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 |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
pBACgus/LRRK2 1867-2176 C-His | 40378 | Michael J Fox Foundation MJFF | Michael J Fox Foundation plasmids (unpublished) | https://media.addgene.org/snapgene-media/v2.0.0/sequences/52055/33e78450-fb17-4c44-8791-65bf64b343dc/addgene-plasmid-40378-sequence-52055.gbk | full | LOCUS sequence-52055-e 8344 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 8344)
AUTHORS MJFF
TITLE Michael J Fox Foundation plasmids
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 8344)
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"Unpublished"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..8344
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_recomb 329..1156
/label=baculovirus recombination region (lef2/ORF603)
/note="contains ORF603 and part of lef2"
CDS 329..453
/codon_start=3
/product="baculovirus late expression factor 2"
/label=lef2
/translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
CDS complement(491..1096)
/codon_start=1
/product="baculovirus ORF603 protein"
/label=ORF603
/translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
CDS complement(1167..2978)
/codon_start=1
/gene="uidA (or gusA) from E. coli"
/product="beta-glucuronidase"
/label=GUS
/translation="MVRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAI
AVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEV
MEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHD
FFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELRDADQQ
VVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGQQF
LINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDW
ADEHGIVVIDETAAVGFSLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARD
KNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAHTDTISD
LFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMY
TDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRK
PKSAAFLLQKRWTGMNFGEKPQQGGKQ"
promoter complement(2987..3302)
/gene="Autographa californica p6.9"
/label=p6.9 promoter
/note="baculovirus late basic protein promoter"
promoter 3317..3408
/gene="polh from Autographa californica"
/label=polyhedrin promoter
/note="promoter for the baculovirus polyhedrin gene"
primer_bind 3333..3350
/label=Polyhedrin-F
/note="Polyhedrin promoter, forward primer"
CDS 4351..4368
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 4394..4411
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
misc_recomb 4448..5805
/label=baculovirus recombination region (ORF1629)
/note="contains part of ORF1629"
CDS complement(4464..5805)
/codon_start=2
/product="baculovirus capsid-associated protein"
/label=ORF1629
/note="required for viral replication"
/translation="PSLRQSLYNTIAYIERLLNIGTVNDSEITMLIADFYDLYSNYNIE
LPPPQALPRSRRPSVVQPAAPAPVPTIVREQTKPEQIIPAAPPPPPSPVPNIPAPPPPP
PPSMSELPPAPPMPTEPQPAAPLDDRQQLLEAIRNEKNRTRLRPVKPKTAPETSTIVEV
PTVLPKETFEPKPPSASPPPPPPPPPPPAPPAPPPMVDLSSAPPPPPLVDLPSEMLPPP
APSLSNVLSELKSGTVRLKPAQKRPQSEIIPKSSTTNLIADVLADTINRRRVAMAKSSS
EATSNDEGWDDDDNRPNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVE
PLLQNKTQTNIDKARLLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDL
IFKFRYKDAENHLIFALTYHPKDYKFNELLKYVQQLSVNQQRTESSA"
primer_bind complement(5897..5914)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(6068..6656)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6148..6167)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6827..7687)
/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 7450..7469
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7688..7792)
/gene="bla"
/label=AmpR promoter
rep_origin 7819..8274
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(7906..7925)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 8116..8137
/label=F1ori-F
/note="F1 origin, forward primer"
ORIGIN
1 ggaacggctc cgcccactat taatgaaatt aaaaattcca attttaaaaa acgcagcaag
61 agaaacattt gtatgaaaga atgcgtagaa ggaaagaaaa atgtcgtcga catgctgaac
121 aacaagatta atatgcctcc gtgtataaaa aaaatattga acgatttgaa agaaaacaat
181 gtaccgcgcg gcggtatgta caggaagagg tttatactaa actgttacat tgcaaacgtg
241 gtttcgtgtg ccaagtgtga aaaccgatgt ttaatcaagg ctctgacgca tttctacaac
301 cacgactcca agtgtgtggg tgaagtcatg catcttttaa tcaaatccca agatgtgtat
361 aaaccaccaa actgccaaaa aatgaaaact gtcgacaagc tctgtccgtt tgctggcaac
421 tgcaagggtc tcaatcctat ttgtaattat tgaataataa aacaattata aatgctaaat
481 ttgtttttta ttaacgatac aaaccaaacg caacaagaac atttgtagta ttatctataa
541 ttgaaaacgc gtagttataa tcgctgaggt aatatttaaa atcattttca aatgattcac
601 agttaatttg cgacaatata attttatttt cacataaact agacgccttg tcgtcttctt
661 cttcgtattc cttctctttt tcatttttct cttcataaaa attaacatag ttattatcgt
721 atccatatat gtatctatcg tatagagtaa attttttgtt gtcataaata tatatgtctt
781 ttttaatggg gtgtatagta ccgctgcgca tagtttttct gtaatttaca acagtgctat
841 tttctggtag ttcttcggag tgtgttgctt taattattaa atttatataa tcaatgaatt
901 tgggatcgtc ggttttgtac aatatgttgc cggcatagta cgcagcttct tctagttcaa
961 ttacaccatt ttttagcagc accggattaa cataactttc caaaatgttg tacgaaccgt
1021 taaacaaaaa cagttcacct cccttttcta tactattgtc tgcgagcagt tgtttgttgt
1081 taaaaataac agccattgta atgagacgca caaactaata tcacaaactg gaaatgtcta
1141 tcaatatata gttgctgatc agatcctcat tgtttgcctc cctgctgcgg tttttcaccg
1201 aagttcatgc cagtccagcg tttttgcagc agaaaagccg ccgacttcgg tttgcggtcg
1261 cgagtgaaga tccctttctt gttaccgcca acgcgcaata tgccttgcga ggtcgcaaaa
1321 tcggcgaaat tccatacctg ttcaccgacg acggcgctga cgcgatcaaa gacgcggtga
1381 tacatatcca gccatgcaca ctgatactct tcactccaca tgtcggtgta cattgagtgc
1441 agcccggcta acgtatccac gccgtattcg gtgatgataa tcggctgatg cagtttctcc
1501 tgccaggcca gaagttcttt ttccagtacc ttctctgccg tttccaaatc gccgctttgg
1561 acataccatc cgtaataacg gttcaggcac agcacatcaa agagatcgct gatggtatcg
1621 gtgtgagcgt cgcagaacat tacattgacg caggtgatcg gacgcgtcgg gtcgagttta
1681 cgcgttgctt ccgccagtgg cgcgaaatat tcccgtgcac cttgcggacg ggtatccggt
1741 tcgttggcaa tactccacat caccacgctt gggtggtttt tgtcacgcgc tatcagctct
1801 ttaatcgcct gtaagtgcgc ttgctgagtt tccccgttga ctgcctcttc gctgtacagt
1861 tctttcggct tgttgcccgc ttcgaaacca atgcctaaag agagcgaaaa gccgacagca
1921 gcagtttcat caatcaccac gatgccatgt tcatctgccc agtcgagcat ctcttcagcg
1981 taagggtaat gcgaggtacg gtaggagttg gccccaatcc agtccattaa tgcgtggtcg
2041 tgcaccatca gcacgttatc gaatcctttg ccacgtaagt ccgcatcttc atgacgacca
2101 aagccagtaa agtagaacgg tttgtggtta atcaggaact gttggccctt cactgccact
2161 gaccggatgc cgacgcgaag cgggtagata tcacactctg tctggctttt ggctgtgacg
2221 cacagttcat agagataacc ttcacccggt tgccagaggt gcggattcac cacttgcaaa
2281 gtcccgctag tgccttgtcc agttgcaacc acctgttgat ccgcatcacg cagttcaacg
2341 ctgacatcac cattggccac cacctgccag tcaacagacg cgtggttaca gtcttgcgcg
2401 acatgcgtca ccacggtgat atcgtccacc caggtgttcg gcgtggtgta gagcattacg
2461 ctgcgatgga ttccggcata gttaaagaaa tcatggaagt aagactgctt tttcttgccg
2521 ttttcgtcgg taatcaccat tcccggcggg atagtctgcc agttcagttc gttgttcaca
2581 caaacggtga tacgtacact tttcccggca ataacatacg gcgtgacatc ggcttcaaat
2641 ggcgtatagc cgccctgatg ctccatcact tcctgattat tgacccacac tttgccgtaa
2701 tgagtgaccg catcgaaacg cagcacgata cgctggcctg cccaaccttt cggtataaag
2761 acttcgcgct gataccagac gttgcccgca taattacgaa tatctgcatc ggcgaactga
2821 tcgttaaaac tgcctggcac agcaattgcc cggctttctt gtaacgcgct ttcccaccaa
2881 cgctgatcaa ttccacagtt ttcgcgatcc agactgaatg cccacaggcc gtcgagtttt
2941 ttgatttcac gggttggggt ttctacagga cggaccatat ttataggtgt aatttatgta
3001 gctgtaattt ttaccttatt aatatttttt acgctttgca ttcgacgact gaactcccaa
3061 atatatgttt aactcgtctt ggtcgtttga atttttgttg ctgtgtttcc taatattttc
3121 catcacctta aatatgttat tgtaatcctc aatgttgaac ttgcaattgg acacggcata
3181 gttttccata gtcgtgtaaa acatggtatt ggctgcattg taatacatcc gactgagcgg
3241 gtacggatct atgtgtttga gcagcctgtt caaaaactct gcatcgtcgc aaaacggaat
3301 ttcgtcgaca gatctgatca tggagataat taaaatgata accatctcgc aaataaataa
3361 gtattttact gttttcgtaa cagttttgta ataaaaaaac ctataaatat aggatccatg
3421 aacatcatgc tcaacaacga tgagctcgaa tttgaacaag ctccagaatt cctgctcggc
3481 gacggctctt tcggctccgt ataccgtgct gcttatgaag gcgaagaagt cgccgtaaag
3541 atcttcaaca agcacacctc gctccgtctg ctccgtcagg aactggtcgt tctgtgtcat
3601 ctgcaccatc cgagcctcat ctccctgctg gccgctggca tccgtcctcg tatgctggtg
3661 atggagctgg catccaaggg ctctctcgac cgcctgctgc agcaggacaa ggcttctctg
3721 acccgtactc tgcagcatcg tatcgctctg catgtagctg acggcctgcg ttatcttcat
3781 tctgcaatga tcatttatcg cgacctgaaa cctcataatg tactgctgtt tactctgtac
3841 ccgaacgctg ccatcatcgc taagattgca gactatggta ttgcgcagta ctgctgccgc
3901 atgggcatta aaacaagcga aggtacgccg ggtttccgtg ctccagaagt ggctcgtggc
3961 aacgttattt acaaccagca agctgacgtt tactcgttcg gtctgctcct gtacgatatc
4021 ctgactaccg gcggccgtat cgtagaaggt ctgaagttcc ctaacgaatt cgacgaactg
4081 gaaatccagg gcaagctgcc agacccggtt aaagaatacg gttgcgctcc gtggccgatg
4141 gtagagaagc tcatcaaaca gtgccttaaa gaaaaccctc aggaacgtcc aacctctgct
4201 caagtgttcg acatccttaa cagcgctgaa ctcgtctgtc tgactcgccg tattctcctg
4261 cctaagaatg ttatcgtgga atgtatggtt gctactcacc ataactctcg caacgcttcc
4321 atctggctcg gctgtggcca cactgaccgt caccaccatc accatcattg ataagcttgc
4381 ggccgcactc gagcaccacc accaccacca ctaacctagg tagctgagcg catgcaagct
4441 gatccgggtt attagtacat ttattaagcg ctagattctg tgcgttgttg atttacagac
4501 aattgttgta cgtattttaa taattcatta aatttataat ctttagggtg gtatgttaga
4561 gcgaaaatca aatgattttc agcgtcttta tatctgaatt taaatattaa atcctcaata
4621 gatttgtaaa ataggtttcg attagtttca aacaagggtt gtttttccga accgatggct
4681 ggactatcta atggattttc gctcaacgcc acaaaacttg ccaaatcttg tagcagcaat
4741 ctagctttgt cgatattcgt ttgtgttttg ttttgtaata aaggttcgac gtcgttcaaa
4801 atattatgcg cttttgtatt tctttcatca ctgtcgttag tgtacaattg actcgacgta
4861 aacacgttaa atagagcttg gacatattta acatcgggcg tgttagcttt attaggccga
4921 ttatcgtcgt cgtcccaacc ctcgtcgtta gaagttgctt ccgaagacga ttttgccata
4981 gccacacgac gcctattaat tgtgtcggct aacacgtccg cgatcaaatt tgtagttgag
5041 ctttttggaa ttatttctga ttgcgggcgt ttttgggcgg gtttcaatct aactgtgccc
5101 gattttaatt cagacaacac gttagaaagc gatggtgcag gcggtggtaa catttcagac
5161 ggcaaatcta ctaatggcgg cggtggtgga gctgatgata aatctaccat cggtggaggc
5221 gcaggcgggg ctggcggcgg aggcggaggc ggaggtggtg gcggtgatgc agacggcggt
5281 ttaggctcaa atgtctcttt aggcaacaca gtcggcacct caactattgt actggtttcg
5341 ggcgccgttt ttggtttgac cggtctgaga cgagtgcgat ttttttcgtt tctaatagct
5401 tccaacaatt gttgtctgtc gtctaaaggt gcagcgggtt gaggttccgt cggcattggt
5461 ggagcgggcg gcaattcaga catcgatggt ggtggtggtg gtggaggcgc tggaatgtta
5521 ggcacgggag aaggtggtgg cggcggtgcc gccggtataa tttgttctgg tttagtttgt
5581 tcgcgcacga ttgtgggcac cggcgcaggc gccgctggct gcacaacgga aggtcgtctg
5641 cttcgaggca gcgcttgggg tggtggcaat tcaatattat aattggaata caaatcgtaa
5701 aaatctgcta taagcattgt aatttcgcta tcgtttaccg tgccgatatt taacaaccgc
5761 tcaatgtaag caattgtatt gtaaagagat tgtctcaagc tcggatcgat cccgcacgcc
5821 gataacaagc cttttcattt ttactacagc attgtagtgg cgagacactt cgctgtcgtc
5881 gaggtttaaa cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
5941 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
6001 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
6061 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
6121 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
6181 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
6241 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
6301 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
6361 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
6421 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
6481 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
6541 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
6601 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
6661 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
6721 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
6781 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
6841 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
6901 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
6961 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
7021 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
7081 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
7141 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
7201 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
7261 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
7321 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
7381 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
7441 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
7501 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
7561 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
7621 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
7681 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
7741 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
7801 cccgaaaagt gccacctgac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
7861 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
7921 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
7981 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
8041 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
8101 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
8161 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
8221 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca atttcccatt
8281 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
8341 gcca
//
| {
"Vector backbone": "pBACgus",
"Backbone size w/o insert (bp)": "",
"Vector type": "Baculovirus",
"Selectable markers": ""
} | {
"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": "Industry MTA"
} | {
"Title": "",
"Publication": "",
"DOI": "",
"Authors": "",
"PubMed Link": "",
"PubMed ID": ""
} | {
"Gene/Insert name": "LRRK2",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "WT aa1867-2176",
"Entrez Gene": "LRRK2",
"Promoter": "polH",
"Tag / Fusion Protein": "6xHis (C terminal on insert)"
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "BamHI (destroyed during cloning)",
"3′ cloning site": "HindIII (destroyed during cloning)",
"5′ sequencing primer": "DSP2466For:GAATTTCGTCGACAGATC",
"3′ sequencing primer": "DSP2466Rev:AATAACCCGGATCAGCTT"
} | {
"Gene/Insert name": "",
"Species": "",
"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": ""
} |
||
Ube2r1 | 31426 | Rachel Klevit | Christensen et al Nat Struct Mol Biol. 2007 Oct . 14(10):941-8. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/37917/caff4ec9-b5e8-4547-9bf4-13320610ed74/addgene-plasmid-31426-sequence-37917.gbk | partial | LOCUS sequence-37917-e 812 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 812)
AUTHORS Christensen DE, Brzovic PS, Klevit RE
TITLE E2-BRCA1 RING interactions dictate synthesis of mono- or specific
polyubiquitin chain linkages.
JOURNAL Nat Struct Mol Biol. 2007 Oct . 14(10):941-8.
PUBMED 17873885
REFERENCE 2 (bases 1 to 812)
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 Struct
Mol Biol. 2007 Oct . 14(10):941-8."
FEATURES Location/Qualifiers
source 1..812
/mol_type="genomic DNA"
/organism="unspecified"
CDS 35..61
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 41..67
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/note="This is a variant of the standard HA tag, which has
sequence YPYDVPDYA. Some anti-HA antibodies may not
recognize this variant."
/translation="YDVPDYASL"
protein_bind 98..122
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
ORIGIN
1 cccaccaaac ccaaaaaaag agatctctat ggcttaccca tacgatgttc cagattacgc
61 tagcttgggt ggtcatatgg ccatggaggc cggatctaca agtttgtaca aaaaagcagg
121 ctccgcggcc gcccccttca ccatggctcg gccgctagtg cccagctcgc agaaggcgct
181 gctgctggag ctcaaggggc tgcaggaaga gccggtcgag ggattccgcg tgacactggt
241 ggacgagggc gatctataca actgggaggt ggccatcttc gggcccccca acacctacta
301 cgagggcggc tacttcaagg cgcgcctcaa gttccccatc gactacccat actctccacc
361 agcctttcgg ttcctgacca agatgtggca ccctaacatc tacgagacgg gggacgtgtg
421 tatctccatc ctccacccgc cggtggacga cccccagagc ggggagctgc cctcagagag
481 gtggaacccc acgcagaacg tcaggaccat tctcctgagt gtgatctccc tcctgaacga
541 gcccaacacc ttctcgcccg caaacgtgga cgcctccgtg atgtacagga agtggaaaga
601 gagcaagggg aaggatcggg agtacacaga catcatccgg aagcaggtcc tggggaccaa
661 ggtggacgcg gagcgtgacg gcgtgaaggt gcccaccacg ctggccgagt actgcgtgaa
721 gaccaaggcg ccggcgcccg acgagggctc agacctcttc tacgacgact actacgagga
781 cggcgaggtg gaggaggagg ccgacagctg ct
//
| {
"Vector backbone": "pACT2",
"Backbone size w/o insert (bp)": "8100",
"Vector type": "Yeast 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": "E2-BRCA1 RING interactions dictate synthesis of mono- or specific polyubiquitin chain linkages",
"Publication": "Nat Struct Mol Biol. 2007 Oct . 14(10):941-8.",
"DOI": "10.1038/nsmb1295",
"Authors": "Christensen DE, Brzovic PS, Klevit RE",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/17873885",
"PubMed ID": "PubMed 17873885"
} | {
"Gene/Insert name": "Ubiquitin Conjugating Enzyme",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "400",
"Mutation": "",
"Entrez Gene": "CDC34",
"Promoter": "",
"Tag / Fusion Protein": "HA (N terminal on backbone)"
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "GAL4 AD",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"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": ""
} |
||
pcDNA3.1hisA cdc25c C330S | 10966 | Toren Finkel | Savitsky et al J Biol Chem. 2002 Jun 7. 277(23):20535-40. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/17935/16b81622-0fe8-4f84-95c8-3c9aee8944cc/addgene-plasmid-10966-sequence-17935.gbk | partial | LOCUS sequence-17935-e 950 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 950)
AUTHORS Savitsky PA, Finkel T
TITLE Redox regulation of Cdc25C.
JOURNAL J Biol Chem. 2002 Jun 7. 277(23):20535-40.
PUBMED 11925443
REFERENCE 2 (bases 1 to 950)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J Biol
Chem. 2002 Jun 7. 277(23):20535-40."
FEATURES Location/Qualifiers
source 1..950
/mol_type="genomic DNA"
/organism="unspecified"
CDS complement(19..60)
/codon_start=1
/product="epitope tag from simian virus 5"
/label=V5 tag
/translation="GKPIPNPLLGLDST"
ORIGIN
1 gtgatgatga ccggtacgcg tagaatcgag accgaggaga gggttaggga taggcttacc
61 ttcgaagggc cctctagatg ggctcatgtc cttcaccaga agggcaatct gctcccgcag
121 ctgccgctcc ccttcctgca ctttgctctg gcttcgacac ctcagcaact cagtcttgtg
181 gtcctgatga tgcatagggc agtagctctg tggttcacac agttccatat attctggaaa
241 gaagtctctg tagccgcctt taaggatata tagctctggg tagtacaatg caggatactg
301 gttcagagac ctgtcctctt cacgcagaca gcggcacatt cgggggcccc tctctgagga
361 gaattcacag tggaacacga tgattattct cttctgggtg tccaaaggga cgatgggctt
421 cttcagaaag aagttaaaca gttcttcctg actatataag tttaaggctc cctggatgtg
481 tcctcccaga tactcataag gatagcgtga atcaatgaca taaaacttct caatcagacc
541 ctggaacttc cccgacagta aggcagccac tgtttctggg ttgacatact tcagatcttg
601 gtgtttccct gacacggttg gcagcgcaca taccttggaa aaatcaccaa tcaggtgccc
661 ctggttagaa tcttcctcca gcatctgagt gatagtaatg tcacacagag agactgtctt
721 ctttaaacat aagcccttcc tgagctttcc ttggccagaa aaatactttt ttttaacttt
781 atctggtatt gtgttgtcct tgaatttttc cacctgcttc agtcttggcc tgttcaagtt
841 ctctggcatc gacggggagc gatataggcc acttctgctc acctttgctt cttgatcttt
901 cagggaaaac tccattaatt catctgaaat ctcttctgcc tggtcttctc
//
| {
"Vector backbone": "pcDNA3.1 His A",
"Backbone size w/o insert (bp)": "5500",
"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": "Redox regulation of Cdc25C",
"Publication": "J Biol Chem. 2002 Jun 7. 277(23):20535-40.",
"DOI": "10.1074/jbc.M201589200",
"Authors": "Savitsky PA, Finkel T",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/11925443",
"PubMed ID": "PubMed 11925443"
} | {
"Gene/Insert name": "cdc25c C330S",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "1450",
"Mutation": "C330S. Cysteine to Serine substitution at aa330.",
"Entrez Gene": "CDC25C",
"Promoter": "",
"Tag / Fusion Protein": "6xHis (N terminal on backbone)"
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "T7",
"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": ""
} |
||
pHR_Gal4UAS_4D5-Low_CAR-mCherry_pGK_4D5-Low_SynNotch_RHL183 | 164827 | Lentiviral vector for Gal4 inducible AntiHER2 4D5-Low CAR-mCherry and constitutive expression of antiHER2 4D5-Low Gal4VP64 Synthetic Notch | Wendell Lim | Hernandez-Lopez et al Science. 2021 Mar 12;371(6534):1166-1171. doi: 10.1126/science.abc1855. Epub 2021 Feb 25. | partial | {
"Vector backbone": "pHR'SIN",
"Backbone size w/o insert (bp)": "",
"Vector type": "Lentiviral",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "NEB Stable",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA ; Takara Bio Limited Use Label License (formerly Clontech)",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "T cell circuits that sense antigen density with an ultrasensitive threshold",
"Publication": "Science. 2021 Mar 12;371(6534):1166-1171. doi: 10.1126/science.abc1855. Epub 2021 Feb 25.",
"DOI": "10.1126/science.abc1855.",
"Authors": "Hernandez-Lopez RA, Yu W, Cabral KA, Creasey OA, Lopez Pazmino MDP, Tonai Y, De Guzman A, Makela A, Saksela K, Gartner ZJ, Lim WA /scienceabc",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/33632893",
"PubMed ID": "PubMed 33632893"
} | {
"Gene/Insert name": "AntiHER2 4D5-Low Chimeric Antigen receptor (CD8alphaTM-41BB-CD3z)",
"Species": "Synthetic",
"Insert Size (bp)": "1542",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "miniCMV",
"Tag / Fusion Protein": "mCherry (C terminal on insert)"
} | {
"Cloning method": "Ligation Independent Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "agagctcgtttagtgaaccg",
"3′ sequencing primer": "CAGCGGGGCTGCTAAAGCGCATGC"
} | {
"Gene/Insert name": "AntiHER2 4D5-Low Gal4VP64 synthetic Notch receptor",
"Species": "Synthetic",
"Insert Size (bp)": "2445",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "pGK",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Ligation Independent Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "GCTTCAAAAGCGCACGTCT",
"3′ sequencing primer": "catagcgtaaaaggagcaaca"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|||
PL2x2_5 | 45491 | David Baker | Mutations_Koga_Nature (unpublished) | https://media.addgene.org/snapgene-media/v2.0.0/sequences/64277/187cc730-3ffb-48fa-8c39-389b2c1a6991/addgene-plasmid-45491-sequence-64277.gbk | partial | LOCUS sequence-64277-e 855 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 855)
TITLE Mutations_Koga_Nature
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 855)
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..855
/mol_type="genomic DNA"
/organism="unspecified"
primer_bind 68..87
/label=T7
/note="T7 promoter, forward primer"
promoter 68..86
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 87..111
/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 126..148
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 474..491
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(544..562)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 558..605
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
primer_bind complement(729..748)
/label=F1ori-R
/note="F1 origin, reverse primer"
ORIGIN
1 gcgccggtga tgccggccac gatgcgtccg gcgtagagga tcgagatcga tctcgatccc
61 gcgaaattaa tacgactcac tataggggaa ttgtgagcgg ataacaattc ccctctagaa
121 ataattttgt ttaactttaa gaaggagata tacatatggg caaagtcctg ctggtgattt
181 ccaccgatac gaacattatc agctctgtcc aggaacgtgc gaaacataat tatccgggtc
241 gttacattcg caccgccacg agttcccaag atatccgcga cattatcaaa agcatgaaag
301 ataacggcaa accgctggtt gtgtttgtga atggtgcatc tcagaacgac gttaacgaat
361 ttcaaaacga agctaaaaaa gaaggcgtta gctatgatgt cctgaaatcg accgacccgg
421 aagaactgac gcagcgtgtt cgcgaatttc tgaaaaccgc cggtagcctc gagcaccacc
481 accaccacca ctgagatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg
541 ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt
601 ttttgctgaa aggaggaact atatccggat tggcgaatgg gacgcgccct gtagcggcgc
661 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
721 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
781 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
841 ccccaaaaac tgata
//
| {
"Vector backbone": "pET29b(+)",
"Backbone size w/o insert (bp)": "5370",
"Vector type": "Bacterial Expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Kanamycin, 50 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "XL10 Gold",
"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": "PL2x2_5",
"Species": "Synthetic",
"Insert Size (bp)": "321",
"Mutation": "changed E30Y of Di-IV_5_PL2x2_6",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": "6xHis (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": ""
} | {
"Gene/Insert name": "",
"Species": "",
"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": ""
} |
||
pBT29 | 138521 | phage cloning | David Liu | Richter et al Nat Biotechnol. 2020 Jul;38(7):883-891. doi: 10.1038/s41587-020-0453-z. Epub 2020 Mar 16. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/273893/5422d40e-9421-4a4e-b7fe-f74178362b09/addgene-plasmid-138521-sequence-273893.gbk | full | LOCUS sequence-273893- 5662 bp DNA circular SYN 26-AUG-2024
DEFINITION phage cloning.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5662)
AUTHORS Richter MF, Zhao KT, Eton E, Lapinaite A, Newby GA, Thuronyi BW,
Wilson C, Koblan LW, Zeng J, Bauer DE, Doudna JA, Liu DR
TITLE Phage-assisted evolution of an adenine base editor with improved Cas
domain compatibility and activity.
JOURNAL Nat Biotechnol. 2020 Jul;38(7):883-891. doi:
10.1038/s41587-020-0453-z. Epub 2020 Mar 16.
