOURNAL bioRxiv 2023.05.19.541510
REFERENCE 2 (bases 1 to 2313)
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
2023.05.19.541510"
FEATURES Location/Qualifiers
source 1..2313
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(357..373)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(357..373)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(370..392)
/label=M13/pUC Reverse
/note="In lacZ gene"
rep_origin complement(545..1133)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(625..644)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
terminator complement(1266..1360)
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
CDS complement(1381..2040)
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind 1955..1974
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
promoter complement(2041..2143)
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
primer_bind 2230..2252
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 2244..2261
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 2245..2261
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
ORIGIN
1 ccggaatcta ttggcctatg tcactgaaac tatccaaaca ccccatgtcg atactgaacg
61 tatcgacgca tacctccttc cttgaaaacg cacaatcaag gtctcggtca gggtcacgcg
121 taggacgcgt cgagtagggt ttggtcatgt gcttttcgtt gttccgcagc cacacgccgt
181 gtgaagctgg ggagtgagac cagtccttac ctgaagaata agtgtcagcc agtgtaaccc
241 gatgaggaac ccaaaaggcg aaccgggcca gacaacccgg cggtatcgca ctcaaagccg
301 aggctaggtg gaggctcagt gatgataagt ctgcgatggt ggatgcatgt gtcatggtca
361 tagctgtttc ctgtgtgaaa ttgttatccg ctcagagggc acaatcctat tccgcgctat
421 ccgacaatct ccaagacatt aggtggagtt cagttcggcg tatggcatat gtcgctggaa
481 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
541 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
601 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
661 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
721 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
781 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
841 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
901 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
961 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
1021 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
1081 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
1141 ctttgatctt ttctacgggg tctgacgctc tattcaacaa agccgccgtc ccgtcaagtc
1201 agcgtaaatg ggtagggggc ttcaaatcgt ccccccatac gatataagtt gttactagtg
1261 cttggattct caccaataaa aaacgcccgg cggcaaccga gcgttctgaa caaatccaga
1321 tggagttctg aggtcattac tggatctatc aacaggagtc caagcgagct cgatatcaaa
1381 ttacgccccg ccctgccact catcgcagta ctgttgtaat tcattaagca ttctgccgac
1441 atggaagcca tcacaaacgg catgatgaac ctgaatcgcc agcggcatca gcaccttgtc
1501 gccttgcgta taatatttgc ccatggtgaa aacgggggcg aagaagttgt ccatattggc
1561 cacgtttaaa tcaaaactgg tgaaactcac ccagggattg gctgagacga aaaacatatt
1621 ctcaataaac cctttaggga aataggccag gttttcaccg taacacgcca catcttgcga
1681 atatatgtgt agaaactgcc ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt
1741 ttcagtttgc tcatggaaaa cggtgtaaca agggtgaaca ctatcccata tcaccagctc
1801 accgtctttc attgccatac gaaattccgg atgagcattc atcaggcggg caagaatgtg
1861 aataaaggcc ggataaaact tgtgcttatt tttctttacg gtctttaaaa aggccgtaat
1921 atccagctga acggtctggt tataggtaca ttgagcaact gactgaaatg cctcaaaatg
1981 ttctttacga tgccattggg atatatcaac ggtggtatat ccagtgattt ttttctccat
2041 tttagcttcc ttagctcctg aaaatctcga taactcaaaa aatacgcccg gtagtgatct
2101 tatttcatta tggtgaaagt tggaacctct tacgtgccga tcaacgtctc attgatacct
2161 gaaacaaaac ccatcgtacg gccaaggaag tctccaataa ctgtgatcca ccacaagcgc
2221 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tcatgcataa tccgcacgca
2281 tctggaataa ggaagtgcca ttccgcctga cct
//
| {
"Vector backbone": "pGLDV_17",
"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)": "E. coli Zymo 10B",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "A scalable framework for high-throughput identification of functional origins of replication in non-model bacteria",
"Publication": "bioRxiv 2023.05.19.541510",
"DOI": "10.1101/2023.05.19.541510",
"Authors": "Gilbert C, Brumwell SL, Crits-Christoph A, Kang SC, Martin-Moldes Z, Alsharif W, Esmurria A, Nguyen M-A, Lee HH, Ostrov N",
"PubMed Link": "",
"PubMed ID": ""
} | {
"Gene/Insert name": "ORI_BC_3",
"Species": "Synthetic",
"Insert Size (bp)": "",
"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": ""
} |
||
pcDNA3-Ro2 | 24503 | Bruce Conklin | Coward et al Proc Natl Acad Sci U S A. 1998 Jan 6;95(1):352-7. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/30253/5188b2c9-702c-492d-bafb-0a70885e6aa4/addgene-plasmid-24503-sequence-30253.gbk | partial | LOCUS sequence-30253-e 911 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 911)
AUTHORS Coward P, Wada HG, Falk MS, Chan SD, Meng F, Akil H, Conklin BR.
TITLE Controlling signaling with a specifically designed Gi-coupled
receptor.
JOURNAL Proc Natl Acad Sci U S A. 1998 Jan 6;95(1):352-7.
PUBMED 9419379
REFERENCE 2 (bases 1 to 911)
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: "1998-01-6"; volume: "95(1)"; pages: "352-7"
FEATURES Location/Qualifiers
source 1..911
/mol_type="genomic DNA"
/organism="unspecified"
ORIGIN
1 tagaataggg ccctctagat catactggtt tattcatccc atcgatgtcc ctcaggtaag
61 caggatcctg aactgtattt cggactctgc tagtgctctg ccgctccatc ctcatcttca
121 gtggaaagca gaagtcccgg aaacaccgct tgaagttttc atcaagaaag gcgtagagaa
181 tgggattcag gctactgttg gtatagccta aggcgatgca gaagtaatag ctggagagag
241 cagctgtgct gtgggaggtg ctccccagag cctgaaccag gatgaatatg tgaatgggag
301 tccagcagac gacgaagact gccaccacca ccaggaccag tctggtgatc ctacgcaggt
361 tgcgatcttt ctctcgggag ccagaaagga gccggacgct cttgagacgc aggatcatca
421 gggtgtagca gacgatgatg atgaggacag ggatcacgaa ggcaaagatg aagacgcaga
481 tcttggtcac agtgtcccag taccagctgg ggctggggaa ctggagcgtg cataccactg
541 ctccatcccg gggttgggtc accccaagga ctatcgctga tataccaaca gatgatgcca
601 gtagccaaat gcagatgttg atgatctttg ctttcaaagg tgttcggaaa tccaaagctt
661 tcacagggtg gcacacggca atgtaacggt ccacactcat catggtcaag gtgaatatgc
721 tggtaaacat gttgtagtag tcaatggaaa tgacaatctt gcacagaaca tctccaaaag
781 gccaagaatt catcaagtag accgtactct gaaagggcat ggttgtagta actaaagcat
841 ctgccaaagc caggttaaat atgtaaatgt tggttgctgt cttcatcttt gtgtatcgga
901 tgatcacgaa c
//
| {
"Vector backbone": "pcDNA3",
"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",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Controlling signaling with a specifically designed Gi-coupled receptor",
"Publication": "Proc Natl Acad Sci U S A. 1998 Jan 6;95(1):352-7.",
"DOI": "10.1073/pnas.95.1.352",
"Authors": "Coward P, Wada HG, Falk MS, Chan SD, Meng F, Akil H, Conklin BR",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/9419379",
"PubMed ID": "PubMed 9419379"
} | {
"Gene/Insert name": "RASSL Ro2",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "mutations in extracellular loops 2 (see comments below) and 3 (E297Q), making it a RASSL.",
"Entrez Gene": "OPRK1",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "KpnI/XhoI (not destroyed)",
"3′ cloning site": "XbaI (destroyed during cloning)",
"5′ sequencing primer": "CMV-F",
"3′ sequencing primer": "BGH-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": ""
} |
||
pET21c(+)-EFG1mt | 31076 | Linda Spremulli | Bhargava et al Protein Expr Purif. 2004 Oct . 37(2):368-76. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/36702/b67efa4c-1197-4892-8443-9632e81025f2/addgene-plasmid-31076-sequence-36702.gbk | partial | LOCUS sequence-36702-e 848 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 848)
AUTHORS Bhargava K, Templeton P, Spremulli LL
TITLE Expression and characterization of isoform 1 of human mitochondrial
elongation factor G.
JOURNAL Protein Expr Purif. 2004 Oct . 37(2):368-76.
PUBMED 15358359
REFERENCE 2 (bases 1 to 848)
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: "Protein
Expr Purif. 2004 Oct . 37(2):368-76."
FEATURES Location/Qualifiers
source 1..848
/mol_type="genomic DNA"
/organism="unspecified"
CDS complement(26..43)
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
ORIGIN
1 gggctttgtt agcagccgga tctcagtggt ggtggtggtg gtgctcgagg ttcttggctt
61 ttcctttttt aacaggaagt tgacctgtag cttccaaata cttattaatg acgtcttctt
121 gtgtggatgg taaacatggc tgatacctgc tatactccat tgtgtattct ccctttccct
181 ctgtgcatga cctaagttca gtggaataac caaacatatc atttagaggg acatctgcat
241 acagtgtaaa atagtcctca actccatctt gcccagtgat taccccatgg cgtcggttaa
301 ttcctgcaat tacttgtccc tgaaattcat ttggagctac aacttccaca gccataatag
361 gttcaagaat acataatgtt gcatttgcca aggcttgttt aagagcacct tctcctgctc
421 ggatgaaaga gatttcatta gaatcaacca tgtggtgtgc tccatcttgc aggacaaacc
481 ggagcccaga gagcttgtga ccagaaagag ggcccttctc gcaggcatct aaaaacccct
541 tttctacagc aggcacaaac tgctttggaa tatttgatcc gaatgtttca tctgaaaatt
601 ccaatttagt gtagtcctct gggtccagag gctccaggac acctattact tttccatact
661 ggcctgcacc acctgattgt tttttatgtg taaagtcaaa cgggacaggg gcagtaatgg
721 tctctcgaaa ggcaactttt ggctttcctg tgatacaagg acagccatac tctctttcca
781 gcctctgagc atagatttcc aggtgtaatt ctcccattcc agatataact gtctctttgt
841 tctcagtg
//
| {
"Vector backbone": "pET-21c(+)",
"Backbone size w/o insert (bp)": "5400",
"Vector type": "Bacterial Expression",
"Selectable markers": "chloramphenicol"
} | {
"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": "Expression and characterization of isoform 1 of human mitochondrial elongation factor G",
"Publication": "Protein Expr Purif. 2004 Oct . 37(2):368-76.",
"DOI": "10.1016/j.pep.2004.06.030",
"Authors": "Bhargava K, Templeton P, Spremulli LL",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/15358359",
"PubMed ID": "PubMed 15358359"
} | {
"Gene/Insert name": "Mature form human mitochondrial elongation factor G isoform 1",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "2145",
"Mutation": "Includes the coding sequence for the mature form. Amino acids 36-751.",
"Entrez Gene": "GFM1",
"Promoter": "",
"Tag / Fusion Protein": "His (C 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-term"
} | {
"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": ""
} |
||
pGL4.26-SS-352 | 68791 | Low | ZF9 Op x6 -- GFP-IRES-NTR | James Collins | Slomovic et al Nat Methods. 2015 Sep 21. doi: 10.1038/nmeth.3585. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/125112/c06303e4-58aa-484e-a547-65598a33ff51/addgene-plasmid-68791-sequence-125112.gbk | full | LOCUS sequence-125112- 6127 bp DNA circular SYN 26-AUG-2024
DEFINITION ZF9 Op x6 -- GFP-IRES-NTR.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6127)
AUTHORS Slomovic S, Collins JJ
TITLE DNA sense-and-respond protein modules for mammalian cells.
JOURNAL Nat Methods. 2015 Sep 21. doi: 10.1038/nmeth.3585.
PUBMED 26389572
REFERENCE 2 (bases 1 to 6127)
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. 2015 Sep 21. doi: 10.1038/nmeth.3585."
FEATURES Location/Qualifiers
source 1..6127
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 126..157
/label=minP
/note="minimal TATA-box promoter with low basal activity"
regulatory 184..193
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 190..909
/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 190..906
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(235..256)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(496..515)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 843..864
/label=EGFP-C
/note="EGFP, forward primer"
polyA_signal 2082..2203
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2093..2112)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 2147..2166
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
promoter 2397..2754
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
enhancer 2397..2588
/label=SV40 enhancer
/note="enhancer for the SV40 early promoter (Herr, 1993)"
primer_bind complement(2420..2440)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 2605..2740
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2667..2686
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2785..3822
/codon_start=1
/product="hygromycin B phosphotransferase"
/label=HygR
/note="confers resistance to hygromycin"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
EVGRV"
RBS 3801..3809
/label=Shine-Dalgarno sequence
/note="full consensus sequence for ribosome-binding sites
upstream of start codons in E. coli; complementary to a
region in the 3' end of the 16S rRNA (Chen et al., 1994)"
polyA_signal 3846..3894
/note="synthetic polyadenylation signal"
primer_bind complement(4050..4067)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4221..4809)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4301..4320)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5009..5869)
/codon_start=1
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
polyA_signal 5974..6022
/note="synthetic polyadenylation signal"
misc_feature 6036..6127
/label=pause site
/note="RNA polymerase II transcriptional pause signal from
the human alpha-2 globin gene"
primer_bind 6076..6095
/label=RVprimer3
/note="pGL3 vector, forward primer"
ORIGIN
1 ggcctaactg gccggtacct gagctcgcta gctgtagatg gagaaatgta gatggagaaa
61 tgtagatgga gaaatgtaga tggagaaatg tagatggaga aatgtagatg gagaagctta
121 gacactagag ggtatataat ggaagctcga cttccagctt ggcaatccgg tactgttggt
181 aaagccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
241 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
301 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
361 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
421 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
481 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
541 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
601 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
661 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
721 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
781 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
841 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
901 tacaagtaaa gcaggtttcc ccaatgacac aaaacgtgca acttgaaact ccgcctggtc
961 tttccaggtc tagaggggta acactttgta ctgcgtttgg ctccacgctc gatccactgg
1021 cgagtgttag taacagcact gttgcttcgt agcggagcat gacggccgtg ggaactcctc
1081 cttggtaaca aggacccacg gggccaaaag ccacgcccac acgggcccgt catgtgtgca
1141 accccagcac ggcgacttta ctgcgaaacc cactttaaag tgacattgaa actggtaccc
1201 acacactggt gacaggctaa ggatgccctt caggtacccc gaggtaacac gcgacactcg
1261 ggatctgaga aggggactgg ggcttctata aaagcgctcg gtttaaaaag cttctatgcc
1321 tgaataggtg accggaggtc ggcacctttc cgttgcaatt actgacccta tgaatacact
1381 cgagatggac atcatcagcg tggctctgaa gaggcactcc accaaggctt tcgacgcttc
1441 caagaaactg acccctgaac aggccgagca gatcaagacc ctgctccagt acagccctag
1501 ctccaccaac agccagcctt ggcacttcat cgtggctagc accgaggaag gcaaagctag
1561 ggtggctaag agcgccgctg gcaactacgt gttcaacgag aggaagatgc tggatgctag
1621 ccacgtggtg gtgttctgcg ctaagaccgc catggacgat gtgtggctga agctggtggt
1681 ggatcaggaa gatgctgatg gcaggttcgc tacccctgaa gctaaggccg ctaacgacaa
1741 gggcaggaag ttcttcgccg acatgcacag gaaggatctg cacgatgatg ctgagtggat
1801 ggccaagcag gtgtacctga acgtgggcaa cttcctgctc ggcgtggctg ccctgggcct
1861 cgatgctgtg cccatcgaag gcttcgatgc tgctatcctg gatgccgagt tcggcctgaa
1921 ggagaaaggc tacaccagcc tggtggtggt gcctgtgggc caccacagcg tggaggactt
1981 caacgctacc ctgcctaaga gcaggctgcc ccagaacatc accctgaccg aggtgtgata
2041 attctagagt cggggcggcc ggccgcttcg agcagacatg ataagataca ttgatgagtt
2101 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
2161 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
2221 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct
2281 ctacaaatgt ggtaaaatcg ataaggatcc gtttgcgtat tgggcgctct tccgctgatc
2341 tgcgcagcac catggcctga aataacctct gaaagaggaa cttggttagc taccttctga
2401 ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
2461 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
2521 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
2581 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
2641 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
2701 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
2761 gattcttctg acactagcgc caccatgaag aagcccgaac tcaccgctac cagcgttgaa
2821 aaatttctca tcgagaagtt cgacagtgtg agcgacctga tgcagttgtc ggagggcgaa
2881 gagagccgag ccttcagctt cgatgtcggc ggacgcggct atgtactgcg ggtgaatagc
2941 tgcgctgatg gcttctacaa agaccgctac gtgtaccgcc acttcgccag cgctgcacta
3001 cccatccccg aagtgttgga catcggcgag ttcagcgaga gcctgacata ctgcatcagt
3061 agacgcgccc aaggcgttac tctccaagac ctccccgaaa cagagctgcc tgctgtgtta
3121 cagcctgtcg ccgaagctat ggatgctatt gccgccgccg acctcagtca aaccagcggc
3181 ttcggcccat tcgggcccca aggcatcggc cagtacacaa cctggcggga tttcatttgc
3241 gccattgctg atccccatgt ctaccactgg cagaccgtga tggacgacac cgtgtccgcc
3301 agcgtagctc aagccctgga cgaactgatg ctgtgggccg aagactgtcc cgaggtgcgc
3361 cacctcgtcc atgccgactt cggcagcaac aacgtcctga ccgacaacgg ccgcatcacc
3421 gccgtaatcg actggtccga agctatgttc ggggacagtc agtacgaggt ggccaacatc
3481 ttcttctggc ggccctggct ggcttgcatg gagcagcaga ctcgctactt cgagcgccgg
3541 catcccgagc tggccggcag ccctcgtctg cgagcctaca tgctgcgcat cggcctggat
3601 cagctctacc agagcctcgt ggacggcaac ttcgacgatg ctgcctgggc tcaaggccgc
3661 tgcgatgcca tcgtccgcag cggggccggc accgtcggtc gcacacaaat cgctcgccgg
3721 agcgcagccg tatggaccga cggctgcgtc gaggtgctgg ccgacagcgg caaccgccgg
3781 cccagtacac gaccgcgcgc taaggaggta ggtcgagttt aaactctaga accggtcatg
3841 gccgcaataa aatatcttta ttttcattac atctgtgtgt tggttttttg tgtgttcgaa
3901 ctagatgctg tcgaccgatg cccttgagag ccttcaaccc agtcagctcc ttccggtggg
3961 cgcggggcat gactatcgtc gccgcactta tgactgtctt ctttatcatg caactcgtag
4021 gacaggtgcc ggcagcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
4081 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
4141 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
4201 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
4261 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
4321 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
4381 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
4441 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
4501 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
4561 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
4621 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
4681 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
4741 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
4801 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
4861 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
4921 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagc
4981 ggccgcaaat gctaaaccac tgcagtggtt accagtgctt gatcagtgag gcaccgatct
5041 cagcgatctg cctatttcgt tcgtccatag tggcctgact ccccgtcgtg tagatcacta
5101 cgattcgtga gggcttacca tcaggcccca gcgcagcaat gatgccgcga gagccgcgtt
5161 caccggcccc cgatttgtca gcaatgaacc agccagcagg gagggccgag cgaagaagtg
5221 gtcctgctac tttgtccgcc tccatccagt ctatgagctg ctgtcgtgat gctagagtaa
5281 gaagttcgcc agtgagtagt ttccgaagag ttgtggccat tgctactggc atcgtggtat
5341 cacgctcgtc gttcggtatg gcttcgttca actctggttc ccagcggtca agccgggtca
5401 catgatcacc catattatga agaaatgcag tcagctcctt agggcctccg atcgttgtca
5461 gaagtaagtt ggccgcggtg ttgtcgctca tggtaatggc agcactacac aattctctta
5521 ccgtcatgcc atccgtaaga tgcttttccg tgaccggcga gtactcaacc aagtcgtttt
5581 gtgagtagtg tatacggcga ccaagctgct cttgcccggc gtctatacgg gacaacaccg
5641 cgccacatag cagtactttg aaagtgctca tcatcgggaa tcgttcttcg gggcggaaag
5701 actcaaggat cttgccgcta ttgagatcca gttcgatata gcccactctt gcacccagtt
5761 gatcttcagc atcttttact ttcaccagcg tttcggggtg tgcaaaaaca ggcaagcaaa
5821 atgccgcaaa gaagggaatg agtgcgacac gaaaatgttg gatgctcata ctcgtccttt
5881 ttcaatatta ttgaagcatt tatcagggtt actagtacgt ctctcaagga taagtaagta
5941 atattaaggt acgggaggta ttggacaggc cgcaataaaa tatctttatt ttcattacat
6001 ctgtgtgttg gttttttgtg tgaatcgata gtactaacat acgctctcca tcaaaacaaa
6061 acgaaacaaa acaaactagc aaaataggct gtccccagtg caagtgcagg tgccagaaca
6121 tttctct
//
| {
"Vector backbone": "pGL4.26 luc2/minp/hygro",
"Backbone size w/o insert (bp)": "4917",
"Vector type": "Mammalian Expression",
"Selectable markers": "Hygromycin"
} | {
"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": "DNA sense-and-respond protein modules for mammalian cells",
"Publication": "Nat Methods. 2015 Sep 21. doi: 10.1038/nmeth.3585.",
"DOI": "10.1038/nmeth.3585.",
"Authors": "Slomovic S, Collins JJ /nmeth",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/26389572",
"PubMed ID": "PubMed 26389572"
} | {
"Gene/Insert name": "ZF9 Op 6x",
"Species": "Synthetic",
"Insert Size (bp)": "81",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "EGFP",
"Species": "Synthetic",
"Insert Size (bp)": "720",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": "IRES (C terminal on insert)"
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "Nitroreductase",
"Species": "E. Coli (human codon optimized)",
"Insert Size (bp)": "654",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": "IRES (N terminal on insert)"
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
pYNL-N1_TUBB5 | 65678 | Expresses beta-tubulin-YNL in mammalian cells | Yasushi Okada | Takai et al Proc Natl Acad Sci U S A. 2015 Apr 7;112(14):4352-6. doi: 10.1073/pnas.1418468112. Epub 2015 Mar 23. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/116525/7470f1e3-b8ea-4b17-a857-4550d9520834/addgene-plasmid-65678-sequence-116525.gbk | partial | LOCUS sequence-116525- 1057 bp DNA linear UNA 26-AUG-2024
DEFINITION Expresses beta-tubulin-YNL in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE natural DNA sequence
ORGANISM unspecified
REFERENCE 1 (bases 1 to 1057)
AUTHORS Takai A, Nakano M, Saito K, Haruno R, Watanabe TM, Ohyanagi T, Jin
T, Okada Y, Nagai T
TITLE Expanded palette of Nano-lanterns for real-time multicolor
luminescence imaging.
JOURNAL Proc Natl Acad Sci U S A. 2015 Apr 7;112(14):4352-6. doi:
10.1073/pnas.1418468112. Epub 2015 Mar 23.