PUBMED 32433547
REFERENCE 2 (bases 1 to 5662)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1038/s41587-020-0453-z"; journalName: "Nat Biotechnol"; date:
"2020-07"; volume: "38"; issue: "7"; pages: "883-891"
FEATURES Location/Qualifiers
source 1..5662
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 23..1255
/codon_start=1
/gene="II"
/product="pII"
/label=M13 gene II
/note="replication"
/translation="MIDMLVLRLPFIDSLVCSRLSGNDLIAFVDLSKIATLSGMNLSAR
TVEYHVDGDLTVSGLSHPFESLPTHYSGIAFKIYEGSKNFYPCVEIKASPAKVLQGHNV
FGTTDLGLCSEALLLNFANSLPCLYDLLDVNATTISRIDAAFSARAPNENIAKQVIDHL
RNVSNGQTKSTRSQNWESTVTWNETSRHRTLVAYLKHVELQHQIQQLSSKPSAKMTSYQ
KEQLKVLSNPDLLEFASGLVRFEARIKTRYLKSFGLPLNLFDAIRFASDYNSQGKDLIF
DLWSFSFSELFKAFEGNSMNIYDDSAVLAAIQSKHFTITPSGKTSFAKASRYFGFYRRL
VNEGYDSVALTMPRNSFWRYVSALVECGIPKSQLMNLSTCNNVVPLVRFINVDFSSQRP
DWYNEPVLKIA"
CDS 1267..1530
/codon_start=1
/gene="V"
/product="pV"
/label=M13 gene V
/note="replication"
/translation="MIKVEIKPSQAQFTTRSGVSRQGKPYSLNEQFCYVDLGNEYPVLV
KITLDEGQPAYAPGLYTVHLSSFKVGQFGSLMIDRLRLVPAK"
CDS 1532..1633
/codon_start=1
/gene="VII"
/product="pVII"
/label=M13 gene VII
/note="minor coat protein"
/translation="MEQVADFDIIYQAMIQISVVLCFALGIIAGGQR"
CDS 1630..1728
/codon_start=1
/gene="IX"
/product="pIX"
/label=M13 gene IX
/note="minor coat protein"
/translation="MSVLVYSFASFVLGWCLRSGITYFTRLMETSS"
CDS 1725..1946
/codon_start=1
/gene="VIII"
/product="pVIII"
/label=M13 gene VIII
/note="major coat protein"
/translation="MKKSLVLKASVAVATLVPMLSFAAEGDDPAKAAFDSLQASATEYI
GYAWAMVVVIVGATIGIKLFKKFTSKAS"
terminator 1953..1992
/label=fd terminator
/note="central terminator from bacteriophage fd (Otsuka and
Kunisawa, 1982)"
protein_bind 2003..2021
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 2028..2046
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
RBS 2064..2075
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 2082..2759
/codon_start=1
/product="monomeric derivative of DsRed (Campbell et al.,
2002)"
/label=mRFP1
/translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
TGA"
terminator 2777..2848
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 2864..2891
/label=T7Te terminator
/note="phage T7 early transcription terminator"
CDS complement(2990..3805)
/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(2990..3805)
/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 3713..3732
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
primer_bind 3797..3816
/label=pENTR-R
/note="pENTR vectors, reverse primer"
CDS complement(3873..4823)
/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 4871..5093
/label=pSC101 ori
/note="low-copy replication origin that requires the Rep101
protein"
ORIGIN
1 cttattatca accggggtac atatgattga catgctagtt ttacggttac cgttcatcga
61 ttctcttgtt tgctccagac tctcaggcaa tgacctgata gcctttgtag atctctcaaa
121 aatagctacc ctctccggca tgaatttatc agctagaacg gttgaatatc atgttgatgg
181 tgatttgact gtctccggcc tttcccaccc ttttgaatct ttacctacac attactcagg
241 cattgcattt aaaatatatg agggttctaa aaatttttat ccttgcgttg aaataaaggc
301 ttctcccgca aaagtattac aggggcataa tgtttttggt acaaccgatt taggtttatg
361 ctctgaggct ttattgctta attttgctaa ttctttgcct tgcctgtatg atttattgga
421 tgttaacgct actactatta gtagaattga tgccgccttt tcagcccgcg ccccaaatga
481 aaatatagct aaacaggtta ttgaccattt gcgaaatgta tctaatggtc aaactaaatc
541 tactcgttcg cagaattggg aatcaactgt tacatggaat gaaacttcca gacaccgtac
601 tttagttgca tatttaaaac atgttgagct acagcaccag attcagcaat taagctctaa
661 gccatccgca aaaatgacct cttatcaaaa ggaacaatta aaggtactct ctaatcctga
721 cctgttggag tttgcttccg gtctggttcg ctttgaggct cgaattaaaa cgagatattt
781 gaagtctttc gggcttcctc ttaatctttt tgatgcaatc cgctttgctt ctgactataa
841 tagtcagggt aaagacctga tttttgattt atggtcattc tcgttttctg aactgtttaa
901 agcatttgag gggaattcaa tgaatattta tgacgattcc gcagtattgg ccgctatcca
961 gtctaaacat tttactatta ccccctctgg caaaacttct tttgcaaaag cctctcgcta
1021 ttttggtttt tatcgtcgtc tggtaaacga gggttatgat agtgttgctc ttactatgcc
1081 tcgtaattcc ttttggcgtt atgtatctgc attagttgaa tgtggtattc ctaaatctca
1141 actgatgaat ctttctacct gtaataatgt tgttccgtta gttcgtttta ttaacgtaga
1201 tttttcttcc caacgtcctg actggtataa tgagccagtt cttaaaatcg cataaggtaa
1261 ttcacaatga ttaaagttga aattaaacca tctcaagccc aatttactac tcgttctggt
1321 gtttctcgtc agggcaagcc ttattcactg aatgagcagt tttgttacgt tgatttgggt
1381 aatgaatatc cggttcttgt caagattact cttgatgaag gtcagccagc ctatgcgccg
1441 ggtctgtaca ccgttcatct gtcctctttc aaagttggtc agttcggttc ccttatgatt
1501 gaccgtctgc gcctcgttcc ggctaagtaa catggagcag gtcgcggatt tcgacataat
1561 ttatcaggcg atgatacaaa tctccgttgt actctgtttc gcgcttggta taatagctgg
1621 gggtcaaaga tgagtgtttt agtgtattct ttcgcctctt tcgttttagg ttggtgcctc
1681 cgtagtggca ttacgtattt tacccgttta atggaaactt cctcatgaaa aagtctttag
1741 tcctcaaagc ctctgtagcc gttgctaccc tcgttccgat gctgtctttc gctgctgagg
1801 gtgacgatcc cgcaaaagcg gcctttgact ccctgcaagc ctcagcgacc gaatatatcg
1861 gttatgcgtg ggcgatggtt gttgtcattg tcggcgcaac tatcggtatc aagctgttta
1921 agaaattcac ctcgaaagca agttgataaa ctgatacaat taaaggctcc ttttggagcc
1981 tttttttttg gagtagaaga gctccctatc agtgatagag attgacatcc ctatcagtga
2041 tagagatact gagcacgaat ctgaaagagg agaaaggatc tatggcgagt agcgaagacg
2101 ttatcaaaga gttcatgcgt ttcaaagttc gtatggaagg ttccgttaac ggtcacgagt
2161 tcgaaatcga aggtgaaggt gaaggtcgtc cgtacgaagg tacccagacc gctaaactga
2221 aagttaccaa aggtggtccg ctgccgttcg cttgggacat cctgtccccg cagttccagt
2281 acggttccaa agcttacgtt aaacacccgg ctgacatccc ggactacctg aaactgtcct
2341 tcccggaagg tttcaaatgg gaacgtgtta tgaacttcga agacggtggt gttgttaccg
2401 ttacccagga ctcctccctg caagacggtg agttcatcta caaagttaaa ctgcgtggta
2461 ccaacttccc gtccgacggt ccggttatgc agaaaaaaac catgggttgg gaagcttcca
2521 ccgaacgtat gtacccggaa gacggtgctc tgaaaggtga aatcaaaatg cgtctgaaac
2581 tgaaagacgg tggtcactac gacgctgaag ttaaaaccac ctacatggct aaaaaaccgg
2641 ttcagctgcc gggtgcttac aaaaccgaca tcaaactgga catcacctcc cacaacgaag
2701 actacaccat cgttgaacag tacgaacgtg ctgaaggtcg tcactccacc ggtgcttaat
2761 aaggatctcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
2821 tatctgttgt ttgtcggtga acgctctcta ctagagtcac actggctcac cttcgggtgg
2881 gcctttctgc gtttatacag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
2941 tttatttttc taaatacatt caaatatgta tccgctcatg aattaattct tagaaaaact
3001 catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt
3061 gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa
3121 gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc
3181 cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg
3241 agaatggcaa aagtttatgc atttctttcc agacttgttc aacaggccag ccattacgct
3301 cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga
3361 gacgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc
3421 gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata
3481 cctggaatgc tgttttcccg gggatcgcag tggtgagtaa ccatgcatca tcaggagtac
3541 ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca
3601 tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg
3661 catcgggctt cccatacaat cgatagattg tcgcacctga ttgcccgaca ttatcgcgag
3721 cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc ctagagcaag
3781 acgtttcccg ttgaatatgg ctcataacac cccttgtatt actgtttatg taagcagaca
3841 gttttattgt tcatgaaggc caaataggcc gttcagatcc ttccgtattt agccagtatg
3901 ttctctagtg tggttcgttg tttttgcgtg agccatgaga acgaaccatt gagatcatgc
3961 ttactttgca tgtcactcaa aaattttgcc tcaaaactgg tgagctgaat ttttgcagtt
4021 aaagcatcgt gtagtgtttt tcttagtccg ttacgtaggt aggaatctga tgtaatggtt
4081 gttggtattt tgtcaccatt catttttatc tggttgttct caagttcggt tacgagatcc
4141 atttgtctat ctagttcaac ttggaaaatc aacgtatcag tcgggcggcc tcgcttatca
4201 accaccaatt tcatattgct gtaagtgttt aaatctttac ttattggttt caaaacccat
4261 tggttaagcc ttttaaactc atggtagtta ttttcaagca ttaacatgaa cttaaattca
4321 tcaaggctaa tctctatatt tgccttgtga gttttctttt gtgttagttc ttttaataac
4381 cactcataaa tcctcataga gtatttgttt tcaaaagact taacatgttc cagattatat
4441 tttatgaatt tttttaactg gaaaagataa ggcaatatct cttcactaaa aactaattct
4501 aatttttcgc ttgagaactt ggcatagttt gtccactgga aaatctcaaa gcctttaacc
4561 aaaggattcc tgatttccac agttctcgtc atcagctctc tggttgcttt agctaataca
4621 ccataagcat tttccctact gatgttcatc atctgagcgt attggttata agtgaacgat
4681 accgtccgtt ctttccttgt agggttttca atcgtggggt tgagtagtgc cacacagcat
4741 aaaattagct tggtttcatg ctccgttaag tcatagcgac taatcgctag ttcatttgct
4801 ttgaaaacaa ctaattcaga catacatctc aattggtcta ggtgatttta atcactatac
4861 caattgagat gggctagtca atgataatta ctagtccttt tcctttgagt tgtgggtatc
4921 tgtaaattct gctagacctt tgctggaaaa cttgtaaatt ctgctagacc ctctgtaaat
4981 tccgctagac ctttgtgtgt tttttttgtt tatattcaag tggttataat ttatagaata
5041 aagaaagaat aaaaaaagat aaaaagaata gatcccagcc ctgtgtataa ctcactactt
5101 tagtcagttc cgcagtatta caaaaggatg tcgcaaacgc tgtttgctcc tctacaaaac
5161 agaccttaaa accctaaagg cttaagtagc accctcgcaa gctcgggcaa atcgctgaat
5221 attccttttg tctccgacca tcaggcacct gagtcgctgt ctttttcgtg acattcagtt
5281 cgctgcgctc acggctctgg cagtgaatgg gggtaaatgg cactacaggc gccttttatg
5341 gattcatgca aggaaactac ccataataca agaaaagccc gtcacgggct tctcagggcg
5401 ttttatggcg ggtctgctat gtggtgctat ctgacttttt gctgttcagc agttcctgcc
5461 ctctgatttt ccagtctgac cacttcggat tatcccgtga caggtcattc agactggcta
5521 atgcacccag taaggcagcg gtatcatcaa caggcttacc cgtcttactg tcaagaggac
5581 atccggtcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttgctgagga
5641 gacttaggga ccctgctctt ct
//
| {
"Vector backbone": "pCMV with a BR322 origin",
"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)": "DH5alpha",
"Copy number": "Unknown"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Phage-assisted evolution of an adenine base editor with improved Cas domain compatibility and activity",
"Publication": "Nat Biotechnol. 2020 Jul;38(7):883-891. doi: 10.1038/s41587-020-0453-z. Epub 2020 Mar 16.",
"DOI": "10.1038/s41587-020-0453-z.",
"Authors": "Richter MF, Zhao KT, Eton E, Lapinaite A, Newby GA, Thuronyi BW, Wilson C, Koblan LW, Zeng J, Bauer DE, Doudna JA, Liu DR /s---z",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/32433547",
"PubMed ID": "PubMed 32433547"
} | {
"Gene/Insert name": "phage genes",
"Species": "Other",
"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": ""
} |
|
EGFP-uTEV1-MAVS-mTA6 | 171004 | HiLITR protease with MAVS [Y536delinsFIVLI, R537A, R538A] C-terminal tmd targeting information (Mitochondria) | Alice Ting | Coukos et al Elife. 2021 Aug 20;10. pii: 69142. doi: 10.7554/eLife.69142. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/335832/5ebb6d5d-f762-48db-9863-1e867104dc79/addgene-plasmid-171004-sequence-335832.gbk | full | LOCUS sequence-335832- 8608 bp DNA circular SYN 26-AUG-2024
DEFINITION HiLITR protease with MAVS [Y536delinsFIVLI, R537A, R538A] C-terminal
tmd targeting information (Mitochondria).
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8608)
AUTHORS Coukos RW, Yao D, Sanchez ML, Strand ET, Olive ME, Udeshi ND,
Weissman JS, Carr SA, Bassik MC, Ting AY
TITLE An engineered transcriptional reporter of protein localization
identifies regulators of mitochondrial and ER membrane protein
trafficking in high-throughput CRISPRi screens.
JOURNAL Elife. 2021 Aug 20;10. pii: 69142. doi: 10.7554/eLife.69142.
PUBMED 34414886
REFERENCE 2 (bases 1 to 8608)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Elife.";
date: "2021-08-20"; volume: "10. pii: 69142. doi"; pages: "
10.7554/eLife.69142"
FEATURES Location/Qualifiers
source 1..8608
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 177..196
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 436..457
/label=EGFP-N
/note="EGFP, reverse primer"
regulatory 499..508
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
promoter complement(525..839)
/label=tight TRE promoter
/note="Tet-responsive promoter PTight, consisting of seven
tet operator sequences followed by the minimal CMV
promoter"
primer_bind complement(528..552)
/label=LNCX
/note="Human CMV promoter, forward primer"
protein_bind 599..617
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 635..653
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 670..688
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 706..724
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 742..760
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 777..795
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 813..831
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
misc_feature 893..1010
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
CDS complement(1223..1267)
/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 1452..1685
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
misc_feature 2178..2303
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
LTR 2350..2530
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
promoter complement(2531..2757)
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
primer_bind complement(2785..2805)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(2785..2803)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(2824..2840)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2824..2840)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(2837..2859)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 2848..2864
/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(2872..2902)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2917..2938
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(3055..3072)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3226..3814)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3306..3325)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3985..4845)
/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 4608..4627
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(4846..4917)
/gene="bla"
/label=AmpR promoter
rep_origin 4917..5052
/label=SV40 ori
/note="SV40 origin of replication"
polyA_signal 5092..5213
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5103..5122)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 5157..5176
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
LTR 5285..5518
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
CDS 5723..5734
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind 6132..6152
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(6192..6938)
/codon_start=1
/product="improved tetracycline-controlled transactivator"
/label=rtTA-Advanced
/note="In the Tet-On(R) system, rtTA-Advanced binds to the
Tet-responsive element and stimulates transcription only in
the presence of tetracycline or doxycycline."
/translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
DALDDFDLDMLPADALDDFDLDMLPG"
promoter complement(6948..7458)
/label=hPGK promoter
/note="human phosphoglycerate kinase 1 promoter"
primer_bind complement(7011..7029)
/label=hPGK-F
/note="Human PGK promoter, forward primer"
CDS complement(join(8394..8608,1..502))
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS complement(join(8394..8608,1..502))
/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(8436..8457)
/label=EGFP-C
/note="EGFP, forward primer"
ORIGIN
1 tcttgaagtt caccttgatg ccgttcttct gcttgtcggc catgatatag acgttgtggc
61 tgttgtagtt gtactccagc ttgtgcccca ggatgttgcc gtcctccttg aagtcgatgc
121 ccttcagctc gatgcggttc accagggtgt cgccctcgaa cttcacctcg gcgcgggtct
181 tgtagttgcc gtcgtccttg aagaagatgg tgcgctcctg gacgtagcct tcgggcatgg
241 cggacttgaa gaagtcgtgc tgcttcatgt ggtcggggta gcggctgaag cactgcacgc
301 cgtaggtcag ggtggtcacg agggtgggcc agggcacggg cagcttgccg gtggtgcaga
361 tgaacttcag ggtcagcttg ccgtaggtgg catcgccctc gccctcgccg gacacgctga
421 acttgtggcc gtttacgtcg ccgtccagct cgaccaggat gggcaccacc ccggtgaaca
481 gctcctcgcc cttgctcacc atggtggcgc tagccaattc tccaggcgat ctgacggttc
541 actaaacgag ctctgcttat ataggcctcc caccgtacac gcctacctcg acatacgttc
601 tctatcactg atagggagta aactcgacat acgttctcta tcactgatag ggataaactc
661 gacatacgtt ctctatcact gatagggagt aaactcgaca tacgttctct atcactgata
721 gggagtaaac tcgacatacg ttctctatca ctgataggga gtaaactcga catcgttctc
781 tatcactgat agggagtaaa ctcgacatac gttctctatc actgataggg agtaaactcg
841 acatatcgat tcgcggccaa agtggatctc tgctgtccct gtaataaacc cgaaaatttt
901 gaatttttgt aatttgtttt tgtaattctt tagtttgtat gtctgttgct attatgtcta
961 ctattctttc ccctgcactg taccccccaa tccccccttt tcttttaaaa ttgtggatga
1021 atactgccat ttgtctcgag gtcgagaatt gtcccctcgg ggttgggagg tgggtctgaa
1081 acgataatgg tgaatatccc tgcctaactc tattcactat agaaagtaca gcaaaaacta
1141 ttcttaaacc taccaagcct cctactatca ttatgaataa ttttatatac cacagccaat
1201 ttgttatgtt aaaccaattc cacaaacttg cccatttatc taattccaat aattcttgtt
1261 cattcttttc ttgctggttt tgcgattctt caattaagga gtgtattaag cttgtgtaat
1321 tgttaatttc tctgtcccac tccatccagg tcgtgtgatt ccaaatctgt tccagagatt
1381 tattactcca actagcattc caaggcacag cagtggtgca aatgagtttt ccagagcaac
1441 cccaaatccc caggagctgt tgatccttta ggtatctttc cacagccagg attcttgcct
1501 ggagctgctt gatgccccag actgtgagtt gcaacagatg ctgttgcgcc tcaatagccc
1561 tcagcaaatt gttctgctgc tgcactatac cagacaataa ttgtctggcc tgtaccgtca
1621 gcgtcattga cgctgcgccc atagtgcttc ctgctgctcc caagaaccca aggaacaaag
1681 ctcctattcc cactgctctt ttttctctct gcaccactct tctctttgcc ttggtgggtg
1741 ctactcctaa tggttcaatt tttactactt tatatttata taattcactt ctccaattgt
1801 ccctcatatc tcctcctcca ggtctgaaga tcagcggccg cttgctgtgc ggtggtctta
1861 cttttgtttt gctcttcctc tatcttgtct aaagcttcct tggtgtcttt tatctctatc
1921 ctttgatgca cacaatagag ggttgctact gtattatata atgatctaag ttcttctgat
1981 cctgtctgaa gggatggttg tagctgtccc agtatttgtc tacagccttc tgatgtttct
2041 aacaggccag gattaactgc gaatcgttct agctccctgc ttgcccatac tatatgtttt
2101 aatttatatt ttttctttcc ccctggcctt aaccgaattt tttcccatcg cgatctaatt
2161 ctcccccgct taatactgac gctctcgcac ccatctctct ccttctagcc tccgctagtc
2221 aaaatttttg gcgtactcac cagtcgccgc ccctcgcctc ttgccgtgcg cgcttcagca
2281 agccgagtcc tgcgtcgaga gagctcctct ggtttccctt tcgctttcaa gtccctgttc
2341 gggcgccact gctagagatt ttccacactg actaaaaggg tctgagggat ctctagttac
2401 cagagtcaca caacagacgg gcacacacta cttgaagcac tcaaggcaag ctttattgag
2461 gcttaagcag tgggttccct agttagccag agagctccca ggctcagatc tggtctaacc
2521 agagagaccc gtttatgtat cgagctaggc acttaaatac aatatctctg caatgcggca
2581 attcagtggt tcgtccaatc catgtcagac ccgtctgttg ccttcctaat aaggcacgat
2641 cgtaccacct tacttccacc aatcggcatg cacggtgctt tttctctcct tgtaaggcat
2701 gttgctaact catcgttacc atgttgcaag actacaagag tattgcataa gactacatta
2761 agcttgcagc tccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc
2821 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
2881 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
2941 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
3001 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
3061 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
3121 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
3181 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
3241 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
3301 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
3361 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
3421 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
3481 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
3541 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
3601 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
3661 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
3721 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
3781 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
3841 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
3901 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
3961 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
4021 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
4081 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
4141 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
4201 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
4261 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
4321 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
4381 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
4441 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
4501 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
4561 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
4621 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
4681 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
4741 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
4801 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
4861 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaaaggcc
4921 tccaaaaaag cctcctcact acttctggaa tagctcagag gccgaggcgg cctcggcctc
4981 tgcataaata aaaaaaatta gtcagccatg gggcggagaa tgggcggaac tgggcggagt
5041 taggggcggg atgggcggag ttaggggcgg gatagctaga gccagacatg ataagataca
5101 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
5161 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttcctctca
5221 ctctctgata ttcatttctt tgcaagttat aaatactgaa taataagatg acatgaacta
5281 ctactgctag agattttcca cactgactaa aagggtctga gggatctcta gttaccagag
5341 tcacacaaca gacgggcaca cactacttga agcactcaag gcaagcttta ttgaggctta
5401 agcagtgggt tccctagtta gccagagagc tcccaggctc agatctggtc taaccagaga
5461 gacccagtac aagcaaaaag cagatcttgt cttcgttggg agtgaattag cccttccagt
5521 cccccctttt cttttaaaaa gtggctaaga tctacagctg ccttgtaagt cattggtctt
5581 aaaggtacct ctagtccgga cgccgaggcg aaacaggcgg ggaggcggcc caaagggaga
5641 tccgactcgt ctgagggcga aggcgaagac gcggaagagg ccgcagagcc ggcagcaggc
5701 cgcgggaagg aaggtccgct ggattgaggg ccgaagggac gtagcagaag gacgtcccgc
5761 gcagaatcca ggtggcaaca caggcgagca gccatggaaa ggacgtcagc ttccccgaca
5821 acaccacgga attgtcagtg cccaacagcc gagcccctgt ccagcagcgg gcaaggcagg
5881 cggcgatgag ttccgccgtg gcaataggga gggggaaagc gaaagtcccg gaaaggagct
5941 gacaggtggt ggcaatgccc caaccagtgg gggttgcgtc agcaaacaca gtgcacacca
6001 cgccacgttg cctgacaacg ggccacaact cctcataaag agacagcaac caggatttat
6061 acaaggagga gaaaatgaaa gccatacggg aagcaatagc atgatacaaa ggcattaaag
6121 cagcgtatcc acatagcgta aaaggagcaa catagttaag aatacgatat cttggatcga
6181 tccttactta gttacccggg gagcatgtca aggtcaaaat cgtcaagagc gtcagcaggc
6241 agcatatcaa ggtcaaagtc gtcaagggca tcggctggga gcatgtctaa gtcaaaatcg
6301 tcaagggcgt cggtcggccc gccgctttcg cactttagct gtttctccag gccacatatg
6361 attagttcca ggccgaaaag gaaggcaggt tcggctccct gccggtcgaa cagctcaatt
6421 gcttgtctca gaagtggggg catagaatcg gtggtaggtg tctctctttc ctcttttgct
6481 acttgatgct cctgttcctc caatacgcag cccagtgtaa agtggcccac ggcggacaga
6541 gcgtacagtg cgttctccag ggagaagcct tgctgacaca ggaacgcgag ctgattttcc
6601 agggtttcgt actgtttctc tgttgggcgg gtgccgagat gcactttagc cccgtcgcga
6661 tgtgagagga gagcacagcg gtatgacttg gcgttgttcc gcagaaagtc ttgccatgac
6721 tcgccttcca gggggcagaa gtgggtatga tgcctgtcca gcatctcgat tggcagggca
6781 tcgagcaggg cccgcttgtt cttcacgtgc cagtacaggg taggctgctc aactcccagc
6841 ttttgagcga gtttccttgt cgtcaggcct tcgataccga caccattgag taattccaga
6901 gctccgttta tgactttgct cttgtccagt ctagacatgg tgaattgctg gggagagagg
6961 tcggtgattc ggtcaacgag ggagccgact gccgacgtgc gctccggagg cttgcagaat
7021 gcggaacacc gcgcgggcag gaacagggcc cacactaccg ccccacaccc cgcctcccgc
7081 accgcccctt cccggccgct gctctcggcg cgccctgctg agcagccgct attggccaca
7141 gcccatcgcg gtcggcgcgc tgccattgct ccctggcgct gtccgtctgc gagggtacta
7201 gtgagacgtg cggcttccgt ttgtcacgtc cggcacgccg cgaaccgcaa ggaaccttcc
7261 cgacttaggg gcggagcagg aagcgtcgcc ggggggccca caagggtagc ggcgaagatc
7321 cgggtgacgc tgcgaacgga cgtgaagaat gtgcgagacc cagggtcggc gccgctgcgt
7381 ttcccggaac cacgcccaga gcagccgcgt ccctgcgcaa acccagggct gccttggaaa
7441 aggcgcaacc ccaaccccgg atgaattcgg ccctagtgca gagccgcccg gatcagtacg
7501 atgaacagca ccaccaggag tgtgactacc agcacccctg tcacagccac ctggagccac
7561 agagccccag gtgagggcct tccactacct gagccggaac tgccgctccc ctctccactc
7621 ccaggcttac cagacccaga agtagagccg ctaccctcgc ctgatccggg cttgccagaa
7681 cctgacgtgg aaccgaattc caccatgaaa actttatggc ccccccacaa tactgagtca
7741 gcatttaatc gccaaccact aacccactgc tgcgcctcct gatttgtcaa caattccatg
7801 aagtttttcg gcacgcttgt gaaataattg tttgtgttgg tgaaattcga tgctgagtgt
7861 ataccaacaa tgaacccatc tctagttgat actaatggat tgccacactg cccatccttg
7921 gtttgaatcc aatgcttcca gaatatgcca tcagatgaag ggaatgtgca actagtgtct
7981 gacaccatgc tagacatgct cttagtttgg aagttggttg tcacaagaca tatgcgctct
8041 tccctttgtg gctctctaaa tttcagcttt tgaggaaatg gtgggaaatc cttaggcatg
8101 cgaataatta tcatgtccct cccatcaatg aggtgttgtt gcaaagtcgt ggtgttcttg
8161 accttgaata caccatgtag tgattggacc aacagtgttc cattatttct tctaaacaag
8221 tgcttgtttg taatgatgaa gggaccaaat ccaataccat acaacgatgt tgtgtgccca
8281 tcagattcat tcgtcaaatg acaaatggtg ctcgatatcg ggttgtaatc acgtggtccc
8341 ttaaacaagc tttctccgcc tgaaccggat ccgcctgtgc cagatccaag gtccttgtac
8401 agctcgtcca tgccgagagt gatcccggcg gcggtcacga actccagcag gaccatgtga
8461 tcgcgcttct cgttggggtc tttgctcagg gcggactggg tgctcaggta gtggttgtcg
8521 ggcagcagca cggggccgtc gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc
8581 acgctgccgt cctcgatgtt gtggcgga
//
| {
"Vector backbone": "pCW-Cas9",
"Backbone size w/o insert (bp)": "8000",
"Vector type": "Mammalian Expression, Lentiviral, Synthetic Biology",
"Selectable markers": "EGFP"
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "NEB Stable",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "An engineered transcriptional reporter of protein localization identifies regulators of mitochondrial and ER membrane protein trafficking in high-throughput CRISPRi screens",
"Publication": "Elife. 2021 Aug 20;10. pii: 69142. doi: 10.7554/eLife.69142.",
"DOI": "10.7554/eLife.69142.",
"Authors": "Coukos RW, Yao D, Sanchez ML, Strand ET, Olive ME, Udeshi ND, Weissman JS, Carr SA, Bassik MC, Ting AY /eLife",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/34414886",
"PubMed ID": "PubMed 34414886"
} | {
"Gene/Insert name": "EGFP-uTEV1-MAVS(tmd)[Y536delinsFIVLI, R537A, R538A]",
"Species": "Synthetic",
"Insert Size (bp)": "1600",
"Mutation": "MAVS(tmd)[Y536delinsFIVLI, R537A, R538A]",
"Entrez Gene": "",
"Promoter": "pTRE-tight (TetOn)",
"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": ""
} |
|
pABY-T1 | 201764 | For C-terminal tagging of a protein-of-interest with the ALFA tag using S2/S3 primers in yeast | Saravanan Palani | Akhuli et al mSphere. 2022 Oct 26;7(5):e0033322. doi: 10.1128/msphere.00333-22. Epub 2022 Oct 3. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/403474/79d6cb09-9c6f-48cd-a477-6d6c8924c7cc/addgene-plasmid-201764-sequence-403474.gbk | full | LOCUS sequence-403474- 3956 bp DNA circular SYN 26-AUG-2024
DEFINITION For C-terminal tagging of a protein-of-interest with the ALFA tag
using S2/S3 primers in yeast.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3956)
AUTHORS Akhuli D, Dhar A, Viji AS, Bhojappa B, Palani S
TITLE ALIBY: ALFA Nanobody-Based Toolkit for Imaging and Biochemistry in
Yeast.
JOURNAL mSphere. 2022 Oct 26;7(5):e0033322. doi: 10.1128/msphere.00333-22.
Epub 2022 Oct 3.
PUBMED 36190134
REFERENCE 2 (bases 1 to 3956)
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: "mSphere.";
date: "2022-10-26"; volume: "7(5):e0033322. doi"; pages: "
10.1128/msphere.00333-22. Epub 2022 Oct 3"
FEATURES Location/Qualifiers
source 1..3956
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(31..50)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(32..50)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(137..154)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(308..896)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(388..407)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(1067..1927)
/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 1690..1709
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(1928..2032)
/gene="bla"
/label=AmpR promoter
primer_bind 2100..2118
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(2156..2178)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 2278..2297
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
promoter 2378..2396
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 2378..2395
/label=SP6
/note="SP6 promoter, forward primer"
CDS 2440..2478
/codon_start=1
/product="rationally designed epitope tag that forms a
stable alpha-helix (Gotzke et al., 2019)"
/label=ALFA-tag
/note="It may be beneficial to flank the ALFA-tag sequence
with proline residues to preserve the helical structure."
/translation="SRLEEELRRRLTE"
gene 2527..3883
/label=kanMX
/note="yeast selectable marker conferring kanamycin
resistance (Wach et al., 1994)"
promoter 2527..2870
/label=TEF promoter
/note="Ashbya gossypii TEF promoter"
CDS 2871..3680
/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 complement(2938..2957)
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
terminator 3686..3883
/label=TEF terminator
/note="Ashbya gossypii TEF terminator"
ORIGIN
1 tggcctatgc ggccgcggat ctgccggtct ccctatagtg agtcgtatta atttcgataa
61 gccaggttaa cctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
121 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
181 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
241 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
301 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
361 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
421 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
481 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
541 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
601 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
661 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
721 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
781 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
841 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
901 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
961 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
1021 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
1081 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
1141 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
1201 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
1261 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
1321 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
1381 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
1441 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
1501 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
1561 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
1621 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
1681 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
1741 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
1801 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
1861 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
1921 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
1981 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
2041 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
2101 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
2161 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
2221 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
2281 catcagagca gattgtactg agagtgcacc atatggacat attgtcgtta gaacgcggct
2341 acaattaata cataacctta tgtatcatac acatacgatt taggtgacac tatagaacgc
2401 ggccgccagc tgaagcttcg tacgctgcag gtcgacccat caagattaga agaggagctg
2461 aggagaagat tgaccgagta aagatctgtt tagcttgcct cgtccccgcc gggtcacccg
2521 gccagcgaca tggaggccca gaataccctc cttgacagtc ttgacgtgcg cagctcaggg
2581 gcatgatgtg actgtcgccc gtacatttag cccatacatc cccatgtata atcatttgca
2641 tccatacatt ttgatggccg cacggcgcga agcaaaaatt acggctcctc gctgcagacc
2701 tgcgagcagg gaaacgctcc cctcacagac gcgttgaatt gtccccacgc cgcgcccctg
2761 tagagaaata taaaaggtta ggatttgcca ctgaggttct tctttcatat acttcctttt
2821 aaaatcttgc taggatacag ttctcacatc acatccgaac ataaacaacc atgggtaagg
2881 aaaagactca cgtttcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt
2941 ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcga ttgtatggga
3001 agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta
3061 cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc
3121 attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc ggcaaaacag
3181 cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag
3241 tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg
3301 tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt
3361 ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataagcttt
3421 tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt
3481 ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat
3541 accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac
3601 ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga
3661 tgctcgatga gtttttctaa tcagtactga caataaaaag attcttgttt tcaagaactt
3721 gtcatttgta tagttttttt atattgtagt tgttctattt taatcaaatg ttagcgtgat
3781 ttatattttt tttcgcctcg acatcatctg cccagatgcg aagttaagtg cgcagaaagt
3841 aatatcatgc gtcaatcgta tgtgaatgct ggtcgctata ctgctgtcga ttcgatacta
3901 acgccgccat ccagtttaaa cgagctcgaa ttcatcgatg atatcagatc cactag
//
| {
"Vector backbone": "pFA6a",
"Backbone size w/o insert (bp)": "3917",
"Vector type": "Yeast 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": "ALIBY: ALFA Nanobody-Based Toolkit for Imaging and Biochemistry in Yeast",
"Publication": "mSphere. 2022 Oct 26;7(5):e0033322. doi: 10.1128/msphere.00333-22. Epub 2022 Oct 3.",
"DOI": "10.1128/msphere.00333-22.",
"Authors": "Akhuli D, Dhar A, Viji AS, Bhojappa B, Palani S /msphere-",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/36190134",
"PubMed ID": "PubMed 36190134"
} | {
"Gene/Insert name": "ALFA tag",
"Species": "Synthetic",
"Insert Size (bp)": "39",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "SalI (not destroyed)",
"3′ cloning site": "BglII (not destroyed)",
"5′ sequencing primer": "SP6",
"3′ sequencing primer": "Kan-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": ""
} |
|
pFTK077 | 171349 | Part Plasmid Entry Vector (LVL0) from the Fungal Modular Cloning ToolKit. | Arnold Driessen | Mozsik et al ACS Synth Biol. 2021 Nov 2. doi: 10.1021/acssynbio.1c00260. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/365609/7ba90bb3-503f-4d36-9af5-05016fc2ba67/addgene-plasmid-171349-sequence-365609.gbk | full | LOCUS sequence-365609- 2753 bp DNA circular SYN 26-AUG-2024
DEFINITION Part Plasmid Entry Vector (LVL0) from the Fungal Modular Cloning
ToolKit..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2753)
AUTHORS Mozsik L, Pohl C, Meyer V, Bovenberg RAL, Nygard Y, Driessen AJM
TITLE Modular Synthetic Biology Toolkit for Filamentous Fungi.
JOURNAL ACS Synth Biol. 2021 Nov 2. doi: 10.1021/acssynbio.1c00260.
PUBMED 34726388
REFERENCE 2 (bases 1 to 2753)
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. 2021 Nov 2. doi: 10.1021/acssynbio.1c00260."
FEATURES Location/Qualifiers
source 1..2753
/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 tcagcttctt caaccgactc cggcttcact tcttcgcccg cactgcctac
2281 cgtttgtacc atctgactca tataaatgtc tagcccctac ctacactata cctaagggag
2341 agaagcgtag agtgattaac gtacgggcct atagtacccc gatctctaga tagaacattt
2401 agtagagatt aggatgccta actaatttaa cttgagcatt gtcccgttca tattgatttt
2461 cagtccatta tacactctta atcgtttccc ggtagaagcc tgatatatac gaccataggg
2521 tgtggagaac agggcttccc gtctgcttgg ccgtacttaa gctatatatt ctacacggcc
2581 aatactcaat gtgcccttag cacctaagcg gcactctagg gtaagtgcgg gtgatatagg
2641 tgagaagtct taagactgat gacaggatat cacgcgttac cctgcaccgt acctactacc
2701 ttcaatatca actctttcag gatggacagg gtcgacacgc ttgagaccac gaa
//
| {
"Vector backbone": "pICH41276",
"Backbone size w/o insert (bp)": "",
"Vector type": "Bacterial Expression, Yeast Expression, 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": "Modular Synthetic Biology Toolkit for Filamentous Fungi",
"Publication": "ACS Synth Biol. 2021 Nov 2. doi: 10.1021/acssynbio.1c00260.",
"DOI": "10.1021/acssynbio.1c00260.",
"Authors": "Mozsik L, Pohl C, Meyer V, Bovenberg RAL, Nygard Y, Driessen AJM /acssynbioc",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/34726388",
"PubMed ID": "PubMed 34726388"
} | {
"Gene/Insert name": "TpenDE Pc21g21370 terminator",
"Species": "Penicillium, Aspergillus, or other fungal sources",
"Insert Size (bp)": "",
"Mutation": "n/a",
"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": ""
} |
|
pM9-BetaGal-8C | 172489 | Plasmid encoding E. coli BetaGalactosidase with an N-terminal M9 nuclear localization sequence | Siegfried Musser | Chowdhury et al Nat Cell Biol. 2022 Jan;24(1):112-122. doi: 10.1038/s41556-021-00815-6. Epub 2022 Jan 10. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/356067/6e117671-b940-434d-80f4-3f8025689df6/addgene-plasmid-172489-sequence-356067.gbk | full | LOCUS sequence-356067- 8613 bp DNA circular SYN 26-AUG-2024
DEFINITION Plasmid encoding E. coli BetaGalactosidase with an N-terminal M9
nuclear localization sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8613)
AUTHORS Chowdhury R, Sau A, Musser SM
TITLE Super-resolved 3D tracking of cargo transport through nuclear pore
complexes.
JOURNAL Nat Cell Biol. 2022 Jan;24(1):112-122. doi:
10.1038/s41556-021-00815-6. Epub 2022 Jan 10.
PUBMED 35013558
REFERENCE 2 (bases 1 to 8613)
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/s41556-021-00815-6"; journalName: "Nat Cell Biol"; date:
"2022-01"; volume: "24"; issue: "1"; pages: "112-122"
FEATURES Location/Qualifiers
source 1..8613
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin 48..503
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(135..154)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 345..366
/label=F1ori-F
/note="F1 origin, forward primer"
CDS complement(595..1410)
/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(595..1410)
/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 1318..1337
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
primer_bind 1402..1421
/label=pENTR-R
/note="pENTR vectors, reverse primer"
rep_origin 1532..2120
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2021..2040
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 2274..2291
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 2306..2445
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(2371..2390)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 2531..2553
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(2547..2738)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(3547..4629)
/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 4591..4610
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(4630..4707)
/gene="lacI"
/label=lacI promoter
primer_bind 4913..4932
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind 5020..5039
/label=T7
/note="T7 promoter, forward primer"
promoter 5020..5038
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 5039..5063
/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 5078..5100
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 5111..5128
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 5138..5155
/codon_start=1
/product="thrombin recognition and cleavage site"
/label=thrombin site
/translation="LVPRGS"
CDS 5162..5206
/codon_start=1
/product="affinity and epitope tag derived from pancreatic
ribonuclease A"
/label=S-Tag
/translation="KETAAAKFERQHMDS"
CDS 5222..5236
/codon_start=1
/product="enterokinase recognition and cleavage site"
/label=enterokinase site
/translation="DDDDK"
CDS 5284..5295
/codon_start=1
/product="WELQut protease recognition and cleavage site"
/label=WELQut site
/translation="WELQ"
CDS 8493..8510
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(8563..8581)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator join(8577..8613,1..11)
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
ORIGIN
1 ggggtttttt gctgaaagga ggaactatat ccggattggc gaatgggacg cgccctgtag
61 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
121 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
181 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
241 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
301 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
361 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
421 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
481 caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct
541 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgaatta attcttagaa
601 aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata
661 tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat
721 ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa
781 tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc
841 cggtgagaat ggcaaaagtt tatgcatttc tttccagact tgttcaacag gccagccatt
901 acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg
961 agcgagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa
1021 ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc
1081 taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg catcatcagg
1141 agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct
1201 gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc
1261 tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc cgacattatc
1321 gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc gcggcctaga
1381 gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt ttatgtaagc
1441 agacagtttt attgttcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
1501 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
1561 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
1621 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
1681 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
1741 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
1801 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
1861 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
1921 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
1981 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
2041 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
2101 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
2161 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
2221 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
2281 cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg
2341 gtatttcaca ccgcaatggt gcactctcag tacaatctgc tctgatgccg catagttaag
2401 ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca
2461 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
2521 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
2581 aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt
2641 tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag
2701 cgggccatgt taagggcggt tttttcctgt ttggtcactg atgcctccgt gtaaggggga
2761 tttctgttca tgggggtaat gataccgatg aaacgagaga ggatgctcac gatacgggtt
2821 actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact ggcggtatgg
2881 atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt taatacagat
2941 gtaggtgttc cacagggtag ccagcagcat cctgcgatgc agatccggaa cataatggtg
3001 cagggcgctg acttccgcgt ttccagactt tacgaaacac ggaaaccgaa gaccattcat
3061 gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc
3121 ggtgattcat tctgctaacc agtaaggcaa ccccgccagc ctagccgggt cctcaacgac
3181 aggagcacga tcatgcgcac ccgtggggcc gccatgccgg cgataatggc ctgcttctcg
3241 ccgaaacgtt tggtggcggg accagtgacg aaggcttgag cgagggcgtg caagattccg
3301 aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa
3361 atgacccaga gcgctgccgg cacctgtcct acgagttgca tgataaagaa gacagtcata
3421 agtgcggcga cgatagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct
3481 ctcaagggca tcggtcgaga tcccggtgcc taatgagtga gctaacttac attaattgcg
3541 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
3601 ggccaacgcg cggggagagg cggtttgcgt attgggcgcc agggtggttt ttcttttcac
3661 cagtgagacg ggcaacagct gattgccctt caccgcctgg ccctgagaga gttgcagcaa
3721 gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg
3781 gatataacat gagctgtctt cggtatcgtc gtatcccact accgagatgt ccgcaccaac
3841 gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc gccatctgat cgttggcaac
3901 cagcatcgca gtgggaacga tgccctcatt cagcatttgc atggtttgtt gaaaaccgga
3961 catggcactc cagtcgcctt cccgttccgc tatcggctga atttgattgc gagtgagata
4021 tttatgccag ccagccagac gcagacgcgc cgagacagaa cttaatgggc ccgctaacag
4081 cgcgatttgc tggtgaccca atgcgaccag atgctccacg cccagtcgcg taccgtcttc
4141 atgggagaaa ataatactgt tgatgggtgt ctggtcagag acatcaagaa ataacgccgg
4201 aacattagtg caggcagctt ccacagcaat ggcatcctgg tcatccagcg gatagttaat
4261 gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac aggcttcgac
4321 gccgcttcgt tctaccatcg acaccaccac gctggcaccc agttgatcgg cgcgagattt
4381 aatcgccgcg acaatttgcg acggcgcgtg cagggccaga ctggaggtgg caacgccaat
4441 cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt aattcagctc
4501 cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa acgtggctgg cctggttcac
4561 cacgcgggaa acggtctgat aagagacacc ggcatactct gcgacatcgt ataacgttac
4621 tggtttcaca ttcaccaccc tgaattgact ctcttccggg cgctatcatg ccataccgcg
4681 aaaggttttg cgccattcga tggtgtccgg gatctcgacg ctctccctta tgcgactcct
4741 gcattaggaa gcagcccagt agtaggttga ggccgttgag caccgccgcc gcaaggaatg
4801 gtgcatgcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca
4861 cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt
4921 cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc
4981 cggcgtagag gatcgagatc gatctcgatc ccgcgaaatt aatacgactc actatagggg
5041 aattgtgagc ggataacaat tcccctctag aaataatttt gtttaacttt aagaaggaga
5101 tatacatatg caccatcatc atcatcattc ttctggtctg gtgccacgcg gttctggtat
5161 gaaagaaacc gctgctgcta aattcgaacg ccagcacatg gacagcccag atctgggtac
5221 cgacgacgac gacaaggcca tggctgatat cggatccgga ggtggtggaa gctacaatga
5281 ttttgggaat tacaacaatc agtcttcaaa ttttggaccc atgaagggag gaaattttgg
5341 aggcagaagc tctggcccct atggcggtgg aggccaatac tttgcaaaac cacgaaacca
5401 aggtggctat ggcggttcca gcagcagcag tagctatggc agtggcagaa gatttggggg
5461 cgggctggta ccgaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
5521 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgctt
5581 tgcctggttt ccggcaccag aagcggtgcc ggaaagctgg ctggaggccg atcttcctga
5641 ggccgatact gtcgtcgtcc cctcaaactg gcagatgcac ggttacgatg cgcccatcta
5701 caccaacgtg acctatccca ttacggtcaa tccgccgttt gttcccacgg agaatccgac
5761 gggttgttac tcgctcacat ttaatgttga tgaaagctgg ctacaggaag gccagacgcg
5821 aattattttt gatggcgtta actcggcgtt tcatctgtgg tgcaacgggc gctgggtcgg
5881 ttacggccag gacagtcgtt tgccgtctga atttgacctg agcgcatttt tacgcgccgg
5941 agaaaaccgc ctcgcggtga tggtgctgcg ctggagtgac ggcagttatc tggaagatca
6001 ggatatgtgg cggatgagcg gcattttccg tgacgtctcg ttgctgcata aaccgactac
6061 acaaatcagc gatttccatg ttgccactcg ctttaatgat gatttcagcc gcgctgtact
6121 ggaggctgaa gttcagatgt gcggcgagtt gcgtgactac ctacgggtaa cagtttcttt
6181 atggcagggt gaaacgcagg tcgccagcgg caccgcgcct ttcggcggtg aaattatcga
6241 tgagcgtggt ggttatgccg atcgcgtcac actacgtctg aacgtcgaaa acccgaaact
6301 gtggagcgcc gaaatcccga atctctatcg tgcggtggtt gaactgcaca ccgccgacgg
6361 cacgctgatt gaagcagaag cctgcgatgt cggtttccgc gaggtgcgga ttgaaaatgg
6421 tctgctgctg ctgaacggca agccgttgct gattcgaggc gttaaccgtc acgagcatca
6481 tcctctgcat ggtcaggtca tggatgagca gacgatggtg caggatatcc tgctgatgaa
6541 gcagaacaac tttaacgccg tgcgctgttc gcattatccg aaccatccgc tgtggtacac
6601 gctgtgcgac cgctacggcc tgtatgtggt ggatgaagcc aatattgaaa cccacggcat
6661 ggtgccaatg aatcgtctga ccgatgatcc gcgctggcta ccggcgatga gcgaacgcgt
6721 aacgcgaatg gtgcagcgcg atcgtaatca cccgagtgtg atcatctggt cgctggggaa
6781 tgaatcaggc cacggcgcta atcacgacgc gctgtatcgc tggatcaaat ctgtcgatcc
6841 ttcccgcccg gtgcagtatg aaggcggcgg agccgacacc acggccaccg atattatttg
6901 cccgatgtac gcgcgcgtgg atgaagacca gcccttcccg gctgtgccga aatggtccat
6961 caaaaaatgg ctttcgctac ctggagagac gcgcccgctg atcctttgcc aatacgccca
7021 cgcgatgggt aacagtcttg gcggtttcgc taaatactgg caggcgtttc gtcagtatcc
7081 ccgtttacag ggcggcttcg tctgggactg ggtggatcag tcgctgatta aatatgatga
7141 aaacggcaac ccgtggtcgg cttacggcgg tgattttggc gatacgccga acgatcgcca
7201 gttctgtatg aacggtctgg tctttgccga ccgcacgccg catccagcgc tgacggaagc
7261 aaaacaccag cagcagtttt tccagttccg tttatccggg caaaccatcg aagtgaccag
7321 cgaatacctg ttccgtcata gcgataacga gctcctgcac tggatggtgg cgctggatgg
7381 taagccgctg gcaagcggtg aagtgcctct ggatgtcgct ccacaaggta aacagttgat
7441 tgaactgcct gaactaccgc agccggagag cgccgggcaa ctctggctca cagtacgcgt
7501 agtgcaaccg aacgcgaccg catggtcaga agccgggcac atcagcgcct ggcagcagtg
7561 gcgtctggcg gaaaacctca gtgtgacgct ccccgccgcg tcccacgcca tcccgcatct
7621 gaccaccagc gaaatggatt tttgcatcga gctgggtaat aagcgttggc aatttaaccg
7681 ccagtcaggc tttctttcac agatgtggat tggcgataaa aaacaactgc tgacgccgct
7741 gcgcgatcag ttcacccgtg caccgctgga taacgacatt ggcgtaagtg aagcgacccg
7801 cattgaccct aacgcctggg tcgaacgctg gaaggcggcg ggccattacc aggccgaagc
7861 agcgttgttg cagtgcacgg cagatacact tgctgatgcg gtgctgatta cgaccgctca
7921 cgcgtggcag catcagggga aaaccttatt tatcagccgg aaaacctacc ggattgatgg
7981 tagtggtcaa atggcgatta ccgttgatgt tgaagtggcg agcgatacac cgcatccggc
8041 gcggattggc ctgaactgcc agctggcgca ggtagcagag cgggtaaact ggctcggatt
8101 agggccgcaa gaaaactatc ccgaccgcct tactgccgcc tgttttgacc gctgggatct
8161 gccattgtca gacatgtata ccccgtacgt cttcccgagc gaaaacggtc tgcgctgcgg
8221 gacgcgcgaa ttgaattatg gcccacacca gtggcgcggc gacttccagt tcaacatcag
8281 ccgctacagt caacagcaac tgatggaaac cagccatcgc catctgctgc acgcggaaga
8341 aggcacatgg ctgaatatcg acggtttcca tatggggatt ggtggcgacg actccctgag
8401 cccgtcagta tcggcggaat tccagctgag cgccggtcgc taccattacc agttggtctg
8461 gtctcaaaaa taagcttgcg gccgcactcg agcaccacca ccaccaccac tgagatccgg
8521 ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag
8581 cataacccct tggggcctct aaacgggtct tga
//
| {
"Vector backbone": "pET30a",
"Backbone size w/o insert (bp)": "5422",
"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": "Super-resolved 3D tracking of cargo transport through nuclear pore complexes",
"Publication": "Nat Cell Biol. 2022 Jan;24(1):112-122. doi: 10.1038/s41556-021-00815-6. Epub 2022 Jan 10.",
"DOI": "10.1038/s41556-021-00815-6.",
"Authors": "Chowdhury R, Sau A, Musser SM /s---",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/35013558",
"PubMed ID": "PubMed 35013558"
} | {
"Gene/Insert name": "M9-BetaGal-8C",
"Species": "Escherichia coli",
"Insert Size (bp)": "3363",
"Mutation": "BetaGal has two cysteine mutations (C77A, C1022S), to reduce the number of maleimide reactive sites, and two enzymatic mutations (E538Q, W999L), to reduce Kcat and sugar binding affinity.",
"Entrez Gene": "",
"Promoter": "T7",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Unknown",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "T7 Promoter primer",
"3′ sequencing primer": "T7 Terminator primer"
} | {
"Gene/Insert name": "",
"Species": "",
"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": ""
} |
|
CMV-d2eGFP-16pf | 21969 | Phil Sharp | Ebert et al Nat Methods. 2007 Sep . 4(9):721-6. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/28266/a21717e9-07bc-423e-9ebd-d515e95cdb87/addgene-plasmid-21969-sequence-28266.gbk | partial | LOCUS sequence-28266-e 1028 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 1028)
AUTHORS Ebert MS, Neilson JR, Sharp PA
TITLE MicroRNA sponges: competitive inhibitors of small RNAs in mammalian
cells.
JOURNAL Nat Methods. 2007 Sep . 4(9):721-6.
PUBMED 17694064
REFERENCE 2 (bases 1 to 1028)
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. 2007 Sep . 4(9):721-6."
FEATURES Location/Qualifiers
source 1..1028
/mol_type="genomic DNA"
/organism="unspecified"
primer_bind 161..182
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(437..454)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 443..667
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(800..819)
/label=F1ori-R
/note="F1 origin, reverse primer"
ORIGIN
1 gacttcagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
61 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
121 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
181 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagaagctt agccatggct
241 tcccgccgga ggtggaggag caggatgatg gcacgctgcc catgtcttgt gcccaggaga
301 gcgggatgga ccgtcaccct gcagcctgtg cttctgctag gatcaatgtg taggcggccg
361 cctcgagcgc caatatttac gtgctgctac gccaatattt acgtgctgct agggcccgtt
421 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
481 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
541 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
601 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
661 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
721 tgtagcggcg cattaancgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
781 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
841 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
901 cggcacctcg accccaaaaa cttgattagg gtgatggnnc gngtactaga agtcctattc
961 cgagttctat ctctagaant ataggacttc ctggnaaaaa gcctgaactc ncgcgacgtc
1021 tgtcanna
//
| {
"Vector backbone": "pcDNA5",
"Backbone size w/o insert (bp)": "5000",
"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": "MicroRNA sponges: competitive inhibitors of small RNAs in mammalian cells",
"Publication": "Nat Methods. 2007 Sep . 4(9):721-6.",
"DOI": "10.1038/nmeth1079",
"Authors": "Ebert MS, Neilson JR, Sharp PA",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/17694064",
"PubMed ID": "PubMed 17694064"
} | {
"Gene/Insert name": "CMV d2eGFP miR-16 perfect",
"Species": "",
"Insert Size (bp)": "50",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "XhoI (not destroyed)",
"3′ cloning site": "ApaI (not destroyed)",
"5′ sequencing primer": "n/a",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"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": ""
} |
||
IOS1 A1_pECIA14 | 114736 | Prey vector IOS1 A1_pECIA14 should be used with bait vector IOS1 A1_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/270064/72e88661-3b0b-42df-96dc-fec4083b1f14/addgene-plasmid-114736-sequence-270064.gbk | full | LOCUS sequence-270064- 6668 bp DNA circular SYN 26-AUG-2024
DEFINITION Prey vector IOS1 A1_pECIA14 should be used with bait vector IOS1
A1_pECIA2..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6668)
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 6668)
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..6668
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(38..60)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 160..179
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 373..395
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 388..404
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 410..836
/label=MT promoter
/note="Drosophila metallothionein promoter"
sig_peptide 854..907
/codon_start=1
/product="Drosophila BiP signal sequence"
/label=BiP signal sequence
/translation="MKLCILLAVVAFVGLSLG"
CDS 2351..2374
/codon_start=1
/product="recognition and cleavage site for human
rhinovirus 3C and PreScission proteases"
/label=HRV 3C site
/translation="LEVLFQGP"
CDS 4049..4072
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS 4088..4105
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(4128..4145)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 4178..4312
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4202..4221)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 4256..4275
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind complement(4456..4472)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4456..4472)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4469..4491)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4480..4496
/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(4504..4534)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4549..4570
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4687..4704)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4858..5446)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4938..4957)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5617..6477)
/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 6240..6259
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6478..6582)
/gene="bla"
/label=AmpR promoter
primer_bind 6650..6668
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
ORIGIN
1 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
61 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
121 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
181 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
241 tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
301 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
361 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaatta attcgttgca
421 ggacaggatg tggtgcccga tgtgactagc tctttgctgc aggccgtcct atcctctggt
481 tccgataaga gacccagaac tccggccccc caccgcccac cgccaccccc atacatatgt
541 ggtacgcaag taagagtgcc tgcgcatgcc ccatgtgccc caccaagagc tttgcatccc
601 atacaagtcc ccaaagtgga gaaccgaacc aattcttcgc gggcagaaca aaagcttctg
661 cacacgtctc cactcgaatt tggagccggc cggcgtgtgc aaaagaggtg aatcgaacga
721 aagacccgtg tgtaaagccg cgtttccaaa atgtataaaa ccgagagcat ctggccaatg
781 tgcatcagtt gtggtcagca gcaaaatcaa gtgaatcatc tcagtgcaac taaagggggg
841 atccgatctc aatatgaagt tatgcatatt actggccgtc gtggcctttg ttggcctctc
901 gctcgggtcg ggattcatca gtttggattg cggttcgcct agagaaacta gctttcgtga
961 gaaaacaaca aacattactt acatttccga tgcgaatttc atcaataccg gtgttggtgg
1021 aagcatcaaa caaggatacc ggactcagtt tcagcaacaa acatggaacc tacggagttt
1081 tccacaaggc ataagaaact gctatacctt aaacctaacc ataggcgacg aatatctaat
1141 cagagccaat ttccttcacg gcggttatga cgacaaaccc tcaacgcaat tcgagctcta
1201 cctcggacct aatctttggt ctacagttac aaccacgaac gagaccgaag catcaatctt
1261 tgagatgatt catatcttga ccactgatcg tttacaaatc tgtctggtca agacaggaaa
1321 cgcaacgcct tttatctccg ctttagagct tcgtaaactc atgaacacaa cttacttgac
1381 tcgacaaggt tccttgcaga ccttcatcag agcagacgtt ggcgcgactg ttaaccaagg
1441 ctacaggtac ggaatcgatg tgtttgatcg tgtctggact ccgtataatt tcggaaactg
1501 gtcacagatt agtaccaatc agtcagtgaa tataaacaat gactatcaac caccagagat
1561 tgctatggtc acagcctctg ttccaacaga cccggacgca gctatgaaca tttcgctagt
1621 cggggtggaa cgtactgtgc agttctacgt ttttatgcat tttgcagaga ttcaagagct
1681 caaatccaat gatacaagag agttcaacat catgtacaat aacaagcata tctatggacc
1741 ttttagacca cttaatttca ccacgtcttc cgtatttaca cctaccgaag ttgtcgccga
1801 tgcaaatggg cagtacatat tttcgcttca aagaacagga aattcgactt tacctcctct
1861 gctcaacgct atggagattt attcggttaa cttgttgccg caacaggaaa ctgatagaaa
1921 agaagttgat gcaatgatga acatcaagtc ggcttacggc gtgaacaaga ttgactggga
1981 aggagatcct tgcgtgccgc tggattacaa gtggtctggt gttaattgca cttatgttga
2041 caatgagacc ccaaagatca tttctttgga tctgtctaca agtggcttga ccggagagat
2101 cttagaattt atatcagacc ttaccagttt agaagttctt gacttgtcta acaattcttt
2161 aactggttca gtcccagagt ttctagctaa tatggaaacc ttaaaactca taaatctatc
2221 tggaaatgag ctaaatggct ctatccctgc aactctcctt gataaagaac gacgaggatc
2281 tattacgtta agtatcgaag gaaataccgg gctttgttca tctacatcat gcgctaccac
2341 gaaaaagaag ctggaggtgc tgttccaggg accaagcggc aatcccggcg gcgatctggc
2401 cccccagatg ctgcgcgagc tgcaggagac caatgccgcc ctgcaggacg tgcgcgagct
2461 gctgcgccac cgcgtgaagg agatcacctt cctgaagaac accgtgatgg agtgcgacgc
2521 ctgcggcatg cagccctcta gaggctccgg ctccatcatc ccagttgagg aggagaaccc
2581 ggacttctgg aaccgcgagg cagccgaggc cctgggtgcc gccaagaagc tgcagcctgc
2641 acagacagcc gccaagaacc tcatcatctt cctgggcgat gggatggggg tgtctacggt
2701 gacagctgcc aggatcctaa aagggcagaa gaaggacaaa ctggggcctg agatacccct
2761 ggccatggac cgcttcccat atgtggctct gtccaagaca tacaatgtag acaaacatgt
2821 gccagacagt ggagccacag ccacggccta cctgtgcggg gtcaagggca acttccagac
2881 cattggcttg agtgcagccg cccgctttaa ccagtgcaac acgacacgcg gcaacgaggt
2941 catctccgtg atgaatcggg ccaagaaagc agggaagtca gtgggagtgg taaccaccac
3001 acgagtgcag cacgcctcgc cagccggcac ctacgcccac acggtgaacc gcaactggta
3061 ctcggacgcc gacgtgcctg cctcggcccg ccaggagggg tgccaggaca tcgctacgca
3121 gctcatctcc aacatggaca ttgacgtgat cctaggtgga ggccgaaagt acatgtttcg
3181 catgggaacc ccagaccctg agtacccaga tgactacagc caaggtggga ccaggctgga
3241 cgggaagaat ctggtgcagg aatggctggc gaagcgccag ggtgcccggt atgtgtggaa
3301 ccgcactgag ctcatgcagg cttccctgga cccgtctgtg acccatctca tgggtctctt
3361 tgagcctgga gacatgaaat acgagatcca ccgagactcc acactggacc cctccctgat
3421 ggagatgaca gaggctgccc tgcgcctgct gagcaggaac ccccgcggct tcttcctctt
3481 cgtggagggt ggtcgcatcg accatggtca tcatgaaagc agggcttacc gggcactgac
3541 tgagacgatc atgttcgacg acgccattga gagggcgggc cagctcacca gcgaggagga
3601 cacgctgagc ctcgtcactg ccgaccactc ccacgtcttc tccttcggag gctaccccct
3661 gcgagggagc tccatcttcg ggctggcccc tggcaaggcc cgggacagga aggcctacac
3721 ggtcctccta tacggaaacg gtccaggcta tgtgctcaag gacggcgccc ggccggatgt
3781 taccgagagc gagagcggga gccccgagta tcggcagcag tcagcagtgc ccctggacga
3841 agagacccac gcaggcgagg acgtggcggt gttcgcgcgc ggcccgcagg cgcacctggt
3901 tcacggcgtg caggagcaga ccttcatagc gcacgtcatg gccttcgccg cctgcctgga
3961 gccctacacc gcctgcgacc tggcgccccc cgccggcacc accgacgccg cgcacccggg
4021 ttctagagga agcacctccg gcgataacga ttacaaggat gatgatgata aggccagcgg
4081 caccggtcat catcaccatc accattgagt ttaaacccgc tgatcagcct cgactgtgcc
4141 ttctaaggcc tgagctcgct gatcagcctc gatcgaggat ccagacatga taagatacat
4201 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
4261 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
4321 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa
4381 gtaaaacctc tacaaatgtg gtatggctga ttatgatcag tcgacctgca ggcatgcaag
4441 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
4501 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
4561 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
4621 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
4681 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
4741 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
4801 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
4861 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
4921 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
4981 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
5041 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
5101 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
5161 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
5221 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
5281 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
5341 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
5401 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
5461 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
5521 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
5581 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
5641 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
5701 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
5761 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
5821 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
5881 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
5941 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
6001 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
6061 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
6121 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
6181 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
6241 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
6301 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
6361 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
6421 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
6481 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
6541 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
6601 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
6661 cacgaggc
//
| {
"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": "AT1G51800",
"Species": "A. thaliana (mustard weed)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "Metallothionein (Copper-Inducible)",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Ligation Independent Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pHH0103_CLTC_Clathrin | 109997 | Protein expression and purification of CLTC_Clathrin | 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/215485/1df52119-1da4-4c40-b9cd-6678718dffc7/addgene-plasmid-109997-sequence-215485.gbk | full | LOCUS sequence-215485- 7815 bp DNA circular SYN 26-AUG-2024
DEFINITION Protein expression and purification of CLTC_Clathrin.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7815)
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 7815)
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..7815
/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 2010..2057
/label=fd terminator
/note="central terminator from bacteriophage fd (Otsuka and
Kunisawa, 1982)"
rep_origin complement(3650..4105)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3787..3808)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 3999..4018
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind complement(4147..4169)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
misc_feature 4255..4395
/label=bom
/note="basis of mobility region from pBR322"
primer_bind 4310..4329
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(4410..4427)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4581..5168)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5339..6199)
/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 5962..5981
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6200..6304)
/gene="bla"
/label=AmpR promoter
primer_bind 6372..6390
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 6413..6490
/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 6491..7573
/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(6510..6529)
/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 cggccatggc gcagattctg ccgatccgtt ttcaagaaca tctgcagctg caaaacctgg
961 gcattaaccc ggccaatatc ggttttagca ccctgacgat ggaatctgat aaattcattt
1021 gcatccgcga aaaagtgggt gaacaggcgc aagtggttat tatcgatatg aacgacccga
1081 gcaatccgat tcgtcgcccg atcagtgcag actccgctat catgaacccg gcatctaaag
1141 tgattgcgct gaaagccggc aaaaccctgc agatcttcaa catcgaaatg aaaagtaaaa
1201 tgaaagcgca caccatgacg gatgacgtta cgttctggaa atggatttcc ctgaacaccg
1261 ttgcactggt cacggataat gctgtttatc attggtcaat ggagggtgaa tcgcagccgg
1321 tcaaaatgtt tgatcgtcac agctctctgg caggctgcca gattatcaac taccgcaccg
1381 acgctaaaca aaaatggctg ctgctgacgg gcatcagtgc acagcaaaat cgtgtcgtgg
1441 gtgctatgca gctgtatagt gtggatcgca aagtttccca accgattgaa ggtcatgcgg
1501 cctcatttgc gcagttcaaa atggaaggca acgccgaaga atcgaccctg ttttgtttcg
1561 cggtccgtgg tcaggccggc ggtaaactgc acattatcga agttggcacc ccgccgacgg
1621 gtaaccaacc gtttccgaaa aaagccgtgg atgtgttttt cccgccggaa gcgcagaatg
1681 actttccggt ggccatgcaa atcagcgaaa aacatgatgt tgtcttcctg atcaccaaat
1741 atggctacat tcacctgtat gacctggaaa ccggcacgtg catttacatg aatcgtatct
1801 ctggcgaaac cattttcgtt acggcaccgc atgaagcaac cgctggcatt atcggtgtca
1861 accgcaaggg tcaggtgctg agcgtctgtg tggaagaaga aaatattatc ccgtacatta
1921 ccaacgtgct gcagaatccg gatctggcac tgcgtatggc tgttcgcaac aatctggcgg
1981 gcgccgaaga actggcggcc gcataataaa ccgatacaat taaaggctcc ttttggagcc
2041 ttttttttgg agattttcaa cgtgaaaaaa ttattattcg caattccttt agttgttcct
2101 ttctattctc actccgctga aactgttgaa agttgtttag caaaacccca tacagaaaat
2161 tcatttacta acgtctggaa agacgacaaa actttagatc gttacgctaa ctatgagggc
2221 tgtctgtgga atgctacagg cgttgtagtt tgtactggtg acgaaactca gtgtctagct
2281 agagtggcgg tggctctggt tccggtgatt ttgattatga aaagatggca aacgctaata
2341 agggggctat gaccgaaaat gccgatgaaa acgcgctaca gtctgacgct aaaggcaaac
2401 ttgattctgt cgctactgat tacggtgctg ctatcgatgg tttcattggt gacgtttccg
2461 gccttgctaa tggtaatggt gctactggtg attttgctgg ctctaattcc caaatggctc
2521 aagtcggtga cggtgataat tcacctttaa tgaataattt ccgtcaatat ttaccttccc
2581 tccctcaatc ggttgaatgt cgcccttttg tctttggcgc tggtaaacca tatgaatttt
2641 ctattgattg tgacaaaata aacttattcc gtggtgtctt tgcgtttctt ttatatgttg
2701 ccacctttat gtatgtattt tctacgtttg ctaacatact gcgtaataag gagtcttaat
2761 catgccagtt cttttggcta gcgccgccct ataccttgtc tgcctccccg cgttgcgtcg
2821 cggtgcatgg agccgggcca cctcgacctg aatggaagcc ggcggcacct cgctaacgga
2881 ttcaccactc caagaattgg agccaatcaa ttcttgcgga gaactgtgaa tgcgcaaacc
2941 aacccttggc agaacatatc catcgcgtcc gccatctcca gcagccgcac gcggcgcatc
3001 tcgggcagcg ttgggtcctg gccacgggtg cgcatgatcg tgctcctgtc gttgaggacc
3061 cggctaggct ggcggggttg ccttactggt tagcagaatg aatcaccgat acgcgagcga
3121 acgtgaagcg actgctgctg caaaacgtct gcgacctgag caacaacatg aatggtcttc
3181 ggtttccgtg tttcgtaaag tctggaaacg cggaagtcag cgccctgcac cattatgttc
3241 cggatctgca tcgcaggatg ctgctggcta ccctgtggaa cacctacatc tgtattaacg
3301 aagcgctggc attgaccctg agtgattttt ctctggtccc gccgcatcca taccgccagt
3361 tgtttaccct cacaacgttc cagtaaccgg gcatgttcat catcagtaac ccgtatcgtg
3421 agcatcctct ctcgtttcat cggtatcatt acccccatga acagaaatcc cccttacacg
3481 gaggcatcag tgaccaaaca ggaaaaaacc gcccttaaca tggcccgctt tatcagaagc
3541 cagacattaa cgcttctgga gaaactcaac gagctggacg cggatgaaca ggcagacatc
3601 tgtgaatcgc ttcacgacca cgctgatgag ctttaccgca ggatccgcga aattgtaagc
3661 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
3721 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
3781 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
3841 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
3901 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
3961 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
4021 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
4081 cttaatgcgc cgctacaggg cgcgtcgcgg atcctgcctc gcgcgtttcg gtgatgacgg
4141 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
4201 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc
4261 catgacccag tcacgtagcg atagcggagt gtatactggc ttaactatgc ggcatcagag
4321 cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga
4381 aaataccgca tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
4441 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
4501 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
4561 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
4621 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca gggtttcccc
4681 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
4741 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
4801 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
4861 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
4921 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
4981 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
5041 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
5101 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
5161 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
5221 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
5281 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
5341 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
5401 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
5461 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
5521 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
5581 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
5641 ttgttgccat tgctgcaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
5701 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
5761 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
5821 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
5881 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
5941 cttgcccggc gtcaacacgg gataataccg cgccacatag cagaacttta aaagtgctca
6001 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
6061 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
6121 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
6181 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
6241 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
6301 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
6361 taacctataa aaataggcgt atcacgaggc cctttcgtct tcaagaattc ccgacaccat
6421 cgaatggtgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga gtcaattcag
6481 ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg gtgtctctta
6541 tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa
6601 agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac aacaactggc
6661 gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc acgcgccgtc
6721 gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc
6781 gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca
6841 acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca ttgctgtgga
6901 agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga cacccatcaa
6961 cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc tggtcgcatt
7021 gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg
7081 tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag cggaacggga
7141 aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga atgagggcat
7201 cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat
7261 taccgagtcc gggctgcgcg ttggtgcgga tatctcggta gtgggatacg acgataccga
7321 agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc gcctgctggg
7381 gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga agggcaatca
7441 gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc
7501 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
7561 aagcgggcag tgagcgcaac gcaattaatg taagttagct cactcattag gcacaattct
7621 catgtttgac agcttatcat cgactgcacg gtgcaccaat gcttctggcg tcaggcagcc
7681 atcggaagct gtggtatggc tgtgcaggtc gtaaatcact gcataattcg tgtcgctcaa
7741 ggcgcactcc cgttctggat aatgtttttt gcgccgacat cataacggtt ctggcaaata
7801 ttctgaaatg agctg
//
| {
"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": "CLTC_Clathrin",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "CLTC",
"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": ""
} |
|
CDS12_mTurquoise2 | 136089 | FP for N-term fusion with a CTAG | Jim Haseloff | Sauret-Gueto et al ACS Synth Biol. 2020 Mar 12. doi: 10.1021/acssynbio.9b00511. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/275020/c70e3173-4f94-417c-b041-bcbf6ec01e77/addgene-plasmid-136089-sequence-275020.gbk | full | LOCUS sequence-275020- 2777 bp DNA circular SYN 26-AUG-2024
DEFINITION FP for N-term fusion with a CTAG.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2777)
AUTHORS Sauret-Gueto S, Frangedakis E, Silvestri L, Rebmann M, Tomaselli M,
Markel K, Delmans M, West A, Patron NJ, Haseloff J
TITLE Systematic tools for reprogramming plant gene expression in a simple
model, Marchantia polymorpha.