PUBMED 25831507
REFERENCE 2 (bases 1 to 1057)
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.1418468112"; journalName: "Proc Natl Acad Sci U S A";
date: "2015-04-7- 7"; volume: "112"; issue: "14"; pages: "4352-6"
FEATURES Location/Qualifiers
source 1..1057
/mol_type="genomic DNA"
/organism="unspecified"
ORIGIN
1 ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc ggactcagat
61 ctcgagctca agcttcgaat tccaccatga gggaaatcgt gcacatccag gccggacagt
121 gtggcaacca gatcggtgct aagttctggg aggtgataag cgatgaacat ggcatcgacc
181 ccaccggtac ctaccacggt gacagcgacc tgcagctgga ccgaatctct gtgtactata
241 atgaagccac aggtggcaag tatgtccctc gagctatctt ggtggatcta gaacctggga
301 ctatggactc cgttcgctca ggtccttttg gccagatctt cagaccagac aacttcgttt
361 tcggtcagtc tggggcaggc aacaactggg ctaaaggcca ctacacagag ggagctgagt
421 tggttgactc tgtcttggat gtggtgcgga aggaggcgga gagctgtgat tgcctgcaag
481 gctttcagct gacccactca ctgggtggag gcactggctc tggcatgggc accctgctca
541 tcagcaagat ccgggaagaa tatcctgacc gtatcatgaa taccttcagt gtggtgccct
601 cgcccaaagt gtctgatacc gtggtcgagc cctacaatgc caccctgtct gtccatcagt
661 tggttgagaa cacggatgag acctactgca tcgacaacga ggccctctac gacatctgct
721 tccgtaccct caagctcacc acgccaacct acggagacct gaaccatctc gtctcggcca
781 ccatgagcgg cgtcaccacc tgcctccgtt tcccgggcca gcttaatgct gaccttcgaa
841 agctggctgt caacatggtg ccattcccac gtctccactt cttcatgcct ggctttgccc
901 ctctcaccag ccgtggaagc cagcagtacc gggccctcac tgtgcctgaa cttacccagc
961 aggtcttcga tgccaagaac atgatggccg cctgcgaccc gcgccacggc cggtacctca
1021 cagttgccgc cgtcttccgt ggacggatgt ccatgaa
//
| {
"Vector backbone": "pYNL-N1",
"Backbone size w/o insert (bp)": "5600",
"Vector type": "Mammalian Expression, Luciferase",
"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": "Expanded palette of Nano-lanterns for real-time multicolor luminescence imaging",
"Publication": "Proc Natl Acad Sci U S A. 2015 Apr 7;112(14):4352-6. doi: 10.1073/pnas.1418468112. Epub 2015 Mar 23.",
"DOI": "10.1073/pnas.1418468112.",
"Authors": "Takai A, Nakano M, Saito K, Haruno R, Watanabe TM, Ohyanagi T, Jin T, Okada Y, Nagai T /pnas",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/25831507",
"PubMed ID": "PubMed 25831507"
} | {
"Gene/Insert name": "beta-tubulin class V",
"Species": "M. musculus (mouse)",
"Insert Size (bp)": "1300",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "CMV",
"Tag / Fusion Protein": "YNL (C terminal on backbone)"
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "EcoRI (not destroyed)",
"3′ cloning site": "SalI (not destroyed)",
"5′ sequencing primer": "CMV Forward",
"3′ sequencing primer": "EGFP-N"
} | {
"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": ""
} |
|
T4 Lysozyme M106A WT* | 18504 | Brian Matthews | T4 Lysozyme Plasmids - Tray 4 (unpublished) | https://media.addgene.org/snapgene-media/v2.0.0/sequences/24290/da8f9bdb-2be2-4c61-803b-34a98858f3d2/addgene-plasmid-18504-sequence-24290.gbk | partial | LOCUS sequence-24290-e 831 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 831)
TITLE T4 Lysozyme Plasmids - Tray 4
REFERENCE 2 (bases 1 to 831)
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..831
/mol_type="genomic DNA"
/organism="unspecified"
primer_bind complement(712..734)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 723..739
/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 747..763
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 747..763
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
ORIGIN
1 tactgaaaag cccgcctcat taggcgggct cagatctgct catgtttgac agcttatcat
61 cgataagctt cggcaataaa ttcttgatat gttttcatat gtttccttta aatattttaa
121 ttagtaattg tctattcaag taattgtgaa tatactatca caattccaag agaaagtaaa
181 cagctttata gatttttata cgcgtcccaa gtgccagttc taaacgttgt aatgactcgt
241 tttgcgcgat taggtgtttg attataccat ctacttttag ctaagttaac tgctgcttca
301 tcccagcgtt tttgttgaag catacgtaaa gagttagtaa atcctgccac accggtttct
361 cccgcttgga aaaccatatt aatcaatgca gcgcgacgaa ccgcatcaag agaatcataa
421 accggtttta atttagcatt tctcagaatt ccgcgaacag cagcatcaac atcctgatta
481 aagagttttt cagcctcatc ttttgtaatt acaccattag tattacgccc aatagcttta
541 tctaattcag atttagcagc attaagtgat ggactttttg taagcaaatg accgatgcca
601 atagtgtaat agccttctgt gtctttatag attttaagtc taaggccttc atctatacgt
661 aacatttcaa atatattcat aatacctcct aagcatcgat ggatcctgtt tcctgtgtga
721 aattgttatc cgctcacaat tccacacagg aaacagctat gaccatgata cggatcactg
781 gcngcgttta cacgtcgtgc tggaaaccct gcgtaccact ancgcctgcg c
//
| {
"Vector backbone": "pHS1403",
"Backbone size w/o insert (bp)": "5000",
"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": "T4 Lysozyme",
"Species": "Bacteriophage T4",
"Insert Size (bp)": "600",
"Mutation": "M106A WT*",
"Entrez Gene": "e",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "BamHI (not destroyed)",
"3′ cloning site": "HindIII (not destroyed)",
"5′ sequencing primer": "na",
"3′ sequencing primer": "GGCGTATCACGAGGCCC"
} | {
"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": ""
} |
||
WIPI2 sgRNA | 207554 | pX330 expressing Cas9 and a sgRNA targeting the WIPI2 locus | Jens Schmidt | Broadbent et al J Cell Biol. 2023 Jul 3;222(7):e202210078. doi: 10.1083/jcb.202210078. Epub 2023 Apr 28. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/426772/1d1619a9-c811-40fa-82c9-4bf70bf28784/addgene-plasmid-207554-sequence-426772.gbk | full | LOCUS sequence-426772- 8507 bp DNA circular SYN 27-AUG-2024
DEFINITION pX330 expressing Cas9 and a sgRNA targeting the WIPI2 locus.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8507)
AUTHORS Broadbent DG, Barnaba C, Perez GI, Schmidt JC
TITLE Quantitative analysis of autophagy reveals the role of ATG9 and ATG2
in autophagosome formation.
JOURNAL J Cell Biol. 2023 Jul 3;222(7):e202210078. doi:
10.1083/jcb.202210078. Epub 2023 Apr 28.
PUBMED 37115157
REFERENCE 2 (bases 1 to 8507)
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: "J Cell
Biol."; date: "2023-07-3"; volume: "222(7):e202210078. doi"; pages:
" 10.1083/jcb.202210078. Epub 2023 Apr 28"
FEATURES Location/Qualifiers
source 1..8507
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 6..254
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA
(Domitrovich & Kunkel, 2003)"
primer_bind 6..26
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 177..196
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
misc_RNA 276..351
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
enhancer 448..733
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer;
contains an 18-bp deletion relative to the standard CMV
enhancer"
promoter 735..1011
/label=chicken beta-actin promoter
intron 1012..1239
/label=hybrid intron
/note="hybrid between chicken beta-actin (CBA) and minute
virus of mice (MMV) introns (Gray et al., 2011)"
regulatory 1251..1260
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 1260..1325
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
CDS 1332..1352
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 1377..5477
/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 1377..5477
/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 5478..5525
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
primer_bind complement(5553..5570)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 5559..5766
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
repeat_region 5775..5915
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 5775..5904
/label=AAV2 ITR
rep_origin 5990..6445
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(6077..6096)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 6287..6308
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(6462..6481)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 6581..6603
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(6641..6659)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 6727..6831
/gene="bla"
/label=AmpR promoter
CDS 6832..7692
/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(7050..7069)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 7863..8451
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 8352..8371
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 catgtgaggg cctatttccc atgattcctt catatttgca tatacgatac aaggctgtta
61 gagagataat tggaattaat ttgactgtaa acacaaagat attagtacaa aatacgtgac
121 gtagaaagta ataatttctt gggtagtttg cagttttaaa attatgtttt aaaatggact
181 atcatatgct taccgtaact tgaaagtatt tcgatttctt ggctttatat atcttgtgga
241 aaggacgaaa caccgcgcgc gcccagccat gaaccgtttt agagctagaa atagcaagtt
301 aaaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg cttttttgtt
361 ttagagctag aaatagcaag ttaaaataag gctagtccgt ttttagcgcg tgcgccaatt
421 ctgcagacaa atggctctag aggtacccgt tacataactt acggtaaatg gcccgcctgg
481 ctgaccgccc aacgaccccc gcccattgac gtcaatagta acgccaatag ggactttcca
541 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
601 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcattg
661 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
721 cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca tctccccccc
781 ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag cgatgggggc
841 gggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg gggcggggcg
901 aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt tccttttatg
961 gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc gggagtcgct
1021 gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc gccccggctc
1081 tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc tccgggctgt
1141 aattagctga gcaagaggta agggtttaag ggatggttgg ttggtggggt attaatgttt
1201 aattacctgg agcacctgcc tgaaatcact ttttttcagg ttggaccggt gccaccatgg
1261 actataagga ccacgacgga gactacaagg atcatgatat tgattacaaa gacgatgacg
1321 ataagatggc cccaaagaag aagcggaagg tcggtatcca cggagtccca gcagccgaca
1381 agaagtacag catcggcctg gacatcggca ccaactctgt gggctgggcc gtgatcaccg
1441 acgagtacaa ggtgcccagc aagaaattca aggtgctggg caacaccgac cggcacagca
1501 tcaagaagaa cctgatcgga gccctgctgt tcgacagcgg cgaaacagcc gaggccaccc
1561 ggctgaagag aaccgccaga agaagataca ccagacggaa gaaccggatc tgctatctgc
1621 aagagatctt cagcaacgag atggccaagg tggacgacag cttcttccac agactggaag
1681 agtccttcct ggtggaagag gataagaagc acgagcggca ccccatcttc ggcaacatcg
1741 tggacgaggt ggcctaccac gagaagtacc ccaccatcta ccacctgaga aagaaactgg
1801 tggacagcac cgacaaggcc gacctgcggc tgatctatct ggccctggcc cacatgatca
1861 agttccgggg ccacttcctg atcgagggcg acctgaaccc cgacaacagc gacgtggaca
1921 agctgttcat ccagctggtg cagacctaca accagctgtt cgaggaaaac cccatcaacg
1981 ccagcggcgt ggacgccaag gccatcctgt ctgccagact gagcaagagc agacggctgg
2041 aaaatctgat cgcccagctg cccggcgaga agaagaatgg cctgttcgga aacctgattg
2101 ccctgagcct gggcctgacc cccaacttca agagcaactt cgacctggcc gaggatgcca
2161 aactgcagct gagcaaggac acctacgacg acgacctgga caacctgctg gcccagatcg
2221 gcgaccagta cgccgacctg tttctggccg ccaagaacct gtccgacgcc atcctgctga
2281 gcgacatcct gagagtgaac accgagatca ccaaggcccc cctgagcgcc tctatgatca
2341 agagatacga cgagcaccac caggacctga ccctgctgaa agctctcgtg cggcagcagc
2401 tgcctgagaa gtacaaagag attttcttcg accagagcaa gaacggctac gccggctaca
2461 ttgacggcgg agccagccag gaagagttct acaagttcat caagcccatc ctggaaaaga
2521 tggacggcac cgaggaactg ctcgtgaagc tgaacagaga ggacctgctg cggaagcagc
2581 ggaccttcga caacggcagc atcccccacc agatccacct gggagagctg cacgccattc
2641 tgcggcggca ggaagatttt tacccattcc tgaaggacaa ccgggaaaag atcgagaaga
2701 tcctgacctt ccgcatcccc tactacgtgg gccctctggc caggggaaac agcagattcg
2761 cctggatgac cagaaagagc gaggaaacca tcaccccctg gaacttcgag gaagtggtgg
2821 acaagggcgc ttccgcccag agcttcatcg agcggatgac caacttcgat aagaacctgc
2881 ccaacgagaa ggtgctgccc aagcacagcc tgctgtacga gtacttcacc gtgtataacg
2941 agctgaccaa agtgaaatac gtgaccgagg gaatgagaaa gcccgccttc ctgagcggcg
3001 agcagaaaaa ggccatcgtg gacctgctgt tcaagaccaa ccggaaagtg accgtgaagc
3061 agctgaaaga ggactacttc aagaaaatcg agtgcttcga ctccgtggaa atctccggcg
3121 tggaagatcg gttcaacgcc tccctgggca cataccacga tctgctgaaa attatcaagg
3181 acaaggactt cctggacaat gaggaaaacg aggacattct ggaagatatc gtgctgaccc
3241 tgacactgtt tgaggacaga gagatgatcg aggaacggct gaaaacctat gcccacctgt
3301 tcgacgacaa agtgatgaag cagctgaagc ggcggagata caccggctgg ggcaggctga
3361 gccggaagct gatcaacggc atccgggaca agcagtccgg caagacaatc ctggatttcc
3421 tgaagtccga cggcttcgcc aacagaaact tcatgcagct gatccacgac gacagcctga
3481 cctttaaaga ggacatccag aaagcccagg tgtccggcca gggcgatagc ctgcacgagc
3541 acattgccaa tctggccggc agccccgcca ttaagaaggg catcctgcag acagtgaagg
3601 tggtggacga gctcgtgaaa gtgatgggcc ggcacaagcc cgagaacatc gtgatcgaaa
3661 tggccagaga gaaccagacc acccagaagg gacagaagaa cagccgcgag agaatgaagc
3721 ggatcgaaga gggcatcaaa gagctgggca gccagatcct gaaagaacac cccgtggaaa
3781 acacccagct gcagaacgag aagctgtacc tgtactacct gcagaatggg cgggatatgt
3841 acgtggacca ggaactggac atcaaccggc tgtccgacta cgatgtggac catatcgtgc
3901 ctcagagctt tctgaaggac gactccatcg acaacaaggt gctgaccaga agcgacaaga
3961 accggggcaa gagcgacaac gtgccctccg aagaggtcgt gaagaagatg aagaactact
4021 ggcggcagct gctgaacgcc aagctgatta cccagagaaa gttcgacaat ctgaccaagg
4081 ccgagagagg cggcctgagc gaactggata aggccggctt catcaagaga cagctggtgg
4141 aaacccggca gatcacaaag cacgtggcac agatcctgga ctcccggatg aacactaagt
4201 acgacgagaa tgacaagctg atccgggaag tgaaagtgat caccctgaag tccaagctgg
4261 tgtccgattt ccggaaggat ttccagtttt acaaagtgcg cgagatcaac aactaccacc
4321 acgcccacga cgcctacctg aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta
4381 agctggaaag cgagttcgtg tacggcgact acaaggtgta cgacgtgcgg aagatgatcg
4441 ccaagagcga gcaggaaatc ggcaaggcta ccgccaagta cttcttctac agcaacatca
4501 tgaacttttt caagaccgag attaccctgg ccaacggcga gatccggaag cggcctctga
4561 tcgagacaaa cggcgaaacc ggggagatcg tgtgggataa gggccgggat tttgccaccg
4621 tgcggaaagt gctgagcatg ccccaagtga atatcgtgaa aaagaccgag gtgcagacag
4681 gcggcttcag caaagagtct atcctgccca agaggaacag cgataagctg atcgccagaa
4741 agaaggactg ggaccctaag aagtacggcg gcttcgacag ccccaccgtg gcctattctg
4801 tgctggtggt ggccaaagtg gaaaagggca agtccaagaa actgaagagt gtgaaagagc
4861 tgctggggat caccatcatg gaaagaagca gcttcgagaa gaatcccatc gactttctgg
4921 aagccaaggg ctacaaagaa gtgaaaaagg acctgatcat caagctgcct aagtactccc
4981 tgttcgagct ggaaaacggc cggaagagaa tgctggcctc tgccggcgaa ctgcagaagg
5041 gaaacgaact ggccctgccc tccaaatatg tgaacttcct gtacctggcc agccactatg
5101 agaagctgaa gggctccccc gaggataatg agcagaaaca gctgtttgtg gaacagcaca
5161 agcactacct ggacgagatc atcgagcaga tcagcgagtt ctccaagaga gtgatcctgg
5221 ccgacgctaa tctggacaaa gtgctgtccg cctacaacaa gcaccgggat aagcccatca
5281 gagagcaggc cgagaatatc atccacctgt ttaccctgac caatctggga gcccctgccg
5341 ccttcaagta ctttgacacc accatcgacc ggaagaggta caccagcacc aaagaggtgc
5401 tggacgccac cctgatccac cagagcatca ccggcctgta cgagacacgg atcgacctgt
5461 ctcagctggg aggcgacaaa aggccggcgg ccacgaaaaa ggccggccag gcaaaaaaga
5521 aaaagtaaga attcctagag ctcgctgatc agcctcgact gtgccttcta gttgccagcc
5581 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
5641 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
5701 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagagaata gcaggcatgc
5761 tggggagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg
5821 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc
5881 ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct
5941 tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt
6001 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
6061 agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
6121 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
6181 cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga
6241 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
6301 caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg
6361 ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
6421 aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc
6481 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
6541 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
6601 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
6661 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
6721 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
6781 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
6841 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
6901 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
6961 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
7021 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
7081 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
7141 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
7201 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
7261 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
7321 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
7381 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
7441 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
7501 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtggaag ccgcggtatc
7561 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
7621 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
7681 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
7741 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
7801 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
7861 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
7921 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
7981 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
8041 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
8101 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
8161 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
8221 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
8281 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
8341 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
8401 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
8461 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctca
//
| {
"Vector backbone": "pX330 Addgene #42230",
"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": "Quantitative analysis of autophagy reveals the role of ATG9 and ATG2 in autophagosome formation",
"Publication": "J Cell Biol. 2023 Jul 3;222(7):e202210078. doi: 10.1083/jcb.202210078. Epub 2023 Apr 28.",
"DOI": "10.1083/jcb.202210078.",
"Authors": "Broadbent DG, Barnaba C, Perez GI, Schmidt JC /jcb",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/37115157",
"PubMed ID": "PubMed 37115157"
} | {
"Gene/Insert name": "CGCGCGCCCAGCCATGAACC",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "CMV and U6",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "BbsI (unknown if destroyed)",
"3′ cloning site": "BbsI (unknown if destroyed)",
"5′ sequencing primer": "U6 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": ""
} |
|
pU2T2U2T2-YFP | 84550 | Hybrid reporter construct with 8 transcription factor binding sites (2 UAS, 2 TetO, 2 UAS, 2 TetO) driving EYFP | Joshua Leonard | Hartfield et al ACS Synth Biol. 2017 Nov 17;6(11):2042-2055. doi: 10.1021/acssynbio.6b00279. Epub 2017 Aug 23. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/169190/81f8021d-2158-4af0-a913-dff25e663a75/addgene-plasmid-84550-sequence-169190.gbk | full | LOCUS sequence-169190- 4727 bp DNA circular SYN 26-AUG-2024
DEFINITION Hybrid reporter construct with 8 transcription factor binding sites
(2 UAS, 2 TetO, 2 UAS, 2 TetO) driving EYFP.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4727)
AUTHORS Hartfield RM, Schwarz KA, Muldoon JJ, Bagheri N, Leonard JN
TITLE Multiplexing Engineered Receptors for Multiparametric Evaluation of
Environmental Ligands.
JOURNAL ACS Synth Biol. 2017 Nov 17;6(11):2042-2055. doi:
10.1021/acssynbio.6b00279. Epub 2017 Aug 23.