JOURNAL ACS Synth Biol. 2020 Mar 12. doi: 10.1021/acssynbio.9b00511.
PUBMED 32163700
REFERENCE 2 (bases 1 to 2777)
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. 2020 Mar 12. doi: 10.1021/acssynbio.9b00511."
FEATURES Location/Qualifiers
source 1..2777
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 83..101
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
terminator complement(105..148)
/label=bacterial terminator
/note="putative bacterial transcription terminator"
CDS 165..881
/codon_start=1
/product="enhanced monomeric variant of CFP (Goedhart et
al., 2012)"
/label=mTurquoise2
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(210..231)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(471..490)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 818..839
/label=EGFP-C
/note="EGFP, forward primer"
terminator 901..958
/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(982..999)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(1153..1741)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1233..1252)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
terminator complement(1924..2018)
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
CDS complement(2039..2698)
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind 2613..2632
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
1 ttggaacctc ttacgtgccc gatcaactcg agtgccacct gacgtctaag aaaccattat
61 tatcatgaca ttaacctata aaaataggcg tatcacgagg cagaatttca gataaaaaaa
121 atccttagct ttcgctaagg atgatttctg gaattcggtc tcaaatggtg agcaagggcg
181 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
241 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
301 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctgt
361 cctggggcgt gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca
421 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
481 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
541 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
601 actttagcga caacgtctat atcaccgccg acaagcagaa gaacggcatc aaggccaact
661 tcaagatccg ccacaacatc gaggacggcg gcgtgcagct cgccgaccac taccagcaga
721 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
781 ccaagctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
841 ccgccgccgg gatcactctc ggcatggacg agctgtacaa ggcttcgcga gaccctgcag
901 tccggcaaaa aagggcaagg tgtcaccacc ctgccctttt tctttaaaac cgaaaagatt
961 acttcgcgtt atgcaggctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
1021 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
1081 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
1141 gttgctggcg tttttccaca ggctccgccc ccctgacgag catcacaaaa atcgacgctc
1201 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
1261 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
1321 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
1381 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
1441 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
1501 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
1561 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
1621 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
1681 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
1741 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
1801 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
1861 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gctcgaggct
1921 tggattctca ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca aatccagatg
1981 gagttctgag gtcattactg gatctatcaa caggagtcca agcgagctcg atatcaaatt
2041 acgccccgcc ctgccactca tcgcagtact gttgtaattc attaagcatt ctgccgacat
2101 ggaagccatc acaaacggca tgatgaacct gaatcgccag cggcatcagc accttgtcgc
2161 cttgcgtata atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc atattggcca
2221 cgtttaaatc aaaactggtg aaactcaccc agggattggc tgagacgaaa aacatattct
2281 caataaaccc tttagggaaa taggccaggt tttcaccgta acacgccaca tcttgcgaat
2341 atatgtgtag aaactgccgg aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt
2401 cagtttgctc atggaaaacg gtgtaacaag ggtgaacact atcccatatc accagctcac
2461 cgtctttcat tgccatacga aattccggat gagcattcat caggcgggca agaatgtgaa
2521 taaaggccgg ataaaacttg tgcttatttt tctttacggt ctttaaaaag gccgtaatat
2581 ccagctgaac ggtctggtta taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt
2641 ctttacgatg ccattgggat atatcaacgg tggtatatcc agtgattttt ttctccattt
2701 tagcttcctt agctcctgaa aatctcgata actcaaaaaa tacgcccggt agtgatctta
2761 tttcattatg gtgaaag
//
| {
"Vector backbone": "pUAP4",
"Backbone size w/o insert (bp)": "",
"Vector type": "Plant Expression, 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": "OpenMTA",
"Industry Terms": "OpenMTA"
} | {
"Title": "Systematic tools for reprogramming plant gene expression in a simple model, Marchantia polymorpha",
"Publication": "ACS Synth Biol. 2020 Mar 12. doi: 10.1021/acssynbio.9b00511.",
"DOI": "10.1021/acssynbio.9b00511.",
"Authors": "Sauret-Gueto S, Frangedakis E, Silvestri L, Rebmann M, Tomaselli M, Markel K, Delmans M, West A, Patron NJ, Haseloff J /acssynbiob",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/32163700",
"PubMed ID": "PubMed 32163700"
} | {
"Gene/Insert name": "CDS12_mTurquoise2(noStop)",
"Species": "Synthetic",
"Insert Size (bp)": "",
"Mutation": "BsaI/ SapI domesticated",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "BsaI (destroyed during cloning)",
"3′ cloning site": "BsaI (destroyed during cloning)",
"5′ sequencing primer": "n/a",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"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-m4134408 | 152305 | 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/313244/d6488cdc-96fe-4c29-b97a-fbd96c23d9dc/addgene-plasmid-152305-sequence-313244.gbk | full | LOCUS sequence-313244- 6485 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 6485)
TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for
protein expression and/or purification
REFERENCE 2 (bases 1 to 6485)
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..6485
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 1..304
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 306..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 925..945
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQS"
CDS 946..972
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
polyA_signal 1025..1146
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1036..1055)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 1090..1109
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
terminator 1261..1307
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
terminator 1337..1473
/label=rrnG terminator
/note="transcription terminator from the E. coli ribosomal
RNA rrnG operon (Albrechtsen et al., 1991)"
misc_feature 1518..1589
/label=3' beta-globin insulator
/note="insulator downstream of the human beta-globin locus
(Farrell et al., 2002)"
rep_origin complement(1815..2403)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1895..1914)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
promoter 2903..4112
/label=UbC promoter
/note="human ubiquitin C promoter"
primer_bind 3949..3968
/label=hUBCpro-F
/note="Human Ubiquitin C (UbC) promoter, forward primer"
CDS 4116..4715
/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(4116..4135)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 4612..4632
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
polyA_signal 4742..4790
/note="synthetic polyadenylation signal"
CDS 5056..5916
/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(5274..5293)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
terminator 5929..6065
/label=rrnG terminator
/note="transcription terminator from the E. coli ribosomal
RNA rrnG operon (Albrechtsen et al., 1991)"
misc_feature 6217..6288
/label=5' beta-globin insulator
/note="insulator upstream of the human beta-globin locus
(Farrell et al., 2002)"
terminator complement(6293..6339)
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
polyA_signal 6357..6405
/note="synthetic polyadenylation signal"
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 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc
541 gatcctacca tccactcgac acacccgcca gcggccgcca ccatgaataa cgagttaagc
601 ccagtagcgc ttcgccagat gagttgcgcc gcaggtacaa cccaaactgc ttgtacggat
661 gacaacgccc tcgcttacta taataccaca aagggaggcc gattcgttct ggccctgttg
721 tcggatctgc aagacttgaa atgggcacgt tttcccaagt ccgacgggac tggcaccatt
781 tacacagaac tagagccgcc atgcaggttc gtgactgata cccctaaagg acccaaggtc
841 aaatatctgt attttatcaa ggggcttaac aatctcaata gaggcatggt gttaggatct
901 ctagccgcaa cggtgcggct gcaagaaaac ctttactttc agtcgtaccc ctatgacgta
961 ccagattatg cgtaagctag cttgactgac tgagatacag cgtaccttca gctcacagac
1021 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
1081 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
1141 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag
1201 gttttttaaa gcaagtaaaa cctctacaaa tgtggtattg gcccatctct atcggtatcg
1261 tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgtgc ccctcgggcc
1321 ggattgctat ctaccggcat tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt
1381 atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg cccacttcca
1441 tacgtgtcct ccttaccaga aatttatcct taaggtcgtc agctatcctg caggcgatct
1501 ctcgatttcg atcaagacat tcctttaatg gtcttttctg gacaccacta ggggtcagaa
1561 gtagttcatc aaactttctt ccctccctaa tctcattggt taccttgggc tatcgaaact
1621 taattaacca gtcaagtcag ctacttggcg agatcgactt gtctgggttt cgactacgct
1681 cagaattgcg tcagtcaagt tcgatctggt ccttgctatt gcacccgttc tccgattacg
1741 agtttcattt aaatcatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
1801 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
1861 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
1921 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
1981 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
2041 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
2101 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
2161 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
2221 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
2281 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
2341 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
2401 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
2461 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
2521 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
2581 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgca
2641 tttaaatttc cgaactctcc aaggccctcg tcggaaaatc ttcaaacctt tcgtccgatc
2701 catcttgcag gctacctctc gaacgaacta tcgcaagtct cttggccggc cttgcgcctt
2761 ggctattgct tggcagcgcc tatcgccagg tattactcca atcccgaata tccgagatcg
2821 ggatcacccg agagaagttc aacctacatc ctcaatcccg atctatccga gatccgagga
2881 atatcgaaat cggggcgcgc ctggcctccg cgccgggttt tggcgcctcc cgcgggcgcc
2941 cccctcgtca cggcgagcgc tgccacgtca gacgaagggc gcaggagcgt cctgatcctt
3001 ccgcccggac gctcaggaca gcggcccgct gctcataaga ctcggcctta gaaccccagt
3061 atcagcagaa ggacatttta ggacgggact tgggtgactc tagggcactg gttttctttc
3121 cagagagcgg aacaggcgag gaaaagtagt cccttctcgg cgattctgcg gagggatctc
3181 cgtggggcgg tgaacgccga tgattatata aggacgcgcc gggtgtggca cagctagttc
3241 cgtcgcagcc gggatttggg tcgcggttct tgtttgtgga tcgctgtgat cgtcacttgg
3301 tgagtagcgg gctgctgggc tggccggggc tttcgtggcc gccgggccgc tcggtgggac
3361 ggaagcgtgt ggagagaccg ccaagggctg tagtctgggt ccgcgagcaa ggttgccctg
3421 aactgggggt tggggggagc gcagcaaaat ggcggctgtt cccgagtctt gaatggaaga
3481 cgcttgtgag gcgggctgtg aggtcgttga aacaaggtgg ggggcatggt gggcggcaag
3541 aacccaaggt cttgagccct tcgctaatgc gggaaagctc ttattcgggt gagatgggct
3601 gggcaccatc tggggaccct gacgtgaagt ttgtcactga ctggagaact cggtttgtcg
3661 tctgttgcgg gggcggcagt tatggcggtg ccgttgggca gtgcacccgt acctttggga
3721 gcgcgcgccc tcgtcgtgtc gtgacgtcac ccgttctgtt ggcttataat gcagggtggg
3781 gccacctgcc ggtaggtgtg cggtaggctt ttctccgtcg caggacgcag ggttcgggcc
3841 tagggtaggc tctcctgaat cgacaggcgc cggacctctg gtgaggggag ggataagtga
3901 ggcgtcagtt tctttggtcg gttttatgta cctatcttct taagtagctg aagctccggt
3961 tttgaactat gcgctcgggg ttggcgagtg tgttttgtga agttttttag gcaccttttg
4021 aaatgtaatc atttgggtca atatgtaatt ttcagtgtta gacttgtaaa ttgtccgcta
4081 aattctggcc gtttttggct tttttgttag acaacatgac cgagtacaag cccacggtgc
4141 gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg
4201 actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc
4261 tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg
4321 acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg
4381 ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga
4441 tggagggcct cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg
4501 gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg
4561 aggccgccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc
4621 ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc
4681 gcacctggtg catgacccgc aagcccggtg cctaggctag tatgtaagcc tagtcttaga
4741 taataaaatc gctatccatc gaagatggat gtgtgttggt tttttgtgtg tgtaacgcta
4801 ggcgcgcctg gtgtaccgag aacgatcctc tcagtgcgag tctcgacgat ccatatcgtt
4861 gcttggcagt cagccagtcg gaatccagct tgggacccag gaagtccaat cgtcagatat
4921 tgtactcaag cctggtcacg gcagcgtacc gatctgttta aacctagata ttgatagtct
4981 gatcggtcaa cgtataatcg agtcctagct tttgcaaaca tctatcaaga gacaggatca
5041 gcaggaggct ttcgcatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
5101 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
5161 gatcagttgg gtgcgcgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
5221 gagagttttc gccccgaaga acgctttcca atgatgagca cttttaaagt tctgctatgt
5281 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
5341 tctcagaatg acttggttga gtattcacca gtcacagaaa agcatcttac ggatggcatg
5401 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
5461 cttctgacaa cgattggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
5521 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
5581 cgtgacacca cgatgcctgt agcaatggca acaaccttgc gtaaactatt aactggcgaa
5641 ctacttactc tagcttcccg gcaacagttg atagactgga tggaggcgga taaagttgca
5701 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
5761 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
5821 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
5881 gctgagatag gtgcctcact gattaagcat tggtaaccga ttctaggtgc attggcgcag
5941 aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc acatatgcca gattcagcaa
6001 cggatacggc ttccccaact tgcccacttc catacgtgtc ctccttacca gaaatttatc
6061 cttaagatcc cgaatcgttt aaactcgact ctggctctat cgaatctccg tcgtttcgag
6121 cttacgcgaa cagccgtggc gctcatttgc tcgtcgggca tcgaatctcg tcagctatcg
6181 tcagcttacc tttttggcag cgatcgcggc tcccgacatc ttggaccatt agctccacag
6241 gtatcttctt ccctctagtg gtcataacag cagcttcagc tacctctcaa ttcaaaaaac
6301 ccctcaagac ccgtttagag gccccaaggg gttatgctat caatcgttgc gttacacaca
6361 caaaaaacca acacacatcc atcttcgatg gatagcgatt ttattatcta actgctgatc
6421 gagtgtagcc agatctagta atcaattacg gggtcattag ttcatagccc atatatggag
6481 ttccg
//
| {
"Vector backbone": "pTwist_CMV_Puro_6",
"Backbone size w/o insert (bp)": "6080",
"Vector type": "Mammalian Expression",
"Selectable markers": "Puromycin"
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "OpenMTA",
"Industry Terms": "OpenMTA"
} | {
"Title": "",
"Publication": "",
"DOI": "",
"Authors": "",
"PubMed Link": "",
"PubMed ID": ""
} | {
"Gene/Insert name": "SARS-CoV-2 nsp9",
"Species": "Severe acute respiratory syndrome coronavirus 2",
"Insert Size (bp)": "338",
"Mutation": "wt",
"Entrez Gene": "ORF1ab",
"Promoter": "CMV",
"Tag / Fusion Protein": "Cleavable TEV;HA (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": ""
} |
||
pAAV-CaMKIIa-eGFP-Cre | 120219 | Expresses cre under the control of CamKIIa promotor | Marie Carlén | Ahrlund-Richter et al Nat Neurosci. 2019 Apr;22(4):657-668. doi: 10.1038/s41593-019-0354-y. Epub 2019 Mar 18. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/235838/d1404462-69c5-4e3e-8ef8-7573532a6078/addgene-plasmid-120219-sequence-235838.gbk | full | LOCUS sequence-235838- 7105 bp DNA circular SYN 25-AUG-2024
DEFINITION Expresses cre under the control of CamKIIa promotor.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7105)
AUTHORS Ahrlund-Richter S, Xuan Y, van Lunteren JA, Kim H, Ortiz C, Pollak
Dorocic I, Meletis K, Carlen M
TITLE A whole-brain atlas of monosynaptic input targeting four different
cell types in the medial prefrontal cortex of the mouse.
JOURNAL Nat Neurosci. 2019 Apr;22(4):657-668. doi:
10.1038/s41593-019-0354-y. Epub 2019 Mar 18.
PUBMED 30886408
REFERENCE 2 (bases 1 to 7105)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1038/s41593-019-0354-y"; journalName: "Nat Neurosci"; date:
"2019-04"; volume: "22"; issue: "4"; pages: "657-668"
FEATURES Location/Qualifiers
source 1..7105
/mol_type="other DNA"
/organism="synthetic DNA construct"
repeat_region 1..130
/label=AAV2 ITR (alternate)
/note="Functional equivalent of wild-type AAV2 ITR"
promoter 149..1437
/label=CaMKII promoter
/note="promoter for the mouse calcium/calmodulin-dependent
kinase II alpha subunit"
CDS 1455..2171
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 1455..2171
/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(1761..1780)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 2108..2129
/label=EGFP-C
/note="EGFP, forward primer"
CDS 2181..3209
/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="SNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLL
SVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLP
RPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFL
GIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVER
WISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQR
YLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLED
GD"
misc_feature 3232..3820
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(3285..3305)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(3703..3714)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
polyA_signal 3852..4328
/label=hGH poly(A) signal
/note="human growth hormone polyadenylation signal"
primer_bind complement(4051..4070)
/label=hGH-PA-R
/note="Human growth hormone terminator, reverse primer"
repeat_region 4368..4508
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 4368..4497
/label=AAV2 ITR
rep_origin 4583..5038
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4670..4689)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4880..4901
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(5055..5074)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 5174..5196
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(5234..5252)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 5320..5424
/gene="bla"
/label=AmpR promoter
CDS 5425..6285
/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(5643..5662)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 6456..7044
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6945..6964
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
121 aggggttcct gcggccgcac gcgtttaaca ttatggcctt aggtcacttc atctccatgg
181 ggttcttctt ctgattttct agaaaatgag atgggggtgc agagagcttc ctcagtgacc
241 tgcccagggt cacatcagaa atgtcagagc tagaacttga actcagatta ctaatcttaa
301 attccatgcc ttgggggcat gcaagtacga tatacagaag gagtgaactc attagggcag
361 atgaccaatg agtttaggaa agaagagtcc agggcagggt acatctacac cacccgccca
421 gccctgggtg agtccagcca cgttcacctc attatagttg cctctctcca gtcctacctt
481 gacgggaagc acaagcagaa actgggacag gagccccagg agaccaaatc ttcatggtcc
541 ctctgggagg atgggtgggg agagctgtgg cagaggcctc aggaggggcc ctgctgctca
601 gtggtgacag ataggggtga gaaagcagac agagtcattc cgtcagcatt ctgggtctgt
661 ttggtacttc ttctcacgct aaggtggcgg tgtgatatgc acaatggcta aaaagcaggg
721 agagctggaa agaaacaagg acagagacag aggccaagtc aaccagacca attcccagag
781 gaagcaaaga aaccattaca gagactacaa gggggaaggg aaggagagat gaattagctt
841 cccctgtaaa ccttagaacc cagctgttgc cagggcaacg gggcaatacc tgtctcttca
901 gaggagatga agttgccagg gtaactacat cctgtctttc tcaaggacca tcccagaatg
961 tggcacccac tagccgttac catagcaact gcctctttgc cccacttaat cccatcccgt
1021 ctgttaaaag ggccctatag ttggaggtgg gggaggtagg aagagcgatg atcacttgtg
1081 gactaagttt gttcgcatcc ccttctccaa ccccctcagt acatcaccct gggggaacag
1141 ggtccacttg ctcctgggcc cacacagtcc tgcagtattg tgtatataag gccagggcaa
1201 agaggagcag gttttaaagt gaaaggcagg caggtgttgg ggaggcagtt accggggcaa
1261 cgggaacagg gcgtttcgga ggtggttgcc atggggacct ggatgctgac gaaggctcgc
1321 gaggctgtga gcagccacag tgccctgctc agaagcccca agctcgtcag tcaagccggt
1381 tctccgtttg cactcaggag cacgggcagg cgagtggccc ctagttctgg gggcagctct
1441 agagcggtac caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
1501 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
1561 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
1621 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
1681 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
1741 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
1801 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
1861 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg
1921 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca
1981 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
2041 tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc
2101 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
2161 agctgtacaa ggctgctgct tccaatttac tgaccgtaca ccaaaatttg cctgcattac
2221 cggtcgatgc aacgagtgat gaggttcgca agaacctgat ggacatgttc agggatcgcc
2281 aggcgttttc tgagcatacc tggaaaatgc ttctgtccgt ttgccggtcg tgggcggcat
2341 ggtgcaagtt gaataaccgg aaatggtttc ccgcagaacc tgaagatgtt cgcgattatc
2401 ttctatatct tcaggcgcgc ggtctggcag taaaaactat ccagcaacat ttgggccagc
2461 taaacatgct tcatcgtcgg tccgggctgc cacgaccaag tgacagcaat gctgtttcac
2521 tggttatgcg gcggatccga aaagaaaacg ttgatgccgg tgaacgtgca aaacaggctc
2581 tagcgttcga acgcactgat ttcgaccagg ttcgttcact catggaaaat agcgatcgct
2641 gccaggatat acgtaatctg gcatttctgg ggattgctta taacaccctg ttacgtatag
2701 ccgaaattgc caggatcagg gttaaagata tctcacgtac tgacggtggg agaatgttaa
2761 tccatattgg cagaacgaaa acgctggtta gcaccgcagg tgtagagaag gcacttagcc
2821 tgggggtaac taaactggtc gagcgatgga tttccgtctc tggtgtagct gatgatccga
2881 ataactacct gttttgccgg gtcagaaaaa atggtgttgc cgcgccatct gccaccagcc
2941 agctatcaac tcgcgccctg gaagggattt ttgaagcaac tcatcgattg atttacggcg
3001 ctaaggatga ctctggtcag agatacctgg cctggtctgg acacagtgcc cgtgtcggag
3061 ccgcgcgaga tatggcccgc gctggagttt caataccgga gatcatgcaa gctggtggct
3121 ggaccaatgt aaatattgtc atgaactata tccgtaacct ggatagtgaa acaggggcaa
3181 tggtgcgcct gctggaagat ggcgattaag aattatatca agcttatcga taatcaacct
3241 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
3301 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
3361 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
3421 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
3481 attgccacca cctgtcagct cctttccgga actttcgctt tccccctccc tattgccacg
3541 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
3601 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctatgtt
3661 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
3721 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
3781 cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg agcgctgctc
3841 gagagatcta cgggtggcat ccctgtgacc cctccccagt gcctctcctg gccctggaag
3901 ttgccactcc agtgcccacc agccttgtcc taataaaatt aagttgcatc attttgtctg
3961 actaggtgtc cttctataat attatggggt ggaggggggt ggtatggagc aaggggcaag
4021 ttgggaagac aacctgtagg gcctgcgggg tctattggga accaagctgg agtgcagtgg
4081 cacaatcttg gctcactgca atctccgcct cctgggttca agcgattctc ctgcctcagc
4141 ctcccgagtt gttgggattc caggcatgca tgaccaggct cagctaattt ttgttttttt
4201 ggtagagacg gggtttcacc atattggcca ggctggtctc caactcctaa tctcaggtga
4261 tctacccacc ttggcctccc aaattgctgg gattacaggc gtgaaccact gctcccttcc
4321 ctgtccttct gattttgtag gtaaccacgt gcggaccgag cggccgcagg aacccctagt
4381 gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa
4441 ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg
4501 cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
4561 catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
4621 gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct
4681 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
4741 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg
4801 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
4861 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc
4921 tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat
4981 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta
5041 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
5101 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
5161 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
5221 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
5281 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
5341 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
5401 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
5461 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
5521 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
5581 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
5641 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
5701 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
5761 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
5821 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
5881 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
5941 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
6001 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
6061 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
6121 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
6181 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
6241 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
6301 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
6361 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
6421 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
6481 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
6541 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
6601 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
6661 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
6721 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
6781 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
6841 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
6901 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
6961 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
7021 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
7081 ttttgctggc cttttgctca catgt
//
| {
"Vector backbone": "pAAV",
"Backbone size w/o insert (bp)": "",
"Vector type": "AAV, Cre/Lox",
"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": "A whole-brain atlas of monosynaptic input targeting four different cell types in the medial prefrontal cortex of the mouse",
"Publication": "Nat Neurosci. 2019 Apr;22(4):657-668. doi: 10.1038/s41593-019-0354-y. Epub 2019 Mar 18.",
"DOI": "10.1038/s41593-019-0354-y.",
"Authors": "Ahrlund-Richter S, Xuan Y, van Lunteren JA, Kim H, Ortiz C, Pollak Dorocic I, Meletis K, Carlen M /s---y",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/30886408",
"PubMed ID": "PubMed 30886408"
} | {
"Gene/Insert name": "cre",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": "EGFP"
} | {
"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": ""
} |
|
GFP-2xPH-TAPP1 WT | 161990 | Low | Expresses two tandem repeats of Tapp1 PH domain that binds to PI(3,4)P2. Fused to eGFP | David Bryant | Roman-Fernandez et al Nat Commun. 2018 Nov 28;9(1):5041. doi: 10.1038/s41467-018-07464-8. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/329343/42da3d87-0993-4617-8fd4-38e09d749c51/addgene-plasmid-161990-sequence-329343.gbk | full | LOCUS sequence-329343- 6747 bp DNA circular SYN 25-AUG-2024
DEFINITION Expresses two tandem repeats of Tapp1 PH domain that binds to
PI(3,4)P2. Fused to eGFP.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6747)
AUTHORS Roman-Fernandez A, Roignot J, Sandilands E, Nacke M, Mansour MA,
McGarry L, Shanks E, Mostov KE, Bryant DM
TITLE The phospholipid PI(3,4)P2 is an apical identity determinant.