PUBMED 28771312
REFERENCE 2 (bases 1 to 4727)
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/acssynbio.6b00279"; journalName: "ACS Synth Biol"; date:
"2017-11-17- 17"; volume: "6"; issue: "11"; pages: "2042-2055"
FEATURES Location/Qualifiers
source 1..4727
/mol_type="other DNA"
/organism="synthetic DNA construct"
intron 1..573
/label=beta-globin intron
/note="intron from rabbit beta-globin gene"
primer_bind complement(709..728)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_signal 774..829
/label=beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(828..847)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
primer_bind complement(1220..1237)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(1391..1979)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1471..1490)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(2150..3010)
/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 2773..2792
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(3011..3115)
/gene="bla"
/label=AmpR promoter
primer_bind 3183..3201
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
polyA_signal 3546..3680
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3583..3602)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3637..3656
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
CDS complement(3705..4424)
/codon_start=1
/product="enhanced YFP"
/label=EYFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(3750..3771)
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind 4099..4118
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind complement(4532..4556)
/label=LNCX
/note="Human CMV promoter, forward primer"
protein_bind 4572..4590
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 4591..4609
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 4650..4668
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 4669..4687
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
1 gtgagtttgg ggacccttga ttgttctttc tttttcgcta ttgaaaaatt catgttatat
61 ggagggggca aagttttcag ggtgttgttt agaatgggaa gatgtccctt gtatcaccat
121 ggaccctcat gataattttg tttctttcac tttctactct gttgacaacc attgtctcct
181 cttattttct tttcattttc tgtaactttt ttcgttaaac tttagcttgc atttgtaacg
241 aatttttaaa ttcactttcg tttatttgtc agattgtaag tactttctct aatcactttt
301 ttttcaaggc aatcagggta attatattgt acttcagcac agttttagag aacaattgtt
361 ataattaaat gataaggtag aatatttctg catataaatt ctggctggcg tggaaatatt
421 cttattggta gaaacaacta catcctggta atcatcctgc ctttctcttt atggttacaa
481 tgatatacac tgtttgagat gaggataaaa tactctgagt ccaaaccggg cccctctgct
541 aaccatgttc atgccttctt ctttttccta cagctcctgg gcaacgtgct ggttgttgtg
601 ctgtctcatc attttggcaa agaattcact cctcaggtgc aggctgccta tcagaaggtg
661 gtggctggtg tggccaatgc cctggctcac aaataccact gagatcgatc tttttccctc
721 tgccaaaaat tatggggaca tcatgaagcc ccttgagcat ctgacttctg ggtaataaag
781 gaaatttatt ttcattgcaa tagtgtgtgg gaattttttg tgtctctcac tcggaaggac
841 atatgggagg gcaaatcatt taaaacatca gaatgagtat ttggtttaga gtttggcaac
901 atatgccata tgctggctgc catgaacaaa ggtggctata aagaggtcat cagtatatga
961 aacagccccc tgctgtccat tccttattcc atagaaaagc cttgacttga ggttagattt
1021 tttttatatt ttgttttgtg ttattttttt ctttaacatc cctaaaattt tccttacatg
1081 ttttactagc cagatttttc ctcctctcct gactactccc agtcatagct gtccctcttc
1141 tcttatgaac tcgactgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
1201 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
1261 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
1321 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
1381 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
1441 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
1501 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
1561 cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg
1621 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
1681 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
1741 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
1801 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
1861 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
1921 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
1981 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
2041 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
2101 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
2161 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
2221 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
2281 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
2341 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
2401 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
2461 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
2521 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
2581 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
2641 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
2701 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
2761 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
2821 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
2881 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
2941 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
3001 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
3061 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
3121 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
3181 aaaataggcg tatcacgagg ccctttcgtc ttcactcgac gctggtcgag ctgatacttc
3241 ccgtccgcca ggggacatgc cggcgatgct gaaggtcgcg cgcattcccg atgaagaggc
3301 cggttaccgc ctgttgacct ggtgggacgg gcagggcgcc gcccgagtct tgcctcggcg
3361 gcgggcgctc tgctcatgga gcgcgcgtcc ggggaccttg cacagatagc gtggtccggc
3421 cagacgacga ggcttgcagg atcataatca gccataccac atttgtagag gttttacttg
3481 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
3541 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
3601 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
3661 tatcttatca tgtctggatc ctctagaact aggtcgacaa gcttttactt gtacagctcg
3721 tccatgccga gagtgatccc ggcggcggtc acgaactcca gcaggaccat gtgatcgcgc
3781 ttctcgttgg ggtctttgct cagggcggac tggtagctca ggtagtggtt gtcgggcagc
3841 agcacggggc cgtcgccgat gggggtgttc tgctggtagt ggtcggcgag ctgcacgctg
3901 ccgtcctcga tgttgtggcg gatcttgaag ttcaccttga tgccgttctt ctgcttgtcg
3961 gccatgatat agacgttgtg gctgttgtag ttgtactcca gcttgtgccc caggatgttg
4021 ccgtcctcct tgaagtcgat gcccttcagc tcgatgcggt tcaccagggt gtcgccctcg
4081 aacttcacct cggcgcgggt cttgtagttg ccgtcgtcct tgaagaagat ggtgcgctcc
4141 tggacgtagc cttcgggcat ggcggacttg aagaagtcgt gctgcttcat gtggtcgggg
4201 tagcgggcga agcactgcag gccgtagccg aaggtggtca cgagggtggg ccagggcacg
4261 ggcagcttgc cggtggtgca gatgaacttc agggtcagct tgccgtaggt ggcatcgccc
4321 tcgccctcgc cggacacgct gaacttgtgg ccgtttacgt cgccgtccag ctcgaccagg
4381 atgggcacca ccccggtgaa cagctcctcg cccttgctca ccatactagc gcggcctcga
4441 cgatatctct agagctgaga acttcaggca ggctggatcg gtcccggtgt cttctatgga
4501 ggtcaaaaca gcgtggatgg cgtctccagg cgatctgacg gttcactaaa cgagctctgc
4561 ttataggtac ctccctatca gtgatagaga tccctatcag tgatagagac ggagtactgt
4621 cctccgcgga gtactgtcct ccgaccggct ccctatcagt gatagagatc cctatcagtg
4681 atagagacgg agtactgtcc tccgcggagt actgtcctcc gaccggt
//
| {
"Vector backbone": "pBI",
"Backbone size w/o insert (bp)": "3722",
"Vector type": "Mammalian Expression",
"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": "Multiplexing Engineered Receptors for Multiparametric Evaluation of Environmental Ligands",
"Publication": "ACS Synth Biol. 2017 Nov 17;6(11):2042-2055. doi: 10.1021/acssynbio.6b00279. Epub 2017 Aug 23.",
"DOI": "10.1021/acssynbio.6b00279.",
"Authors": "Hartfield RM, Schwarz KA, Muldoon JJ, Bagheri N, Leonard JN /acssynbiob",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/28771312",
"PubMed ID": "PubMed 28771312"
} | {
"Gene/Insert name": "U2T2U2T2-EYFP",
"Species": "Synthetic",
"Insert Size (bp)": "1029",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "CMV-min",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "AgeI (not destroyed)",
"3′ cloning site": "HindIII (not destroyed)",
"5′ sequencing primer": "gcaatagcatcacaaatttc",
"3′ sequencing primer": "cccataatttttggcagagg"
} | {
"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": ""
} |
|
pSc-His6-Sem1Sc | 83235 | pGREG600-His6-Sem1Sc, Expresses the His6-Sem1 in yeast | Gali Prag | Levin-Kravets et al Nat Methods. 2016 Nov;13(11):945-952. doi: 10.1038/nmeth.4003. Epub 2016 Oct 3. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/163610/4d902b82-096e-4c3f-bfe7-d8633af9a0a2/addgene-plasmid-83235-sequence-163610.gbk | full | LOCUS sequence-163610- 7475 bp DNA circular SYN 26-AUG-2024
DEFINITION pGREG600-His6-Sem1Sc, Expresses the His6-Sem1 in yeast.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7475)
AUTHORS Levin-Kravets O, Tanner N, Shohat N, Attali I, Keren-Kaplan T,
Shusterman A, Artzi S, Varvak A, Reshef Y, Shi X, Zucker O, Baram T,
Katina C, Pilzer I, Ben-Aroya S, Prag G
TITLE A bacterial genetic selection system for ubiquitylation cascade
discovery.
JOURNAL Nat Methods. 2016 Nov;13(11):945-952. doi: 10.1038/nmeth.4003. Epub
2016 Oct 3.
PUBMED 27694912
REFERENCE 2 (bases 1 to 7475)
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/nmeth.4003"; journalName: "Nat Methods"; date: "2016-11";
volume: "13"; issue: "11"; pages: "945-952"
FEATURES Location/Qualifiers
source 1..7475
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 75..105
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 113..129
/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 118..140
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 137..153
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 137..153
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 172..192
/label=T3
/note="T3 promoter, forward primer"
promoter 174..192
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 232..673
/gene="S. cerevisiae GAL1"
/label=GAL1 promoter
/note="inducible promoter, regulated by Gal4"
protein_bind 232..349
/label=UAS
/bound_moiety="Gal4"
/note="upstream activating sequence mediating
Gal4-dependent induction"
primer_bind 644..667
/label=GAL1
/note="S. cerevisiae GAL1 promoter, forward primer"
CDS 770..787
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 1015..1026
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
terminator 1067..1314
/gene="S. cerevisiae CYC1"
/label=CYC1 terminator
/note="transcription terminator for CYC1"
primer_bind complement(1083..1101)
/label=CYC1
/note="CYC1 transcription termination signal, reverse
primer"
protein_bind complement(1327..1360)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
gene complement(1418..2774)
/label=kanMX
/note="yeast selectable marker conferring kanamycin
resistance (Wach et al., 1994)"
terminator complement(1418..1615)
/label=TEF terminator
/note="Ashbya gossypii TEF terminator"
CDS complement(1621..2430)
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin"
/translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind 2344..2363
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
promoter complement(2431..2774)
/label=TEF promoter
/note="Ashbya gossypii TEF promoter"
protein_bind complement(2829..2862)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
primer_bind complement(2898..2917)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(2899..2917)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(2927..2944)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(2927..2943)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2936..2958)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 3088..3543
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3175..3194)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3385..3406
/label=F1ori-F
/note="F1 origin, forward primer"
CDS complement(3674..4477)
/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
YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
promoter complement(4478..4698)
/gene="S. cerevisiae URA3"
/label=URA3 promoter
primer_bind complement(4724..4743)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 4843..4865
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(4903..4921)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
misc_feature 4963..5466
/label=CEN/ARS
/note="S. cerevisiae CEN6 centromere fused to an
autonomously replicating sequence"
promoter 5503..5607
/gene="bla"
/label=AmpR promoter
CDS 5792..6169
/label=ISS
/note="immunostimulatory sequence from the AmpR gene;
contains unmethylated CpG dinucleotides in the context of
5'-AACGTT-3' (Sato et al., 1996)"
primer_bind complement(5826..5845)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 6638..7226
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 7127..7146
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 7380..7397
/label=L4440
/note="L4440 vector, forward primer"
ORIGIN
1 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta cctcactcat
61 taggcacccc aggctttaca ctttatgctt ccggctccta tgttgtgtgg aattgtgagc
121 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac
181 cctcactaaa gggaacaaaa gctggagctc gtttaaacgg cgcgcctagt acggattaga
241 agccgccgag cgggtgacag ccctccgaag gaagactctc ctccgtgcgt cctcgtcttc
301 accggtcgcg ttcctgaaac gcagatgtgc ctcgcgccgc actgctccga acaataaaga
361 ttctacaata ctagctttta tggttatgaa gaggaaaaat tggcagtaac ctggccccac
421 aaaccttcaa atgaacgaat caaattaaca accataggat gataatgcga ttagtttttt
481 agccttattt ctggggtaat taatcagcga agcgatgatt tttgatctat taacagatat
541 ataaatgcaa aaactgcata accactttaa ctaatacttt caacattttc ggtttgtatt
601 acttcttatt caaatgtaat aaaagtatca acaaaaaatt gttaatatac ctctatactt
661 taacgtcaag gagaaaaacc ccggattctg aagcggccgc actagtggat cccccgggct
721 gcaggaattc gatatcaagc ttatcgatac cgtcgacatg ggcagcagcc atcaccatca
781 tcaccacatg agtacggacg ttgctgctgc acaagcccaa agtaaaatag atttaacaaa
841 aaaaaaaaat gaggaaataa ataagaagtc tctagaggaa gacgatgaat ttgaagattt
901 ccccattgac acatgggcca atggtgaaac aataaaaagc aatgctgtaa cgcaaacaaa
961 catatgggaa gaaaactggg atgatgttga agttgacgac gatttcacca atgaattgaa
1021 ggccgaattg gatagataca aacgggaaaa tcagtgataa ctcgagtcat gtaattagtt
1081 atgtcacgct tacattcacg ccctcccccc acatccgctc taaccgaaaa ggaaggagtt
1141 agacaacctg aagtctaggt ccctatttat ttttttatag ttatgttagt attaagaacg
1201 ttatttatat ttcaaatttt tctttttttt ctgtacagac gcgtgtacgc atgtaacatt
1261 atactgaaaa ccttgcttga gaaggttttg ggacgctcga aggctttaat ttgcggccgg
1321 tacctaataa cttcgtatag catacattat acgaagttat attaagggtt ctcgacgttt
1381 tcgacactgg atggcggcgt tagtatcgaa tcgacagcag tatagcgacc agcattcaca
1441 tacgattgac gcatgatatt actttctgcg cacttaactt cgcatctggg cagatgatgt
1501 cgaggcgaaa aaaaatataa atcacgctaa catttgatta aaatagaaca actacaatat
1561 aaaaaaacta tacaaatgac aagttcttga aaacaagaat ctttttattg tcagtactga
1621 ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat
1681 accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca
1741 taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc
1801 tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac
1861 tgaatccggt gagaatggca aaagcttatg catttctttc cagacttgtt caacaggcca
1921 gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg
1981 cgcctgagcg agacgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga
2041 atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata
2101 ttcttctaat acctggaatg ctgttttgcc ggggatcgca gtggtgagta accatgcatc
2161 atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt
2221 tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa
2281 caactctggc gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac
2341 attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg
2401 cctcgaaacg tgagtctttt ccttacccat ggttgtttat gttcggatgt gatgtgagaa
2461 ctgtatccta gcaagatttt aaaaggaagt atatgaaaga agaacctcag tggcaaatcc
2521 taacctttta tatttctcta caggggcgcg gcgtggggac aattcaacgc gtctgtgagg
2581 ggagcgtttc cctgctcgca ggtctgcagc gaggagccgt aatttttgct tcgcgccgtg
2641 cggccatcaa aatgtatgga tgcaaatgat tatacatggg gatgtatggg ctaaatgtac
2701 gggcgacagt cacatcatgc ccctgagctg cgcacgtcaa gactgtcaag gagggtattc
2761 tgggcctcca tgtcgctggc cgggtgaccc ggcggggacg aggcaagcta aacagatctc
2821 tagacctaat aacttcgtat agcatacatt atacgaagtt atattaaggg ttgtcttaat
2881 taagggtacc caattcgccc tatagtgagt cgtattacgc gcgctcactg gccgtcgttt
2941 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
3001 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
3061 tgcgcagcct gaatggcgaa tggcgcgacg cgccctgtag cggcgcatta agcgcggcgg
3121 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
3181 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
3241 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
3301 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
3361 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
3421 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
3481 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa
3541 tttcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatagggta
3601 ataactgata taattaaatt gaagctctaa tttgtgagtt tagtatacat gcatttactt
3661 ataatacagt tttttagttt tgctggccgc atcttctcaa atatgcttcc cagcctgctt
3721 ttctgtaacg ttcaccctct accttagcat cccttccctt tgcaaatagt cctcttccaa
3781 caataataat gtcagatcct gtagagacca catcatccac ggttctatac tgttgaccca
3841 atgcgtctcc cttgtcatct aaacccacac cgggtgtcat aatcaaccaa tcgtaacctt
3901 catctcttcc acccatgtct ctttgagcaa taaagccgat aacaaaatct ttgtcgctct
3961 tcgcaatgtc aacagtaccc ttagtatatt ctccagtaga tagggagccc ttgcatgaca
4021 attctgctaa catcaaaagg cctctaggtt cctttgttac ttcttctgcc gcctgcttca
4081 aaccgctaac aatacctggg cccaccacac cgtgtgcatt cgtaatgtct gcccattctg
4141 ctattctgta tacacccgca gagtactgca atttgactgt attaccaatg tcagcaaatt
4201 ttctgtcttc gaagagtaaa aaattgtact tggcggataa tgcctttagc ggcttaactg
4261 tgccctccat ggaaaaatca gtcaagatat ccacatgtgt ttttagtaaa caaattttgg
4321 gacctaatgc ttcaactaac tccagtaatt ccttggtggt acgaacatcc aatgaagcac
4381 acaagtttgt ttgcttttcg tgcatgatat taaatagctt ggcagcaaca ggactaggat
4441 gagtagcagc acgttcctta tatgtagctt tcgacatgat ttatcttcgt ttcctgcagg
4501 tttttgttct gtgcagttgg gttaagaata ctgggcaatt tcatgtttct tcaacactac
4561 atatgcgtat atataccaat ctaagtctgt gctccttcct tcgttcttcc ttctgttcgg
4621 agattaccga atcaaaaaaa tttcaaagaa accgaaatca aaaaaaagaa taaaaaaaaa
4681 atgatgaatt gaattgaaaa gctgtggtat ggtgcactct cagtacaatc tgctctgatg
4741 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
4801 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
4861 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
4921 ttttataggt taatgtcatg ataataatgg tttcttagga cggatcgctt gcctgtaact
4981 tacacgcgcc tcgtatcttt taatgatgga ataatttggg aatttactct gtgtttattt
5041 atttttatgt tttgtatttg gattttagaa agtaaataaa gaaggtagaa gagttacgga
5101 atgaagaaaa aaaaataaac aaaggtttaa aaaatttcaa caaaaagcgt actttacata
5161 tatatttatt agacaagaaa agcagattaa atagatatac attcgattaa cgataagtaa
5221 aatgtaaaat cacaggattt tcgtgtgtgg tcttctacac agacaagatg aaacaattcg
5281 gcattaatac ctgagagcag gaagagcaag ataaaaggta gtatttgttg gcgatccccc
5341 tagagtcttt tacatcttcg gaaaacaaaa actatttttt ctttaatttc tttttttact
5401 ttctattttt aatttatata tttatattaa aaaatttaaa ttataattat ttttatagca
5461 cgtgatgaaa aggacccagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
5521 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
5581 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
5641 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
5701 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
5761 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
5821 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
5881 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
5941 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
6001 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
6061 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctaat gaagccatac
6121 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
6181 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
6241 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
6301 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
6361 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
6421 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
6481 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
6541 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
6601 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
6661 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
6721 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
6781 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
6841 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
6901 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
6961 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
7021 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
7081 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
7141 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
7201 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
7261 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
7321 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
7381 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
7441 gttggccgat tcattaatgc agctggcacg acagg
//
| {
"Vector backbone": "pGREG600",
"Backbone size w/o insert (bp)": "7176",
"Vector type": "Yeast Expression",
"Selectable markers": "Neomycin (select with G418), URA3"
} | {
"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": "A bacterial genetic selection system for ubiquitylation cascade discovery",
"Publication": "Nat Methods. 2016 Nov;13(11):945-952. doi: 10.1038/nmeth.4003. Epub 2016 Oct 3.",
"DOI": "10.1038/nmeth.4003.",
"Authors": "Levin-Kravets O, Tanner N, Shohat N, Attali I, Keren-Kaplan T, Shusterman A, Artzi S, Varvak A, Reshef Y, Shi X, Zucker O, Baram T, Katina C, Pilzer I, Ben-Aroya S, Prag G /nmeth",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/27694912",
"PubMed ID": "PubMed 27694912"
} | {
"Gene/Insert name": "Sem 1",
"Species": "S. cerevisiae (budding yeast)",
"Insert Size (bp)": "270",
"Mutation": "",
"Entrez Gene": "SEM1",
"Promoter": "",
"Tag / Fusion Protein": "His6 (N terminal on insert)"
} | {
"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": ""
} |
|
pMK-PpsbA (-42 to +9) | 123480 | MoChlo kit | Occhialini et al Plant Physiol. 2019 Jan 24. pii: pp.18.01220. doi: 10.1104/pp.18.01220. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/247677/3f7b8466-97fe-4e1f-9c80-122a0acf9de3/addgene-plasmid-123480-sequence-247677.gbk | full | LOCUS sequence-247677- 2421 bp DNA circular SYN 26-AUG-2024
DEFINITION MoChlo kit.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2421)
AUTHORS Occhialini A, Piatek A, Pfotenhauer AC, Frazier TP, Stewart CN,
Lenaghan S
TITLE MoChlo: A versatile modular cloning toolbox for chloroplast
biotechnology.
JOURNAL Plant Physiol. 2019 Jan 24. pii: pp.18.01220. doi:
10.1104/pp.18.01220.
PUBMED 30679266
REFERENCE 2 (bases 1 to 2421)
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: "Plant
Physiol. 2019 Jan 24. pii: pp.18.01220. doi: 10.1104/pp.18.01220."
FEATURES Location/Qualifiers
source 1..2421
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 278..300
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 292..309
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 293..309
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 320..339
/label=T7
/note="T7 promoter, forward primer"
promoter 320..338
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 343..354
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(627..644)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(737..1325)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(817..836)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(1499..2293)
/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"
promoter complement(2294..2398)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
121 gatagggttg agtggccgct acagggcgct cccattcgcc attcaggctg cgcaactgtt
181 gggaagggcg tttcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
241 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
301 acggccagtg agcgcgacgt aatacgactc actatagggc gaattgaagg aaggccgtca
361 aggccacgtg tcttgtccag agctcggtct caggagaata aaaagccttc cattttctat
421 tttgatttgt agaaaactag tgtgcttggg agtccctgat gattaaataa accaagattt
481 tacctacttg agaccggtac ctggagcaca agactggcct catgggcctt cctttcactg
541 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aacatggtca tagctgtttc
601 cttgcgtatt gggcgctctc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
661 gtaaagcctg gggtgcctaa tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
721 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
781 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
841 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
901 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
961 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
1021 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
1081 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
1141 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
1201 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
1261 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
1321 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
1381 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
1441 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttatt
1501 agaaaaattc atccagcaga cgataaaacg caatacgctg gctatccggt gccgcaatgc
1561 catacagcac cagaaaacga tccgcccatt cgccgcccag ttcttccgca atatcacggg
1621 tggccagcgc aatatcctga taacgatccg ccacgcccag acggccgcaa tcaataaagc
1681 cgctaaaacg gccattttcc accataatgt tcggcaggca cgcatcacca tgggtcacca
1741 ccagatcttc gccatccggc atgctcgctt tcagacgcgc aaacagctct gccggtgcca
1801 ggccctgatg ttcttcatcc agatcatcct gatccaccag gcccgcttcc atacgggtac
1861 gcgcacgttc aatacgatgt ttcgcctgat gatcaaacgg acaggtcgcc gggtccaggg
1921 tatgcagacg acgcatggca tccgccataa tgctcacttt ttctgccggc gccagatggc
1981 tagacagcag atcctgaccc ggcacttcgc ccagcagcag ccaatcacgg cccgcttcgg
2041 tcaccacatc cagcaccgcc gcacacggaa caccggtggt ggccagccag ctcagacgcg
2101 ccgcttcatc ctgcagctcg ttcagcgcac cgctcagatc ggttttcaca aacagcaccg
2161 gacgaccctg cgcgctcaga cgaaacaccg ccgcatcaga gcagccaatg gtctgctgcg
2221 cccaatcata gccaaacaga cgttccaccc acgctgccgg gctacccgca tgcaggccat
2281 cctgttcaat catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
2341 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
2401 catttccccg aaaagtgcca c
//
| {
"Vector backbone": "pMK-T",
"Backbone size w/o insert (bp)": "",
"Vector type": "Synthetic Biology",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Kanamycin, 50 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "Top10",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "MoChlo: A versatile modular cloning toolbox for chloroplast biotechnology",
"Publication": "Plant Physiol. 2019 Jan 24. pii: pp.18.01220. doi: 10.1104/pp.18.01220.",
"DOI": "10.1104/pp.18.01220.",
"Authors": "Occhialini A, Piatek A, Pfotenhauer AC, Frazier TP, Stewart CN, Lenaghan S /pp",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/30679266",
"PubMed ID": "PubMed 30679266"
} | {
"Gene/Insert name": "PpsbA (-42 to +9)",
"Species": "",
"Insert Size (bp)": "",
"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": ""
} |
||
FER gRNA (BRDN0001146110) | 76802 | 3rd generation lentiviral gRNA plasmid targeting human FER | Doench et al Nat Biotechnol. 2016 Jan 18. doi: 10.1038/nbt.3437. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/186534/fe8767d3-a421-4169-a5f8-ce527776972a/addgene-plasmid-76802-sequence-186534.gbk | full | LOCUS sequence-186534- 8323 bp DNA circular SYN 26-AUG-2024
DEFINITION 3rd generation lentiviral gRNA plasmid targeting human FER.
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 cggataacca gtcttgtcac aggttttaga 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": "FER",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "FER",
"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": ""
} |
||
pRAJ31m | 39256 | Alfonso Jaramillo | Rodrigo et al Proc Natl Acad Sci U S A. 2012 Sep 18;109(38):15271-6. Epub 2012 Sep 4. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/49421/d24c48f4-41c6-4379-a33b-6627ca3c3c83/addgene-plasmid-39256-sequence-49421.gbk | full | LOCUS sequence-49421-e 4976 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 4976)
AUTHORS Rodrigo G, Landrain TE, Jaramillo A
TITLE De novo automated design of small RNA circuits for engineering
synthetic riboregulation in living cells.
JOURNAL Proc Natl Acad Sci U S A. 2012 Sep 18;109(38):15271-6. Epub 2012 Sep
4.
PUBMED 22949707
REFERENCE 2 (bases 1 to 4976)
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: "2012-09-18"; volume: "109(38)"; pages:
"15271-6. Epub 2012 Sep 4"
FEATURES Location/Qualifiers
source 1..4976
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 1..22
/label=BioBrick prefix
/note="BioBrick prefix for parts that do not start with
""ATG"""
CDS complement(26..703)
/codon_start=1
/product="monomeric derivative of DsRed (Campbell et al.,
2002)"
/label=mRFP1
/translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
TGA"
RBS 710..721
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
protein_bind 773..789
/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)."
RBS 836..847
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 863..1573
/codon_start=1
/product="GFP variant that folds robustly even when fused
to poorly folded proteins (Pedelacq et al., 2006)"
/label=superfolder GFP
/translation="RKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
ICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDGT
YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF
VTAAGITHGMDELYK"
misc_feature 1580..1600
/label=BioBrick suffix
/note="universal suffix for all parts"
terminator 1636..1667
/label=tonB terminator
/note="bidirectional E. coli tonB-P14 transcription
terminator"
CDS complement(1766..2581)
/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"
primer_bind 2489..2508
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
primer_bind 2573..2592
/label=pENTR-R
/note="pENTR vectors, reverse primer"
CDS complement(2939..3889)
/codon_start=1
/gene="rep101"
/product="RepA protein needed for replication with the
pSC101 origin"
/label=Rep101
/translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
rep_origin 3937..4159
/label=pSC101 ori
/note="low-copy replication origin that requires the Rep101
protein"
terminator complement(4651..4745)
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
terminator complement(4901..4930)
/label=T3Te terminator
/note="phage T3 early transcription terminator"
ORIGIN
1 gaattcgcgg ccgcttctag agttattaag caccggtgga gtgacgacct tcagcacgtt
61 cgtactgttc aacgatggtg tagtcttcgt tgtgggaggt gatgtccagt ttgatgtcgg
121 ttttgtaagc acccggcagc tgaaccggtt ttttagccat gtaggtggtt ttaacttcag
181 cgtcgtagtg accaccgtct ttcagtttca gacgcatttt gatttcacct ttcagagcac
241 cgtcttccgg gtacatacgt tcggtggaag cttcccaacc catggttttt ttctgcataa
301 ccggaccgtc ggacgggaag ttggtaccac gcagtttaac tttgtagatg aactcaccgt
361 cttgcaggga ggagtcctgg gtaacggtaa caacaccacc gtcttcgaag ttcataacac
421 gttcccattt gaaaccttcc gggaaggaca gtttcaggta gtccgggatg tcagccgggt
481 gtttaacgta agctttggaa ccgtactgga actgcgggga caggatgtcc caagcgaacg
541 gcagcggacc acctttggta actttcagtt tagcggtctg ggtaccttcg tacggacgac
601 cttcaccttc accttcgatt tcgaactcgt gaccgttaac ggaaccttcc atacgaactt
661 tgaaacgcat gaactctttg ataacgtctt cggaggaagc catctagtat ttctcctctt
721 tctctagtgg atccgctagg atgcatataa atgtgagcgg ataacattga cattgtgagc
781 ggataacaag atactgagca cagtggatat gctttacctc ttggtgagac gacataaaga
841 ggagaaacat aatatgacta gccgtaaagg cgaagagctg ttcactggtg tcgtccctat
901 tctggtggaa ctggatggtg atgtcaacgg tcataagttt tccgtgcgtg gcgagggtga
961 aggtgacgca actaatggta aactgacgct gaagttcatc tgtactactg gtaaactgcc
1021 ggtaccttgg ccgactctgg taacgacgct gacttatggt gttcagtgct ttgctcgtta
1081 tccggaccat atgaagcagc atgacttctt caagtccgcc atgccggaag gctatgtgca
1141 ggaacgcacg atttccttta aggatgacgg cacgtacaaa acgcgtgcgg aagtgaaatt
1201 tgaaggcgat accctggtaa accgcattga gctgaaaggc attgacttta aagaagacgg
1261 caatatcctg ggccataagc tggaatacaa ttttaacagc cacaatgttt acatcaccgc
1321 cgataaacaa aaaaatggca ttaaagcgaa ttttaaaatt cgccacaacg tggaggatgg
1381 cagcgtgcag ctggctgatc actaccagca aaacactcca atcggtgatg gtcctgttct
1441 gctgccagac aatcactatc tgagcacgca aagcgttctg tctaaagatc cgaacgagaa
1501 acgcgatcat atggttctgc tggagttcgt aaccgcagcg ggcatcacgc atggtatgga
1561 tgaactgtac aaatgatgat actagtagcg gccgctgcag gagtcactaa gggttagtta
1621 gttagattag cagaaagtca aaagcctccg accggaggct tttgactaaa acttcccttg
1681 gggttatcat tggggctcac tcaaaggcgg taatcagata aaaaaaatcc ttagctttcg
1741 ctaaggatga tttctgctag tattattaga aaaactcatc gagcatcaaa tgaaactgca
1801 atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag
1861 gagaaaactc accgaggcag ttccaaagaa tggcaaggtc ctggtaacgg tctgcgattc
1921 cgacccgtcc aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa
1981 gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaagagc ttgtgcattt
2041 ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa
2101 ccaaaccgtt attcatgcgt gattgcgcct gagcaagacg aaatacacga tcgctgttaa
2161 aaggacaatt acaaacagga atcgaatgta accggcgcag gaacacggcc agcgcatcaa
2221 caatattttc acctgaatca ggatattctt ctaatacctg gaaggctgtt ttcccaggaa
2281 tcgcggtggt gagtaaccac gcatcatcag gagtacggat aaaatgcttg atggtcggga
2341 gaggcataaa ctccgtcagc cagttgagac ggaccatctc atctgtaaca tcattggcaa
2401 cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaagcgat
2461 agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag
2521 cgtccatgtt ggagtttaag cgcggacggg agcaagacgt ttcccgttga atatggctca
2581 taacacccct tgtattactg tttatgtaag cagacagttt tattgttcat gatgatatat
2641 ttttatcttg tgcaatgtaa catcagagat tttgagacac aacgtggctt tgttgaataa
2701 atcgaacttt tgctgagttg aaggatcagc tctagtagtt acattgtcga tctgttcatg
2761 gtgaacagct ttaaatgcac caaaaactcg taaaagctct gatgtatcta tcttttttac
2821 accgttttca tctgtgcata tggacagttt tccctttgat atctaacggt gaacagttgt
2881 tctacttttg tttgttagtc ttgatgcttc actgatagat acaagagcca taagaacctc
2941 agatccttcc gtatttagcc agtatgttct ctagtgtggt tcgttgtttt tgcgtgagcc
3001 atgagaacga accattgaga tcatgcttac tttgcatgtc actcaaaaat tttgcctcaa
3061 aactggtgag ctgaattttt gcagttaaag catcgtgtag tgtttttctt agtccgttac
3121 gtaggtagga atctgatgta atggttgttg gtattttgtc accattcatt tttatctggt
3181 tgttctcaag ttcggttacg agatccattt gtctatctag ttcaacttgg aaaatcaacg
3241 tatcagtcgg gcggcctcgc ttatcaacca ccaatttcat attgctgtaa gtgtttaaat
3301 ctttacttat tggtttcaaa acccattggt taagcctttt aaactcatgg tagttatttt
3361 caagcattaa catgaactta aattcatcaa ggctaatctc tatatttgcc ttgtgagttt
3421 tcttttgtgt tagttctttt aataaccact cataaatcct catagagtat ttgttttcaa
3481 aagacttaac atgttccaga ttatatttta tgaatttttt taactggaaa agataaggca
3541 atatctcttc actaaaaact aattctaatt tttcgcttga gaacttggca tagtttgtcc
3601 actggaaaat ctcaaagcct ttaaccaaag gattcctgat ttccacagtt ctcgtcatca
3661 gctctctggt tgctttagct aatacaccat aagcattttc cctactgatg ttcatcatct
3721 gagcgtattg gttataagtg aacgataccg tccgttcttt ccttgtaggg ttttcaatcg
3781 tggggttgag tagtgccaca cagcataaaa ttagcttggt ttcatgctcc gttaagtcat
3841 agcgactaat cgctagttca tttgctttga aaacaactaa ttcagacata catctcaatt
3901 ggtctaggtg attttaatca ctataccaat tgagatgggc tagtcaatga taattacatg
3961 tccttttcct ttgagttgtg ggtatctgta aattctgcta gacctttgct ggaaaacttg
4021 taaattctgc tagaccctct gtaaattccg ctagaccttt gtgtgttttt tttgtttata
4081 ttcaagtggt tataatttat agaataaaga aagaataaaa aaagataaaa agaatagatc
4141 ccagccctgt gtataactca ctactttagt cagttccgca gtattacaaa aggatgtcgc
4201 aaacgctgtt tgctcctcta caaaacagac cttaaaaccc taaaggctta agtagcaccc
4261 tcgcaagctc gggcaaatcg ctgaatattc cttttgtctc cgaccatcag gcacctgagt
4321 cgctgtcttt ttcgtgacat tcagttcgct gcgctcacgg ctctggcagt gaatgggggt
4381 aaatggcact acaggcgcct tttatggatt catgcaagga aactacccat aatacaagaa
4441 aagcccgtca cgggcttctc agggcgtttt atggcgggtc tgctatgtgg tgctatctga
4501 ctttttgctg ttcagcagtt cctgccctct gattttccag tctgaccact tcggattatc
4561 ccgtgacagg tcattcagac tggctaatgc acccagtaag gcagcggtat catcaacagg
4621 cttacccgtc ttactgtccc tagtgcttgg attctcacca ataaaaaacg cccggcggca
4681 accgagcgtt ctgaacaaat ccagatggag ttctgaggtc attactggat ctatcaacag
4741 gagtccaagc gagctcgtaa acttggtctg acagctctag ctccggcaaa aaaacgggca
4801 aggtgtcacc accctgccct ttttctttaa aaccgaaaag attacttcgc gtttgccacc
4861 tgacgtctaa gaaaaggaat attcagcaat ttgcccgtgc cgaagaaagg cccacccgtg
4921 aaggtgagcc agtgagttga ttgctacgta attagttagt tagcccttag tgactc
//
| {
"Vector backbone": "pSTC1",
"Backbone size w/o insert (bp)": "4860",
"Vector type": "Bacterial Expression, Synthetic Biology",
"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": "De novo automated design of small RNA circuits for engineering synthetic riboregulation in living cells",
"Publication": "Proc Natl Acad Sci U S A. 2012 Sep 18;109(38):15271-6. Epub 2012 Sep 4.",
"DOI": "10.1073/pnas.1203831109",
"Authors": "Rodrigo G, Landrain TE, Jaramillo A",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/22949707",
"PubMed ID": "PubMed 22949707"
} | {
"Gene/Insert name": "RAJ31 system (no transRNA)",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "pLlacO-1",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "NheI (destroyed during cloning)",
"3′ cloning site": "AvrII (destroyed during cloning)",
"5′ sequencing primer": "tgccacctgacgtctaagaa",
"3′ sequencing primer": "attaccgcctttgagtgagc"
} | {
"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": ""
} |
||
mEmerald-Inversin-C-18 | 54131 | Low | Localization: Cilium, Excitation: 487, Emission: 509 | Michael Davidson | Davidson mEmerald (unpublished) | https://media.addgene.org/snapgene-media/v2.0.0/sequences/91797/4abf02c1-0fc3-46a6-b3b4-414b027e291a/addgene-plasmid-54131-sequence-91797.gbk | partial | LOCUS sequence-91797-e 852 bp DNA linear UNA 26-AUG-2024
DEFINITION Localization: Cilium, Excitation: 487, Emission: 509.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE natural DNA sequence
ORGANISM unspecified
REFERENCE 1 (bases 1 to 852)
TITLE Davidson mEmerald
REFERENCE 2 (bases 1 to 852)
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..852
/mol_type="genomic DNA"
/organism="unspecified"
ORIGIN
1 gacgagctgt acaagtccgg aagtgctggc agcggtggag gcagcgcatc cggcggaggc
61 agcggaagca acaagtcaga gaacctgctg tttgctggtt catcattagc atcacaagtc
121 catgctgctg ccgttaatgg agataagggt gctctacaga ggctcatcgt aggaaactct
181 gctcttaaag acaaagaaga tcagtttggg agaacaccac ttatgtattg cgtgttggct
241 gacagattgg attgtgcaga tgctcttctg aaggcaggag cagatgtgaa taaaactgac
301 catagccaga gaacagccct ccatcttgca gcccagaagg gaaattatcg tttcatgaaa
361 ctcttactta cacgcagagc aaactggatg caaaaggatc tggaagagat gactcctttg
421 cacttgacca cccggcacag gagccctaag tgtttggcac ttctgctgaa gtttatggca
481 ccaggagaag tggatacaca ggataaaaac aagcaaacag ctctgcattg gagtgcctac
541 tacaataacc ctgagcatgt gaagctgctc atcaagcatg attctaacat tgggattcct
601 gatgttgaag gcaagatccc acttcactgg gcagccaacc ataaagatcc aagtgctgtt
661 cacacagtga gatgcattct ggatgctgct ccaacagagt ctttactgaa ctggcaagac
721 tacgagggtc gaactcctct tcactttgca gttgctgatg ggaatgtgac cgtggttgat
781 gtcttgacct catatgaaag ctgcaatata acgtcttatg ataacttatt tcgaacccca
841 ctgcactggg ca
//
| {
"Vector backbone": "mEmerald-C1",
"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": "Inversin",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "INVS",
"Promoter": "CMV",
"Tag / Fusion Protein": "mEmerald (N 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": ""
} |
pMC-6-R-9 | 197726 | Phytobrick (MoClo) Level 0 Part | Daniel Voytas | Chamness et al Plant Genome. 2023 Mar 9:e20312. doi: 10.1002/tpg2.20312. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/391076/3188b191-c1f8-4e44-9dc7-5b548cf5a7fe/addgene-plasmid-197726-sequence-391076.gbk | full | LOCUS sequence-391076- 3743 bp DNA circular SYN 27-AUG-2024
DEFINITION Phytobrick (MoClo) Level 0 Part.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3743)
AUTHORS Chamness JC, Kumar J, Cruz AJ, Rhuby E, Holum MJ, Cody JP, Tibebu R,
Gamo ME, Starker CG, Zhang F, Voytas DF
TITLE An extensible vector toolkit and parts library for advanced
engineering of plant genomes.
JOURNAL Plant Genome. 2023 Mar 9:e20312. doi: 10.1002/tpg2.20312.
PUBMED 36896468
REFERENCE 2 (bases 1 to 3743)
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: "Plant
Genome. 2023 Mar 9:e20312. doi: 10.1002/tpg2.20312."
FEATURES Location/Qualifiers
source 1..3743
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 1561..1581
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
primer_bind complement(1623..1641)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 1709..1813
/gene="bla"
/label=AmpR promoter
CDS 2035..2826
/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 2997..3585
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3486..3505
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind join(3739..3743,1..13)
/label=L4440
/note="L4440 vector, forward primer"
ORIGIN
1 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
61 gccgattcat taatcactct gtggtctcaa atggctctca ccaaagacac agaaattagt
121 acaataaacc gtcaaatgtc agacttcagc gagctatcac agatcttacc actccaccaa
181 ataagcaaaa tcaaagatat actggagaac gaaaatccac ttccaaaaga gaagcttgcc
241 agccacctca ctatgattat actaatggca aaccttgcat cccagaagcg aaaagatgtt
301 ccagtgaaaa ggagcacatt tttgaaatat cagcgaagta tcagtaaaac actgcagtat
361 gatagctcca ccaaaactgt ctccttcgag tatcacttga aagatccatc caaattgatt
421 aagggcctag aagatgtagt aagcccttat aggttcgtgg ttggggtaca cgaaaaaccc
481 gacgatgtaa tgtcccatct cagtgctgtc cacatgcgaa aggaagcagg acgtaagagg
541 gatctgggga ataagatcaa tgatgaaata actaaaatag cagagacaca agagactatt
601 tggggattcg tgggcaaaac tatggacctt atcgaggcac gaacaacccg accaacaact
661 aaggcagctt ataacctttt gttacaagcc acattcatga actgttgtag agctgatgac
721 ctcaaaaaca cagatattaa gacttttgag gtaatccccg ataaacacct gggaaggatg
781 ctacgtgcat tcgtgcccga aaccaaaacc ggtaccaggt tcgtatattt cttcccttgc
841 aaaggaaggt gtgatcctct cttggccctg gatagctact tacagtggac cgaccccatc
901 cccaaaacca gaactaccga tgaagatgca cgatatgatt accagctctt gagaaatagt
961 ttgcttggca gttacgatgg ctttatatcc aagcagagcg acgagtctat ttttaagatc
1021 cccaacgggc ctaaagctca ccttggcagg cacgtaacag catcatactt gagtaacaac
1081 gagatggata aggaggccac tttatacggg aattggtcag ccgcacgtga agagggtgtg
1141 agtcgagttg ccaaagctcg ttatatgcat accatcgaga aatccccccc aagctatttg
1201 ttcgcatttt tgagcgggtt ttacaatatc actgctgaaa gggcttgcga gttagtagac
1261 ccaaacagca acccctgcga acaagacaag aatatcccca tgatttctga catagagaca
1321 ctaatggcac gatacggtaa aaacgccgag attattccta tggacgttct cgtattcctg
1381 agttcatacg caaggttcaa gaataacgaa ggtaaggagt ataaactcca agctcgttca
1441 agtcgtggag ttccagattt tccagataat ggtagaactg ccctgtataa tgccttaact
1501 gccgcacacg taaaaagaag gaaaatatca atcgtggtcg gtaggtctat tgacacttca
1561 ccaaagaaga agcgaaaagt atagctttga gaccacgaag tggctcttca gtggacgaaa
1621 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
1681 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
1741 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
1801 aaaaaggaag agtatgcgct cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg
1861 taacggcgca gtggcggttt tcatggcttg ttatgactgt ttttttttgg ggtacagtct
1921 atgcctcggg catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg
1981 agcagcaacg atgttacgca gcagggcagt cgccctaaaa caaagttaaa catcatgagg
2041 gaagcggtga tcgccgaagt atcgactcaa ctatcagagg tagttggcgt catcgagcgc
2101 catctcgaac cgacgttgct ggccgtacat ttgtacggct ccgcagtgga tggcggcctg
2161 aagccacaca gcgatattga tttgctggtt acggtgaccg taaggcttga tgaaacaacg
2221 cggcgagctt tgatcaacga ccttttggaa acttcggctt cccctggaga gagcgagatt
2281 ctccgcgctg tagaagtcac cattgttgtg cacgacgaca tcattccgtg gcgttatcca
2341 gctaagcgcg aactgcaatt tggagaatgg cagcgcaatg acattcttgc aggtatcttc
2401 gagccagcca cgatcgacat tgatctggct atcttgctga caaaagcaag agaacatagc
2461 gttgccttgg taggtccagc ggcggaggaa ctctttgatc cggttcctga acaggatcta
2521 tttgaggcgc taaatgaaac cttaacgcta tggaactcgc cgcccgactg ggctggcgat
2581 gagcgaaatg tagtgcttac gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc
2641 gcgccgaagg atgtcgctgc cgactgggca atggagcgcc tgccggccca gtatcagccc
2701 gtcatacttg aagctagaca ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc
2761 gcagatcagt tggaagaatt tgtccattac gtaaaaggcg agatcaccaa ggtagtcggc
2821 aaataactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
2881 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
2941 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
3001 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
3061 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
3121 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
3181 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
3241 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
3301 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
3361 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
3421 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
3481 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
3541 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
3601 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
3661 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
3721 agccgaacga ccgagcgcag cga
//
| {
"Vector backbone": "pMC-6-ccdB-9",
"Backbone size w/o insert (bp)": "",
"Vector type": "Plant Expression",
"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": "An extensible vector toolkit and parts library for advanced engineering of plant genomes",
"Publication": "Plant Genome. 2023 Mar 9:e20312. doi: 10.1002/tpg2.20312.",
"DOI": "10.1002/tpg2.20312.",
"Authors": "Chamness JC, Kumar J, Cruz AJ, Rhuby E, Holum MJ, Cody JP, Tibebu R, Gamo ME, Starker CG, Zhang F, Voytas DF /tpg",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/36896468",
"PubMed ID": "PubMed 36896468"
} | {
"Gene/Insert name": "R recombinase (tomato optimized)",
"Species": "Zygosaccharomyces rouxii",
"Insert Size (bp)": "1494",
"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": ""
} |
|
pQE-WT-CTGA-R13-AKSRV | 63622 | CTGA-R13-AKSRV RNAP mutant | Andrew Ellington | Meyer et al ACS Synth Biol. 2014 Oct 7. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/105987/77c2c6bb-8bc9-4c06-8b79-91fb20bc9d58/addgene-plasmid-63622-sequence-105987.gbk | full | LOCUS sequence-105987- 7154 bp DNA circular SYN 26-AUG-2024
DEFINITION CTGA-R13-AKSRV RNAP mutant.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7154)
AUTHORS Meyer AJ, Ellefson JW, Ellington AD
TITLE Directed Evolution of a Panel of Orthogonal T7 RNA Polymerase
Variants for in Vivo or in Vitro Synthetic Circuitry.
JOURNAL ACS Synth Biol. 2014 Oct 7.
PUBMED 25279711
REFERENCE 2 (bases 1 to 7154)
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. 2014 Oct 7."
FEATURES Location/Qualifiers
source 1..7154
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 10..54
/label=T5 promoter
/note="bacteriophage T5 promoter for E. coli RNA
polymerase, with embedded lac operator"
protein_bind 30..46
/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)."
protein_bind 62..78
/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)."