JOURNAL Nat Commun. 2018 Nov 28;9(1):5041. doi: 10.1038/s41467-018-07464-8.
PUBMED 30487552
REFERENCE 2 (bases 1 to 6747)
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: "2018-11-28"; volume: "9(1):5041. doi"; pages: "
10.1038/s41467-018-07464-8"
FEATURES Location/Qualifiers
source 1..6747
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 61..364
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 365..568
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 519..539
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 607..616
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 613..1329
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 613..1329
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(658..679)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(919..938)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1266..1287
/label=EGFP-C
/note="EGFP, forward primer"
regulatory 2302..2311
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 2311..2340
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
polyA_signal 3535..3656
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3572..3591)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3626..3645
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(3663..4118)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3800..3821)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 4012..4031
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 4145..4249
/gene="bla"
/label=AmpR promoter
promoter 4251..4608
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
enhancer 4251..4442
/label=SV40 enhancer
/note="enhancer for the SV40 early promoter (Herr, 1993)"
primer_bind complement(4274..4294)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 4459..4594
/label=SV40 ori
/note="SV40 origin of replication"
CDS 4643..5437
/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 4643..5437
/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(4697..4716)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 5307..5326
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(5625..5644)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 5669..5716
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 6045..6633
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6534..6553
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
1321 ctgtacaagt ccggactcag atctcgagtg aatgatcagc aggacctagt ggaatgggta
1381 aatgtgttaa acaaagctat aaaaattaca gtaccaaagc agtcagactc acagcctaat
1441 tctgataacc taagtcgcca tggtgaatgt gggaaaaagc aagtgtctta cagaactgat
1501 attgttggtg gcgtacccat cattactccc actcagaaag aagaagtaaa tgaatgtggt
1561 gaaagtattg acagaaataa tctgaaacgg tcacaaagcc atcttcctta ctttactcct
1621 aaaccacctc aagatagtgc ggttatcaaa gctggatatt gtgtaaaaca aggagcagtg
1681 atgaaaaact ggaagagaag atattttcaa ttggatgaaa acacaatagg ctacttcaaa
1741 tctgaactgg aaaaggaacc tcttcgcgta ataccactta aagaggttca taaagtccag
1801 gaatgtaagc aaagcgacat aatgatgagg gacaacctct ttgaaattgt aacaacgtct
1861 cgaactttct atgtgcaggc tgatagccct gaagagatgc acagttggat taaagcagtc
1921 tctggcgcca ttgtagcaca gcggggtccc ggcagatctg cgtcttctga gcatcccccc
1981 ggtccttcag aatccaaaca cgctttccgt cctaccaacg cagccgccgc cacctcacat
2041 tccacagcct ctcgcagcaa ctctttggtc tcaaccttta ccatggagaa gcgaggattt
2101 tacgagtctc ttgccaaggt caagccaggg aacttcaagg tccagactgt ctctccaaga
2161 gaaccagctt ccaaagtgac tgaacaagct ctgttaagac ctcaaagtaa aaatggccct
2221 caggaaaaag attgtgacct agtagacttg gacgatgcga gccttccggt cagtgacgtg
2281 ggtaccgcgg gcccgggatc cgccaccatg gaacaaaaac tcatctcaga agaggatctg
2341 aatgatcagc aggacctagt ggaatgggta aatgtgttaa acaaagctat aaaaattaca
2401 gtaccaaagc agtcagactc acagcctaat tctgataacc taagtcgcca tggtgaatgt
2461 gggaaaaagc aagtgtctta cagaactgat attgttggtg gcgtacccat cattactccc
2521 actcagaaag aagaagtaaa tgaatgtggt gaaagtattg acagaaataa tctgaaacgg
2581 tcacaaagcc atcttcctta ctttactcct aaaccacctc aagatagtgc ggttatcaaa
2641 gctggatatt gtgtaaaaca aggagcagtg atgaaaaact ggaagagaag atattttcaa
2701 ttggatgaaa acacaatagg ctacttcaaa tctgaactgg aaaaggaacc tcttcgcgta
2761 ataccactta aagaggttca taaagtccag gaatgtaagc aaagcgacat aatgatgagg
2821 gacaacctct ttgaaattgt aacaacgtct cgaactttct atgtgcaggc tgatagccct
2881 gaagagatgc acagttggat taaagcagtc tctggcgcca ttgtagcaca gcggggtccc
2941 ggcagatctg cgtcttctga gcatcccccc ggtccttcag aatccaaaca cgctttccgt
3001 cctaccaacg cagccgccgc cacctcacat tccacagcct ctcgcagcaa ctctttggtc
3061 tcaaccttta ccatggagaa gcgaggattt tacgagtctc ttgccaaggt caagccaggg
3121 aacttcaagg tccagactgt ctctccaaga gaaccagctt ccaaagtgac tgaacaagct
3181 ctgttaagac ctcaaagtaa aaatggccct caggaaaaag attgtgacct agtagacttg
3241 gacgatgcga gccttccggt cagtgacgtg tgaggcagaa gcgcacggag cctgcctgcc
3301 tctgccgtcc tcagttacct ttcatgaggc ttctagccaa agatgataaa gggggaaatg
3361 gtttttagtg cgtatattat actgcctctt aggtgtactc tttgaggatc caccggatct
3421 agataactga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
3481 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
3541 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
3601 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc
3661 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
3721 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
3781 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
3841 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
3901 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
3961 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
4021 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
4081 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
4141 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
4201 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
4261 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
4321 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
4381 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
4441 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
4501 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
4561 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga
4621 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
4681 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
4741 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
4801 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
4861 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
4921 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
4981 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
5041 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
5101 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
5161 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
5221 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
5281 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
5341 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
5401 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
5461 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
5521 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
5581 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag
5641 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt
5701 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
5761 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
5821 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
5881 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
5941 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
6001 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
6061 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
6121 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
6181 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
6241 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
6301 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
6361 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
6421 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
6481 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
6541 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
6601 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
6661 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
6721 gtggataacc gtattaccgc catgcat
//
| {
"Vector backbone": "eGFP-C1",
"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)": "NEB Stable",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "The phospholipid PI(3,4)P2 is an apical identity determinant",
"Publication": "Nat Commun. 2018 Nov 28;9(1):5041. doi: 10.1038/s41467-018-07464-8.",
"DOI": "10.1038/s41467-018-07464-8.",
"Authors": "Roman-Fernandez A, Roignot J, Sandilands E, Nacke M, Mansour MA, McGarry L, Shanks E, Mostov KE, Bryant DM /s---",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/30487552",
"PubMed ID": "PubMed 30487552"
} | {
"Gene/Insert name": "PH-TAPP1",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "PLEKHA1",
"Promoter": "",
"Tag / Fusion Protein": "eGFP (N terminal on backbone)"
} | {
"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": ""
} |
EC67844 | 209514 | (Empty Backbone)Cloning in complete guide expression cassettes, with Cas9 | Lawrenson et al Research Square, 12 March 2024, PREPRINT | https://media.addgene.org/snapgene-media/v2.0.0/sequences/413950/b65852ce-a82e-4072-8e10-abea77c5e78a/addgene-plasmid-209514-sequence-413950.gbk | full | LOCUS sequence-413950- 17181 bp DNA circular SYN 27-AUG-2024
DEFINITION Cloning in complete guide expression cassettes, with Cas9.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 17181)
AUTHORS Lawrenson T, Clarke M, Kirby R, Forner M, Steuernagel B, Harwood W
TITLE An Optimised CRISPR Cas9 and Cas12a Toolkit for Barley and Wheat
JOURNAL Research Square, 12 March 2024, PREPRINT
REFERENCE 2 (bases 1 to 17181)
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: "Research
Square, 12 March 2024, PREPRINT"
FEATURES Location/Qualifiers
source 1..17181
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 32..56
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
CDS 365..994
/codon_start=1
/product="stability protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/label=pVS1 StaA
/translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
CDS 1423..2496
/codon_start=1
/product="replication protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/label=pVS1 RepA
/translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
QAAADRIRKESRQPPAAGASSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
LVMRYRNLIEGEASAGS"
rep_origin 2562..2756
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
promoter 2818..2922
/gene="bla"
/label=AmpR promoter
CDS 2923..3717
/codon_start=1
/gene="aphA-3"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin"
/translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
rep_origin 3888..4476
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4377..4396
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
misc_feature 4612..4636
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
polyA_signal 4724..4898
/label=CaMV poly(A) signal
/note="cauliflower mosaic virus polyadenylation signal"
intron 5620..5809
/label=cat1 intron
/note="castor bean catalase intron, modified"
terminator complement(8037..8289)
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
CDS complement(8305..8325)
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS complement(14058..14078)
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
protein_bind 16202..16223
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 16238..16268
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 16276..16292
/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 16310..16321
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 16328..17005
/codon_start=1
/product="monomeric derivative of DsRed (Campbell et al.,
2002)"
/label=mRFP1
/translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
TGA"
terminator 17050..17121
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 17137..17164
/label=T7Te terminator
/note="phage T7 early transcription terminator"
ORIGIN
1 cacgaagtga tccgtttaaa ctatcagtgt ttgacaggat atattggcgg gtaaacctaa
61 gagaaaagag cgtttattag aataatcgga tatttaaaag ggcgtgaaaa ggtttatccg
121 ttcgtccatt tgtatgtgca tgccaaccac agggttcccc tcgggagtca gccgtgcggc
181 tgcatgaaat cctggccggt ttgtctgatg ccaagctggc ggcctggccg gccagcttgg
241 ccgctgaaga aaccgagcgc cgccgtctaa aaaggtgatg tgtatttgag taaaacagct
301 tgcgtcatgc ggtcgctgcg tatatgatgc gatgagtaaa taaacaaata cgcaagggga
361 acgcatgaag gttatcgctg tacttaacca gaaaggcggg tcaggcaaga cgaccatcgc
421 aacccatcta gcccgcgccc tgcaactcgc cggggccgat gttctgttag tcgattccga
481 tccccagggc agtgcccgcg attgggcggc cgtgcgggaa gatcaaccgc taaccgttgt
541 cggcatcgac cgcccgacga ttgaccgcga cgtgaaggcc atcggccggc gcgacttcgt
601 agtgatcgac ggagcgcccc aggcggcgga cttggctgtg tccgcgatca aggcagccga
661 cttcgtgctg attccggtgc agccaagccc ttacgacata tgggccaccg ccgacctggt
721 ggagctggtt aagcagcgca ttgaggtcac ggatggaagg ctacaagcgg cctttgtcgt
781 gtcgcgggcg atcaaaggca cgcgcatcgg cggtgaggtt gccgaggcgc tggccgggta
841 cgagctgccc attcttgagt cccgtatcac gcagcgcgtg agctacccag gcactgccgc
901 cgccggcaca accgttcttg aatcagaacc cgagggcgac gctgcccgcg aggtccaggc
961 gctggccgct gaaattaaat caaaactcat ttgagttaat gaggtaaaga gaaaatgagc
1021 aaaagcacaa acacgctaag tgccggccgt ccgagcgcac gcagcagcaa ggctgcaacg
1081 ttggccagcc tggcagacac gccagccatg aagcgggtca actttcagtt gccggcggag
1141 gatcacacca agctgaagat gtacgcggta cgccaaggca agaccattac cgagctgcta
1201 tctgaataca tcgcgcagct accagagtaa atgagcaaat gaataaatga gtagatgaat
1261 tttagcggct aaaggaggcg gcatggaaaa tcaagaacaa ccaggcaccg acgccgtgga
1321 atgccccatg tgtggaggaa cgggcggttg gccaggcgta agcggctggg ttgtctgccg
1381 gccctgcaat ggcactggaa cccccaagcc cgaggaatcg gcgtgagcgg tcgcaaacca
1441 tccggcccgg tacaaatcgg cgcggcgctg ggtgatgacc tggtggagaa gttgaaggcc
1501 gcgcaggccg cccagcggca acgcatcgag gcagaagcac gccccggtga atcgtggcaa
1561 gcggccgctg atcgaatccg caaagaatcc cggcaaccgc cggcagccgg tgcgtcgtcg
1621 attaggaagc cgcccaaggg cgacgagcaa ccagattttt tcgttccgat gctctatgac
1681 gtgggcaccc gcgatagtcg cagcatcatg gacgtggccg ttttccgtct gtcgaagcgt
1741 gaccgacgag ctggcgaggt gatccgctac gagcttccag acgggcacgt agaggtttcc
1801 gcagggccgg ccggcatggc cagtgtgtgg gattacgacc tggtactgat ggcggtttcc
1861 catctaaccg aatccatgaa ccgataccgg gaagggaagg gagacaagcc cggccgcgtg
1921 ttccgtccac acgttgcgga cgtactcaag ttctgccggc gagccgatgg cggaaagcag
1981 aaagacgacc tggtagaaac ctgcattcgg ttaaacacca cgcacgttgc catgcagcgt
2041 acgaagaagg ccaagaacgg ccgcctggtg acggtatccg agggtgaagc cttgattagc
2101 cgctacaaga tcgtaaagag cgaaaccggg cggccggagt acatcgagat cgagctagct
2161 gattggatgt accgcgagat tacagaaggc aagaacccgg acgtgctgac ggttcacccc
2221 gattactttt tgatcgatcc cggcatcggc cgttttctct accgcctggc acgccgcgcc
2281 gcaggcaagg cagaagccag atggttgttc aagacgatct acgaacgcag tggcagcgcc
2341 ggagagttca agaagttctg tttcaccgtg cgcaagctga tcgggtcaaa tgacctgccg
2401 gagtacgatt tgaaggagga ggcggggcag gctggcccga tcctagtcat gcgctaccgc
2461 aacctgatcg agggcgaagc atccgccggt tcctaatgta cggagcagat gctagggcaa
2521 attgccctag caggggaaaa aggtcgaaaa agcttctttc ctgtggatag cacgtacatt
2581 gggaacccaa agccgtacat tgggaaccgg aacccgtaca ttgggaaccc aaagccgtac
2641 attgggaacc ggtcacacat gtaagtgact gatataaaag agaaaaaagg cgatttttcc
2701 gcctaaaact ctttaaaact tattaaaact cttaaaaccc gcctggcctg tgcataactg
2761 tctggccagc gcacagccga acagctgcaa aaagcgccca catcaaggct ccgagtgcgc
2821 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
2881 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatggctaa aatgagaata
2941 tcaccggaat tgaaaaaact gatcgaaaaa taccgctgcg taaaagatac ggaaggaatg
3001 tctcctgcta aggtatataa gctggtggga gaaaatgaaa acctatattt aaaaatgacg
3061 gacagccggt ataaagggac cacctatgat gtggaacggg aaaaggacat gatgctatgg
3121 ctggaaggaa agctgcctgt tccaaaggtc ctgcactttg aacggcatga tggctggagc
3181 aatctgctca tgagtgaggc cgatggcgtc ctttgctcgg aagagtatga agatgaacaa
3241 agccctgaaa agattatcga gctgtatgcg gagtgcatca ggctctttca ctccatcgac
3301 atatcggatt gtccctatac gaatagctta gacagccgct tagccgaatt ggattactta
3361 ctgaataacg atctggccga tgtggattgc gaaaactggg aagaggacac tccatttaaa
3421 gatccgcgcg agctgtatga ttttttaaag acggaaaagc ccgaagagga acttgtcttt
3481 tcccacggcg acctgggaga cagcaacatc tttgtgaaag atggcaaagt aagtggcttt
3541 attgatcttg ggagaagcgg cagggcggac aagtggtatg acattgcctt ctgcgtccgg
3601 tcgctcaggg aggatatcgg ggaagaacag tatgtcgagc tattttttga cttactgggg
3661 atcaagcctg attgggagaa aataaaatat tatattttac tggatgaatt gttttagctg
3721 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
3781 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
3841 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
3901 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
3961 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
4021 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
4081 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
4141 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
4201 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
4261 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
4321 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
4381 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
4441 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
4501 cggttcctgc tcggatctgt tggaccggac agtagtcatg gttgatgggc tgcctgtatc
4561 gagtggtgat tttgtgccga gctgccggtc ggggagctgt tggctggctg gtggcaggat
4621 atattgtggt gtaaacaaat tgacgcttag acaacttaat aacacattgc ggacgttttt
4681 aatgtactgg ggttgaacac tctgtgccga attcggatcc agcgatctgg attttagtac
4741 tggattttgg ttttaggaat tagaaatttt attgatagaa gtattttaca aatacaaata
4801 catactaagg gtttcttata tgctcaacac atgagcgaaa ccctatagga accctaattc
4861 ccttatctgg gaactactca cacattatta tggagaaact cgagcttgtc gatcgactct
4921 agctagagaa gcctattcct ttgccctcgg acgagtgctg gggcgtcggt ttccactatc
4981 ggcgagtact tctacacagc catcggtcca gacggccgcg cttctgcggg cgatttgtgt
5041 acgcccgaca gtcccggctc cggatcggac gattgcgtcg catcgaccct gcgcccaagc
5101 tgcatcatcg aaattgccgt caaccaagct ctgatagagt tggtcaagac caatgcggag
5161 catatacgcc cggagccgcg gcgatcctgc aagctccgga tgcctccgct cgaagtagcg
5221 cgtctgctgc tccatacaag ccaaccacgg cctccagaag aagatgttgg cgacctcgta
5281 ttgggaatcc ccgaacatcg cctcgctcca gtcaatgacc gctgttatgc ggccattgtc
5341 cgtcaggaca ttgttggagc cgaaatccgc gtgcacgagg tgccggactt cggggcagtc
5401 ctcggcccaa agcatcagct catcgagagc ctgcgcgacg gacgcactga cggtgtcgtc
5461 catcacagtt tgccagtgat acacatgggg atcagcaatc gcgcatatga aatcacgcca
5521 tgtagtgtat tgaccgattc cttgcggtcc gaatgggccg aacccgctcg tctggctaag
5581 atcggccgca gcgatcgcat ccatggcctc cgcgaccggc tgcagttatc atcatcatca
5641 tagacacacg aaataaagta atcagattat cagttaaagc tatgtaatat ttacaccata
5701 accaatcaat taaaaaatag atcagtttaa agaaagatca aagctcaaaa aaataaaaag
5761 agaaaagggt cctaaccaag aaaatgaagg agaaaaacta gaaatttacc tgcagaacag
5821 cgggcagttc ggtttcaggc aggtcttgca acgtgacacc ctgtgcacgg cgggagatgc
5881 aataggtcag gctctcgctg aattccccaa tgtcaagcac ttccggaatc gggagcgcgg
5941 ccgatgcaaa gtgccgataa acataacgat ctttgtagaa accatcggcg cagctattta
6001 cccgcaggac atatccacgc cctcctacat cgaagctgaa agcacgagat tcttcgccct
6061 ccgagagctg catcaggtcg gagacgctgt cgaacttttc gatcagaaac ttctcgacag
6121 acgtcgcggt gagttcaggc tttttcattc tgcaaaagag aaccagacaa cagggtaagt
6181 gcctagcagt aaacaaacag aactcatcac aagcaaacag caacatcata ttcataccaa
6241 caggtcatgt gtgttcatca catcattagt actaagcatg ccatcatcca agtatatcaa
6301 agtaagggca aagagcatgc atgatcatca ggtgcacaaa agaatcatca aattgtagca
6361 gtacatatct tcatctatca tgcatatcta tccataacag gacgatgcat gttgaccagg
6421 taaaagctac aggatcctat aggaacagca gcgtatatat cttcaccaat cttagcatct
6481 taatcatgtg gcacatgcag tttcaattta agcacatgag ctagttgatt atgaggtacc
6541 agagaatcat caaactgatg tagcagcata tatcctcatc tatcatgcaa tctaatctaa
6601 tctaactaaa caggaaaggt gtgctattca gttaaaagct accgcatcat atacaaacgg
6661 cagcataaga aaaagcataa tcatcttaat catgcaacaa acgcagattc ataataagcc
6721 caagagctag cttgtgatga tcttattcta ctctgatcta cagcaatcag ataacgacct
6781 aaccttgcac atggcaacaa aacaatcgat cggacgaatc agttgtttgt tcctagctag
6841 caccatcgaa ccagataata gatgcacgta cagatcccga aaacgaaccc aaaaacaggg
6901 cagacctagc tgaacctagg cagcgaccca gcagatcgtg agaacgatct catctacgaa
6961 cagcctagaa gcaaccccac gattcccgga caaacgacct aaaatcccca caaatcacat
7021 gagcatgaca ggataaacag cggaaccgat cagatctaca cgaaaacccc acctcccagc
7081 cacccacgat caggaaacac gcggatctag catgatttcg tcaacgcctc agcctagttc
7141 ctagccacag accaagcaga accaccaaac cacgccgagc gaggagatgg ggcaagagga
7201 cgggggacac gatcgccgta ccttgaagcg ggggaaggat cgccgagggt cgcgagaaga
7261 gcaatttgga tttggaaccc gggggttgtg cgctccgaac gatcagacga tgtgagattg
7321 tgggaagagg cgcggagggc cctgtattta tgggctgcga cggggggagg agaggtgggg
7381 agggttgggg aaggaatccc ccacccgtgc cgtgacggtt ccgggccgtg tgagaggagc
7441 ccgctcgtgt ccgccacgca atttccgcga tcggagcgga gctttcgaga ggcggctgga
7501 tggttggtgg ccgttagatt tgtagacgcc gttaacgcct cgcctccacc gggaagagtt
7561 ttgagcagcc gcttatgaca atggcttaac gacgttagac ggagcgttag tggcaggcca
7621 tgatagaata gacgtattgc aatgggatta taattaaata aataagaata taataagata
7681 tggcaagtcg gcactcatga catggtgtcc gaaatgatag tgctcacttt cttagccgag
7741 aaagttgacg cgactgattt agaagttaag atattatttc tctcttcttt tctttcctcg
7801 tcatataagg atgaaataaa ctttagagat tgccggtagt gattttggat ttcggcgatc
7861 aggcttggtt tgccggtttc ggacggtgtg ccttaggcca cccgcagtgt atcttgtaat
7921 gttcaaccga taagcaaggg tggggctcaa gcaagtagta aacaactatg tcaaatgtca
7981 ccatggttat ggtcttgttt agttggcttc ctccgcaaga attcaagctt agcgtcgatc
8041 tagtaacata gatgacaccg cgcgcgataa tttatcctag tttgcgcgct atattttgtt
8101 ttctatcgcg tattaaatgt ataattgcgg gactctaatc ataaaaaccc atctcataaa
8161 taacgtcatg cattacatgt taattattac atgcttaacg taattcaaca gaaattatat
8221 gataatcatc gcaagaccgg caacaggatt caatcttaag aaactttatt gccaaatgtt
8281 tgaacgatct gcttgacaag ctcagacttt acgcttcttc tttgggtcag ccctagagtc
8341 accacccaac tgcgacagat ctatccttgt ctcatagagc cctgtaatag attggtgaat
8401 aagggtagcg tccaacactt ccttggttga tgtgtagcgc tttctgtcga tagtggtatc
8461 gaagtactta aaggctgctg gggctcccag attggtcagt gtgaacaggt ggatgatgtt
8521 ttccgcttgt tcccttatcg gcttatctct gtgcttgttg tacgcggaaa gcacctttaa
8581 ttgtagaaaa ccagtaaaac atggacaatt aggaggcatc gattatctaa ccaaagactt
8641 taacaacact aatctgccac taatattcat gagaagtcct taccttatcg aggttagcgt
8701 ctgcaagaat gacccgcttg gaaaattcgc tgatctgctc tatgatctca tcaagatagt
8761 gcttgtgctg ctcgacaaac aactgcttct gctcgttgtc ctctggtgac cccttcagct
8821 tctcatagtg tgaggccaag tacaggaagt tgacgtactt actgggcaat gccagctcgt
8881 tgcccttctg cagttctccg gcagatgcca gcatgcgttt tctgccattt tccaattcga
8941 aaagcgaata cttgggcagt ttgataatga ggtctttctt cacctctttg taacctggtt
9001 tgtataattt tggttcagtt aattaggaaa attacataat ggctccaaaa aaaaattatc
9061 gaccttttac caaaaaaaat aaaatttatc gaccttgaca ggaaaaagtt acaaatcaaa
9121 agaaagaata tcgtctggca tccaatattt tacctttagc ctcgaggaag tcgatcggat
9181 tcttctcgaa cgaggacctt tccattatgg taattccaag cagttccttg acgcttttca
9241 gtttcttgct cttgcccttc tccaccttgg ccactaccag tacgctgtat gccacggtag
9301 ggctatcaaa gcctccgtat ttcttcggat cccaatcctt cttcctcgca atcagtttgt
9361 cggagttgcg cttagggaga atggactcct tggagaaacc accggtttga acctccgttt
9421 tcttgactat gttgacttgg ggcatgctga gaaccttacg cacagtcgca aaatccctgc
9481 ccttatccca gacaatctca cctgcaagaa acagcaaaga gagagttaag atataagtga
9541 acaagaaagt cttgggatcg acatacatac aatcaattga tcaaattcct aggattaaac
9601 atatgaaata ggaaagacag acctgtttcg ccatttgtct cgatcagagg cctcttccgg
9661 atttcaccat tcgccagagt gatctctgtc ttgaagaagt tcatgatatt cgagtaaaag
9721 aagtatttcg cggtagcctt gccgatctct tgttctgact tcgcgatcat cttcctaacg
9781 tcgtacacct tgtaatcccc gtacacaaac tcagactcca gtttcggata cttcttgatg
9841 agagcagtgc caacaacggc gttcaggtag gcgtcatggg catgatggta gttgttgatc
9901 tcgcgaacct tgtaaaactg gaagtccttc cggaagtcac taaccagttt gctcttgagg
9961 gtgatcacct gcgtaaccat caaaagtcaa tatggattca tcacaaatta atcaggaata
10021 gaaagatcgg gtggccttga agttgatcaa ataaaaagtt tcgaacttta gtttgatatg
10081 gtaaaggaaa gttggaaact ttacctttac ctccctgata agtttgtcat tctcatcata
10141 cttcgtattc atacgcgaat cgagtatctg ggccacatgt ttcgtaatct gacgagtttc
10201 aaccaactgg cgcttgatga agccggcttt gtcgagctca gacagtccgc ccctttctgc
10261 cttcgtgaga ttgtcgaatt tccgttgggt tatgagctta gcattcaaaa gctgcctcca
10321 atagttcttc atcttcttca caacctcctc gctgggtaca ttgtcggatt ttccgcggtt
10381 cttgtctgag cgcgtaagga cctataacac gggagaaacc aaaaagcaaa ctcatttcac
10441 gccaaatact acagatttct caaaaattaa gttataatcc atatatcctg acaagaaatt
10501 gattaaaaca cagttgcttt accttgttgt caatactatc gtccttcagg aaagattgag
10561 ggacaatgtg atccacatca tagtcggaga gtctgttaat gtcgagctct tggtcgacgt
10621 acatatctct cccattctgg aggtaataga ggtacagctt ctcgttctgg agttgagtat
10681 tttcgactgg gtgctctttc agaatctgac tcccaagctc cttgatgcct tcctctatcc
10741 gcttcatgcg ctcccgtgag ttcttctgcc ctttctgagt cgtctggttt tccctcgcca
10801 tctcgatgac gatgttctcc ggcttatggc ggcccatgac ctgtagcaat aattaaaaca
10861 aaaataccac aaacagttag aactgaggca agaagaatga atgcagagaa aaaaaatctt
10921 agctattcat ggaaatggaa gtgcgacgaa cacctaaaat gcaaggagca taaccaaatg
10981 cagaacttac cttcaccagc tcatccacaa ccttcactgt ctgcagtatg cccttcttta