RBS 97..108
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 127..144
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
terminator 2821..2915
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
CDS 2959..3618
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind complement(3025..3044)
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
terminator 3683..3769
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
CDS complement(3862..4944)
/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 4906..4925
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(4945..5022)
/gene="lacI (mutant)"
/label=lacIq promoter
/note="In the lacIq allele, a single base change in the
promoter boosts expression of the lacI gene about 10-fold."
primer_bind complement(5106..5128)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
misc_feature 5214..5354
/label=bom
/note="basis of mobility region from pBR322"
primer_bind 5269..5288
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(5369..5386)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5540..6128)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5620..5639)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6235..7050)
/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(6235..7050)
/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 6958..6977
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
primer_bind 7042..7061
/label=pENTR-R
/note="pENTR vectors, reverse primer"
primer_bind 7122..7140
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
ORIGIN
1 ctcgagaaat cataaaaaat ttatttgctt tgtgagcgga taacaattat aatagattca
61 attgtgagcg gataacaatt tcacacagaa ttcattaaag aggagaaatt aactatgaga
121 ggatcgcatc accatcacca tcacggatcc aacacgatta acatcgctaa gaacgacttc
181 tctgacatcg aactggctgc tatcccgttc aacactctgg ctgaccatta cggtgagcgt
241 ttagctcgcg aacagttggc ccttgagcat gagtcttacg agatgggtga agcacgcttc
301 cgcaagatgt ttgagcgtca acttaaagct ggtgaggttg cggataacgc tgccgccaag
361 cctctcatca ctaccctact ccctaagatg attgcacgca tcaacgactg gtttgaggaa
421 gtgaaagcta agcgcggcaa gcgcccgaca gccttccagt tcctgcaaga aatcaagccg
481 gaagccgtag cgtacatcac cattaagacc actctggctt gcctaaccag tgctgacaat
541 acaaccgttc aggctgtagc aagcgcaatc ggtcgggcca ttgaggacga ggctcgcttc
601 ggtcgtatcc gtgaccttga agctaagcac ttcaagaaaa acgttgagga acaactcaac
661 aagcgcgtag ggcacgtcta caagaaagca tttatgcaag ttgtcgaggc tgacatgctc
721 tctaagggtc tactcggtgg cgaggcgtgg tcttcgtggc ataaggaaga ctctattcat
781 gtaggagtac gctgcatcga gatgctcatt gagtcaaccg gaatggttag cttacaccgc
841 caaaatgctg gcgtagtagg tcaagactct gagactatcg aactcgcacc tgaatacgct
901 gaggctatcg caacccgtgc aggtgcgctg gctggcatct ctccgatgtt ccaaccttgc
961 gtagttcctc ctaagccgtg gactggcatt actggtggtg gctattgggc taacggtcgt
1021 cgtcctctgg cgctggtgcg tactcacagt aagaaagcac tgatgcgcta cgaagacgtt
1081 tacatgcctg aggtgtacaa agcgattaac attgcgcaaa acaccgcatg gaaaatcaac
1141 aagaaagtcc tagcggtcgc caacgtaatc accaagtgga agcattgtcc ggtcgaggac
1201 atccctgcga ttgagcgtga agaactcccg atgaaaccgg aagacatcga catgaatcct
1261 gaggctctca ccgcgtggaa acgtgctgcc gctgctgtgt accgcaagga caaggctcgc
1321 aagtctcgcc gtatcagcct tgagttcatg cttgagcaag ccaataagtt tgctaaccat
1381 aaggccatct ggttccctta caacatggac tggcgcggtc gtgtttacgc tgtgtcaatg
1441 ttcaacccgc aaggtaacga tatgaccaaa ggactgctta cgctggcgaa aggtaaacca
1501 atcggtaagg aaggttacta ctggctgaaa atccacggtg caaactgtgc gggtgtcgat
1561 aaggttccgt tccctgagcg catcaagttc attgaggaaa accacgagaa catcatggct
1621 tgcgctaagt ctccactgga gaacacttgg tgggctgagc aagattctcc gttctgcttc
1681 cttgcattct gctttgagta cgctggggta cagcaccacg gcctgagcta taactgctcc
1741 cttccgctgg cgtttgacgg gtcttgctct ggcatccagc acttctccgc gatgctccga
1801 gatgaggtag gtggtcgcgc ggttaacttg cttcctagtg aaaccgttca ggacatctac
1861 gggattgttg ctaagaaagt caacgagatt ctacaagcag acgcaatcaa tgggaccgat
1921 aacgaagtag ttaccgtgac cgatgagaac actggtgaaa tctctgagaa agtcaagctg
1981 ggcactaagg cactggctgg tcaatggctg gcttacggtg ttactcgcag tgtgactaag
2041 cgttcagtca tgacgctggc ttacgggtcc aaagagttcg gcttccgtca acaagtgctg
2101 gaagatacca ttcagccagc tattgattcc ggcaagggtc tgatgttcac tcagccgaat
2161 caggctgctg gatacatggc taagctgatt tgggaatctg tgagcgtgac ggtggtagct
2221 gcggttgaag caatgaactg gcttaagtct gctgctaagc tgctggctgc tgaggtcaaa
2281 gataagaaga ctggagagat tcttcgcaag cgttgcgctg cgcattgggt aactcctgat
2341 ggtttccctg tgtggcagga atacaagaag cctattaaaa cgcgcatctc tctgatgttc
2401 ctcggtcagt tcaccttaaa gcctaccatt aacaccaaca aagatagcga gattgatgca
2461 cgcaaacagg tgtctggtat cgctcctaac tttgtacaca gccaagacgg tagccacctt
2521 cgtaagactg tagtgtgggc acacgagaag tacggaatcg aatcttttgc actgattcac
2581 gactccttcg gtaccattcc ggctgacgct gcgaacctgt tcaaagcagt gcgcgaaact
2641 atggttgaca catatgagtc ttgtgatgta ctggctgatt tctacgacca gttcgctgac
2701 cagttgcacg agtctcaatt ggacaaaatg ccagcacttc cggctaaagg taacttgaac
2761 ctccgtgaca tcttagagtc ggacttcgcg ttcgcgtaaa agcttaatta gctgagcttg
2821 gactcctgtt gatagatcca gtaatgacct cagaactcca tctggatttg ttcagaacgc
2881 tcggttgccg ccgggcgttt tttattggtg agaatccaag ctagcttggc gagattttca
2941 ggagctaagg aagctaaaat ggagaaaaaa atcactggat ataccaccgt tgatatatcc
3001 caatggcatc gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg tacctataac
3061 cagaccgttc agctggatat tacggccttt ttaaagaccg taaagaaaaa taagcacaag
3121 ttttatccgg cctttattca cattcttgcc cgcctgatga atgctcatcc ggaatttcgt
3181 atggcaatga aagacggtga gctggtgata tgggatagtg ttcacccttg ttacaccgtt
3241 ttccatgagc aaactgaaac gttttcatcg ctctggagtg aataccacga cgatttccgg
3301 cagtttctac acatatattc gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc
3361 cctaaagggt ttattgagaa tatgtttttc gtctcagcca atccctgggt gagtttcacc
3421 agttttgatt taaacgtggc caatatggac aacttcttcg cccccgtttt caccatgggc
3481 aaatattata cgcaaggcga caaggtgctg atgccgctgg cgattcaggt tcatcatgcc
3541 gtttgtgatg gcttccatgt cggcagaatg cttaatgaat tacaacagta ctgcgatgag
3601 tggcagggcg gggcgtaatt tttttaaggc agttattggt gcccttaaac gcctggggta
3661 atgactctct agcttgaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt
3721 tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccctctag
3781 attacgtgca gtcgatgata agctgtcaaa catgagaatt gtgcctaatg agtgagctaa
3841 cttacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
3901 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt
3961 ggtttttctt ttcaccagtg agacgggcaa cagctgattg cccttcaccg cctggccctg
4021 agagagttgc agcaagcggt ccacgctggt ttgccccagc aggcgaaaat cctgtttgat
4081 ggtggttaac ggcgggatat aacatgagct gtcttcggta tcgtcgtatc ccactaccga
4141 gatatccgca ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc ccagcgccat
4201 ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc tcattcagca tttgcatggt
4261 ttgttgaaaa ccggacatgg cactccagtc gccttcccgt tccgctatcg gctgaatttg
4321 attgcgagtg agatatttat gccagccagc cagacgcaga cgcgccgaga cagaacttaa
4381 tgggcccgct aacagcgcga tttgctggtg acccaatgcg accagatgct ccacgcccag
4441 tcgcgtaccg tcttcatggg agaaaataat actgttgatg ggtgtctggt cagagacatc
4501 aagaaataac gccggaacat tagtgcaggc agcttccaca gcaatggcat cctggtcatc
4561 cagcggatag ttaatgatca gcccactgac gcgttgcgcg agaagattgt gcaccgccgc
4621 tttacaggct tcgacgccgc ttcgttctac catcgacacc accacgctgg cacccagttg
4681 atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga
4741 ggtggcaacg ccaatcagca acgactgttt gcccgccagt tgttgtgcca cgcggttggg
4801 aatgtaattc agctccgcca tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg
4861 gctggcctgg ttcaccacgc gggaaacggt ctgataagag acaccggcat actctgcgac
4921 atcgtataac gttactggtt tcacattcac caccctgaat tgactctctt ccgggcgcta
4981 tcatgccata ccgcgaaagg ttttgcacca ttcgatggtg tcggaatttc gggcagcgtt
5041 gggtcctggc cacgggtgcg catgatctag agctgcctcg cgcgtttcgg tgatgacggt
5101 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
5161 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc
5221 atgacccagt cacgtagcga tagcggagtg tatactggct taactatgcg gcatcagagc
5281 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa
5341 aataccgcat caggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
5401 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
5461 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
5521 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
5581 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
5641 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
5701 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
5761 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
5821 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
5881 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
5941 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
6001 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
6061 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
6121 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
6181 cacgttaagg gattttggtc atggcgttaa gggattttgg tcatgaatta attcttagaa
6241 aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata
6301 tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat
6361 ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa
6421 tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc
6481 cggtgagaat ggcaaaagtt tatgcatttc tttccagact tgttcaacag gccagccatt
6541 acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg
6601 agcgagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa
6661 ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc
6721 taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg catcatcagg
6781 agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct
6841 gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc
6901 tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc cgacattatc
6961 gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc gcggcctaga
7021 gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt ttatgtaagc
7081 agacagtttt attgttcatg accatgacat taacctataa aaataggcgt atcacgaggc
7141 cctttcgtct tcac
//
| {
"Vector backbone": "pQE",
"Backbone size w/o insert (bp)": "",
"Vector type": "Bacterial Expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Kanamycin, 50 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH10B",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Industry MTA"
} | {
"Title": "Directed Evolution of a Panel of Orthogonal T7 RNA Polymerase Variants for in Vivo or in Vitro Synthetic Circuitry",
"Publication": "ACS Synth Biol. 2014 Oct 7.",
"DOI": "10.1021/sb500299c",
"Authors": "Meyer AJ, Ellefson JW, Ellington AD",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/25279711",
"PubMed ID": "PubMed 25279711"
} | {
"Gene/Insert name": "CTGA-R13-AKSRV RNAP",
"Species": "T7 bacteriophage",
"Insert Size (bp)": "2685",
"Mutation": "V725A, Q744K, L747I, N748S, R756T, Q758K, H772R, E775V",
"Entrez Gene": "",
"Promoter": "T5/Lac",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Gibson Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "TTGTGAGCGGATAACAATT",
"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": ""
} |
|
pCRRNA-G11-R0 | 101805 | Repress mCherry and GFP independently, with different strengths | Vigouroux et al Mol Syst Biol. 2018 Mar 8;14(3):e7899. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/205489/28435274-06fb-4d19-806a-c43e00545f94/addgene-plasmid-101805-sequence-205489.gbk | full | LOCUS sequence-205489- 2936 bp DNA circular SYN 26-AUG-2024
DEFINITION Repress mCherry and GFP independently, with different strengths.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2936)
AUTHORS Vigouroux A, Oldewurtel E, Cui L, Bikard D, van Teeffelen S
TITLE Tuning dCas9's ability to block transcription enables robust,
noiseless knockdown of bacterial genes.
JOURNAL Mol Syst Biol. 2018 Mar 8;14(3):e7899.
PUBMED 29519933
REFERENCE 2 (bases 1 to 2936)
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 Syst
Biol."; date: "2018-03-8"; volume: "14(3)"; pages: "e7899"
FEATURES Location/Qualifiers
source 1..2936
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(14..32)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
CDS 211..1005
/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(265..284)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 875..894
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
terminator 1036..1130
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
rep_origin 1218..1806
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 1707..1726
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
terminator complement(2191..2277)
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
misc_feature 2307..2438
/label=crRNA leader
/note="crRNA leader sequence for the Streptococcus pyogenes
CRISPR/Cas system"
repeat_region 2439..2474
/label=DR
/note="direct repeat for the Streptococcus pyogenes
CRISPR/Cas system"
repeat_region 2505..2540
/label=DR
/note="direct repeat for the Streptococcus pyogenes
CRISPR/Cas system"
repeat_region 2571..2606
/label=DR
/note="direct repeat for the Streptococcus pyogenes
CRISPR/Cas system"
misc_RNA 2727..2805
/label=tracrRNA
/note="trans-activating CRISPR RNA for the Streptococcus
pyogenes CRISPR/Cas9 system"
ORIGIN
1 ttaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
61 gtttcttaga cgtcggaatt gccagctggg gcgccctctg gtaaggttgg gaagccctgc
121 aaagtaaact ggatggcttt cttgccgcca aggatctgat ggcgcagggg atcaagatct
181 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
241 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
301 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
361 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
421 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
481 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
541 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
601 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
661 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
721 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
781 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
841 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
901 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
961 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
1021 tcgagagctc gcttggactc ctgttgatag atccagtaat gacctcagaa ctccatctgg
1081 atttgttcag aacgctcggt tgccgccggg cgttttttat tggtgagaat ccaagcacta
1141 gtcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
1201 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
1261 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
1321 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
1381 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
1441 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
1501 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
1561 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
1621 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
1681 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
1741 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
1801 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
1861 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
1921 tgagctgata ccgctcgccg cagccgaacg ccccaaaaag cctcgctttc agcacctgtc
1981 gtttcctttc ttttcagagg gtattttaaa taaaaacatt aagttatgac gaagaagaac
2041 gggaacgcct taaaccggaa aattttcata aatagcgaaa acccgcgagg tcgccgcccc
2101 gtaacctgtc ggatcaccgg aaaggacccg taaagtgata atgattatca tctacatatc
2161 acaacgtgcg tggagggtct agggcggcgg atttgtccta ctcaggagag cgttcaccga
2221 caaacaacag ataaaacgaa aggcccagtc tttcgactga gcctttcgtt ttatttgatg
2281 cctctagcac gcgtaccatg ggatcctatt tcttaataac taaaaatatg gtataatact
2341 cttaataaat gcagtaatac aggggctttt caagactgaa gtctagctga gacaaatagt
2401 gcgattacga aattttttag acaaaaatag tctacgaggt tttagagcta tgctgttttg
2461 aatggtccca aaacccttca ccttgtggtg cttcagagaa tttggtttta gagctatgct
2521 gttttgaatg gtcccaaaac atcgcacatc ctggtcgcga cattaagagt gttttagagc
2581 tatgctgttt tgaatggtcc caaaacttca gcacactgag acttgttgag ttgaattcat
2641 gagtattaag tattgtttta tggctgataa atttctttga atttctcctt gattatttgt
2701 tataaaagtt ataaaataat cttgttggaa ccattcaaaa cagcatagca agttaaaata
2761 aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt ttgatacttc
2821 tattctactc tgactgcaaa ccaaaaaaac aagcgctttc aaaacgcttg ttttatcatt
2881 tttagggaaa ttaatctctt aatcctttta tcattctaca tttaggcgct gccatc
//
| {
"Vector backbone": "pCRRNAcos",
"Backbone size w/o insert (bp)": "2870",
"Vector type": "CRISPR, Synthetic Biology",
"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": "Tuning dCas9's ability to block transcription enables robust, noiseless knockdown of bacterial genes",
"Publication": "Mol Syst Biol. 2018 Mar 8;14(3):e7899.",
"DOI": "",
"Authors": "Vigouroux A, Oldewurtel E, Cui L, Bikard D, van Teeffelen S",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/29519933",
"PubMed ID": "PubMed 29519933"
} | {
"Gene/Insert name": "crRNAs targeting GFP and mCherry",
"Species": "Synthetic",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "Spy 1049",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "Unknown (unknown if destroyed)",
"3′ cloning site": "Unknown (unknown if destroyed)",
"5′ sequencing primer": "TTACTTTGCAGGGCTTCCCAACC",
"3′ sequencing primer": "TTTGATGCCTCTAGCACGCG"
} | {
"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": ""
} |
||
LPAR4-DuET | 213331 | The DuET construct encodes the named human GPCR engineered with a cleaved hemagglutinin signal peptide, a 5' FLAG and a 3' 1D4 epitope tag optimized for expression in mammalian cells and proteomics studies. | Thomas Sakmar | Dahl et al Sci Adv. 2023 May 3;9(18):eadf9297. doi: 10.1126/sciadv.adf9297. Epub 2023 May 3. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/430865/065b72fc-38ad-4293-9fe4-3121f3ab40ea/addgene-plasmid-213331-sequence-430865.gbk | full | LOCUS sequence-430865- 6650 bp DNA circular SYN 27-AUG-2024
DEFINITION The DuET construct encodes the named human GPCR engineered with a
cleaved hemagglutinin signal peptide, a 5' FLAG and a 3' 1D4 epitope
tag optimized for expression in mammalian cells and proteomics
studies..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6650)
AUTHORS Dahl L, Kotliar IB, Bendes A, Dodig-Crnkovic T, Fromm S, Elofsson A,
Uhlen M, Sakmar TP, Schwenk JM
TITLE Multiplexed selectivity screening of anti-GPCR antibodies.
JOURNAL Sci Adv. 2023 May 3;9(18):eadf9297. doi: 10.1126/sciadv.adf9297.
Epub 2023 May 3.
PUBMED 37134173
REFERENCE 2 (bases 1 to 6650)
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: "2023-05-3"; volume: "9(18):eadf9297. doi"; pages: "
10.1126/sciadv.adf9297. Epub 2023 May 3"
FEATURES Location/Qualifiers
source 1..6650
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(1198..1217)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(1199..1217)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter complement(1262..1465)
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind complement(1291..1311)
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
enhancer 1466..1845
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
primer_bind 2017..2036
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
promoter 2111..2215
/gene="bla"
/label=AmpR promoter
CDS 2216..3076
/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(2434..2453)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 3247..3835
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3736..3755
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 3989..4006
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 4123..4144
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 4159..4189
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4197..4213
/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 4202..4224
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 4221..4237
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 4221..4237
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
polyA_signal 4286..4407
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4297..4316)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 4351..4370
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
CDS complement(4581..5375)
/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(4692..4711)
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind 5302..5321
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
promoter complement(5442..5771)
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 5456..5591
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(5510..5529)
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind 5756..5776
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin complement(5785..6213)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5895..5916)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 6107..6126
/label=F1ori-R
/note="F1 origin, reverse primer"
polyA_signal 6259..6483
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind 6472..6489
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
ORIGIN
1 gttgtctggt cggacacttc ttcctgaatc gcaggcagag agggtttagt tgtcagtggg
61 gtttctgtct taaacagtga ttccattcgg atgtgcgcgt taatgtagaa gctcttctga
121 aaactctcca gtgtgaaata atagatgaaa gggtcgaagc agcagttcag ggtagcgagg
181 cacaatgtaa taggatacat aatcttagcg aacctttcca ggaagcagtt ggtgatggcc
241 tgactccgca ccagtgcata caggaaaagc acagaattat acggcacgaa gcacacaacg
301 aacaccgcca tatggactgt gatcatcttg agcactttct tcttatttgt gccgatctgg
361 gagagcgtcg ctggtttccg caaagtcctc aagaccacgg atgaacaact cacattcaga
421 atcagtggga tgataaaccc taccacttct ataaaaattg taatctttga caggtacgtc
481 ttccagactc gctttgaaaa accctcgaaa catgtagtgg ttgcgttatt cacattcgtg
541 gtgctaaaca gagaagcaga aataccccca gagagcacga gaatccacac tcccgcacac
601 actatagcgg agttccttct ggtgcggatc gttcgtgacc gaaatggata gactatggcc
661 aggaacctgt ccacgctgat acatgtcagg aacagcatgg acccgtagat attggtcaga
721 aaggccgttc cactgatttt gcacagggta tccccaaatg gccagtgccg gttgaaattg
781 tagaagatct tgaagggcaa ggtacaaacg aacagcaaat ctgagacggc aaggttggta
841 ataaaaatgg cggtttcgga ccgcattttc atcctaaagc agaacacgaa caaggacacg
901 gaatttgtaa taagcccgag gatgaaaacc acggaataaa cggcgccgtt cagattgtac
961 ttaaaggaat catcgactat acaagtgttg ttagctgtgg cattgcccaa tcgaggccgc
1021 agggagctat tggaatcctg gaattgaaag tcaatgaatc gccggtcacc catatcgatg
1081 ctggcgtcat catcgtcctt gtagtcggcg aataccaggc agaagatgta gctcagggcg
1141 atgatcgtct tcatggtggc gctaagctta agtttaaacg ctagccagct tgggtctccc
1201 tatagtgagt cgtattaatt tcgataagcc agtaagcagt gggttctcta gttagccaga
1261 gagctctgct tatatagacc tcccaccgta cacgcctacc gcccatttgc gtcaatgggg
1321 cggagttgtt acgacatttt ggaaagtccc gttgattttg gtgccaaaac aaactcccat
1381 tgacgtcaat ggggtggaga cttggaaatc cccgtgagtc aaaccgctat ccacgcccat
1441 tgatgtactg ccaaaaccgc atcaccatgg taatagcgat gactaatacg tagatgtact
1501 gccaagtagg aaagtcccat aaggtcatgt actgggcata atgccaggcg ggccatttac
1561 cgtcattgac gtcaataggg ggcgtacttg gcatatgata cacttgatgt actgccaagt
1621 gggcagttta ccgtaaatac tccacccatt gacgtcaatg gaaagtccct attggcgtta
1681 ctatgggaac atacgtcatt attgacgtca atgggcgggg gtcgttgggc ggtcagccag
1741 gcgggccatt taccgtaagt tatgtaacgc ggaactccat atatgggcta tgaactaatg
1801 accccgtaat tgattactat taataactag tcaataatca atgtcaacgc gtatatctgg
1861 cccgtacatc gcgaagcagc gcaaaacgcc taaccctaag cagattcttc atgcaaatgt
1921 cggtcaagcc ttgccttgtt gtagcttaaa ttttgctcgc gcactactca gcgacctcca
1981 acacacaagc agggagcaga tactggctta actatgcggc atcagagcag attgtactga
2041 gagtgcacca taggggatcg ggagatctcc cgatccgtcg acgtcaggtg gcacttttcg
2101 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
2161 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
2221 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
2281 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
2341 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
2401 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
2461 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
2521 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
2581 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
2641 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
2701 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
2761 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
2821 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
2881 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
2941 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
3001 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
3061 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
3121 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
3181 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
3241 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
3301 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
3361 tggcttcagc agagcgcaga taccaaatac cgttcttcta gtgtagccgt agttaggcca
3421 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
3481 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
3541 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
3601 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
3661 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
3721 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
3781 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
3841 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
3901 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
3961 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
4021 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
4081 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
4141 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
4201 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca agctctagct
4261 agaggtcgac ggtatacaga catgataaga tacattgatg agtttggaca aaccacaact
4321 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
4381 accattataa gctgcaataa acaagttggg gtgggcgaag aactccagca tgagatcccc
4441 gcgctggagg atcatccagc cggcgtcccg gaaaacgatt ccgaagccca acctttcata
4501 gaaggcggcg gtggaatcga aatctcgtga tggcaggttg ggcgtcgctt ggtcggtcat
4561 ttcgaacccc agagtcccgc tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc
4621 tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc ggtcagccca ttcgccgcca
4681 agctcttcag caatatcacg ggtagccaac gctatgtcct gatagcggtc cgccacaccc
4741 agccggccac agtcgatgaa tccagaaaag cggccatttt ccaccatgat attcggcaag
4801 caggcatcgc catgggtcac gacgagatcc tcgccgtcgg gcatgcgcgc cttgagcctg
4861 gcgaacagtt cggctggcgc gagcccctga tgctcttcgt ccagatcatc ctgatcgaca
4921 agaccggctt ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat
4981 gggcaggtag ccggatcaag cgtatgcagc cgccgcattg catcagccat gatggatact
5041 ttctcggcag gagcaaggtg agatgacagg agatcctgcc ccggcacttc gcccaatagc
5101 agccagtccc ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc
5161 gtggccagcc acgatagccg cgctgcctcg tcctgcagtt cattcagggc accggacagg
5221 tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca gccggaacac ggcggcatca
5281 gagcagccga ttgtctgttg tgcccagtca tagccgaata gcctctccac ccaagcggcc
5341 ggagaacctg cgtgcaatcc atcttgttca atcatgcgaa acgatcctca tcctgtctct
5401 tgatcagatc cgaaaatgga tatacaagct cccgggagct ttttgcaaaa gcctaggcct
5461 ccaaaaaagc ctcctcacta cttctggaat agctcagagg cagaggcggc ctcggcctct
5521 gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact gggcggagtt
5581 aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt gagatgcatg
5641 ctttgcatac ttctgcctgc tggggagcct ggggactttc cacacctggt tgctgactaa
5701 ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt tccacaccct
5761 aactgacaca cattccacag aattaattcg cgttaaattt ttgttaaatc agctcatttt
5821 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
5881 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
5941 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
6001 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
6061 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
6121 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
6181 ccgccgcgct taatgcgccg ctacagggcg cgtggggata ccccctagag ccccagctgg
6241 ttctttccgc ctcagaagcc atagagccca ccgcatcccc agcatgcctg ctattgtctt
6301 cccaatcctc ccccttgctg tcctgcccca ccccaccccc cagaatagaa tgacacctac
6361 tcagacaatg cgatgcaatt tcctcatttt attaggaaag gacagtggga gtggcacctt
6421 ccagggtcaa ggaaggcacg ggggaggggc aaacaacaga tggctggcaa ctagaaggca
6481 cagtcgaggc tgatcagcgg gtttaaacgg gccctctaga ctcgagttag gcaggggcca
6541 cctgagaagt ctcggtctta gacacggtgg tgctggcctc atcgaattca ccggtgcgtc
6601 caccggtatc gataaaggta ctttccagca tcagttcacc gccattgttt
//
| {
"Vector backbone": "pcDNA 3.1 (+)",
"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)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Multiplexed selectivity screening of anti-GPCR antibodies",
"Publication": "Sci Adv. 2023 May 3;9(18):eadf9297. doi: 10.1126/sciadv.adf9297. Epub 2023 May 3.",
"DOI": "10.1126/sciadv.adf9297.",
"Authors": "Dahl L, Kotliar IB, Bendes A, Dodig-Crnkovic T, Fromm S, Elofsson A, Uhlen M, Sakmar TP, Schwenk JM /sciadvadf",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/37134173",
"PubMed ID": "PubMed 37134173"
} | {
"Gene/Insert name": "LPAR4",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "LPAR4",
"Promoter": "CMV, T7",
"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": ""
} |
|
MG15-luc (mut p53 binding sites) | 16443 | High | Bert Vogelstein | el-Deiry et al Cell. 1993 Nov 19. 75(4):817-25. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/220477/2291ff3a-ab37-4c32-9a95-7a4f0e1370b7/addgene-plasmid-16443-sequence-220477.gbk | full | LOCUS sequence-220477- 5961 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 5961)
AUTHORS el-Deiry WS, Tokino T, Velculescu VE, Levy DB, Parsons R, Trent JM,
Lin D, Mercer WE, Kinzler KW, Vogelstein B
TITLE WAF1, a potential mediator of p53 tumor suppression.
JOURNAL Cell. 1993 Nov 19. 75(4):817-25.
PUBMED 8242752
REFERENCE 2 (bases 1 to 5961)
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. 1993
Nov 19. 75(4):817-25."
FEATURES Location/Qualifiers
source 1..5961
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(410..426)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
CDS 480..2132
/codon_start=1
/gene="luc"
/product="firefly luciferase"
/label=luciferase
/translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
primer_bind complement(546..564)
/label=LucNrev
/note="Firefly luciferase, reverse primer"
intron 2263..2328
/label=small t intron
/note="SV40 (simian virus 40) small t antigen intron"
CDS 2458..2478
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
polyA_signal 2903..3037
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2940..2959)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2994..3013
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(3034..3050)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
promoter complement(3087..3105)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(3126..3142)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3126..3142)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3139..3161)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3150..3166
/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(3174..3204)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3219..3240
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(3357..3374)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3528..4116)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3608..3627)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4287..5147)
/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 4910..4929
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5148..5252)
/gene="bla"
/label=AmpR promoter
rep_origin complement(5278..5733)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5415..5436)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 5627..5646
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 5860..5882
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 5874..5891
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 5875..5891
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 5898..5917
/label=T7
/note="T7 promoter, forward primer"
promoter 5898..5916
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind 5941..5957
/label=pBluescriptKS
/note="For pBluescript vector"
primer_bind 5942..5958
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
ORIGIN
1 agcttgatcc ttaatggact ttaatggcct tatggacttt aatggcctta atggacttta
61 atggccttaa tggactttaa tggccttaat ggactttaat ggccttaatg gactttaatg
121 gccttaatgg actttaatgg ccttaatgga ctttaatggc cttaatggac tttaatggcc
181 atcgaattcg atctgttttt tttagtatta agcagaggcc gggggcccct ggcctccgct
241 tactctggag aaaaagaaga gaggcattcc agaggcaact tgtcaaaaca ggactggcgc
301 cttggaggcg ctgtggggcc acccaaattg atataattaa gccccaaccg cctcttcccg
361 cctcatttca gcctcaccac cctcgacaag cttatcgata ccgtcggggg atccactagt
421 tctagagcgg ccgccaccgc ggtggagctc gagcttggca ttccggtact gttggtaaaa
481 tggaagacgc caaaaacata aagaaaggcc cggcgccatt ctatcctcta gaggatggaa
541 ccgctggaga gcaactgcat aaggctatga agagatacgc cctggttcct ggaacaattg
601 cttttacaga tgcacatatc gaggtgaaca tcacgtacgc ggaatacttc gaaatgtccg
661 ttcggttggc agaagctatg aaacgatatg ggctgaatac aaatcacaga atcgtcgtat
721 gcagtgaaaa ctctcttcaa ttctttatgc cggtgttggg cgcgttattt atcggagttg
781 cagttgcgcc cgcgaacgac atttataatg aacgtgaatt gctcaacagt atgaacattt
841 cgcagcctac cgtagtgttt gtttccaaaa aggggttgca aaaaattttg aacgtgcaaa
901 aaaaattacc aataatccag aaaattatta tcatggattc taaaacggat taccagggat
961 ttcagtcgat gtacacgttc gtcacatctc atctacctcc cggttttaat gaatacgatt
1021 ttgtaccaga gtcctttgat cgtgacaaaa caattgcact gataatgaat tcctctggat
1081 ctactgggtt acctaagggt gtggcccttc cgcatagaac tgcctgcgtc agattctcgc
1141 atgccagaga tcctattttt ggcaatcaaa tcattccgga tactgcgatt ttaagtgttg
1201 ttccattcca tcacggtttt ggaatgttta ctacactcgg atatttgata tgtggatttc
1261 gagtcgtctt aatgtataga tttgaagaag agctgttttt acgatccctt caggattaca
1321 aaattcaaag tgcgttgcta gtaccaaccc tattttcatt cttcgccaaa agcactctga
1381 ttgacaaata cgatttatct aatttacacg aaattgcttc tgggggcgca cctctttcga
1441 aagaagtcgg ggaagcggtt gcaaaacgct tccatcttcc agggatacga caaggatatg
1501 ggctcactga gactacatca gctattctga ttacacccga gggggatgat aaaccgggcg
1561 cggtcggtaa agttgttcca ttttttgaag cgaaggttgt ggatctggat accgggaaaa
1621 cgctgggcgt taatcagaga ggcgaattat gtgtcagagg acctatgatt atgtccggtt
1681 atgtaaacaa tccggaagcg accaacgcct tgattgacaa ggatggatgg ctacattctg
1741 gagacatagc ttactgggac gaagacgaac acttcttcat agttgaccgc ttgaagtctt
1801 taattaaata caaaggatat caggtggccc ccgctgaatt ggaatcgata ttgttacaac
1861 accccaacat cttcgacgcg ggcgtggcag gtcttcccga cgatgacgcc ggtgaacttc
1921 ccgccgccgt tgttgttttg gagcacggaa agacgatgac ggaaaaagag atcgtggatt
1981 acgtcgccag tcaagtaaca accgcgaaaa agttgcgcgg aggagttgtg tttgtggacg
2041 aagtaccgaa aggtcttacc ggaaaactcg acgcaagaaa aatcagagag atcctcataa
2101 aggccaagaa gggcggaaag tccaaattgt aaaatgtaac tgtattcagc gatgacgaaa
2161 ttcttagcta ttgtaatact ctagaggatc tttgtgaagg aaccttactt ctgtggtgtg
2221 acataattgg acaaactacc tacagagatt taaagctcta aggtaaatat aaaattttta
2281 agtgtataat gtgttaaact actgattcta attgtttgtg tattttagat tccaacctat
2341 ggaactgatg aatgggagca gtggtggaat gcctttaatg aggaaaacct gttttgctca
2401 gaagaaatgc catctagtga tgatgaggct actgctgact ctcaacattc tactcctcca
2461 aaaaagaaga gaaaggtaga agaccccaag gactttcctt cagaattgct aagttttttg
2521 agtcatgctg tgtttagtaa tagaactctt gcttgctttg ctatttacac cacaaaggaa
2581 aaagctgcac tgctatacaa gaaaattatg gaaaaatatt ctgtaacctt tataagtagg
2641 cataacagtt ataatcataa catactgttt tttcttactc cacacaggca tagagtgtct
2701 gctattaata actatgctca aaaattgtgt acctttagct ttttaatttg taaaggggtt
2761 aataaggaat atttgatgta tagtgccttg actagagatc ataatcagcc ataccacatt
2821 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
2881 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
2941 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
3001 gtccaaactc atcaatgtat cttatcatgt ctggatccac tagttctaga gcggccgcca
3061 ccgcggtgga gctccagctt ttgttccctt tagtgagggt taatttcgag cttggcgtaa
3121 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
3181 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
3241 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
3301 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
3361 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
3421 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
3481 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
3541 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
3601 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
3661 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
3721 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
3781 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
3841 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
3901 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
3961 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
4021 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
4081 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
4141 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
4201 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
4261 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
4321 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
4381 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
4441 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
4501 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
4561 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
4621 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
4681 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
4741 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
4801 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
4861 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
4921 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
4981 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
5041 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
5101 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
5161 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
5221 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctaaa
5281 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
5341 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
5401 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
5461 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
5521 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
5581 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
5641 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
5701 ccgccgcgct taatgcgccg ctacagggcg cgtcccattc gccattcagg ctgcgcaact
5761 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat
5821 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa
5881 cgacggccag tgaattgtaa tacgactcac tatagggcga attgggtacc gggccccccc
5941 tcgaggtcga cggtatcgat a
//
| {
"Vector backbone": "n/a",
"Backbone size w/o insert (bp)": "",
"Vector type": "Mammalian Expression, Luciferase",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "Unknown"
} | {
"Academic/Nonprofit Terms": "UBMTA ; Luciferase Limited Use Label License",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "WAF1, a potential mediator of p53 tumor suppression",
"Publication": "Cell. 1993 Nov 19. 75(4):817-25.",
"DOI": "10.1016/0092-8674(93)90500-P",
"Authors": "el-Deiry WS, Tokino T, Velculescu VE, Levy DB, Parsons R, Trent JM, Lin D, Mercer WE, Kinzler KW, Vogelstein B",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/8242752",
"PubMed ID": "PubMed 8242752"
} | {
"Gene/Insert name": "p53-binding site (mutant)",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "TP53",
"Promoter": "",
"Tag / Fusion Protein": "luciferase (C terminal on backbone)"
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "EcoRV (destroyed during cloning)",
"3′ cloning site": "EcoRV (destroyed during cloning)",
"5′ sequencing primer": "M13pUC-Fwd",
"3′ sequencing primer": "LucNRev"
} | {
"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": ""
} |
|
D12_Ld_040370 | 135828 | 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/274527/fe198c24-d9bc-4cfc-ae57-3c4c8e9cf92c/addgene-plasmid-135828-sequence-274527.gbk | full | LOCUS sequence-274527- 6555 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 6555)
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 6555)
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..6555
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 1..204
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 155..175
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 201..225
/label=LNCX
/note="Human CMV promoter, forward primer"
primer_bind 706..725
/label=pMT2-F
/note="Synthetic intron, forward primer"
regulatory 748..757
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 1333..1350
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
polyA_signal 1402..1457
/label=beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(1456..1475)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
primer_bind complement(3693..3711)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 3779..3883
/gene="bla"
/label=AmpR promoter
CDS 3884..4744
/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(4102..4121)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 4915..5503
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5404..5423
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 5657..5674
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 5791..5812
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 5827..5857
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5865..5881
/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 5870..5892
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 5889..5905
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 5889..5905
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
enhancer 6176..6555
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
ORIGIN
1 gtgatgcggt tttggcagta caccaatggg cgtggatagc ggtttgactc acggggattt
61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
121 tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa tgggcggtag gcgtgtacgg
181 tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcctcac tctcttccgc
241 atcgctgtct gcgagggcca gctgttgggc tcgcggttga ggacaaactc ttcgcggtct
301 ttccagtact cttggatcgg aaacccgtcg gcctccgaac ggtactccgc caccgaggga
361 cctgagcgag tccgcatcga ccggatcgga aaacctctcg agaaaggcgt ctaaccagtc
421 acagtcgcaa ggtaggctga gcaccgtggc gggcggcagc gggtggcggt cggggttgtt
481 tctggcggag gtgctgctga tgatgtaatt aaagtaggcg gtcttgagac ggcggatggt
541 cgaggtgagg tgtggcaggc ttgagatcca gctgttgggg tgagtactcc ctctcaaaag
601 cgggcattac ttctgcgcta agattgtcag tttccaaaaa cgaggaggat ttgatattca
661 cctggcccga tctggccata cacttgagtg acaatgacat ccactttgcc tttctctcca
721 caggtgtcca ctcccaggtc caagtttgcc accatggagt ttcagaccca ggtactcatg
781 tccctgctgc tctgcatgtc tggtgcggcc gccaccgata tgctgctggc ccccagcgac
841 agcatcccct tcaccacctt cggccagctg cagcaggtct gcgccgccac acaggccgga
901 agcacccctg tggccatgta caccgccacc atgaccgccc tggtggccaa cctgatgagc
961 gagaagaaag tgacccacgc ctacgtgtcc gcctacctga acgccgaccg gcggtggact
1021 tggtttgtgt acgagggcga cgaagtgatc gagatgctga tgaacagcag ccagtgggcc
1081 accggcaacc ccacaggcgc ccacaccacc taccggcacg tggtgtacag caccaccgac
1141 aagggcctga tcagcgccac aggctacgac aactaccccg tcgtgtgcaa atacgagaac
1201 cagaccgact acagcccccc tggcaagcgg ttccctggcg cgcccggcag cctgcaccac
1261 atcctggacg cccagaagat gctgtggaac cacagggaca ggaacctgcc tcctctggcc
1321 cctctgggcc cccaccacca tcaccaccat tgagtctaga cacgtgatta attaaggatc
1381 ccccgacctc gacctctggc taataaagga aatttatttt cattgcaata gtgtgttgga
1441 attttttgtg tctctcactc ggaaggacat atgggagggc aaatcatttg gtcgagatcc
1501 ctcggagatc tctagctaga ggatcgatcc ccgccccgga cgaactaaac ctgactacga
1561 catctctgcc ccttcttcgc ggggcagtgc atgtaatccc ttcagttggt tggtacaact
1621 tgccaactgg gccctgttcc acatgtgaca cgggggggga ccaaacacaa aggggttctc
1681 tgactgtagt tgacatcctt ataaatggat gtgcacattt gccaacactg agtggctttc
1741 atcctggagc agactttgca gtctgtggac tgcaacacaa cattgccttt atgtgtaact
1801 cttggctgaa gctcttacac caatgctggg ggacatgtac ctcccagggg cccaggaaga
1861 ctacgggagg ctacaccaac gtcaatcaga ggggcctgtg tagctaccga taagcggacc
1921 ctcaagaggg cattagcaat agtgtttata aggccccctt gttaacccta aacgggtagc
1981 atatgcttcc cgggtagtag tatatactat ccagactaac cctaattcaa tagcatatgt
2041 tacccaacgg gaagcatatg ctatcgaatt agggttagta aaagggtcct aaggaacagc
2101 gatatctccc accccatgag ctgtcacggt tttatttaca tggggtcagg attccacgag
2161 ggtagtgaac cattttagtc acaagggcag tggctgaaga tcaaggagcg ggcagtgaac
2221 tctcctgaat cttcgcctgc ttcttcattc tccttcgttt agctaataga ataactgctg
2281 agttgtgaac agtaaggtgt atgtgaggtg ctcgaaaaca aggtttcagg tgacgccccc
2341 agaataaaat ttggacgggg ggttcagtgg tggcattgtg ctatgacacc aatataaccc
2401 tcacaaaccc cttgggcaat aaatactagt gtaggaatga aacattctga atatctttaa
2461 caatagaaat ccatggggtg gggacaagcc gtaaagactg gatgtccatc tcacacgaat
2521 ttatggctat gggcaacaca taatcctagt gcaatatgat actggggtta ttaagatgtg
2581 tcccaggcag ggaccaagac aggtgaacca tgttgttaca ctctatttgt aacaagggga
2641 aagagagtgg acgccgacag cagcggactc cactggttgt ctctaacacc cccgaaaatt
2701 aaacggggct ccacgccaat ggggcccata aacaaagaca agtggccact cttttttttg
2761 aaattgtgga gtgggggcac gcgtcagccc ccacacgccg ccctgcggtt ttggactgta
2821 aaataagggt gtaataactt ggctgattgt aaccccgcta accactgcgg tcaaaccact
2881 tgcccacaaa accactaatg gcaccccggg gaatacctgc ataagtaggt gggcgggcca
2941 agataggggc gcgattgctg cgatctggag gacaaattac acacacttgc gcctgagcgc
3001 caagcacagg gttgttggtc ctcatattca cgaggtcgct gagagcacgg tgggctaatg
3061 ttgccatggg tagcatatac tacccaaata tctggatagc atatgctatc ctaatctata
3121 tctgggtagc ataggctatc ctaatctata tctgggtagc atatgctatc ctaatctata
3181 tctgggtagt atatgctatc ctaatttata tctgggtagc ataggctatc ctaatctata
3241 tctgggtagc atatgctatc ctaatctata tctgggtagt atatgctatc ctaatctgta
3301 tccgggtagc atatgctatc ctaatagaga ttagggtagt atatgctatc ctaatttata
3361 tctgggtagc atatactacc caaatatctg gatagcatat gctatcctaa tctatatctg
3421 ggtagcatat gctatcctaa tctatatctg ggtagcatag gctatcctaa tctatatctg
3481 ggtagcatat gctatcctaa tctatatctg ggtagtatat gctatcctaa tttatatctg
3541 ggtagcatag gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg
3601 ggtagtatat gctatcctaa tctgtatccg ggtagcatat gctatcctca tgataagctg
3661 tcaaacatga gaattttctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt
3721 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
3781 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
3841 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
3901 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
3961 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
4021 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
4081 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca
4141 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
4201 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
4261 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
4321 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
4381 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac
4441 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
4501 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
4561 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
4621 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
4681 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
4741 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
4801 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
4861 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
4921 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
4981 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
5041 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
5101 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
5161 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
5221 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
5281 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
5341 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
5401 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
5461 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
5521 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
5581 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
5641 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
5701 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
5761 ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc
5821 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca
5881 atttcacaca ggaaacagct atgaccatga ttacgccaag ctctagctag aggtcgacca
5941 attctcatgt ttgacagctt atcatcgcag atccgggcaa cgttgttgcc attgctgcag
6001 gcgcagaact ggtaggtatg gaagatctat acattgaatc aatattggca attagccata
6061 ttagtcattg gttatatagc ataaatcaat attggctatt ggccattgca tacgttgtat
6121 ctatatcata atatgtacat ttatattggc tcatgtccaa tatgaccgcc atgttgacat
6181 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
6241 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
6301 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
6361 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
6421 tatcatatgc caagtccgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
6481 tatgcccagt acatgacctt acgggacttt cctacttggc agtacatcta cgtattagtc
6541 atcgctatta ccatg
//
| {
"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",
"Species": "Leishmania donovani",
"Insert Size (bp)": "423",
"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": ""
} |
|
pKDM | 110740 | (Empty Backbone)Plasmid for the insect cell expression system, two promoters, Multiplication Module M, Tn7 transposition | Tim Richmond | Fitzgerald et al Nat Methods. 2006 Dec;3(12):1021-32. doi: 10.1038/nmeth983. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/213586/c3563ee1-5174-44e9-9d27-5099cffc1753/addgene-plasmid-110740-sequence-213586.gbk | full | LOCUS sequence-213586- 4760 bp DNA circular SYN 26-AUG-2024
DEFINITION Plasmid for the insect cell expression system, two promoters,
Multiplication Module M, Tn7 transposition.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4760)
AUTHORS Fitzgerald DJ, Berger P, Schaffitzel C, Yamada K, Richmond TJ,
Berger I
TITLE Protein complex expression by using multigene baculoviral vectors.
JOURNAL Nat Methods. 2006 Dec;3(12):1021-32. doi: 10.1038/nmeth983.