11041 tagctggcga tccagccagg ttggctatgt gttcgtgaag gctatcgccc tgacctgaca
11101 cttgggcctt ctggatatct tctttgaagg tcagtgagtc atcgtgaata agctgcatga
11161 agttccgatt cgcaaaaccg tcggatttga gaaagtccag tatagtcttg ccggattgct
11221 tgtcgcggat gccgttgatg agcttcctgg agagccttcc ccaacctaaa acatagaaca
11281 aacatttgtc tcatcacaaa tatgcatcaa ggaacactca gaagttagtc atatacccaa
11341 attcttgaaa ctagagaaac aaaaaaagag ggatcattgc ggaaaccttt tgacctctta
11401 cctgtatatc tcctcctctt caactgcttc atgaccttgt catcgaacag atgcgcataa
11461 gtcttaaggc gctcctcgat catttctctg tcctcaaaca gcgtgagagt cagcacgatg
11521 tcctcgagga tgtcctcgtt ctcctcgttg tcaaggaagt ccttgtcctt gatgattttc
11581 aacaggtcat ggtatgtgcc aagagaggca ttgaacctat cctcaacgcc gctgatttcc
11641 acagaatcaa agcattcgat cttcttaaag tagtcctctt taagctgttt cacagtgacc
11701 ttccgattgg ttttgaacaa gagatcgaca atggccttct tctgctcacc tgttataaca
11761 gtaaaaatta ctttagaaac aaaacaaaag tgtggtcttt taggaatgat tattgatcag
11821 ttcatagttt aaaatcttgt gtttttatcg taaaagtgtg gaccaggatt tacctgacag
11881 gaaggccggt ttgcgcatcc cctccgtcac atatttcacc ttcgtcagct cgttgtagac
11941 cgtgaagtat tcgtacagga gactatgctt gggcaggact ttctcatttg ggagattctt
12001 atcgaagttg gtcatccttt cgatgaagga ctgtgcactc gcacccttgt ccacgacctc
12061 ttcgaagttc caaggcgtga tagtctcctc ggatttgcgt gtcatccacg caaaccgcga
12121 gttcccgcgt gcgagtggac ctaatgaatt aatagaaaaa tcaggcaaat tagaaaagaa
12181 ataattctca aaagagaaca actgaggtca gaaaatagca ttcatcaact gaaactaggt
12241 atttcttcat atgatactaa cctacatagt agggaatgcg gaacgtgagg atcttttcaa
12301 tcttctctct gttgtctttc aggaacggat agaagtcctc ttgcctcctc aagatagcat
12361 gcagctcacc gaggtggatc tggtgcggta tgctgccatt gtcgaaggta cgctgcttcc
12421 gaagcaaatc ttcgcggttc aatttaacga gcagctcctc tgtaccgtcc atcttttcca
12481 gaataggctt gatgaatttg tagaattcct cctgacttgc acctaataat atcatcagag
12541 gaaaactcaa ttcttatatc ttacagaaat gaacaacaaa atacatgtag cagagaggaa
12601 aacaatcata agtaacaact tacctccatc aatgtaccct gcgtagccat tcttcgattg
12661 gtcaaagaaa atctccttgt acttctcggg gagttgctgg cgaaccaaag ccttcagaag
12721 ggtcagatcc tggtgatgct cgtcgtacct cttgatcatg gaggcgctaa gtggcgcttt
12781 ggttatctcc gtgttgaccc gcagaatgtc actgagcaag atagcgtctg acagattctt
12841 ggcagccaag aacaggtcgg catactgatc acctacaaaa gtttaaaaga ctcaatttta
12901 actctacagc ttcaaataaa tttccgcaaa gaaattgata ggtttagaca acaaaagaag
12961 agttccaatt tcaagagcac ctatttgggc gagcagattg tccagatcat catcgtatgt
13021 atcctttgac agctgcagct tcgcgtcctc ggcgaggtca aagttgcttt tgaagttagg
13081 ggtgaggccc agagacagtg caatcagatt cccgaacaag ccgttcttct tctcgcctgg
13141 caattgagcg atcagattct caagccgtct ggacttcgaa agtctggcag acagtattgc
13201 cttggcgtcg acacctgaga ttgtttgcaa agaaaaatct caaagcatgt acaatctata
13261 aacttatgat ggaaaaagaa aagaaaggta actaaggaat gttacctgat gcattgatgg
13321 ggttctcctc gaacagctga ttgtaggttt ggacgagctg aatgaacagt ttatccacat
13381 cggagttatc cgggttgagg tccccttcga tcaagaaatg gccacgaaac tttatcatgt
13441 gcgcgagagc caggtatatg agccgaaggt ctgccttatc ggttgagtcc accagcttct
13501 tgcgaaggtg gtatatagtc ggatactttt catgataggc tacttcgtca acaatattcc
13561 caaagatggg gtgacgctcg tgtttcttat cttcctctac caggaaggat tcctcgaggc
13621 gatggaagaa cgagtcgtca acctgcaaaa tcaagtttat tgatcatcaa gtgagtttac
13681 atgtagttaa atgctttcag taatatccca atgaagcagg atatgctttc agtacaaaat
13741 acataagcat actatatcat aaaaatcctt accttcgcca tctcgttgct aaagatctct
13801 tggagataac agattctgtt cttcctgcga gtgtaccttc tcctcgcagt acgtttaagc
13861 cttgtcgcct ccgcggtttc tccagagtca aagagcagag ctccgatcaa gttcttctta
13921 atgctgtgcc tatctgtgtt gcccagcact ttaaatttct tcgaaggtac cttgtattca
13981 tcggtgataa cggcccaccc aactgagttc gtaccaatat caagtccaat gctgtacttc
14041 ttgtccattt tccagctgac cttccgcttc ttcttcggtg ggctagaagc cattcctgca
14101 gaagtaacac caaacaacag ggtgagcatc gacaaaagaa acagtaccaa gcaaataaat
14161 agcgtatgaa ggcagggcta aaaaaatcca catatagctg ctgcatatgc catcatccaa
14221 gtatatcaag atcaaaataa ttataaaaca tacttgttta ttataataga taggtactca
14281 aggttagagc atatgaatag atgctgcata tgccatcatg tatatgcatc agtaaaaccc
14341 acatcaacat gtatacctat cctagatcga tatttccatc catcttaaac tcgtaactat
14401 gaagatgtat gacacacaca tacagttcca aaattaataa atacaccagg tagtttgaaa
14461 cagtattcta ctccgatcta gaacgaatga acgaccgccc aaccacacca catcatcaca
14521 accaagcgaa caaaaagcat ctctgtatat gcatcagtaa aacccgcatc aacatgtata
14581 cctatcctag atcgatattt ccatccatca tcttcaattc gtaactatga atatgtatgg
14641 cacacacata cagatccaaa attaataaat ccaccaggta gtttgaaaca gaattctact
14701 ccgatctaga acgaccgccc aaccagacca catcatcaca accaagacaa aaaaaagcat
14761 gaaaagatga cccgacaaac aagtgcacgg catatattga aataaaggaa aagggcaaac
14821 caaaccctat gcaacgaaac aaaaaaaatc atgaaatcga tcccgtctgc ggaacggcta
14881 gagccatccc aggattcccc aaagagaaac actggcaagt tagcaatcag aacgtgtctg
14941 acgtacaggt cgcatccgtg tacgaacgct agcagcacgg atctaacaca aacacggatc
15001 taacacaaac atgaacagaa gtagaactac cgggccctaa ccatggaccg gaacgccgat
15061 ctagagaagg tagagagggg ggggggggga ggacgagcgg cgtaccttga agcggaggtg
15121 ccgacgggtg gatttggggg agatctggtt gtgtgtgtgt gcgctccgaa caacacgagg
15181 ttggggaaag agggtgtgga gggggtgtct atttattacg gcgggcgagg aagggaaagc
15241 gaaggagcgg tgggaaagga atcccccgta gctgccgtgc cgtgagagga ggaggaggcc
15301 gcctgccgtg ccggctcacg tctgccgctc cgccacgcaa tttctggatg ccgacagcgg
15361 agcaagtcca acggtggagc ggaactctcg agaggggtcc agaggcagcg acagagatgc
15421 cgtgccgtct gcttcgcttg gcccgacgcg acgctgctgg ttcgctggtt ggtgtccgtt
15481 agactcgtcg acggcgttta acaggctggc attatctact cgaaacaaga aaaatgtttc
15541 cttagttttt ttaatttctt aaagggtatt tgtttaattt ttagtcactt tattttattc
15601 tattttatat ctaaattatt aaataaaaaa actaaaatag agttttagtt ttcttaattt
15661 agaggctaaa atagaataaa atagatgtac taaaaaaatt agtctataaa aaccattaac
15721 cctaaaccct aaatggatgt actaataaaa tggatgaagt attatatagg tgaagctatt
15781 tgcaaaaaaa aaggagaaca catgcacact aaaaagataa aactgtagag tcctgttgtc
15841 aaaatactca attgtccttt agaccatgtc taactgttca tttatatgat tctctaaaac
15901 actgatatta ttgtagtact atagattata ttattcgtag agtaaagttt aaatatatgt
15961 ataaagatag ataaactgca cttcaaacaa gtgtgacaaa aaaaatatgt ggtaattttt
16021 tataacttag acatgcaatg ctcattatct ctagagaggg gcacgaccgg gtcacgctgc
16081 acctccacta tgcctgagac ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
16141 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
16201 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
16261 gtatgttgtg tggaattgtg agcggataac aatttcacac atactagaga aagaggagaa
16321 atactagatg gcttcctccg aggacgttat caaagagttc atgcgtttca aagttcgtat
16381 ggaaggttcc gttaacggtc acgagttcga aatcgaaggt gaaggtgaag gtcgtccgta
16441 cgaaggtacc cagaccgcta aactgaaagt taccaaaggt ggtccgctgc cgttcgcttg
16501 ggacatcctg tccccgcagt tccagtacgg ttccaaagct tacgttaaac acccggctga
16561 catcccggac tacctgaaac tgtccttccc ggaaggtttc aaatgggaac gtgttatgaa
16621 cttcgaggac ggtggtgttg ttaccgttac ccaggactcc tccctgcaag acggtgagtt
16681 catctacaaa gttaaactgc gtggtaccaa cttcccgtcc gacggtccgg ttatgcagaa
16741 aaaaaccatg ggttgggaag cttccaccga acgtatgtac ccggaggacg gtgctctgaa
16801 aggtgaaatc aaaatgcgtc tgaaactgaa agacggtggt cactacgacg ctgaagttaa
16861 aaccacctac atggctaaaa aaccggttca gctgccgggt gcttacaaaa ccgacatcaa
16921 actggacatc acctcccaca acgaggacta caccatcgtt gaacagtacg aacgtgctga
16981 aggtcgtcac tccaccggtg cttaataacg ctgatagtgc tagtgtagat cgctactaga
17041 gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
17101 tgtttgtcgg tgaacgctct ctactagagt cacactggct caccttcggg tgggcctttc
17161 tgcgtttata ggtctcaggg a
//
| {
"Vector backbone": "Unknown",
"Backbone size w/o insert (bp)": "",
"Vector type": "CRISPR",
"Selectable markers": "RFP & hygromycin for plant"
} | {
"Bacterial Resistance(s)": "Kanamycin, 50 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA ; Takara Bio Limited Use Label License (formerly Clontech)",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "An Optimised CRISPR Cas9 and Cas12a Toolkit for Barley and Wheat",
"Publication": "Research Square, 12 March 2024, PREPRINT",
"DOI": "10.21203/rs.3.rs-4024107/v1",
"Authors": "Lawrenson T, Clarke M, Kirby R, Forner M, Steuernagel B, Harwood W",
"PubMed Link": "",
"PubMed ID": ""
} | {
"Gene/Insert name": "None",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
||
pCIG3 N | 132941 | MLV packaging plasmid for N-tropic MLV gag-pol expression | Jeremy Luban | Kim et al Nat Microbiol. 2019 Dec;4(12):2044-2051. doi: 10.1038/s41564-019-0592-5. Epub 2019 Oct 21. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/258740/25bea6ce-e50b-447b-803d-c34851cf84b1/addgene-plasmid-132941-sequence-258740.gbk | full | LOCUS sequence-258740- 11093 bp DNA circular SYN 26-AUG-2024
DEFINITION MLV packaging plasmid for N-tropic MLV gag-pol expression.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11093)
AUTHORS Kim K, Dauphin A, Komurlu S, McCauley SM, Yurkovetskiy L, Carbone C,
Diehl WE, Strambio-De-Castillia C, Campbell EM, Luban J
TITLE Cyclophilin A protects HIV-1 from restriction by human TRIM5alpha.
JOURNAL Nat Microbiol. 2019 Dec;4(12):2044-2051. doi:
10.1038/s41564-019-0592-5. Epub 2019 Oct 21.
PUBMED 31636416
REFERENCE 2 (bases 1 to 11093)
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/s41564-019-0592-5"; journalName: "Nat Microbiol"; date:
"2019-12"; volume: "4"; issue: "12"; pages: "2044-2051"
FEATURES Location/Qualifiers
source 1..11093
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 77..380
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 381..592
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 543..563
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 589..613
/label=LNCX
/note="Human CMV promoter, forward primer"
intron 753..885
/label=chimeric intron
/note="chimera between introns from human beta-globin and
immunoglobulin heavy chain genes"
primer_bind 868..887
/label=pMT2-F
/note="Synthetic intron, forward primer"
promoter 930..948
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
misc_feature 5837..6211
/label=pol region
/note="Moloney murine leukemia virus (MMLV) pol region
containing the splice acceptor site"
primer_bind 6149..6165
/label=pBMN 5'
/note="MMLV sequence, for inserts in pBMN retroviral
vector"
polyA_signal 7063..7197
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(7100..7119)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 7154..7173
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(8001..8020)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 8611..8630
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 8805..8853
/note="synthetic polyadenylation signal"
primer_bind complement(8894..8913)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 9013..9035
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(9073..9091)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 9159..9263
/gene="bla"
/label=AmpR promoter
CDS 9264..10124
/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(9482..9501)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 10295..10883
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 10784..10803
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 ggcattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctgcgatcgc ccgccccgtt
541 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcgtttagt
601 gaaccgtcag atcactagaa gctttattgc ggtagtttat cacagttaaa ttgctaacgc
661 agtcagtgct tctgacacaa cagtctcgaa cttaagctgc agtgactctc ttaaggtagc
721 cttgcagaag ttggtcgtga ggcactgggc aggtaagtat caaggttaca agacaggttt
781 aaggagacca atagaaactg ggcttgtcga gacagagaag actcttgcgt ttctgatagg
841 cacctattgg tcttactgac atccactttg cctttctctc cacaggtgtc cactcccagt
901 tcaattacag ctcttaaggc tagagtactt aatacgactc actataggct agcctcgaga
961 attcgagctc gcccgccgaa accgcgccgc gcgtcttgtc tgtctcagta ttgttttgtt
1021 atttgtctgt tcgttattgt tttggaccgc ttctaaaaac atgggacaaa ccgtaaccac
1081 ccctctgagt ctgaccctag aacactggaa agatgtccag cgcatcgcgt ccaatcagtc
1141 cgtagatgtc aagaagagac gctgggtcac cttctgctct gccgagtggc caactttcgg
1201 tgtagggtgg ccacaagatg gtacttttaa cttggatatt attctacagg ttaaatctaa
1261 ggttttctct cctggtcccc acggacaccc ggatcaggtc ccatatattg tcacctggga
1321 ggctattgcc tatgaacccc ctccgtgggt aaaacctttt gtctctccca aactctcccc
1381 ctctccaacc gctcccatcc ttccatccgg tccttcgacc caacctccgc cccgatctgc
1441 cctttaccct gctcttactc cctctataaa acccagacct tctaaacctc aggttctctc
1501 cgataatggc ggacctctca ttgaccttct ctcagaagac cctccgccgt acggaggaca
1561 gggaccgtcc tcctctgacg gagatggcga cagagaagag gccacctcca cttctgagat
1621 tcctgccccc tctcccatag tgtctcgcct gcggggcaaa agagaccccc ccgcggcaga
1681 ttccaccacc tctcgggctt tcccactccg tttggggggt aatggtcagt tgcagtactg
1741 gccgttttcc tcctctgatc tatataactg gaaaaataat aatccttcct tctctgagga
1801 tccaggtaaa ctgactgcat tgattgaatc tgtcctcacc acccaccagc ccacctggga
1861 tgattgccag caattattag ggactctgct taccggggaa gagaagcagc gggtgctcct
1921 ggaagcccga aaagctgtcc ggggcaacga tgggcgcccc acccaactgc ccaacgaggt
1981 tgacgctgct tttccccttg aacgtcccga ttgggattac accacccaaa gaggtaggaa
2041 ccacctagtt ctctatcgcc agttgctttt agcgggtctc caaaacgcgg gccgaagccc
2101 caccaatttg gccaaggtaa aaggaataac ccagggacct gatgagtccc cctcagcctt
2161 cctagagaga ctcaaggaag cctatcgcag atacactcct tatgatcctg aggaccctgg
2221 gcaagaaacg aatgtatcta tgtcattcat ctggcagtcc gctccagaca ttggtcgaaa
2281 gttagagcgg ttagaagact taaaaagtaa aactttagga gatttagtga gagaagccga
2341 aaggatcttt aataaaagag aaactccaga agaaagagaa gagcgcgtta ggagggagac
2401 agaggaaaag gaagagcggc gtagggcaga agaggagcag aaagagaaag agagggaccg
2461 taggagacat agagaaatga gcaaactttt ggccaccgta gttagtggac agagacagga
2521 tagacagggg ggagagcgaa ggaggcccca acttgacaag gatcaatgtg cctactgtaa
2581 agaaaaagga cactgggcta aagactgccc aaagaagcca cggggtcccc gaggaccgcg
2641 accccagacc tccctcctga ctttagacga ctaggggggt cagggtcagg agcccccccc
2701 tgaacccagg ataaccctca ctgtcggggg gcaacccgtc accttcctgg tggatactgg
2761 ggcccaacac tccgtgctga ctcaaaatcc tgggccccta agtgacaggt cggcttgggt
2821 ccaaggggct actggaggaa agcggtatcg ctggaccaca gatcgcaagg tacacctagc
2881 taccggtaag gtcactcact ctttcctcca tgtgccagac tgcccctacc ccttgctagg
2941 aagagacttg ttaactaaac taaaagccca gatccacttt gagggatcag gagcccaggt
3001 tgtgggacca aaaggacagc ctctacaggt gttgacccta aacctagaag atgagtatcg
3061 gcttcatgag acctcagcag agccggaagc ttctcctggg tccacctggc tttccgactt
3121 tccccaggcc tgggcagaaa ccgggggcat gggactggca gttcgccagg cgcctctaat
3181 catacctctg aaggcaacct ccacccctgt gtccataaaa caatacccca tgtcacagga
3241 agccagactg gggatcaagc cccacataca gaggctgttg gaccagggaa tactggtacc
3301 ctgccagtcc ccctggaata cacccctgct acccgttaag aaaccaggaa ctaacgatta
3361 taggcctgtc caggatctga gagaagtcaa caagcgggtg gaagacatcc accccaccgt
3421 gcccaatcct tataacctcc taagtgggct cccaccgtcc caccggtggt atactgtgct
3481 tgatttaaag gatgcctttt tctgcctgag actccacccc accagtcagc ctctcttcgc
3541 cttcgagtgg agagatccag ggatgggaat ctcaggacaa ttaacctgga ccagactccc
3601 acagggtttc aaaaacagtc ccaccctgtt tgatgaggca ctacacagag acctagcaga
3661 cttccggatc cagcacccag acttgatcct gctacagtac gtggatgaca tactactggc
3721 cgccacttct gagctcgact gccaacaagg tactcgggcc ctgttactaa ccctaggaaa
3781 cctcgggtat cgggcctcgg ccaagaaagc ccaactttgc cagaaacagg tcaagtacct
3841 ggggtatctc ctaaaagagg gtcagagatg gctgactgag gccagaaaag agactgtgat
3901 ggggcagcct actccgaaga cccctcgaca actaagggag ttcctaggga cggcaggctt
3961 ctgtcgcctc tggatccctg ggtttgcgga aatggcagcc cccttgtatc ctcttaccaa
4021 aacggggact ctgttcaatt ggggcccaga ccagcaaaag gcctatcaag aaatcaaaca
4081 ggccctccta actgcccccg ccctgggatt gccagatttg actaagccct ttgaactctt
4141 tgtcgacgag aagcagggct acgccaaagg tgtcctaacg caaaaactgg gaccttggcg
4201 tcggccggtg gcctacctgt ccaaaaagct agacccagta gcagctgggt ggcccccttg
4261 cctacggatg gtagcagcca ttgccgtact gacaaaggat gcaggcaagc taaccatggg
4321 acagccacta gtcattctgg ccccccatgc agtagaggca ctagtcaaac aaccccccga
4381 ccgctggctt tccaacgccc ggatgactca ctatcaggcc ttgcttttgg acacggaccg
4441 ggtccagttc ggaccggtgg tagccctgaa cccggctacg ctgctcccac tgcctgagga
4501 agggctgcaa cacaactgcc ttgatatcct ggccgaagcc cacggaaccc gacccgacct
4561 aacggaccag ccgctcccag acgccgacca cacctggtac acggatggaa gcagtctctt
4621 acaagaggga cagcgtaagg cgggagctgc ggtgaccacc gagaccgagg taatctgggc
4681 taaagccctg ccagccggga catccgctca gcgggctgaa ctgatagcac tcacccaggc
4741 cctaaagatg gcagaaggta agaagctaaa tgtttatact gatagccgtt atgcttttgc
4801 tactgcccat atccatggag aaatatacag aaggcgtggg ttgctcacat cagaaggcaa
4861 agagatcaaa aataaagacg agatcttggc cctactaaaa gccctctttc tgcccaaaag
4921 acttagcata atccattgtc caggacatca aaagggacac agcgccgagg ctagaggcaa
4981 ccggatggct gaccaagcgg cccgaaaggc agccatcaca gagactccag acacctctac
5041 cctcctcata gaaaattcat caccctacac ctcagaacat tttcattaca cagtgactga
5101 tataaaggac ctaaccaagt tgggggccat ttatgataaa acaaagaagt attgggtcta
5161 ccaaggaaaa cctgtgatgc ctgaccagtt tacttttgaa ttattagact ttcttcatca
5221 gctgactcac ctcagcttct caaaaatgaa ggctctccta gagagaagcc acagtcccta
5281 ctacatgctg aaccgggatc gaacactcaa aaatatcact gagacctgca aagcttgtgc
5341 acaagtcaac gccagcaagt ctgccgttaa acagggaact agggtccgcg ggcatcggcc
5401 cggcactcat tgggagatcg atttcaccga gataaagccc ggattgtatg gctataaata
5461 tcttctagtt tttatagata ccttttctgg ctggatagaa gccttcccaa ccaagaaaga
5521 aaccgccaag gtcgtaacca agaagctact agaggagatc ttccccaggt tcggcatgcc
5581 tcaggtattg ggaactgaca atgggcctgc cttcgtctcc aaggtgagtc agacagtggc
5641 cgatctgttg gggattgatt ggaaattaca ttgtgcatac agaccccaaa gctcaggcca
5701 ggtagaaaga atgaatagaa ccatcaagga gactttaact aaattaacgc ttgcaactgg
5761 ctctagagac tgggtgctcc tactcccctt agccctgtac cgagcccgca acacgccggg
5821 cccccatggc ctcaccccat atgagatctt atatggggca cccccgcccc ttgtaaactt
5881 ccctgaccct gacatgacaa gagttactaa cagcccctct ctccaagctc acttacaggc
5941 tctctactta gtccagcacg aagtctggag acctctggcg gcagcctacc aagaacaact
6001 ggaccgaccg gtggtacctc acccttaccg agtcggcgac acagtgtggg tccgccgaca
6061 ccagactaag aacctagaac ctcgctggaa aggaccttac acagtcctgc tgaccacccc
6121 caccgccctc aaagtagacg gcatcgcagc ttggatacac gccgcccacg tgaaggctgc
6181 cgaccccggg ggtggaccat cctctagact gacatggcgc gttcaacgct ctcaaaaccc
6241 cttaaaaata aggttaaccc gcgaggcccc ctaatcccct taattcttct gatgctcaga
6301 ggggtcagta ctgcttcgcc cggctccagt gcggcccagc cggccaccat gaaaacattt
6361 aacatttctc aacaagatct agaattagta gaagtagcga cagagaagat tacaatgctt
6421 tatgaggata ataaacatca tgtgggagcg gcaattcgta cgaaaacagg agaaatcatt
6481 tcggcagtac atattgaagc gtatatagga cgagtaactg tttgtgcaga agccattgcg
6541 attggtagtg cagtttcgaa tggacaaaag gattttgaca cgattgtagc tgttagacac
6601 ccttattctg acgaagtaga tagaagtatt cgagtggtaa gtccttgtgg tatgtgtagg
6661 gagttgattt cagactatgc accagattgt tttgtgttaa tagaaatgaa tggcaagtta
6721 gtcaaaacta cgattgaaga actcattcca ctcaaatata cccgaaatta aaagttttac
6781 caccaagctt atcgattagt ccaatttgtt aaagacagga tatcagtggt ccaggctcta
6841 gttttgactc aacaatatca ccagctgaag cctatagagt acgagccata gataaaataa
6901 aagattttat ttagtctcca gaaaaagggg ggaatgaaag accccacctg taggtttggc
6961 aagctagctt aagtaacgcc attttgcaag gcatggaaaa atacataact gagaatagaa
7021 aagttcagat caaggtcagg aacagatgga acagggtcga ggaacttgtt tattgcagct
7081 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
7141 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatcccc
7201 aggaagctcc tctgtgtcct cataaaccct aacctcctct acttgagagg acattccaat
7261 cataggctgc ccatccaccc tctgtgtcct cctgttaatt aggtcactta acaaaaagga
7321 aattgggtag gggtttttca cagaccgctt tctaagggta attttaaaat atctgggaag
7381 tcccttccac tgctgtgttc cagaagtgtt ggtaaacagc ccacaaatgt caacagcaga
7441 aacatacaag ctgtcagctt tgcacaaggg cccaacaccc tgctcatcaa gaagcactgt
7501 ggttgctgtg ttagtaatgt gcaaaacagg aggcacattt tccccacctg tgtaggttcc
7561 aaaatatcta gtgttttcat ttttacttgg atcaggaacc cagcactcca ctggataagc
7621 attatcctta tccaaaacag ccttgtggtc agtgttcatc tgctgactgt caactgtagc
7681 attttttggg gttacagttt gagcaggata tttggtcctg tagtttgcta acacaccctg
7741 cagctccaaa ggttccccac caacagcaaa aaaatgaaaa tttgaccctt gaatgggttt
7801 tccagcacca ttttcatgag ttttttgtgt ccctgaatgc aagtttaaca tagcagttac
7861 cccaataacc tcagttttaa cagtaacagc ttcccacatc aaaatatttc cacaggttaa
7921 gtcctcattt aaattaggca aaggaattcc tgcagcccgg gggatccgcc cgggctagag
7981 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
8041 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
8101 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
8161 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
8221 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
8281 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
8341 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
8401 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
8461 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
8521 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
8581 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
8641 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
8701 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
8761 aatgaccgac caagcgacgc ccaacctgcc atcacgatgg ccgcaataaa atatctttat
8821 tttcattaca tctgtgtgtt ggttttttgt gtgaatcgat agcgataagg atccgcgtat
8881 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
8941 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
9001 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
9061 cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
9121 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
9181 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
9241 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
9301 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
9361 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
9421 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
9481 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
9541 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
9601 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
9661 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
9721 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
9781 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
9841 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
9901 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
9961 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
10021 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
10081 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
10141 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
10201 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
10261 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
10321 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
10381 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
10441 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
10501 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
10561 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
10621 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
10681 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
10741 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
10801 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
10861 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
10921 tttgctggcc ttttgctcac atggctcgac agatcttcaa tattggccat tagccatatt
10981 attcattggt tatatagcat aaatcaatat tggctattgg ccattgcata cgttgtatct
11041 atatcataat atgtacattt atattggctc atgtccaata tgaccgccat gtt
//
| {
"Vector backbone": "pCIG3 N",
"Backbone size w/o insert (bp)": "",
"Vector type": "Retroviral",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "30°C",
"Growth Strain(s)": "NEB Stable",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Cyclophilin A protects HIV-1 from restriction by human TRIM5alpha",
"Publication": "Nat Microbiol. 2019 Dec;4(12):2044-2051. doi: 10.1038/s41564-019-0592-5. Epub 2019 Oct 21.",
"DOI": "10.1038/s41564-019-0592-5.",
"Authors": "Kim K, Dauphin A, Komurlu S, McCauley SM, Yurkovetskiy L, Carbone C, Diehl WE, Strambio-De-Castillia C, Campbell EM, Luban J /s---",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/31636416",
"PubMed ID": "PubMed 31636416"
} | {
"Gene/Insert name": "N-tropic MLV gag-pol",
"Species": "MLV",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "unknown (unknown if destroyed)",
"3′ cloning site": "unknown (unknown if destroyed)",
"5′ sequencing primer": "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": ""
} |
|
pMYT009 | 180662 | Encodes pLacSpTDH3 as a Type 2 part to be used in the Dueber YTK and Ellis MYT systems | Tom Ellis | Shaw et al ACS Synth Biol. 2023 Nov 6. doi: 10.1021/acssynbio.3c00423. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/369267/efa905b1-ae63-41ea-8ccc-728c36fb833a/addgene-plasmid-180662-sequence-369267.gbk | full | LOCUS sequence-369267- 2111 bp DNA circular SYN 26-AUG-2024
DEFINITION Encodes pLacSpTDH3 as a Type 2 part to be used in the Dueber YTK and
Ellis MYT systems.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2111)
AUTHORS Shaw WM, Khalil AS, Ellis T
TITLE A Multiplex MoClo Toolkit for Extensive and Flexible Engineering of
Saccharomyces cerevisiae.
JOURNAL ACS Synth Biol. 2023 Nov 6. doi: 10.1021/acssynbio.3c00423.
PUBMED 37930278
REFERENCE 2 (bases 1 to 2111)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth
Biol. 2023 Nov 6. doi: 10.1021/acssynbio.3c00423."