PUBMED 17117155
REFERENCE 2 (bases 1 to 4760)
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 Methods"; date: "2006-12"; volume: "3"; issue: "12"; pages:
"1021-32"
FEATURES Location/Qualifiers
source 1..4760
/mol_type="other DNA"
/organism="synthetic DNA construct"
mobile_element 74..239
/mobile_element_type="transposon:Tn7"
/label=Tn7L
/note="mini-Tn7 element (left end of the Tn7 transposon)"
polyA_signal 268..402
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(292..311)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 346..365
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
promoter complement(661..752)
/gene="polh from Autographa californica"
/label=polyhedrin promoter
/note="promoter for the baculovirus polyhedrin gene"
primer_bind complement(719..736)
/label=Polyhedrin-F
/note="Polyhedrin promoter, forward primer"
promoter 797..906
/label=p10 promoter
/note="baculovirus promoter for expression in insect cells"
primer_bind complement(998..1017)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 1042..1090
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
promoter 1607..1635
/gene="intI1 (promoter lies within the coding sequence)"
/label=Pc promoter
/note="class 1 integron promoter"
CDS 1824..2357
/codon_start=1
/gene="aacC1"
/product="gentamycin acetyltransferase"
/label=GmR
/note="confers resistance to gentamycin"
/translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
EEVMHFDIDPSTAT"
mobile_element 2424..2648
/mobile_element_type="transposon:Tn7"
/label=Tn7R
/note="mini-Tn7 element (right end of the Tn7 transposon)"
primer_bind complement(2782..2799)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2953..3541)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3033..3052)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind complement(3652..3671)
/label=pENTR-R
/note="pENTR vectors, reverse primer"
CDS 3663..4478
/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 3663..4478
/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 4544..4563
/label=F1ori-R
/note="F1 origin, reverse primer"
ORIGIN
1 gaagatgacg gtttgtcaca tggagttggc aggatgtttg attaaaaaca taacaggaag
61 aaaaatgccc cgctgtgggc ggacaaaata gttgggaact gggaggggtg gaaatggagt
121 ttttaaggat tatttaggga agagtgacaa aatagatggg aactgggtgt agcgtcgtaa
181 gctaatacga aaattaaaaa tgacaaaata gtttggaact agatttcact tatctggttc
241 ggatctccta ggctcaagca gtgatcagat ccagacatga taagatacat tgatgagttt
301 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
361 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
421 cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc
481 tacaaatgtg gtatggctga ttatgatcct ctagtacttc tcgacaagct tgtcgagact
541 gcaggctcta gattcgaaag cggccgcgac tagtgagctc gtcgacgtag gcctttgaat
601 tccgcgcgct tcggaccggg atccgcgccc gatggtggga cggtatgaat aatccggaat
661 atttataggt ttttttatta caaaactgtt acgaaaacag taaaatactt atttatttgc
721 gagatggtta tcattttaat tatctccatg atctattaat attccggagt aggtcgcgaa
781 tcgatactag tatacggacc tttaattcaa cccaacacaa tatattatag ttaaataaga
841 attattatca aatcatttgt atattaatta aaatactata ctgtaaatta cattttattt
901 acaatcactc gacgaagact tgatcacccg ggatctcgag ccatggtgct agcagctgat
961 gcatagcatg cggtaccggg agatggggga ggctaactga aacacggaag gagacaatac
1021 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac gggtgttggg
1081 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
1141 agaccccatt gggaccaata cgcccgcgtt tcttcctttt ccccacccca acccccaagt
1201 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caagccctgc catagccagc
1261 tttgtttaaa caaagctgta gccaaccact agaactatag ctagagtcct gggcgaacaa
1321 acgatgctcg ccttccagaa aaccgaggat gcgaaccact tcatccgggg tcagcaccac
1381 cggcaagcgc cgcgacggcc gaggtctacc gatctcctga agcaagggca gatccgtgca
1441 cagcaccttg ccgtagaaga acagcaaggc cgccaatgcc tgacgatgcg tggagaccga
1501 aaccttgcgc tcgttcgcca gccaggacag aaatgcctcg acttcgctgc tgcccaaggt
1561 tgccgggtga cgcacaccgt ggaaacggat gaaggcacga acccagttga cataagcctg
1621 ttcggttcgt aaactgtaat gcaagtagcg tatgcgctca cgcaactggt ccagaacctt
1681 gaccgaacgc agcggtggta acggcgcagt ggcggttttc atggcttgtt atgactgttt
1741 ttttgtacag tctatgcctc gggcatccaa gcagcaagcg cgttacgccg tgggtcgatg
1801 tttgatgtta tggagcagca acgatgttac gcagcagcaa cgatgttacg cagcagggca
1861 gtcgccctaa aacaaagtta ggtggctcaa gtatgggcat cattcgcaca tgtaggctcg
1921 gccctgacca agtcaaatcc atgcgggctg ctcttgatct tttcggtcgt gagttcggag
1981 acgtagccac ctactcccaa catcagccgg actccgatta cctcgggaac ttgctccgta
2041 gtaagacatt catcgcgctt gctgccttcg accaagaagc ggttgttggc gctctcgcgg
2101 cttacgttct gcccaagttt gagcagccgc gtagtgagat ctatatctat gatctcgcag
2161 tctccggcga gcaccggagg cagggcattg ccaccgcgct catcaatctc ctcaagcatg
2221 aggccaacgc gcttggtgct tatgtgatct acgtgcaagc agattacggt gacgatcccg
2281 cagtggctct ctatacaaag ttgggcatac gggaagaagt gatgcacttt gatatcgacc
2341 caagtaccgc cacctaacaa ttcgttcaag ccgagatcgg cttcccggcc gcggagttgt
2401 tcggtaaatt gtcacaacgc cgcgaatata gtctttacca tgcccttggc cacgcccctc
2461 tttaatacga cgggcaattt gcacttcaga aaatgaagag tttgctttag ccataacaaa
2521 agtccagtat gctttttcac agcataactg gactgatttc agtttacaac tattctgtct
2581 agtttaagac tttattgtca tagtttagat ctattttgtt cagtttaaga ctttattgtc
2641 cgcccacacc cgcttacgca gggcatccat ttattactca accgtaaccg attttgccag
2701 gttacgcggc tggtctatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
2761 catcaggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
2821 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
2881 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
2941 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
3001 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
3061 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
3121 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
3181 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
3241 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
3301 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
3361 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
3421 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
3481 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
3541 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
3601 agggattttg gtcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg
3661 ttatgagcca tattcaacgg gaaacgtctt gctctaggcc gcgattaaat tccaacatgg
3721 atgctgattt atatgggtat aaatgggctc gagataatgt cgggcaatca ggtgcgacaa
3781 tctatcgctt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta
3841 gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc
3901 ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg
3961 cgatccccgg gaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata
4021 ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc
4081 cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg aataacggtt
4141 tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga
4201 aagaaatgca taaacttttg ccattctcac cggattcagt cgtcactcat ggtgatttct
4261 cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag
4321 tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt
4381 ctccttcatt acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata
4441 aattgcagtt tcatttgatg ctcgatgagt ttttctaaga attcttcacg accacgctga
4501 tgagctttac cgcagtacgt ttggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
4561 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
4621 ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc attcaggctg caaataagcg
4681 ttgatattca gtcaattaca aacattaata acgaagagat gacagaaaaa ttttcattct
4741 gtgacagaga aaaagtagcc
//
| {
"Vector backbone": "pKDM",
"Backbone size w/o insert (bp)": "",
"Vector type": "Insect Expression",
"Selectable markers": "Gentamicin"
} | {
"Bacterial Resistance(s)": "Kanamycin and Gentamicin, 50 & 10 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": "Protein complex expression by using multigene baculoviral vectors",
"Publication": "Nat Methods. 2006 Dec;3(12):1021-32. doi: 10.1038/nmeth983.",
"DOI": "10.1038/nmeth983.",
"Authors": "Fitzgerald DJ, Berger P, Schaffitzel C, Yamada K, Richmond TJ, Berger I /nmeth",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/17117155",
"PubMed ID": "PubMed 17117155"
} | {
"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": ""
} |
|
mPapaya1-Alpha-Actinin-19 | 56648 | Localization: Alpha Actinin, Excitation: 530, Emission: 541 | Michael Davidson | Hoi et al Chem Biol. 2013 Oct 24;20(10):1296-304. doi: 10.1016/j.chembiol.2013.08.008. Epub 2013 Oct 3. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/147103/810fb93b-88ae-42d9-aecb-826490fd8bdd/addgene-plasmid-56648-sequence-147103.gbk | partial | LOCUS sequence-147103- 775 bp DNA linear UNA 26-AUG-2024
DEFINITION Localization: Alpha Actinin, Excitation: 530, Emission: 541.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE natural DNA sequence
ORGANISM unspecified
REFERENCE 1 (bases 1 to 775)
AUTHORS Hoi H, Howe ES, Ding Y, Zhang W, Baird MA, Sell BR, Allen JR,
Davidson MW, Campbell RE
TITLE An engineered monomeric Zoanthus sp. yellow fluorescent protein.
JOURNAL Chem Biol. 2013 Oct 24;20(10):1296-304. doi:
10.1016/j.chembiol.2013.08.008. Epub 2013 Oct 3.
PUBMED 24094838
REFERENCE 2 (bases 1 to 775)
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.chembiol.2013.08.008"; journalName: "Chem Biol"; date:
"2013-10-24- 24"; volume: "20"; issue: "10"; pages: "1296-304"
FEATURES Location/Qualifiers
source 1..775
/mol_type="genomic DNA"
/organism="unspecified"
ORIGIN
1 gggccgcggt accgtcgact gcagaattcg aagcttgagg tcactctcgc cgtacagcgc
61 cgtggagaag gacatgtagt ccagagcacc tggcacggag tcggggccgg tgtagggggc
121 catccgcgcg atgcagtact cagcctggtc gggtggcagc tcgcggcgca gctcgtccat
181 ggtaatgtag ttcttgtccc cagccaggat cttgaaggaa gccatgactt ggtctgctgt
241 atctgtgtcg gctgtctcgc gggacatgaa gtcaatgaag gcctggaatg tcactacccc
301 caggcggttg gggtccacaa tgctcatgat gcgggcaaat tctgcttctc cctgggggtc
361 gttgccaata tcataaccca agctgatgag gcaggctttg aactcctcgg gacccagtgt
421 gccggagtga tcccggtcaa agtggttgaa ggaggcccgg aactcattca tctgctcctg
481 gctgatgccc ttggcatccc gggtcaggat ctggttctct acctcattga tggtcctggc
541 gatggtggtg agcagctgct cccagcccac acggatgtgc tccatggtgt agttggtgtg
601 cttgttgtcg aagatgagcg cctcctggat gagctggtgg tcgccctcca gctgatcaat
661 ctttggcttg tagttgacga tgctcttctc atactgccgc aggtggctga gctggtcctc
721 cagggtccca tgcatctcaa tggagatcct cccgatctcc tccatcttgg tctgg
//
| {
"Vector backbone": "mPapaya1",
"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": "Not Available to Industry"
} | {
"Title": "An engineered monomeric Zoanthus sp. yellow fluorescent protein",
"Publication": "Chem Biol. 2013 Oct 24;20(10):1296-304. doi: 10.1016/j.chembiol.2013.08.008. Epub 2013 Oct 3.",
"DOI": "10.1016/j.chembiol.2013.08.008.",
"Authors": "Hoi H, Howe ES, Ding Y, Zhang W, Baird MA, Sell BR, Allen JR, Davidson MW, Campbell RE /jchembiol",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/24094838",
"PubMed ID": "PubMed 24094838"
} | {
"Gene/Insert name": "Alpha-Actinin",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "ACTN1",
"Promoter": "CMV",
"Tag / Fusion Protein": "mPapaya1"
} | {
"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": ""
} |
|
pCI-FLAG-mTUT4(D1028A) | 60220 | Mammalian expression of FLAG-tagged catalytic mutant of TUT4 | Zissimos Mourelatos | Liu et al Mol Cell. 2014 Aug 27. pii: S1097-2765(14)00610-8. doi: 10.1016/j.molcel.2014.07.017. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/99717/b9d473ad-8f98-47b6-8abe-c634ea665c7d/addgene-plasmid-60220-sequence-99717.gbk | partial | LOCUS sequence-99717-e 850 bp DNA linear UNA 26-AUG-2024
DEFINITION Mammalian expression of FLAG-tagged catalytic mutant of TUT4 .
ACCESSION .
VERSION .
KEYWORDS .
SOURCE natural DNA sequence
ORGANISM unspecified
REFERENCE 1 (bases 1 to 850)
AUTHORS Liu X, Zheng Q, Vrettos N, Maragkakis M, Alexiou P, Gregory BD,
Mourelatos Z
TITLE A MicroRNA Precursor Surveillance System in Quality Control of
MicroRNA Synthesis.
JOURNAL Mol Cell. 2014 Aug 27. pii: S1097-2765(14)00610-8. doi:
10.1016/j.molcel.2014.07.017.
PUBMED 25175028
REFERENCE 2 (bases 1 to 850)
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.
2014 Aug 27. pii: S1097-2765(14)00610-8. doi:
10.1016/j.molcel.2014.07.017."
FEATURES Location/Qualifiers
source 1..850
/mol_type="genomic DNA"
/organism="unspecified"
promoter 21..39
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 57..80
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
ORIGIN
1 cgctcttagg ctagagtact taatacgact cactataggc tagcctcgag accatggact
61 acaaagacga tgacgacaag gaagagccca agacttctaa aaacgaaaat catgaaccaa
121 agaagaatat tatttgtgaa gaaagcaaag cagttaaaat tatatctaat caaaccttga
181 agcctagaaa tgataagtca gaaattggga ccagctctct gaatagaaat agtagtaaaa
241 aaaccaagca aaatgatatt tgtatagaaa aaacagaagc taaatcatgt aaagtaaacg
301 ctgccagtgt tccaggccct aaagatttgg ggttagtaca tcgagatcaa agtcattgca
361 aaatgaaaaa gttaccaaat tcacccatga aagcccagaa gggatcttca cagacaaaat
421 tagaaaagac acctagttta cagacaaaag cagaaaaagt accaaagtct ccaaatttac
481 cagtaaaagc agaaaaagcg ccctgtacaa cagcagaagc aacaacagaa aaagcactta
541 attcacagag aaaagaagaa aacacaccca cttctcagat gaaattacag aagacaccca
601 gatcaccttt agagccagaa aatgtgccca gtttactatt gaaagaaaat gtgaaacaga
661 cagaatcaca acagactggg aagaaactta caagctcctt tgtttctatg gataaaagga
721 agagtgaagc tctacaggga gaaaagtcag ctctggaaaa ctcatcacta tctcagaagc
781 agcagactca gacagataat attgctgatt ctgatgacag tgcttcaggg attgaagata
841 cagcagatga
//
| {
"Vector backbone": "pCI",
"Backbone size w/o insert (bp)": "4000",
"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 MicroRNA Precursor Surveillance System in Quality Control of MicroRNA Synthesis",
"Publication": "Mol Cell. 2014 Aug 27. pii: S1097-2765(14)00610-8. doi: 10.1016/j.molcel.2014.07.017.",
"DOI": "10.1016/j.molcel.2014.07.017.",
"Authors": "Liu X, Zheng Q, Vrettos N, Maragkakis M, Alexiou P, Gregory BD, Mourelatos Z /jmolcel",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/25175028",
"PubMed ID": "PubMed 25175028"
} | {
"Gene/Insert name": "TUT4",
"Species": "M. musculus (mouse)",
"Insert Size (bp)": "4900",
"Mutation": "D1028A",
"Entrez Gene": "Tut4",
"Promoter": "CMV",
"Tag / Fusion Protein": "FLAG (N terminal on insert)"
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "BclI (destroyed during cloning)",
"3′ cloning site": "NotI (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": ""
} |
|
pGL3-ACSL4-(-706) | 202513 | Expressing luciferase driven by human ACSL4 promoter (bases -1 to -706 with respect to exon1 start site) | Ming Chen | Wang et al J Clin Invest. 2023 Mar 16:e166647. doi: 10.1172/JCI166647. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/396602/a2fad9e6-4902-467b-8a07-4774795d9fa1/addgene-plasmid-202513-sequence-396602.gbk | full | LOCUS sequence-396602- 5509 bp DNA circular SYN 27-AUG-2024
DEFINITION Expressing luciferase driven by human ACSL4 promoter (bases -1 to
-706 with respect to exon1 start site).
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5509)
AUTHORS Wang ME, Chen J, Lu Y, Bawcom AR, Wu J, Ou J, Asara JM, Armstrong
AJ, Wang Q, Li L, Wang Y, Huang J, Chen M
TITLE RB1-deficient prostate tumor growth and metastasis are vulnerable to
ferroptosis induction via the E2F/ACSL4 axis.
JOURNAL J Clin Invest. 2023 Mar 16:e166647. doi: 10.1172/JCI166647.
PUBMED 36928314
REFERENCE 2 (bases 1 to 5509)
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: "J Clin
Invest. 2023 Mar 16:e166647. doi: 10.1172/JCI166647."
FEATURES Location/Qualifiers
source 1..5509
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 61..82
/label=F1ori-F
/note="F1 origin, forward primer"
polyA_signal 350..398
/note="synthetic polyadenylation signal"
misc_feature 412..503
/label=pause site
/note="RNA polymerase II transcriptional pause signal from
the human alpha-2 globin gene"
primer_bind 452..471
/label=RVprimer3
/note="pGL3 vector, forward primer"
regulatory 1283..1292
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 1289..2941
/codon_start=1
/gene="luc+"
/product="firefly luciferase"
/label=luciferase
/note="enhanced luc+ version of the luciferase gene"
/translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
primer_bind complement(1355..1373)
/label=LucNrev
/note="Firefly luciferase, reverse primer"
polyA_signal 2982..3103
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2993..3012)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 3047..3066
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind complement(3351..3368)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3522..4110)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3602..3621)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4281..5141)
/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 4904..4923
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5142..5246)
/gene="bla"
/label=AmpR promoter
rep_origin join(5273..5509,1..219)
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5360..5379)
/label=F1ori-R
/note="F1 origin, reverse primer"
ORIGIN
1 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
61 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
121 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
181 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttg ccattcgcca ttcaggctgc
241 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag cccaagctac
301 catgataagt aagtaatatt aaggtacggg aggtacttgg agcggccgca ataaaatatc
361 tttattttca ttacatctgt gtgttggttt tttgtgtgaa tcgatagtac taacatacgc
421 tctccatcaa aacaaaacga aacaaaacaa actagcaaaa taggctgtcc ccagtgcaag
481 tgcaggtgcc agaacatttc tctatcgata ggtaccgagc tcttacgcgt gctagcccgg
541 gctcgagtaa gaggcaagat aaacggatga gaggcattag ttcaggctta caggagcaag
601 atgcttttag tcaaagtgtt caagatagct ctcttaaaga ggagcagtga aacccaaaaa
661 tgagtgtctt ttgagggcag ctggaagatg acccgagggg aacttttatc ctggggtgga
721 gtcctgcgaa gcaagccgca ggtgagggcg tgggccaatt ctgcgcctca ctgctgttag
781 gcgcaaggac atcccgaaac acactcaacg acaccagcag cgagactgac agaagcggat
841 cgccagcggg agcaacaact tgaggcgagc cgttagccgc ctcgccttcc tcgcagcgcc
901 cctcccgcgc cccgggaagc ccgttcggct ccgccttttc gactgacgtc ccgggcgagc
961 gggggcgggc gcgtgggtgg gccgagccgt ctgcagccag cgattcggct ggctctgcca
1021 caccaccgcg cgcccccgct ccgcccgccc ctccgggcgc gtcttttccg ggctcgcgct
1081 gagtcccgcc tccgccggct gtccgggtgc gcgcgcgccg ctgcggcttt ttctctggcc
1141 tccgccgcgc gctcctcctc gtcccagcgc tagcgggcac gcggttcctt tttgcgagct
1201 ttccgagtgc caggcgccgg ccggctgcga agacgcggtg ggccgcccct ccgaagcttg
1261 gcattccggt actgttggta aagccaccat ggaagacgcc aaaaacataa agaaaggccc
1321 ggcgccattc tatccgctgg aagatggaac cgctggagag caactgcata aggctatgaa
1381 gagatacgcc ctggttcctg gaacaattgc ttttacagat gcacatatcg aggtggacat
1441 cacttacgct gagtacttcg aaatgtccgt tcggttggca gaagctatga aacgatatgg
1501 gctgaataca aatcacagaa tcgtcgtatg cagtgaaaac tctcttcaat tctttatgcc
1561 ggtgttgggc gcgttattta tcggagttgc agttgcgccc gcgaacgaca tttataatga
1621 acgtgaattg ctcaacagta tgggcatttc gcagcctacc gtggtgttcg tttccaaaaa
1681 ggggttgcaa aaaattttga acgtgcaaaa aaagctccca atcatccaaa aaattattat
1741 catggattct aaaacggatt accagggatt tcagtcgatg tacacgttcg tcacatctca
1801 tctacctccc ggttttaatg aatacgattt tgtgccagag tccttcgata gggacaagac
1861 aattgcactg atcatgaact cctctggatc tactggtctg cctaaaggtg tcgctctgcc
1921 tcatagaact gcctgcgtga gattctcgca tgccagagat cctatttttg gcaatcaaat
1981 cattccggat actgcgattt taagtgttgt tccattccat cacggttttg gaatgtttac
2041 tacactcgga tatttgatat gtggatttcg agtcgtctta atgtatagat ttgaagaaga
2101 gctgtttctg aggagccttc aggattacaa gattcaaagt gcgctgctgg tgccaaccct
2161 attctccttc ttcgccaaaa gcactctgat tgacaaatac gatttatcta atttacacga
2221 aattgcttct ggtggcgctc ccctctctaa ggaagtcggg gaagcggttg ccaagaggtt
2281 ccatctgcca ggtatcaggc aaggatatgg gctcactgag actacatcag ctattctgat
2341 tacacccgag ggggatgata aaccgggcgc ggtcggtaaa gttgttccat tttttgaagc
2401 gaaggttgtg gatctggata ccgggaaaac gctgggcgtt aatcaaagag gcgaactgtg
2461 tgtgagaggt cctatgatta tgtccggtta tgtaaacaat ccggaagcga ccaacgcctt
2521 gattgacaag gatggatggc tacattctgg agacatagct tactgggacg aagacgaaca
2581 cttcttcatc gttgaccgcc tgaagtctct gattaagtac aaaggctatc aggtggctcc
2641 cgctgaattg gaatccatct tgctccaaca ccccaacatc ttcgacgcag gtgtcgcagg
2701 tcttcccgac gatgacgccg gtgaacttcc cgccgccgtt gttgttttgg agcacggaaa
2761 gacgatgacg gaaaaagaga tcgtggatta cgtcgccagt caagtaacaa ccgcgaaaaa
2821 gttgcgcgga ggagttgtgt ttgtggacga agtaccgaaa ggtcttaccg gaaaactcga
2881 cgcaagaaaa atcagagaga tcctcataaa ggccaagaag ggcggaaaga tcgccgtgta
2941 attctagagt cggggcggcc ggccgcttcg agcagacatg ataagataca ttgatgagtt
3001 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
3061 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
3121 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct
3181 ctacaaatgt ggtaaaatcg ataaggatcc gtcgaccgat gcccttgaga gccttcaacc
3241 cagtcagctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt atgactgtct
3301 tctttatcat gcaactcgta ggacaggtgc cggcagcgct cttccgcttc ctcgctcact
3361 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
3421 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
3481 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
3541 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
3601 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
3661 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
3721 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
3781 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
3841 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
3901 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
3961 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
4021 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
4081 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
4141 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
4201 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
4261 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
4321 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
4381 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
4441 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
4501 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
4561 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
4621 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
4681 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
4741 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
4801 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
4861 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
4921 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
4981 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
5041 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
5101 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
5161 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
5221 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgcgccc
5281 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
5341 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
5401 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
5461 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtg
//
| {
"Vector backbone": "pGL3 basic",
"Backbone size w/o insert (bp)": "4818",
"Vector type": "Mammalian Expression, Luciferase",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "Unknown"
} | {
"Academic/Nonprofit Terms": "UBMTA ; Luciferase Limited Use Label License",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "RB1-deficient prostate tumor growth and metastasis are vulnerable to ferroptosis induction via the E2F/ACSL4 axis",
"Publication": "J Clin Invest. 2023 Mar 16:e166647. doi: 10.1172/JCI166647.",
"DOI": "10.1172/JCI166647.",
"Authors": "Wang ME, Chen J, Lu Y, Bawcom AR, Wu J, Ou J, Asara JM, Armstrong AJ, Wang Q, Li L, Wang Y, Huang J, Chen M /JCI",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/36928314",
"PubMed ID": "PubMed 36928314"
} | {
"Gene/Insert name": "ACSL4 promoter (partial)",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "ACSL4",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "XhoI (not destroyed)",
"3′ cloning site": "HindIII (not destroyed)",
"5′ sequencing primer": "RVprimer3",
"3′ sequencing primer": "GLprimer2"
} | {
"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": ""
} |
|
pHES943 | 112641 | yeast mating pheromone response pathway reporter | Hana El-Samad | Harrigan et al Cell. 2018 Oct 18;175(3):877-886.e10. doi: 10.1016/j.cell.2018.09.044. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/224838/6bcd8a99-0aee-426d-95c0-e0b163dce9c2/addgene-plasmid-112641-sequence-224838.gbk | full | LOCUS sequence-224838- 8387 bp DNA circular SYN 26-AUG-2024
DEFINITION yeast mating pheromone response pathway reporter.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8387)
AUTHORS Harrigan P, Madhani HD, El-Samad H
TITLE Real-Time Genetic Compensation Defines the Dynamic Demands of
Feedback Control.
JOURNAL Cell. 2018 Oct 18;175(3):877-886.e10. doi:
10.1016/j.cell.2018.09.044.
PUBMED 30340045
REFERENCE 2 (bases 1 to 8387)
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.2018.09"; journalName: "Cell"; date: "2018-10-18-
18"; volume: "175"; issue: "3"; pages: "877-886.e10"
FEATURES Location/Qualifiers
source 1..8387
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind 91..112
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 127..157
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 165..181
/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 170..192
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 189..205
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 189..205
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
promoter 226..244
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 501..715
/gene="S. cerevisiae URA3"
/label=URA3 promoter
CDS 2110..2826
/codon_start=1
/product="yeast-enhanced green fluorescent protein"
/label=yeGFP
/note="codons optimized for translation in S. cerevisiae
and C. albicans"
/translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
FVTAAGITHGMDELYK"
primer_bind complement(2828..2844)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2828..2844)
/label=pBluescriptSK
/note="For pBluescript vector"
promoter 3409..3752
/label=TEF promoter
/note="Ashbya gossypii TEF promoter"
terminator 4571..4768
/label=TEF terminator
/note="Ashbya gossypii TEF terminator"
primer_bind complement(5190..5209)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(5191..5209)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(5216..5233)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(5216..5232)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5225..5247)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 5373..5828
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5460..5479)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 5670..5691
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(6184..6203)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 6303..6325
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(6363..6381)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 6466..6570
/gene="bla"
/label=AmpR promoter
CDS 6571..7431
/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(6789..6808)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 7602..8190
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 8091..8110
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 8344..8361
/label=L4440
/note="L4440 vector, forward primer"
ORIGIN
1 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
61 ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc
121 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca
181 atttcacaca ggaaacagct atgaccatga ttacgccaag ctcggaatta accctcacta
241 aagggaacaa aagctgggcc ggccatttaa atgtttaaac ctactgcgcc aattgatgac
301 aatacagacg atgataacaa accgaagtta tctgatgtag aaaaggatta aagatgctaa
361 gagatagtga tgatatttca taaataatgt aattctatat atgttaatta ccttttttgc
421 gaggcatatt tatggtgaag gataagtttt gaccatcaaa gaaggttaat gtggctgtgg
481 tttcagggtc cataaagctt ttcaattcat cttttttttt tgttcttttt tttgattccg
541 gtttctttga aatttttttg attcggtaat ctccgagcag aaggaagaac gaaggaagga
601 gcacagactt agattggtat atatacgcat atgtggtgtt gaagaaacat gagattgccc
661 agtattctta acccaactgc acagaacaaa aacctgcagg aaacgaagat aaatcatgtc
721 gaaagctaca tataaggaac gtgctgctac tcatcctagt cctgttgctg ccaagctatt
781 taatatcatg cacgaaaagc aaacaaactt gtgtgcttca ttggatgttc gtaccaccaa
841 ggaattactg gagttagttg aagcattagg tcccaaaatt tgtttactaa aaacacatgt
901 ggatatcttg actgattttt ccatggaggg cacagttaag ccgctaaagg cattatccgc
961 caagtacaat tttttactct tcgaagacag aaaatttgct gacattggta atacagtcaa
1021 attgcagtac tctgcgggtg tatacagaat agcagaatgg gcagacatta cgaatgcaca
1081 cggtgtggtg ggccccctcc aatattctac atgaatgaca cttccaagcg tatcatcagt
1141 tggtgccaca ccattaatca gttttacgga gaaacaatcg ttgcatacac gtttgatgca
1201 ggtccaaatg ctgtgttgta ctacttagct gaaaatgagt cgaaactctt tgcatttatc
1261 tataaattgt ttggctctgt tcctggatgg gacaagaaat ttactactga gcagcttgag
1321 gctttcaacc atcaatttga atcatctaac tttactgcac gtgaattgga tcttgagttg
1381 caaaaggatg ttgccagagt gattttaact caagtcggtt caggcccaca agaaacaaac
1441 gaatctttga ttgacgcaaa gactggtcta ccaaaggaat aagatcaatt cgatatgtaa
1501 catttttctt ttcttttctt ttcctttttt tacaatagct aatttacgtt tccctacggt
1561 attggtcgga acgaccaagc ttcaatttat aaatatctta attttaacag cagttaccac
1621 ttgaatgaga aactttttga tgcctactct tttgcttttt catatcatga cctccctaca
1681 tgcttcggaa tttccttatt cgtagctgtc taacagcacc acttagcaca agtacgtcca
1741 cccgtttcat acttaaagat ctcagcagcc acccaatcta catttagggt acctgtcaca
1801 tatcttctca aattttaggt tagctttcaa ttgtaaagtc tctttttgga tattgcacgt
1861 ttcaagtcaa aacgcggctg taacataatt ttctgaaaca atattaaaac atagtaatga
1921 cttaaaacaa gcgactatcg caaataagca gttcccagca agcaattaaa acgtatataa
1981 gcgcagaata atgatctatc atcaataatt atcgtgaact tttccagtat ttacgttcat
2041 ctttgaaaaa cggcgagcat taacaataaa agctaaaaaa accaaaaaat ttcagctcga
2101 gaattaataa tgtctaaagg tgaagaatta ttcactggtg ttgtcccaat tttggttgaa
2161 ttagatggtg atgttaatgg tcacaaattt tctgtctccg gtgaaggtga aggtgatgct
2221 acttacggta aattgacctt aaaattgatt tgtactactg gtaaattgcc agttccatgg
2281 ccaaccttag tcactacttt aggttatggt ttgcaatgtt ttgctagata cccagatcat
2341 atgaaacaac atgacttttt caagtctgcc atgccagaag gttatgttca agaaagaact
2401 atttttttca aagatgacgg taactacaag accagagctg aagtcaagtt tgaaggtgat
2461 accttagtta atagaatcga attaaaaggt attgatttta aagaagatgg taacatttta
2521 ggtcacaaat tggaatacaa ctataactct cacaatgttt acatcactgc tgacaaacaa
2581 aagaatggta tcaaagctaa cttcaaaatt agacacaaca ttgaagatgg tggtgttcaa
2641 ttagctgacc attatcaaca aaatactcca attggtgatg gtccagtctt gttaccagac
2701 aaccattact tatcctatca atctgcctta tccaaagatc caaacgaaaa gagagaccac
2761 atggtcttgt tagaatttgt tactgctgct ggtattaccc atggtatgga tgaattgtac
2821 aaataaggat ccactagttc tagacacctg caacatgcgt gagtaaataa gcggccgcca
2881 ccgcggtgga gctctaagca aatagctaaa ttatatacga attaatatta tgattaagtg
2941 tttacgtgag tgcgatattt ttattactat cttatacagt tgtatatact ctataaaatg
3001 agttgtctat taattaacgc gatgaatgct ttctgggttt acctctccaa caactctagt
3061 ttacttctca atacattcaa ttgtatttga tttgtcaata cttcatcatt aatcaattct
3121 atagttttgt ttttctcgtt tatttccaaa tttaatgcat caattttatt attcaatttg
3181 tcgttgattt tggttaatga ttttatggtt tgatctctgg cattgattgt ttgtgttagt
3241 ttttcattat tgataattaa attatttaag ttagttatca actcggtgtt ttcaagtttc
3301 aagttttcaa tttctttaga gtttattaga tttgtcaaag tttctgaatt gcttgattgg
3361 tccagatctg tttagcttgc ctcgtccccg ccgggtcacc cggccagcga catggaggcc
3421 cagaataccc tccttgacag tcttgacgtg cgcagctcag gggcatgatg tgactgtcgc
3481 ccgtacattt agcccataca tccccatgta taatcatttg catccataca ttttgatggc
3541 cgcacggcgc gaagcaaaaa ttacggctcc tcgctgcgga cctgcgagca gggaaacgct
3601 cccctcacag acgcgttgaa ttgtccccac gccgcgcccc tgtagagaaa tataaaaggt
3661 taggatttgc cactgaggtt cttctttcat atacttcctt ttaaaatctt gctaggatac
3721 agttctcaca tcacatccga acataaacaa ccatgacagt caacactaag acctatagtg
3781 ggagagcaga aactcatgcc tcaccagtag cacaacgatt atttcgatta atggaactga
3841 agaaaaccaa tttatgtgca tcaattgatg ttgataccac taaggaattc cttgaattaa
3901 ttgataaatt gggtccttat gtatgcttaa tcaagactca tattgatata atcaatgatt
3961 tttcctatga atccactatt gaaccattat tagaactttc acgtaaacat caatttatga
4021 tttttgaaga tagaaaattt gctgatattg gtaataccgt gaagaaacaa tatattggtg
4081 gagtttataa aattagtagt tgggcagata ttactaatgc tcatggtgtc actgggaatg
4141 gagtagttga aggattaaaa cagggagcta aagaaaccac caccaaccaa gagccaagag
4201 ggttattgat gttagctgaa ttatcatcag tgggatcatt agcatatgga gaatattctc
4261 aaaaaactgt tgaaattgct aaatccgata aggaatttgt tattggattt attgcccaac
4321 gtgatatggg tggacaagaa gaaggatttg attggcttat tatgacacct ggagttggat
4381 tagatgataa aggtgatgga ttaggacaac aatatagaac tgttgatgaa gttgttagca
4441 ctggaactga tattatcatt gttggtagag gattgtttgg taaaggaaga gatccagata
4501 ttgaaggtaa aaggtataga gatgctggtt ggaatgctta tttgaaaaag actggccaat
4561 tataatcagt actgacaata aaaagattct tgttttcaag aacttgtcat ttgtatagtt
4621 tttttatatt gtagttgttc tattttaatc aaatgttagc gtgatttata ttttttttcg
4681 cctcgacatc atctgcccag atgcgaagtt aagtgcgcag aaagtaatat catgcgtcaa
4741 tcgtatgtga atgctggtcg ctatactgct gtcgattcga tactaacgcc gccatccagt
4801 gtcgaaaacg agctcgaatt catcgacccg ggaatctcgg tcgtaatgat ttctataatg
4861 acgaaaaaaa aaaaattgga aagaaaaagc ttcatggcct ttataaaaag gaactatcca
4921 atacctcgcc agaaccaagt aacagtattt tacggggcac aaatcaagaa caataagaca
4981 ggactgtaaa gatggacgca ttgaactcca aagaacaaca agagttccaa aaagtagtgg
5041 aacaaaagca aatgaaggat ttcatgcgtt tgtactctaa tctggtagaa agatgtttca
5101 cagactgtgt caatgacttc acaacatcaa agctaaccaa taaggaacaa acatgcatca
5161 tgaagtgcgt ttaaacggcc acgctggccc cctatagtga gtcgtattac aattcactgg
5221 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
5281 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
5341 cccaacagtt gcgcagcctg aatggcgaat ggacgcgccc tgtagcggcg cattaagcgc
5401 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
5461 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
5521 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
5581 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
5641 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
5701 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
5761 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
5821 tacaatttcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
5881 gggtaataac tgatataatt aaattgaagc tctaatttgt gagtttagta ttaggatctc
5941 tacccttggc gaaaagtcct ctgccaacaa tgatgatatc tgatccacca cttacaactt
6001 cgtcgacggt tctgtactgc tgacccaata tgcgtatata taccaatcta agtctgtgct
6061 ccttccttcg ttcttccttc tgttcggaga ttaccgaatc aaaaaaattt caaggaaacc
6121 gaaatcaaaa aaaagaataa aaaaaaaatg atgaattgaa aaggtggtat ggtgcactct
6181 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
6241 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
6301 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
6361 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
6421 gtcggcgcgc cgaggacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
6481 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
6541 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
6601 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
6661 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
6721 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
6781 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
6841 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
6901 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
6961 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
7021 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
7081 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
7141 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
7201 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
7261 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
7321 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
7381 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
7441 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
7501 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
7561 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
7621 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
7681 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
7741 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
7801 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
7861 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
7921 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
7981 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
8041 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
8101 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
8161 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
8221 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
8281 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
8341 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaa
//
| {
"Vector backbone": "pSV606",
"Backbone size w/o insert (bp)": "8000",
"Vector type": "Yeast Expression",
"Selectable markers": "URA3"
} | {
"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": "Real-Time Genetic Compensation Defines the Dynamic Demands of Feedback Control",
"Publication": "Cell. 2018 Oct 18;175(3):877-886.e10. doi: 10.1016/j.cell.2018.09.044.",
"DOI": "10.1016/j.cell.2018.09.044.",
"Authors": "Harrigan P, Madhani HD, El-Samad H /jcell",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/30340045",
"PubMed ID": "PubMed 30340045"
} | {
"Gene/Insert name": "yeGFP",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "AGA1",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "XhoI (not destroyed)",
"3′ cloning site": "BamHI (not destroyed)",
"5′ sequencing primer": "CAGAATAGCAGAATGGGCAGAC",
"3′ sequencing primer": "ATC GCA CTC ACG TAA ACA CTT"
} | {
"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": ""
} |
|
pRS414 Hsh155 Y292C | 87267 | Encodes Hsh155 Y292C for expression in yeast | Aaron Hoskins | Carrocci et al Nucleic Acids Res. 2017 Jan 6. pii: gkw1349. doi: 10.1093/nar/gkw1349. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/189171/c5e1fb2e-8a20-4b4f-9bf1-5962f31feca5/addgene-plasmid-87267-sequence-189171.gbk | full | LOCUS sequence-189171- 8127 bp DNA circular SYN 26-AUG-2024
DEFINITION Encodes Hsh155 Y292C for expression in yeast.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8127)
AUTHORS Carrocci TJ, Zoerner DM, Paulson JC, Hoskins AA
TITLE SF3b1 mutations associated with myelodysplastic syndromes alter the
fidelity of branchsite selection in yeast.
JOURNAL Nucleic Acids Res. 2017 Jan 6. pii: gkw1349. doi:
10.1093/nar/gkw1349.
PUBMED 28062854
REFERENCE 2 (bases 1 to 8127)
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: "Nucleic
Acids Res. 2017 Jan 6. pii: gkw1349. doi: 10.1093/nar/gkw1349."
FEATURES Location/Qualifiers
source 1..8127
/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"
promoter 191..471
/gene="S. cerevisiae TRP1"
/label=TRP1 promoter
CDS 472..1146
/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 complement(1249..1704)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1386..1407)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 1598..1617
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 1830..1852
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 1844..1861
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 1845..1861
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 1871..1890
/label=T7
/note="T7 promoter, forward primer"
promoter 1871..1889
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(5354..5374)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(5354..5372)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(5393..5409)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5393..5409)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(5406..5428)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 5417..5433
/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(5441..5471)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5486..5507
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(5624..5641)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5795..6383)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5875..5894)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6554..7414)
/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 7177..7196
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7415..7519)
/gene="bla"
/label=AmpR promoter
misc_feature 7556..8059
/label=CEN/ARS
/note="S. cerevisiae CEN6 centromere fused to an
autonomously replicating sequence"
primer_bind 8100..8118
/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 accatagatc acgacattac tatatatata atataggaag catttaatag aacagcatcg
241 taatatatgt gtactttgca gttatgacgc cagatggcag tagtggaaga tattctttat
301 tgaaaaatag cttgtcacct tacgtacaat cttgatccgg agcttttctt tttttgccga
361 ttaagaatta attcggtcga aaaaagaaaa ggagagggcc aagagggagg gcattggtga
421 ctattgagca cgtgagtata cgtgattaag cacacaaagg cagcttggag tatgtctgtt
481 attaatttca caggtagttc tggtccattg gtgaaagttt gcggcttgca gagcacagag
541 gccgcagaat gtgctctaga ttccgatgct gacttgctgg gtattatatg tgtgcccaat
601 agaaagagaa caattgaccc ggttattgca aggaaaattt caagtcttgt aaaagcatat
661 aaaaatagtt caggcactcc gaaatacttg gttggcgtgt ttcgtaatca acctaaggag
721 gatgttttgg ctctggtcaa tgattacggc attgatatcg tccaactgca tggagatgag
781 tcgtggcaag aataccaaga gttcctcggt ttgccagtta ttaaaagact cgtatttcca
841 aaagactgca acatactact cagtgcagct tcacagaaac ctcattcgtt tattcccttg
901 tttgattcag aagcaggtgg gacaggtgaa cttttggatt ggaactcgat ttctgactgg
961 gttggaaggc aagagagccc cgaaagctta cattttatgt tagctggtgg actgacgcca
1021 gaaaatgttg gtgatgcgct tagattaaat ggcgttattg gtgttgatgt aagcggaggt
1081 gtggagacaa atggtgtaaa agactctaac aaaatagcaa atttcgtcaa aaatgctaag
1141 aaataggtta ttactgagta gtatttattt aagtattgtt tgtgcacttg ccgatctatg
1201 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggaa attgtaagcg
1261 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
1321 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
1381 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
1441 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
1501 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
1561 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
1621 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
1681 ttaatgcgcc gctacagggc gcgtccattc gccattcagg ctgcgcaact gttgggaagg
1741 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
1801 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
1861 tgagcgcgcg taatacgact cactataggg cgaattgggt accgggcccc cccctcgagc
1921 atgtactttc cgcctaacta ccatcacctg tctttattta ttccgctgac agatatgcac
1981 aaccaattat cgtctcttga aagcttttta gacgtgattg ctgttatttg gcacgttgtt
2041 agtatttaag cttattcggg tttgcattga tatttgcccg agcagaaatc tccttctcga
2101 tgagttagaa ctgaaacatc tcttcaatta cttatacata ataaacatag aaagccagga
2161 cctgtcatca tgagtcatcc gattcaattt gtaaacgcta ataatagcga taagagccac
2221 caattaggcg gtcaatattc tattcctcag gatttgaggg aaaacctgca gaaagaagct
2281 gcacgtattg gtgagaacga aaaagatgtg ctacaggaaa aaatggaaac taggacagtg
2341 caaaatcgag aggattcata ccacaaaaga aggtttgata tgaagtttga gccggattcc
2401 gatacacaaa ctgttacgtc ttcagaaaat acccaagacg ctgtcgtgcc aaggaaaaga
2461 aaatctcgtt gggatgtaaa agggtacgag ccacctgatg aatcttcaac agcggtcaaa
2521 gaaaactctg atagtgcttt ggttaatgta gaaggaatcc atgatttgat gttttttaaa
2581 ccttctgacc ataaatattt cgccgacgtt atatccaaaa aacccataga tgagttaaat
2641 aaggatgaaa agaaagaaag aaccctgtcg atgttacttc tgaaaataaa aaatggcaat
2701 acagcgagta ggaggacttc tatgcggatt ttaaccgata aagctgttac atttggtcca
2761 gaaatgatat ttaaccgctt attgcccata ttactagaca gaagtttgga agaccaggaa
2821 agacatctaa tgatcaagac catagatcgc gtcctttacc agctaggaga tttaacaaaa
2881 ccttacgttc acaaaatact tgtagtagct gccccattgc taatagatga agatccaatg
2941 gttcgttcaa caggccagga gataatcacc aatctgtcta cagtcgctgg cctaaaaacc
3001 attctaacag taatgagacc tgatattgag aatgaggatg agtgtgttag aaatgtaact
3061 tcaagagcgg cggctgtggt ggcaaaagcc cttggtgtta atcaattgct tcctttcatc
3121 aatgcagcat gccactcacg gaagtcttgg aaagcaaggc atactggcat aaaaattgtg
3181 cagcaaatag gtatactttt gggaataggt gtcttgaatc atcttactgg gctgatgagt
3241 tgtatcaaag actgcctaat ggatgatcac gttccggtgc gaattgttac agcacatact
3301 ttatcaacac tagcagaaaa ttcttatcca tatggtatcg aagttttcaa cgttgtactg
3361 gagccattat ggaagggtat aagaagtcat cgtggtaaag tgctatcttc ttttttaaaa
3421 gctgttggat ctatgatccc attgatggac ccagagtatg ccggttatta taccacagaa
3481 gcaatgagaa taattagacg tgaattcgac tctcccgatg atgaaatgaa aaagaccatt
3541 cttttagtat tacaaaaatg tagcgctgta gaatccataa cgccaaaatt tctgagagaa
3601 gaaatcgcgc cagaattctt tcaaaaattt tgggtcagac gagtggcatt ggatagacca
3661 ttaaacaaag ttgttacata tactactgtt acattagcaa agaaattagg ttgttcttat
3721 accattgata aattattaac accattgaga gatgaagctg agccgttcag aacaatggct
3781 gtccatgctg ttactagaac agtaaactta ttagggacag cagatttaga tgagagatta
3841 gaaacgaggc ttattgatgc gctcttgata gcatttcaag aacaaacaaa tagtgattct
3901 ataatcttta aagggtttgg ggcagtgacg gtgtcacttg acatccgtat gaaaccgttt
3961 ttagcaccca ttgtgagtac gattttaaat catttaaagc ataaaactcc cttggttcgt
4021 caacatgcag cagatctatg tgccatacta ataccagtca tcaagaactg tcacgaattc
4081 gaaatgttga ataaattgaa cattatattg tatgaatcct taggcgaagt atatccagag
4141 gtattaggtt ctataatcaa cgctatgtac tgtattacta gcgtaatgga tttggataaa
4201 ctacaacctc ctattaatca aattttacca accttaactc caattttaag aaacaagcat
4261 agaaaagtcg aagtaaacac tatcaaattt gttggactta ttggtaaatt agcacctacc
4321 tatgcacctc ctaaagaatg gatgagaata tgttttgaac tattagaact tttgaaaagt
4381 acaaataaag aaataagaag atcagcaaat gccacgtttg gattcattgc tgaagcaatt
4441 ggacctcacg atgttcttgt tgcgttattg aacaatttaa aggttcaaga acgtcagtta
4501 cgtgtatgca cggctgttgc tatcggtatc gttgctaaag tgtgtggtcc ttacaacgtg
4561 ctgccagtaa tcatgaatga atacacaaca ccagagacta atgttcaaaa cggtgttctc
4621 aaggccatgt cattcatgtt tgagtacatt ggtaacatgt ctaaggatta catatacttc
4681 ataacaccat tattagaaga tgcactcacc gatagggatt tagttcatcg tcagacagcg
4741 tcgaacgtaa taacccactt agctttaaac tgttcaggta caggtcacga agatgccttc
4801 attcatttaa tgaatcttct aatacccaat atttttgaaa cttcaccaca tgctattatg
4861 cgtattttag aaggtttgga ggcactgagt caagctcttg gccctggact attcatgaac
4921 tacatatggg caggattgtt ccatccagcg aagaatgtaa ggaaagcatt ttggagggtg
4981 tataataaca tgtacgtaat gtaccaggac gccatggtac ctttttaccc cgttacacca
5041 gacaacaatg aagaatatat agaagaactg gatttagttc tgtgaatata tatagagcat
5101 ataaataacc acaacttgta agaatatttt acttgacaca cctatatcat cacgtcacga
5161 tgccttcata gattatgatg aaatataaac agaacctgct attggaaaaa aaagggcagg
5221 aaaaaatttt gtgaaagtga gtaaaagaaa gatgaagagc gagacatgga taataagaaa
5281 tagatgtcac taatggacaa ttggaagact gatatggaaa gttacgatga aggaggagct
5341 ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc
5401 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
5461 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
5521 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
5581 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
5641 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
5701 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
5761 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
5821 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
5881 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
5941 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
6001 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
6061 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
6121 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
6181 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
6241 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
6301 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
6361 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
6421 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
6481 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
6541 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
6601 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
6661 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
6721 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
6781 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
6841 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
6901 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
6961 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
7021 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
7081 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
7141 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
7201 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
7261 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
7321 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
7381 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
7441 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
7501 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgggtcc ttttcatcac
7561 gtgctataaa aataattata atttaaattt tttaatataa atatataaat taaaaataga
7621 aagtaaaaaa agaaattaaa gaaaaaatag tttttgtttt ccgaagatgt aaaagactct
7681 agggggatcg ccaacaaata ctacctttta tcttgctctt cctgctctca ggtattaatg
7741 ccgaattgtt tcatcttgtc tgtgtagaag accacacacg aaaatcctgt gattttacat
7801 tttacttatc gttaatcgaa tgtatatcta tttaatctgc ttttcttgtc taataaatat
7861 atatgtaaag tacgcttttt gttgaaattt tttaaacctt tgtttatttt tttttcttca
7921 ttccgtaact cttctacctt ctttatttac tttctaaaat ccaaatacaa aacataaaaa
7981 taaataaaca cagagtaaat tcccaaatta ttccatcatt aaaagatacg aggcgcgtgt
8041 aagttacagg caagcgatcc gtctaagaaa ccattattat catgacatta acctataaaa
8101 ataggcgtat cacgaggccc tttcgtc
//
| {
"Vector backbone": "pRS414",
"Backbone size w/o insert (bp)": "4788",
"Vector type": "Bacterial Expression, 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": "SF3b1 mutations associated with myelodysplastic syndromes alter the fidelity of branchsite selection in yeast",
"Publication": "Nucleic Acids Res. 2017 Jan 6. pii: gkw1349. doi: 10.1093/nar/gkw1349.",
"DOI": "10.1093/nar/gkw1349.",
"Authors": "Carrocci TJ, Zoerner DM, Paulson JC, Hoskins AA /nar/gkw",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/28062854",
"PubMed ID": "PubMed 28062854"
} | {
"Gene/Insert name": "HSH155",
"Species": "S. cerevisiae (budding yeast)",
"Insert Size (bp)": "3426",
"Mutation": "Y292C",
"Entrez Gene": "HSH155",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "SacI (not destroyed)",
"3′ cloning site": "XhoI (not destroyed)",
"5′ sequencing primer": "TGTAAAACGACGGCCAGT",
"3′ sequencing primer": "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": ""
} |