FEATURES Location/Qualifiers
source 1..2111
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind 356..372
/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 409..425
/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)."
terminator complement(478..512)
/label=lambda t0 terminator
/note="minimal transcription terminator from phage lambda
(Scholtissek and Grosse, 1987)"
CDS complement(583..1242)
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind 1157..1176
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
rep_origin 1424..2012
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 1913..1932
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 agtcggtctc aaacggtttt atttctgctg ccatccgtaa atgccaggat ttgagcgggt
61 tacacaatat atctcatatt ttcggtgtct gggtcattac tttactcttg gcatccacta
121 aatatattgg agcctgcttt ttaaactggc ttccagaaaa aaatcaatgg agtgatgcaa
181 actgcctgga gtaaaagatg acacaaggcg attgacctac gcatgtatct atctgatttt
241 cttacacctt ctatttcatt ctaactcttt gatttggaaa acacctaaga aaaaaaaggt
301 tgaaatcagt tccctgaaat tgtcccccta cttgactaat aaatataaaa ggtaattgtg
361 agcggataac aacaagcact gttgggcgtg agtggaggcg ccggataatt gtgagcggat
421 aacaatttaa acacacataa acaaacaaaa aaaaaagatc tatgtgagac cagaccaata
481 aaaaacgccc ggcggcaacc gagcgttctg aacaaatcca gatggagttc tgaggtcatt
541 actggatcta tcaacaggag tccaagcgag ctcgatatca aattacgccc cgccctgcca
601 ctcatcgcag tactgttgta attcattaag cattctgccg acatggaagc catcacaaac
661 ggcatgatga acctgaatcg ccagcggcat cagcaccttg tcgccttgcg tataatattt
721 gcccatggtg aaaacggggg cgaagaagtt gtccatattg gccacgttta aatcaaaact
781 ggtgaaactc acccagggat tggctgaaac gaaaaacata ttctcaataa accctttagg
841 gaaataggcc aggttttcac cgtaacacgc cacatcttgc gaatatatgt gtagaaactg
901 ccggaaatcg tcgtggtatt cactccagag cgatgaaaac gtttcagttt gctcatggaa
961 aacggtgtaa caagggtgaa cactatccca tatcaccagc tcaccgtctt tcattgccat
1021 acgaaattcc ggatgagcat tcatcaggcg ggcaagaatg tgaataaagg ccggataaaa
1081 cttgtgctta tttttcttta cggtctttaa aaaggccgta atatccagct gaacggtctg
1141 gttataggta cattgagcaa ctgactgaaa tgcctcaaaa tgttctttac gatgccattg
1201 ggatatatca acggtggtat atccagtgat ttttttctcc attttagctt ccttagctcc
1261 tgaaaatctc gataactcaa aaaatacgcc cggtagtgat cttatttcat tatggtgaaa
1321 gttggaacct cttacgtgcc cgatcaatca tgaccaaaat cccttaacgt gagttttcgt
1381 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
1441 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
1501 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
1561 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
1621 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
1681 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
1741 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
1801 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
1861 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
1921 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
1981 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
2041 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
2101 tggataaccg t
//
| {
"Vector backbone": "pYTK001",
"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)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "A Multiplex MoClo Toolkit for Extensive and Flexible Engineering of Saccharomyces cerevisiae",
"Publication": "ACS Synth Biol. 2023 Nov 6. doi: 10.1021/acssynbio.3c00423.",
"DOI": "10.1021/acssynbio.3c00423.",
"Authors": "Shaw WM, Khalil AS, Ellis T /acssynbioc",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/37930278",
"PubMed ID": "PubMed 37930278"
} | {
"Gene/Insert name": "pLacSpTDH3",
"Species": "Synthetic",
"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": ""
} |
|
pAc5.1B-lambdaN-HA-DmeIF5B_Q | 147275 | Insect Expression of DmeIF5B | Elisa Izaurralde | Izaurralde lab plasmids (unpublished) | https://media.addgene.org/snapgene-media/v2.0.0/sequences/289722/83a1407b-606c-42c9-9d9a-a19196a2d221/addgene-plasmid-147275-sequence-289722.gbk | full | LOCUS sequence-289722- 8909 bp DNA circular SYN 26-AUG-2024
DEFINITION Insect Expression of DmeIF5B.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8909)
TITLE Izaurralde lab plasmids
REFERENCE 2 (bases 1 to 8909)
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..8909
/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"
regulatory 3636..3645
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 6194..6235
/codon_start=1
/product="epitope tag from simian virus 5"
/label=V5 tag
/translation="GKPIPNPLLGLDST"
CDS 6245..6262
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(6285..6302)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 6335..6469
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(6359..6378)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 6413..6432
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
rep_origin complement(7091..7679)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(7171..7190)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(7850..8710)
/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 8473..8492
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(8711..8815)
/gene="bla"
/label=AmpR promoter
primer_bind 8883..8901
/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 agcttgaatt ctgcagatat catggttaaa gctaaaaagg gcaagaaaga
2761 gatactgacc aaggtcgaag gcggttcctc ggtggacgaa atctccgatg tggacagcga
2821 ccagttgagc ctcaacaaca agcagaatca tgccctgaag ggcaatcaag ctggcgatga
2881 cgatgttaaa gccgtgacca agggggtcaa gaagcttaat gtcaagggaa aaggcaaagc
2941 aggcaaaaat gacgactccg atgaggagga ggtggtgccc gccaagggca aagcttctaa
3001 gaagtcggct tttgagctgc taatggacga cgacgagcag gacgagccag ctgcccagga
3061 aagccagtct gagaaggagg aaaaggtagt ggtctccaag ccacagaaga acgaaaagaa
3121 gggcaaaaag gccaaacgca aggggaagga cgacgacgat gaagatctgg acaaggtttt
3181 agctgagctt caagcggaat acgccggcga ggctgctcct gctgccacta ctgttgtttc
3241 ccccgaagaa ttggccgatg agttctccaa gaaaaaaaag aacaagaagc aggccaagca
3301 ggctgcagtg gcagaaaacg ccggagagga tgaggccagc gatgacgagg agggcggcag
3361 cactgtgaag tcggcagccc agaaaaagaa ggaaaagaag gagcgccaaa agagggaggc
3421 tgccctagcc aagcaaaagg cagctactga acccaaaccg aagcccgtgg agaaacctgc
3481 gccagaaccc gagcctgtgg ctacggaaga agtccagccc gccgccgagg aagaggagga
3541 aaagtccagc aaaaacaaga agaagggcaa gaaagacaaa aaggccgagc ctgaggagga
3601 gaagaaggac gccaagaaga agggtatgtc tgcctccatg gtggcggcca tgcaggagca
3661 gctaagaaag cgaaaggagg aggaggagcg cctggagcgg gaggaggcag agcgaattcg
3721 cctggaggat gagcgcgaag aagcccgcct ggaggcagtg cgtctggagg cagagaggaa
3781 ggagaaaaag aagcagcgcg aggcagagcg caaggctcgc ttgaaggcgg aaggaaaact
3841 gctcaccaag aagcagaagg aggatagggc tagggcacag gctctcctgg agtctctaaa
3901 ggctcagggt ctcgagattc ccgatccgaa cgacaagcga cccacgcggc caggaactcg
3961 tatccgtccg aacaaaaaga agggccctaa ggaagaggcc gccgaagagg aggccaaggc
4021 agcggaagcg gaagcccaat cggccgccgc tgatgccgct gcagctgctg ccgaaaaggc
4081 cgaggaagag agggtcaagg agagttggga tgccaccgat agtgaggctg agccagagcc
4141 cgaggaagaa acgtcgcagg ccactaacaa tggcaagacc gaggctgccg aggaagagaa
4201 cgaagataca gatgacgatg aagatgacga cgatgatgaa gatgacagcg atgactccga
4261 tggggatagt gacgattccc gtgaagtatc ggtgttggcc aatgatccag agtcacgacg
4321 actgcgtgcg gaagccagaa tcctaaaacg gcaagcagag gcggaaaaga agcgctccac
4381 cgacgaactc agagctggag tggtttgcgt gctgggtcat gtagatacag gaaaaaccaa
4441 gatcctggac aaactgcggc gcacacacgt tcaggattcc gaagcgggtg gcatcactca
4501 gcagattgga gccacaaacg ttcccatcga agcaattaag gagcagacta agtacgtaaa
4561 ggcggccgct ggatttgaac atcgcttgcc tggtctattg atcatcgaca cacccggcca
4621 cgagtccttc agcaacctta ggaaccgagg atcatccctc tgcgacatag ccatcctggt
4681 ggtggacatt atgcacggct tggagccaca aaccatcgag tccattcagc tgctcaagaa
4741 gaagaaatgt ccgttcattg tggccctgaa caagatcgat cgtttgtacg attggaagca
4801 gttggccaga cgggatgtga gggatgttct caaggagcag cagagcaaca cacagctgga
4861 gtttcagcag cgcacaaagg acgtgatcct gcagttcgcc gagcagggct tgaatgctgc
4921 tctattctac gaaaacaccg atccgaagac atacatttcc ctggtgccca ccagtgccat
4981 ttcgggcgag ggtatgggca acctgctctt tatgatcgcc gacttctgtc agaatatgct
5041 ggccaagcgc ctgatgtact ccgaggaact gcaggccacc gtgttggagg tgaaggcgct
5101 gccaggtctt ggaaccacca tcgatgccat tctgatcaat ggaaagctcc gcgagggtca
5161 gaccatggtt gtggccggca ctgatggtcc catagtcacc cagatccgat ccctgctgat
5221 gccgcagccc atgaaggagc tgcgtgtgaa gaacgcctat gtggagtaca aggaggttaa
5281 ggctgcgcag ggcgttaaaa ccgccgccaa ggatctggag aaggctattg cgggcatcaa
5341 tctgctgatt gcccataagc ccgatgaagt cgagatttgc acagaggaag ttgcccggga
5401 gctcaagagt gcccttagtc atattaaact ggctcagaca ggcgttcatg tccaggcctc
5461 cacgctgggt tcgttggagg ctcttctgga gttcttgcgc acatctaaga ttccttactc
5521 tgctatacga attggacccg tggtgaaacg cgatgtgatg aaggcctcca caatgttgga
5581 acacgaggcg cagtatgcaa cgattctggc attcgacgtg aagatcgagc gggaggcaca
5641 ggaaatggcc gattccctgg gcgtaaaaat attccaagcg gacattatct accatctctt
5701 cgacaagttc accgcctatc gcgaggagct gaagcagaag aagcgcgagg agttccgatc
5761 cgtagccgtc tttccctgca agttaaggat actgccacag ttcgtattca acagccgaga
5821 tccgatcgtg atgggcgtga tggtcgagaa cggcattgtc aaagtgggca ctccaatttg
5881 cgtgcccagc aaggagttcg tggacattgg aatcgtgact tccatcgaat cgaatcacaa
5941 gcaaatcgaa ttcgcgcgca agggtcaaga gatttgcgtg aaaatcgatc cgattcccgg
6001 tgaatcgccc aaaatgtttg gtcgccactt cgaggccgat gatatgctga ttagcaagat
6061 ctctcgccag agcattgatg cctgcaagga ttacttccgt gacgatctga tcaaggcgga
6121 ttgggcgctg atggtggagc tgaagaagct cttcgagatt ctctaatcga gtctagaggg
6181 cccgcggttc gaaggtaagc ctatccctaa ccctctcctc ggtctcgatt ctacgcgtac
6241 cggtcatcat caccatcacc attgagttta aacccgctga tcagcctcga ctgtgccttc
6301 taaggcctga gctcgctgat cagcctcgat cgaggatcca gacatgataa gatacattga
6361 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
6421 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
6481 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta
6541 aaacctctac aaatgtggta tggctgatta tgatcagtcg accgatgccc ttgagagcct
6601 tcaacccagt cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga
6661 ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc agcgctctgg gtcattttcg
6721 gcgaggaccg ctttcgctgg agcgcgacga tgatcggcct gtcgcttgcg gtattcggaa
6781 tcttgcacgc cctcgctcaa gccttcgtca ctggtcccgc caccaaacgt ttcggcgaga
6841 agcaggccat tatcgccggc atggcggccg acgcgctggg ctacgtcttg ctggcgttcg
6901 cgacgcgagg ctggatggcc ttccccatta tgattcttct cgcttccggc ggcatcggga
6961 tgcccgcgtt gcaggccatg ctgtccaggc aggtagatga cgaccatcag ggacagcttc
7021 aaggatcgct cgcggctctt accagccagc aaaaggccag gaaccgtaaa aaggccgcgt
7081 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
7141 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
7201 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
7261 cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg
7321 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
7381 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
7441 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
7501 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
7561 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
7621 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
7681 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
7741 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
7801 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
7861 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
7921 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
7981 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
8041 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
8101 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
8161 ttgctgcagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
8221 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
8281 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
8341 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
8401 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
8461 cgtcaacacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
8521 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
8581 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
8641 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
8701 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
8761 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
8821 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
8881 aaaataggcg tatcacgagg ccctttcgt
//
| {
"Vector backbone": "Unknown",
"Backbone size w/o insert (bp)": "",
"Vector type": "Insect Expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "Low Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "",
"Publication": "",
"DOI": "",
"Authors": "",
"PubMed Link": "",
"PubMed ID": ""
} | {
"Gene/Insert name": "DmeIF5B",
"Species": "D. melanogaster (fly)",
"Insert Size (bp)": "",
"Mutation": "two non silent mutations A441D and I857T compared to the sequence given by NCBI: NM_143717.1",
"Entrez Gene": "eIF5B",
"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": ""
} |
|
pOPINS-AREL1 | 66710 | Low | Expresses AREL1 HECT domain in E. coli | David Komander | Michel et al Mol Cell. 2015 Apr 2;58(1):95-109. doi: 10.1016/j.molcel.2015.01.042. Epub 2015 Mar 5. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/118782/d1926d64-5173-4ba9-acff-ca0a6344550b/addgene-plasmid-66710-sequence-118782.gbk | partial | LOCUS sequence-118782- 692 bp DNA linear UNA 25-AUG-2024
DEFINITION Expresses AREL1 HECT domain in E. coli.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE natural DNA sequence
ORGANISM unspecified
REFERENCE 1 (bases 1 to 692)
AUTHORS Michel MA, Elliott PR, Swatek KN, Simicek M, Pruneda JN, Wagstaff
JL, Freund SM, Komander D
TITLE Assembly and specific recognition of k29- and k33-linked
polyubiquitin.
JOURNAL Mol Cell. 2015 Apr 2;58(1):95-109. doi:
10.1016/j.molcel.2015.01.042. Epub 2015 Mar 5.
PUBMED 25752577
REFERENCE 2 (bases 1 to 692)
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.molcel.2015.01.042"; journalName: "Mol Cell"; date:
"2015-04-2- 2"; volume: "58"; issue: "1"; pages: "95-109"
FEATURES Location/Qualifiers
source 1..692
/mol_type="genomic DNA"
/organism="unspecified"
CDS complement(16..33)
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
ORIGIN
1 agcagccgga tctcagtggt ggtggtggtg gtgtttaaat ggtctagaaa gctttatcag
61 agcatgccaa agccctcgca accctcgctg atggccagct gcagcatcct gtgcacctct
121 tcataggagt catatgtagg gaggcacagc tggttaaaac atgtgtgtgc agtaggcagc
181 gtgctatggg tcggagcggc aataatctga aatgagggac agagggcggc aaagcctcca
241 ggtggtagct gagaggagcc tgttgtgaac tgaagtagcc gagccaactc ctcctgggtc
301 agactggaaa ccacagtcca aaaccacctc atgacctttt ctctgaaatg ccatgagcca
361 ccaacaacta ctgcatgggc tttgaagtca gacacactga tgtctccagt cccacacatc
421 agcagctcaa gctcattctc atcaaaaata gccaaaaggt tctcagggac caattcattc
481 aggcctttta ggaaatgttc cacctcctct ttcacttgac tggccagccg atattgggcc
541 agcaaattta aatagaagat tttattcgca ttggtgactg gagtttgagc tccacctgtc
601 atgagttcta caaccttatc caattgacct gatttattat atttctcttc tgcaaagacc
661 agctccatct cactcatgtc attgttgagg at
//
| {
"Vector backbone": "pOPINS",
"Backbone size w/o insert (bp)": "5906",
"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": "Assembly and specific recognition of k29- and k33-linked polyubiquitin",
"Publication": "Mol Cell. 2015 Apr 2;58(1):95-109. doi: 10.1016/j.molcel.2015.01.042. Epub 2015 Mar 5.",
"DOI": "10.1016/j.molcel.2015.01.042.",
"Authors": "Michel MA, Elliott PR, Swatek KN, Simicek M, Pruneda JN, Wagstaff JL, Freund SM, Komander D /jmolcel",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/25752577",
"PubMed ID": "PubMed 25752577"
} | {
"Gene/Insert name": "KIAA0317",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "AREL1",
"Promoter": "T7",
"Tag / Fusion Protein": "His-SUMO (N terminal on backbone)"
} | {
"Cloning method": "Ligation Independent Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"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": ""
} |
pMOD_D4910 | 127220 | Module D with ZmUbi1::BBM | Daniel Voytas | Maher et al Nat Biotechnol. 2019 Dec 16. pii: 10.1038/s41587-019-0337-2. doi: 10.1038/s41587-019-0337-2. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/254522/6b546de2-9f56-4f77-91e6-cefd6098f219/addgene-plasmid-127220-sequence-254522.gbk | full | LOCUS sequence-254522- 6832 bp DNA circular SYN 25-AUG-2024
DEFINITION Module D with ZmUbi1::BBM.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6832)
AUTHORS Maher MF, Nasti RA, Vollbrecht M, Starker CG, Clark MD, Voytas DF
TITLE Plant gene editing through de novo induction of meristems.
JOURNAL Nat Biotechnol. 2019 Dec 16. pii: 10.1038/s41587-019-0337-2. doi:
10.1038/s41587-019-0337-2.
PUBMED 31844292
REFERENCE 2 (bases 1 to 6832)
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. 2019 Dec 16. pii: 10.1038/s41587-019-0337-2. doi:
10.1038/s41587-019-0337-2."
FEATURES Location/Qualifiers
source 1..6832
/mol_type="other DNA"
/organism="synthetic DNA construct"
RBS 943..954
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
promoter 2739..2843
/gene="bla"
/label=AmpR promoter
rep_origin 3911..4499
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4400..4419
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 4653..4670
/label=L4440
/note="L4440 vector, forward primer"
ORIGIN
1 atggccactg tgaacaactg gctcgctttc tccctctccc cgcaggagct gccgccctcc
61 cagacgacgg actccacact catctcggcc gccaccgccg accatgtctc cggcgatgtc
121 tgcttcaaca tcccccaaga ttggagcatg aggggatcag agctttcggc gctcgtcgcg
181 gagccgaagc tggaggactt cctcggcggc atctccttct ccgagcagca tcacaaggcc
241 aactgcaaca tgatacccag cactagcagc acagtttgct acgccagctc aggtgctagc
301 accggctacc atcaccagct gtaccaccag cccaccagct cagcgctcca cttcgcggac
361 tccgtaatgg tggcctcctc ggccggtgtc cacgacggcg gtgccatgct cagcgcggcc
421 gccgctaacg gtgtcgctgg cgctgccagt gccaacggcg gcggcatcgg gctgtccatg
481 attaagaact ggctgcggag ccaaccggcg cccatgcagc cgagggtggc ggcggctgag
541 ggcgcgcagg ggctctcttt gtccatgaac atggcgggga cgacccaagg cgctgctggc
601 atgccacttc tcgctggaga gcgcgcacgg gcgcccgaga gtgtatccac gtcagcacag
661 ggtggagccg tcgtcgtcac ggcgccgaag gaggatagcg gtggcagcgg tgttgccggc
721 gctctagtag ccgtgagcac ggacacgggt ggcagcggcg gcgcgtcggc tgacaacacg
781 gcaaggaaga cggtggacac gttcgggcag cgcacgtcga tttaccgtgg cgtgacaagg
841 catagatgga ctgggagata tgaggcacat ctttgggata acagttgcag aagggaaggg
901 caaactcgta agggtcgtca agtctattta ggtggctatg ataaagagga gaaagctgct
961 agggcttatg atcttgctgc tctgaagtac tggggtgcca caacaacaac aaattttcca
1021 gtgagtaact acgaaaagga gctggaggac atgaagcaca tgacaaggca ggagtttgta
1081 gcgcctctga gaaggaagtc cagtggtttc tccagaggtg catccattta caggggagtg
1141 actaggcatc accaacatgg aagatggcaa gcacggattg gacgagttgc agggaacaag
1201 gatctttact tgggcacctt cagcacccag gaggaggcag cggaggcgta cgacatcgcg
1261 gcgatcaagt tccgcggcct caacgccgtc accaacttcg acatgagccg ctacgacgtg
1321 aagtccatcc tggacagcag cgccctcccc atcggcagcg ccgccaagcg cctcaaggag
1381 gccgaggccg cagcgtccgc gcagcaccac catgcgggtg tcgtttccta tgacgttggg
1441 aggattgcca gccaactggg agatggcggt gccctcgctg cggcctatgg tgctcactat
1501 cacggtgccg cgtggccaac gattgcattc cagccgggcg cggcgtccac cggactgtac
1561 catccttacg cgcagcagcc tatgcgcggc ggtggatggt gtaaacaaga gcaagatcac
1621 gctgtgatag cagcggcaca ctccttgcag gatcttcatc atttgaatct cggagccgcc
1681 ggggcccacg actttttctc ggcagggcag caggccgccg ccgctgcgat gcacggcctg
1741 ggtagcatcg acagtgcgtc gctggagcac agcaccggct ccaactccgt cgtctacaac
1801 ggcggggtcg gcgacagcaa cggcgccagc gccgtcggcg gcagtggcgg tggctacatg
1861 atgccgatga gcgctgccgg agcaaccact acatcggcaa tggtgagcca cgagcaggtc
1921 catgcacggg cctacgacga agccaagcag gctgctcaga tggggtacga gagctacctg
1981 gtgaacgcgg agaacaatgg tggcggaagg atgtctgcat gggggactgt cgtgtctgca
2041 gccgcggcgg cagcagcaag cagcaacgac aacatggccg ccgacgtggg ccacggcggc
2101 gcgcagctgt tcagtgtctg gaacgacact taactcgagc cctagacttg tccatcttct
2161 ggattggcca agttaattaa tgtatgaaat aaaaggatgc acacatagtg acatgctaat
2221 cactataatg tgggcatcaa agttgtgtgt tatgtgtaat tactaattat ctgaataaga
2281 gaaagagatc atccatattt cttatcctaa atgaatgtca cgtgtcttta taattctttg
2341 atgaaccaga tgcattttat taaccaattc catatacata taaatattaa tcatatataa
2401 ttaatatcaa ttgggttagc aaaacaaatc tagtctaggt gtgttttgct aattattggg
2461 ggatagtgca aaaagaaatc tacgttctca ataattcaga tagaaaactt aataaagtga
2521 gataatttac atagattgct tttatccttt gatatatgtg aaaccatgca tgatataagg
2581 aaaatagata gagaaataat tttttacatc gttgaatatg taaacaattt aattcaagaa
2641 gctaggaata taaatattga ggagtttatg attagagctc agtgactagc aggtgatgaa
2701 aagccttagg gtcaggtggc acttttcggg gaaatgtgcg atgaacccct atttgtttat
2761 ttttctaaat acattcaaat atgtatccgc taatgagaca ataaccctga taaatgcttc
2821 aattatattg aaaaaggaag agtatgagta tccaacattt ccgtgtcgcc cttattccct
2881 tttttgcggc cttctgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
2941 atgctgaaga tcagttgggt gcccgagtgg gttacattga actggatctc aacagcggca
3001 aaatcttaga gagttttcgc cccgaagaac gcttcccaat gatgagcact ttcaaagttc
3061 tgctatgtgg cgcggtgtta tcccgtattg atgctgggca agagcaactc ggtcgccgca
3121 tacactattc tcagaatgac ttggttgagt attcaccagt cacagaaaag catcttacgg
3181 atggcatgac agtaagagaa ttatgtagtg ctgccataac catgagtgat aacactgcgg
3241 ccaacttact tctgacaacg attggaggac cgaaggagct aaccgctttt ttgcacaaca
3301 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
3361 acgacgagcg tgacaccacg atgcctgtag caatggcaac aactctccgc aaactattaa
3421 ctggcgaact acttactctt gcttcacgcc agcaactcat tgactggatg gaggcggaca
3481 aagttgcagg accacttctg cgctcggcac ttccggctgg ctggtttatt gctgataaat
3541 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
3601 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
3661 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gacgaaggga
3721 aataaatagt agcccgcctg atgtgcgggc ttttttttgg tggtggttac cagtgtcgct
3781 tagctaccgt aggtcctctt aggcggtccg attagcgaca tagtcatcaa tgccttgacg
3841 gcaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
3901 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
3961 caccgctacc agcggtggtt tgtttgccgg atcaagagct actaactctt tttccgaagg
4021 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
4081 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
4141 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
4201 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgctcacag cccagcttgg
4261 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
4321 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
4381 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
4441 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
4501 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
4561 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
4621 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
4681 agcgcccacc tgcaacgtcc cggcgcgcct gcagtgcagc gtgacccggt cgtgcccctc
4741 tctagagata atgagcattg catgtctaag ttataaaaaa ttaccacata ttttttttgt
4801 cacacttgtt tgaagtgcag tttatctatc tttatacata tatttaaact ttactctacg
4861 aataatataa tctatagtac tacaataata tcagtgtttt agagaatcat ataaatgaac
4921 agttagacat ggtctaaagg acaattgagt attttgacaa caggactcta cagttttatc
4981 tttttagtgt gcatgtgttc tccttttttt ttgcaaatag cttcacctat ataatacttc
5041 atccatttta ttagtacatc catttagggt ttagggttaa tggtttttat agactaattt
5101 ttttagtaca tctattttat tctattttag cctctaaatt aagaaaacta aaactctatt
5161 ttagtttttt tatttaataa tttagatata aaatagaata aaataaagtg actaaaaatt
5221 aaacaaatac cctttaagaa attaaaaaaa ctaaggaaac atttttcttg tttcgagtag
5281 ataatgccag cctgttaaac gccgtcgacg agtctaacgg acaccaacca gcgaaccagc
5341 agcgtcgcgt cgggccaagc gaagcagacg gcacggcatc tctgtcgctg cctctggacc
5401 cctctcgaga gttccgctcc accgttggac ttgctccgct gtcggcatcc agaaattgcg
5461 tggcggagcg gcagacgtga gccggcacgg caggcggcct cctcctcctc tcacggcacc
5521 ggcagctacg ggggattcct ttcccaccgc tccttcgctt tcccttcctc gcccgccgta
5581 ataaatagac accccctcca caccctcttt ccccaacctc gtgttgttcg gagcgcacac
5641 acacacaacc agatctcccc caaatccacc cgtcggcacc tccgcttcaa ggtacgccgc
5701 tcgtcctccc cccccccctc tctaccttct ctagatcggc gttccggtcc atggttaggg
5761 cccggtagtt ctacttctgt tcatgtttgt gttagatccg tgtttgtgtt agatccgtgc
5821 tgctagcgtt cgtacacgga tgcgacctgt acgtcagaca cgttctgatt gctaacttgc
5881 cagtgtttct ctttggggaa tcctgggatg gctctagccg ttccgcagac gggatcgatt
5941 tcatgatttt ttttgtttcg ttgcataggg tttggtttgc ccttttcctt tatttcaata
6001 tatgccgtgc acttgtttgt cgggtcatct tttcatgctt ttttttgtct tggttgtgat
6061 gatgtggtct ggttgggcgg tcgttctaga tcggagtaga attaattctg tttcaaacta
6121 cctggtggat ttattaattt tggatctgta tgtgtgtgcc atacatattc atagttacga
6181 attgaagatg atggatggaa atatcgatct aggataggta tacatgttga tgcgggtttt
6241 actgatgcat atacagagat gctttttgtt cgcttggttg tgatgatgtg gtgtggttgg
6301 gcggtcgttc attcgttcta gatcggagta gaatactgtt tcaaactacc tggtgtattt
6361 attaattttg gaactgtatg tgtgtgtcat acatcttcat agttacgagt ttaagatgga
6421 tggaaatatc gatctaggat aggtatacat gttgatgtgg gttttactga tgcatataca
6481 tgatggcata tgcagcatct attcatatgc tctaaccttg agtacctatc tattataata
6541 aacaagtatg ttttataatt attttgatct tgatatactt ggatgatggc atatgcagca
6601 gctatatgtg gattttttta gccctgcctt catacgctat ttatttgctt ggtactgttt
6661 cttttgtcga tgctcaccct gttgtttggt gttacttcct gcaggcttcc ctaacctttg
6721 cactgtccaa aatggcttcc tgatcccctc acttcctcga atcaatctaa gaagaaactc
6781 aagccgcaac cattaggggc agattaattg ctgcactttc agataatcaa gc
//
| {
"Vector backbone": "pMOD_D4800EC",
"Backbone size w/o insert (bp)": "",
"Vector type": "Plant 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": "Plant gene editing through de novo induction of meristems",
"Publication": "Nat Biotechnol. 2019 Dec 16. pii: 10.1038/s41587-019-0337-2. doi: 10.1038/s41587-019-0337-2.",
"DOI": "10.1038/s41587-019-0337-2.",
"Authors": "Maher MF, Nasti RA, Vollbrecht M, Starker CG, Clark MD, Voytas DF /s---",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/31844292",
"PubMed ID": "PubMed 31844292"
} | {
"Gene/Insert name": "BBM",
"Species": "Maize",
"Insert Size (bp)": "",
"Mutation": "codon optimized CDSs",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "unknown (unknown if destroyed)",
"3′ cloning site": "unknown (unknown if destroyed)",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
T4 Lysozyme M120A WT* | 18545 | Brian Matthews | T4 Lysozyme Plasmids - Tray 4 (unpublished) | https://media.addgene.org/snapgene-media/v2.0.0/sequences/24331/f4ff93e2-93c1-4d7e-b1b2-6a5c4d91d26f/addgene-plasmid-18545-sequence-24331.gbk | partial | LOCUS sequence-24331-e 817 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 817)
TITLE T4 Lysozyme Plasmids - Tray 4
REFERENCE 2 (bases 1 to 817)
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..817
/mol_type="genomic DNA"
/organism="unspecified"
primer_bind complement(711..733)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 722..738
/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)."
ORIGIN
1 tncgaaaaag cccgctcatt aggcgggctc agatctgctc atgtttgaca gcttatcatc
61 gataagcttc ggcaataaat tcttgatatg ttttcatatg tttcctttaa atattttaat
121 tagtaattgt ctattcaagt aattgtgaat atactatcac aattccaaga gaaagtaaac
181 agctttatag atttttatac gcgtcccaag tgccagttct aaacgttgta atgactcgtt
241 ttgcgcgatt aggtgtttga ttataccatc tacttttagc taagttaact gctgcttcat
301 cccagcgttt ttgttgaagc gcacgtaaag agttagtaaa tcctgccaca ccggtttctc
361 ccatttggaa aaccatatta atcaatgcag cgcgacgaac cgcatcaaga gaatcataaa
421 ccggttttaa tttagcattt ctcagaattc cgcgaacagc agcatcaaca tcctgattaa
481 agagtttttc agcctcatct tttgtaatta caccattagt attacgccca atagctttat
541 ctaattcaga tttagcagca ttaagtgatg gactttttgt aagcaaatga ccgatgccaa
601 tagtgtaata gccttctgtg tctttataga ttttaagtct aagaccttca tctatacgta
661 acatttcaaa tatattcata atacctccta agcatcgatg gatcctgttt cctgtgtgaa
721 attgttatcc gctcacaatt ccacacatta tacgagccga tgattaatgt caacaggggg
781 angggagtaa gctgatcngn ttcngtgtga atgtatc
//
| {
"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": "M120A 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": ""
} |
||
pLC-Flag-CSNK1A1 G40N-P2A-Blast | 123321 | Lentiviral vector for expression of Flag tagged CK1alpha G40N-P2A-Blast casette from a CMV promoter | Eva Gottwein | Patil et al Blood. 2018 Aug 9;132(6):577-586. doi: 10.1182/blood-2018-01-828418. Epub 2018 Jun 28. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/239599/7fc86592-997d-4320-b52f-2f5df28546c6/addgene-plasmid-123321-sequence-239599.gbk | full | LOCUS sequence-239599- 7954 bp DNA circular SYN 26-AUG-2024
DEFINITION Lentiviral vector for expression of Flag tagged CK1alpha
G40N-P2A-Blast casette from a CMV promoter.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7954)
AUTHORS Patil A, Manzano M, Gottwein E
TITLE CK1alpha and IRF4 are essential and independent effectors of
immunomodulatory drugs in primary effusion lymphoma.
JOURNAL Blood. 2018 Aug 9;132(6):577-586. doi: 10.1182/blood-2018-01-828418.
Epub 2018 Jun 28.
PUBMED 29954751
REFERENCE 2 (bases 1 to 7954)
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:
"10.1182/blood-2018-01-828418"; journalName: "Blood"; date:
"2018-08-9- 9"; volume: "132"; issue: "6"; pages: "577-586"
FEATURES Location/Qualifiers
source 1..7954
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 1..396
/codon_start=1
/gene="Aspergillus terreus BSD"
/product="blasticidin S deaminase"
/label=BSD
/note="confers resistance to blasticidin"
/translation="AKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
AIVKDSDGQPTAVGIRELLPSGYVWEG"
protein_bind complement(416..449)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
misc_feature 505..1093
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(558..578)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(976..987)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(1096..1112)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1097..1113)
/label=pBluescriptKS
/note="For pBluescript vector"
LTR 1622..1802
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
rep_origin complement(1864..2452)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1944..1963)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(2623..3483)
/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 3246..3265
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(3484..3588)
/gene="bla"
/label=AmpR promoter
primer_bind complement(3663..3682)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 3854..4233
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 4235..4433
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
LTR 4451..4631
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 4678..4803
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 5300..5533
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 5718..5762
/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 6060..6177
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
enhancer 6302..6605
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 6606..6809
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
regulatory 6848..6857
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 6857..6880
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS 7898..7954
/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"
ORIGIN
1 gccaagcctt tgtctcaaga agaatccacc ctcattgaaa gagcaacggc tacaatcaac
61 agcatcccca tctctgaaga ctacagcgtc gccagcgcag ctctctctag cgacggccgc
121 atcttcactg gtgtcaatgt atatcatttt actgggggac cttgtgcaga actcgtggtg
181 ctgggcactg ctgctgctgc ggcagctggc aacctgactt gtatcgtcgc gatcggaaat
241 gagaacaggg gcatcttgag cccctgcgga cggtgccgac aggtgcttct cgatctgcat
301 cctgggatca aagccatagt gaaggacagt gatggacagc cgacggcagt tgggattcgt
361 gaattgctgc cctctggtta tgtgtgggag ggctaagaat tcgtcgaggg acctaataac
421 ttcgtatagc atacattata cgaagttata catgtttaag ggttccggtt ccactaggta
481 caattcgata tcaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg
541 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
601 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
661 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
721 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
781 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
841 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
901 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
961 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
1021 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
1081 gccgcctccc cgcatcgata ccgtcgacct cgatcgagac ctagaaaaac atggagcaat
1141 cacaagtagc aatacagcag ctaccaatgc tgattgtgcc tggctagaag cacaagagga
1201 ggaggaggtg ggttttccag tcacacctca ggtaccttta agaccaatga cttacaaggc
1261 agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc
1321 ccaacgaaga caagatatcc ttgatctgtg gatctaccac acacaaggct acttccctga
1381 ttggcagaac tacacaccag ggccagggat cagatatcca ctgacctttg gatggtgcta
1441 caagctagta ccagttgagc aagagaaggt agaagaagcc aatgaaggag agaacacccg
1501 cttgttacac cctgtgagcc tgcatgggat ggatgacccg gagagagaag tattagagtg
1561 gaggtttgac agccgcctag catttcatca catggcccga gagctgcatc cggactgtac
1621 tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
1681 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
1741 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
1801 cagcatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
1861 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
1921 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
1981 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
2041 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
2101 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
2161 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
2221 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
2281 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
2341 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
2401 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
2461 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
2521 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
2581 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
2641 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
2701 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
2761 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
2821 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
2881 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
2941 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
3001 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
3061 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
3121 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
3181 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
3241 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
3301 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
3361 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
3421 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
3481 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
3541 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
3601 aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc
3661 agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg
3721 gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac
3781 aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc
3841 agatatacgc gttgacattg attattgact agttattaat agtaatcaat tacggggtca
3901 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
3961 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
4021 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
4081 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
4141 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
4201 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
4261 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
4321 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
4381 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagcgcgttt
4441 tgcctgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
4501 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
4561 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
4621 aatctctagc agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc
4681 tctcgacgca ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg
4741 gtgagtacgc caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc
4801 gtcagtatta agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg
4861 ggaaagaaaa aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc
4921 gcagttaatc ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta
4981 caaccatccc ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc
5041 ctctattgtg tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata
5101 gaggaagagc aaaacaaaag taagaccacc gcacagcaag cggccggccg ctgatcttca
5161 gacctggagg aggagatatg agggacaatt ggagaagtga attatataaa tataaagtag
5221 taaaaattga accattagga gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag
5281 aaaaaagagc agtgggaata ggagctttgt tccttgggtt cttgggagca gcaggaagca
5341 ctatgggcgc agcgtcaatg acgctgacgg tacaggccag acaattattg tctggtatag
5401 tgcagcagca gaacaatttg ctgagggcta ttgaggcgca acagcatctg ttgcaactca
5461 cagtctgggg catcaagcag ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg
5521 atcaacagct cctggggatt tggggttgct ctggaaaact catttgcacc actgctgtgc
5581 cttggaatgc tagttggagt aataaatctc tggaacagat ttggaatcac acgacctgga
5641 tggagtggga cagagaaatt aacaattaca caagcttaat acactcctta attgaagaat
5701 cgcaaaacca gcaagaaaag aatgaacaag aattattgga attagataaa tgggcaagtt
5761 tgtggaattg gtttaacata acaaattggc tgtggtatat aaaattattc ataatgatag
5821 taggaggctt ggtaggttta agaatagttt ttgctgtact ttctatagtg aatagagtta
5881 ggcagggata ttcaccatta tcgtttcaga cccacctccc aaccccgagg ggacccgaca
5941 ggcccgaagg aatagaagaa gaaggtggag agagagacag agacagatcc attcgattag
6001 tgaacggatc ggcactgcgt gcgccaattc tgcagacaaa tggcagtatt catccacaat
6061 tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata
6121 gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaaattca aaattttcgg
6181 gtttattaca gggacagcag agatccagtt tggttagtac cgggcccgag agttaattaa
6241 ttagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
6301 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
6361 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
6421 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
6481 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
6541 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
6601 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
6661 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
6721 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
6781 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgct
6841 accggtcgcc accatggatt acaaagacga tgacgataag gcgagtagca gcggctccaa
6901 ggctgaattc attgtcggag ggaaatataa actggtacgg aagatcgggt ctggctcctt
6961 cggggacatc tatttggcga tcaacatcac caacaacgag gaagtggcag tgaagctaga
7021 atctcagaag gccaggcatc cccagttgct gtacgagagc aagctctata agattcttca
7081 aggtggggtt ggcatccccc acatacggtg gtatggtcag gaaaaagact acaatgtact
7141 agtcatggat cttctgggac ctagcctcga agacctcttc aatttctgtt caagaaggtt
7201 cacaatgaaa actgtactta tgttagctga ccagatgatc agtagaattg aatatgtgca
7261 tacaaagaat tttatacaca gagacattaa accagataac ttcctaatgg gtattgggcg
7321 tcactgtaat aagttattcc ttattgattt tggtttggcc aaaaagtaca gagacaacag
7381 gacaaggcaa cacataccat acagagaaga taaaaacctc actggcactg cccgatatgc
7441 tagcatcaat gcacatcttg gtattgagca gagtcgccga gatgacatgg aatcattagg
7501 atatgttttg atgtatttta atagaaccag cctgccatgg caagggctaa aggctgcaac
7561 aaagaaacaa aaatatgaaa agattagtga aaagaagatg tccacgcctg ttgaagtttt
7621 atgtaagggg tttcctgcag aatttgcgat gtacttaaac tattgtcgtg ggctacgctt
7681 tgaggaagcc ccagattaca tgtatctgag gcagctattc cgcattcttt tcaggaccct
7741 gaaccatcaa tatgactaca catttgattg gacaatgtta aagcagaaag cagcacagca
7801 ggcagcctct tccagtgggc agggtcagca ggcccaaacc cccacaggca agcaaactga
7861 caaaaccaag agtaacatga aaggtttcgg atccggcgca acaaacttct ctctgctgaa
7921 acaagccgga gatgtcgaag agaatcctgg accg
//
| {
"Vector backbone": "pLC-ZsGreen-P2A-Blast",
"Backbone size w/o insert (bp)": "",
"Vector type": "Mammalian Expression, Lentiviral",
"Selectable markers": "Blasticidin"
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "Stbl3",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "CK1alpha and IRF4 are essential and independent effectors of immunomodulatory drugs in primary effusion lymphoma",
"Publication": "Blood. 2018 Aug 9;132(6):577-586. doi: 10.1182/blood-2018-01-828418. Epub 2018 Jun 28.",
"DOI": "10.1182/blood-2018-01-828418.",
"Authors": "Patil A, Manzano M, Gottwein E /blood---",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/29954751",
"PubMed ID": "PubMed 29954751"
} | {
"Gene/Insert name": "CSNK1A1",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "1035",
"Mutation": "changed Glycine 40 to Asparagine",
"Entrez Gene": "CSNK1A1",
"Promoter": "CMV",
"Tag / Fusion Protein": "Flag (N 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": ""
} |
|
pGAD-Atg17(F317H) | 122269 | For Yeast-two-hybrid analysis | Daniel Klionsky | Liu et al Curr Biol. 2016 Jan 25;26(2):150-160. doi: 10.1016/j.cub.2015.11.054. Epub 2016 Jan 7. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/240231/e77d6f7e-b255-475c-9093-f01afa40e010/addgene-plasmid-122269-sequence-240231.gbk | full | LOCUS sequence-240231- 7933 bp DNA circular SYN 26-AUG-2024
DEFINITION For Yeast-two-hybrid analysis.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7933)
AUTHORS Liu X, Mao K, Yu AYH, Omairi-Nasser A, Austin J 2nd, Glick BS, Yip
CK, Klionsky DJ
TITLE The Atg17-Atg31-Atg29 Complex Coordinates with Atg11 to Recruit the
Vam7 SNARE and Mediate Autophagosome-Vacuole Fusion.
JOURNAL Curr Biol. 2016 Jan 25;26(2):150-160. doi:
10.1016/j.cub.2015.11.054. Epub 2016 Jan 7.
PUBMED 26774783
REFERENCE 2 (bases 1 to 7933)
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.cub.2015.11.054"; journalName: "Curr Biol"; date:
"2016-01-25- 25"; volume: "26"; issue: "2"; pages: "150-160"
FEATURES Location/Qualifiers
source 1..7933
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 10..411
/gene="S. cerevisiae ADH1"
/label=ADH1 promoter
/note="promoter for alcohol dehydrogenase 1"
CDS 457..477
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 493..834
/codon_start=1
/gene="Saccharomyces cerevisiae GAL4 (truncated)"
/product="activation domain of the GAL4 transcriptional
activator"
/label=GAL4 activation domain
/translation="ANFNQSGNIADSSLSFTFTNSSNGPNLITTQTNSQALSQPIASSN
VHDNFMNNEITASKIDDGNNSKPLSPGWTDQTAYNAFGITTGMFNTTTMDDVYNYLFDD
EDTPPNPKKE"
RBS 1905..1916
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
terminator 2516..2703
/gene="S. cerevisiae ADH1"
/label=ADH1 terminator
/note="transcription terminator for the S. cerevisiae
alcohol dehydrogenase 1 (ADH1) gene"
primer_bind complement(2571..2595)
/label=pHybLex-R
/note="ADH terminator, reverse primer"
CDS complement(2824..3918)
/codon_start=1
/gene="S. cerevisiae LEU2"
/product="3-isopropylmalate dehydrogenase, required for
leucine biosynthesis"
/label=LEU2
/note="yeast auxotrophic marker"
/translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
IGGAAIEATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
LIDSAAMILVRNPTHLNGIIITSNMFGDIVSDEASVIPGSLGLLPSASLASLPDKNTAF
GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
DLGGSNSTTEVGDAVAEEVKKILA"
promoter complement(3919..4324)
/gene="S. cerevisiae LEU2"
/label=LEU2 promoter
primer_bind complement(4366..4382)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4366..4382)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4379..4401)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4390..4406
/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(4414..4444)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4459..4480
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4597..4614)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4768..5356)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4848..4867)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5527..6387)
/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 6150..6169
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6388..6492)
/gene="bla"
/label=AmpR promoter
primer_bind 6560..6578
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(6616..6638)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 6738..6757
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
rep_origin join(6774..7933,1..5)
/label=2u ori
/note="yeast 2u plasmid origin of replication"
ORIGIN
1 tcgttgcttg catgcaactt cttttctttt tttttctttt ctctctcccc cgttgttgtc
61 tcaccatatc cgcaatgaca aaaaaatgat ggaagacact aaaggaaaaa attaacgaca
121 aagacagcac caacagatgt cgttgttcca gagctgatga ggggtatctc gaagcacacg
181 aaactttttc cttccttcat tcacgcacac tactctctaa tgagcaacgg tatacggcct
241 tccttccagt tacttgaatt tgaaataaaa aaaagtttgc tgtcttgcta tcaagtataa
301 atagacctgc aattattaat cttttgtttc ctcgtcattg ttctcgttcc ctttcttcct
361 tgtttctttt tctgcacaat atttcaagct ataccaagca tacaatcaac tccaagcttt
421 gcaaagatgg ataaagcgga attaattccc gagcctccaa aaaagaagag aaaggtcgaa
481 ttgggtaccg ccgccaattt taatcaaagt gggaatattg ctgatagctc attgtccttc
541 actttcacta acagtagcaa cggtccgaac ctcataacaa ctcaaacaaa ttctcaagcg
601 ctttcacaac caattgcctc ctctaacgtt catgataact tcatgaataa tgaaatcacg
661 gctagtaaaa ttgatgatgg taataattca aaaccactgt cacctggttg gacggaccaa
721 actgcgtata acgcgtttgg aatcactaca gggatgttta ataccactac aatggatgat
781 gtatataact atctattcga tgatgaagat accccaccaa acccaaaaaa agagatcgaa
841 ttccccgggg gatccatcga taacgaagca gatgttacaa aatttgttaa taatgccagg
901 aaaacgctga ccgatgctca acttttatgt tcaagtgcta atttaaggat tgtagatatt
961 aagaaaaaat tgtcatcttg gcagttgagt atttcaaaac tcaattttct aatagttggc
1021 ttgagacagc aaggaaagtt tctttacact attttaaagg aaggcattgg gacaaagcta
1081 atccagaaac aatggaatca agctgtctta gtggtcttag ttgatgagat gaagtactgg
1141 caatatgaaa tcacttctaa agtacaaagg ttggatggta tagtaaatga attgagcata
1201 tcagaaaaag atgataccga tccctctaaa ctaggagatt acatctcgag agacaacgtt
1261 aacttattga atgacaaact gaaagaagtg ccagtaattg agcgtcaaat tgaaaacatt
1321 aagcttcaat acgaaaatat ggtcagaaaa gttaacaaag aactgattga taccaagttg
1381 acggacgtaa ctcaaaaatt ccaaagtaaa tttggtatag ataacctgat ggaaacaaat
1441 gttgcagagc agtttagcag ggaactaacg gaccttgaaa aagatttagc agagataatg
1501 aattcattga cacagcactt tgataagaca ttgctgttac aagataaaaa aattgataac
1561 gatgaacgtg aggagctgtt taaggtggta caaggcgacg acaaagaact atacaacatt
1621 ttcaaaactc tgcatgaggt aattgatgac gtggacaaaa caattcttaa cttgggtcaa
1681 tttttgcagg caaaaataaa ggaaaagaca gaactacaca gcgaagtttc tgaaataata
1741 aacgatttca atagaaattt ggaatatcta ttaatcttta aagatatttc caatctgatt
1801 gatagccata aaaattcctg tacacaagat attcaaacaa ctaaggaact ttgtgaattt
1861 tatgataatt ttgaagaaag ctacggtaac ttagttctag aagcaaagag gagaaaggat
1921 gtggcaaaca gaatgaaaac tatattgaaa gattgtgaaa agcagttgca gaatttagat
1981 gctcaggacc aggaagaacg tcagaatttt atagcggaaa atggaactta tcttcctgag
2041 acaatctggc ccggtaaaat tgacgatttt tcttccctgt acactttaaa ttacaacgtg
2101 aagaatcctt agaaaaaagg aaaaaaaggc tgcagagatc tatgaatcgt agatactgaa
2161 aaaccccgca agttcacttc aactgtgcat cgtgcaccat ctcaatttct ttcatttata
2221 catcgttttg ccttctttta tgtaactata ctcctctaag tttcaatctt ggccatgtaa
2281 cctctgatct atagaatttt ttaaatgact agaattaatg cccatctttt ttttggacct
2341 aaattcttca tgaaaatata ttacgagggc ttattcagaa gctttggact tcttcgccag
2401 aggtttggtc aagtctccaa tcaaggttgt cggcttgtct accttgccag aaatttacga
2461 aaagatggaa aagggtcaaa tcgttggtag atacgttgtt gacacttcta aataagcgaa
2521 tttcttatga tttatgattt ttattattaa ataagttata aaaaaaataa gtgtatacaa
2581 attttaaagt gactcttagg ttttaaaacg aaaattctta ttcttgagta actctttcct
2641 gtaggtcagg ttgctttctc aggtatagca tgaggtcgct cttattgacc acacctctac
2701 cggcatgccg gtcgaaattc ccctacccta tgaacatatt ccattttgta atttcgtgtc
2761 gtttctatta tgaatttcat ttataaagtt tatgtacaaa tatcataaaa aaagagaatc
2821 tttttaagca aggattttct taacctcttc ggcgacagca tcaccgactt cggtggtact
2881 gttggaacca cctaaatcac cagttctgat acctgcatcc aaaacctttt taactgcatc
2941 ttcaatggcc ttaccttctt caggcaagtt caatgacaat ttcaacatca ttgcagcaga
3001 caagatagtg gcgatagggt tgaccttatt ctttggcaaa tctggagcag aaccgtggca
3061 tggttcgtac aaaccaaatg cggtgttctt gtctggcaaa gaggccaagg acgcagatgg
3121 caacaaaccc aaggaacctg ggataacgga ggcttcatcg gagacgatat caccaaacat
3181 gttgctggtg attataatac catttaggtg ggttgggttc ctaactagga tcatggcggc
3241 agaatcaatc aattgatgtt gaaccttcaa tgtaggaaat tcgttcttga tggtttcctc
3301 cacagttttt ctccataatc ttgaagaggc caaaacatta gctttatcca aggaccaaat
3361 aggcaatggt ggctcatgtt gtagggccat gaaagcggcc attcttgtga ttctttgcac
3421 ttctggaacg gtgtattgtt cactatccca agcgacacca tcaccatcgt cttcctttct
3481 cttaccaaag taaatacctc ccactaattc tctgacaaca acgaagtcag tacctttagc
3541 aaattgtggc ttgattggag ataagtctaa aagagagtcg gatgcaaagt tacatggtct
3601 taagttggcg tacaattgaa gttctttacg gatttttagt aaaccttgtt caggtctaac
3661 actacctgta ccccatttag gaccacccac agcacctaac aaaacggcat cagccttctt
3721 ggaggcttcc agcgcctcat ctggaagtgg gacacctgta gcttcgatag cagcaccacc
3781 aattaaatga ttttcgaaat cgaacttgac attggaacga acatcagaaa tagctttaag
3841 aaccttaatg gcttcggctg tgatttcttg accaacgtgg tcacctggca aaacgacgat
3901 cttcttaggg gcagacatta gaatggtata tccttgaaat atatatatat attgctgaaa
3961 tgtaaaaggt aagaaaagtt agaaagtaag acgattgcta accacctatt ggaaaaaaca
4021 ataggtcctt aaataatatt gtcaacttca agtattgtga tgcaagcatt tagtcatgaa
4081 cgcttctcta ttctatatga aaagccggtt ccggcgctct cacctttcct ttttctccca
4141 atttttcagt tgaaaaaggt atatgcgtca ggcgacctct gaaattaaca aaaaatttcc
4201 agtcatcgaa tttgattctg tgcgatagcg cccctgtgtg ttctcgttat gttgaggaaa
4261 aaaataatgg ttgctaagag attcgaactc ttgcatctta cgatacctga gtattcccac
4321 agttggggga tctcgactct agctagagga tcaattcgta atcatgtcat agctgtttcc
4381 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
4441 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
4501 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
4561 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
4621 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
4681 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
4741 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
4801 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
4861 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
4921 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
4981 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
5041 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
5101 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
5161 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
5221 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
5281 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
5341 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
5401 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
5461 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
5521 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
5581 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
5641 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
5701 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
5761 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
5821 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
5881 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
5941 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
6001 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
6061 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
6121 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
6181 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
6241 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
6301 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
6361 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
6421 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
6481 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
6541 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
6601 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
6661 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
6721 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc ataacgcatt
6781 taagcataaa cacgcactat gccgttcttc tcatgtatat atatatacag gcaacacgca
6841 gatataggtg cgacgtgaac agtgagctgt atgtgcgcag ctcgcgttgc attttcggaa
6901 gcgctcgttt tcggaaacgc tttgaagttc ctattccgaa gttcctattc tctagctaga
6961 aagtatagga acttcagagc gcttttgaaa accaaaagcg ctctgaagac gcactttcaa
7021 aaaaccaaaa acgcaccgga ctgtaacgag ctactaaaat attgcgaata ccgcttccac
7081 aaacattgct caaaagtatc tctttgctat atatctctgt gctatatccc tatataacct
7141 acccatccac ctttcgctcc ttgaacttgc atctaaactc gacctctaca ttttttatgt
7201 ttatctctag tattactctt tagacaaaaa aattgtagta agaactattc atagagtgaa
7261 tcgaaaacaa tacgaaaatg taaacatttc ctatacgtag tatatagaga caaaatagaa
7321 gaaaccgttc ataattttct gaccaatgaa gaatcatcaa cgctatcact ttctgttcac
7381 aaagtatgcg caatccacat cggtatagaa tataatcggg gatgccttta tcttgaaaaa
7441 atgcacccgc agcttcgcta gtaatcagta aacgcgggaa gtggagtcag gcttttttta
7501 tggaagagaa aatagacacc aaagtagcct tcttctaacc ttaacggacc tacagtgcaa
7561 aaagttatca agagactgca ttatagagcg cacaaaggag aaaaaaagta atctaagatg
7621 ctttgttaga aaaatagcgc tctcgggatg catttttgta gaacaaaaaa gaagtataga
7681 ttctttgttg gtaaaatagc gctctcgcgt tgcatttctg ttctgtaaaa atgcagctca
7741 gattctttgt ttgaaaaatt agcgctctcg cgttgcattt ttgttttaca aaaatgaagc
7801 acagattctt cgttggtaaa atagcgcttt cgcgttgcat ttctgttctg taaaaatgca
7861 gctcagattc tttgtttgaa aaattagcgc tctcgcgttg catttttgtt ctacaaaatg
7921 aagcacagat gct
//
| {
"Vector backbone": "pGAD-c1",
"Backbone size w/o insert (bp)": "",
"Vector type": "Yeast Expression",
"Selectable markers": "LEU2"
} | {
"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": "The Atg17-Atg31-Atg29 Complex Coordinates with Atg11 to Recruit the Vam7 SNARE and Mediate Autophagosome-Vacuole Fusion",
"Publication": "Curr Biol. 2016 Jan 25;26(2):150-160. doi: 10.1016/j.cub.2015.11.054. Epub 2016 Jan 7.",
"DOI": "10.1016/j.cub.2015.11.054.",
"Authors": "Liu X, Mao K, Yu AYH, Omairi-Nasser A, Austin J nd, Glick BS, Yip CK, Klionsky DJ /jcub",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/26774783",
"PubMed ID": "PubMed 26774783"
} | {
"Gene/Insert name": "ATG17",
"Species": "S. cerevisiae (budding yeast)",
"Insert Size (bp)": "",
"Mutation": "Phenylalanine 317 to Histidine",
"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": ""
} |
|
1531_pAAV-U6-SA-Self1-gRNA-HLP-EmGFP-spA | 109318 | Plasmid for liver-specific expression of EmGFP with a gRNA against SaCas9 | William Lagor | Li et al Mol Ther Methods Clin Dev. 2018 Dec 6;12:111-122. doi: 10.1016/j.omtm.2018.11.009. eCollection 2019 Mar 15. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/213163/3ce664f6-d46b-4395-8a87-765e73141a8d/addgene-plasmid-109318-sequence-213163.gbk | full | LOCUS sequence-213163- 4352 bp DNA circular SYN 26-AUG-2024
DEFINITION Plasmid for liver-specific expression of EmGFP with a gRNA against
SaCas9.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4352)
AUTHORS Li A, Lee CM, Hurley AE, Jarrett KE, De Giorgi M, Lu W, Balderrama
KS, Doerfler AM, Deshmukh H, Ray A, Bao G, Lagor WR
TITLE A Self-Deleting AAV-CRISPR System for In Vivo Genome Editing.
JOURNAL Mol Ther Methods Clin Dev. 2018 Dec 6;12:111-122. doi:
10.1016/j.omtm.2018.11.009. eCollection 2019 Mar 15.
PUBMED 30619914
REFERENCE 2 (bases 1 to 4352)
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 Ther
Methods Clin Dev."; date: "2018-12-6"; volume: "12:111-122. doi";
pages: " 10.1016/j.omtm.2018.11.009. eCollection 2019 Mar 15"
FEATURES Location/Qualifiers
source 1..4352
/mol_type="other DNA"
/organism="synthetic DNA construct"
repeat_region 4..133
/label=AAV2 ITR
promoter 255..503
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA
(Domitrovich & Kunkel, 2003)"
primer_bind 255..275
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 426..445
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
misc_RNA 525..600
/label=Sa gRNA scaffold
/note="guide RNA scaffold for the Staphylococcus aureus
CRISPR/Cas9 system"
regulatory 877..886
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 883..1602
/codon_start=1
/product="Emerald GFP"
/label=EmGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(928..949)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1189..1208)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1536..1557
/label=EGFP-C
/note="EGFP, forward primer"
polyA_signal 1617..1665
/note="synthetic polyadenylation signal"
repeat_region 1713..1842
/label=AAV2 ITR
rep_origin complement(2079..2667)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2159..2178)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(2838..3698)
/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 3461..3480
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(3699..3803)
/gene="bla"
/label=AmpR promoter
rep_origin complement(3829..4284)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3966..3987)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 4178..4197
/label=F1ori-R
/note="F1 origin, reverse primer"
ORIGIN
1 cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc
61 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc
121 actaggggtt ccttgtagtt aatgattaac ccgccatgct acttatctac gtagccatgc
181 tctggtgtac aaaaaagcag gctttaaagg aaccaattca gtcgactgga tccggtacca
241 aggtcgggca ggaagagggc ctatttccca tgattccttc atatttgcat atacgataca
301 aggctgttag agagataatt agaattaatt tgactgtaaa cacaaagata ttagtacaaa
361 atacgtgacg tagaaagtaa taatttcttg ggtagtttgc agttttaaaa ttatgtttta
421 aaatggacta tcatatgctt accgtaactt gaaagtattt cgatttcttg gctttatata
481 tcttgtggaa aggacgaaac accggtgatg ccgatgtcca ggccgtttta gtactctgga
541 aacagaatct actaaaacaa ggcaaaatgc cgtgtttatc tcgtcaactt gttggcgaga
601 tttttttcta gaaagctttg tttgctgctt gcaatgtttg cccattttag ggtggacaca
661 ggacgctgtg gtttctgagc cagggggcga ctcagatccc agccagtgga cttagcccct
721 gtttgctcct ccgataactg gggtgacctt ggttaatatt caccagcagc ctcccccgtt
781 gcccctctgg atccactgct taaatacgga cgaggacagg gccctgtctc ctcagcttca
841 ggcaccacca ctgacctggg acagtgaatc accggtgcca ccatggtgag caagggcgag
901 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
961 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
1021 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccttcacc
1081 tacggcgtgc agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag
1141 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
1201 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
1261 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
1321 aacagccaca aggtctatat caccgccgac aagcagaaga acggcatcaa ggtgaacttc
1381 aagacccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
1441 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
1501 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
1561 gccgccggga tcactctcgg catggacgag ctgtacaagt aagaattcct agagcgaata
1621 aaagatcttt attttcatta gatctgtgtg ttggtttttt gtgtgatgca gctacgtaga
1681 taagtagcat ggcgggttaa tcattaacta caaggaaccc ctagtgatgg agttggccac
1741 tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc
1801 gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcatta atgaatcggc
1861 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
1921 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
1981 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
2041 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
2101 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
2161 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
2221 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
2281 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
2341 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
2401 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
2461 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
2521 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
2581 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
2641 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
2701 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
2761 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
2821 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
2881 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
2941 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
3001 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
3061 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
3121 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
3181 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
3241 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
3301 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
3361 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
3421 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
3481 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
3541 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
3601 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgctgcaaaa
3661 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
3721 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
3781 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg
3841 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
3901 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
3961 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
4021 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
4081 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
4141 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
4201 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
4261 ttaatgcgcc gctacagggc gcgtcccatt cgccattcag gctgcgcaac tgttgggaag
4321 ggcgatcggt gcgggcctct tcgctattac gc
//
| {
"Vector backbone": "pEMBL8",
"Backbone size w/o insert (bp)": "4362",
"Vector type": "Mammalian Expression, AAV, 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": "A Self-Deleting AAV-CRISPR System for In Vivo Genome Editing",
"Publication": "Mol Ther Methods Clin Dev. 2018 Dec 6;12:111-122. doi: 10.1016/j.omtm.2018.11.009. eCollection 2019 Mar 15.",
"DOI": "10.1016/j.omtm.2018.11.009.",
"Authors": "Li A, Lee CM, Hurley AE, Jarrett KE, De Giorgi M, Lu W, Balderrama KS, Doerfler AM, Deshmukh H, Ray A, Bao G, Lagor WR /jomtm",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/30619914",
"PubMed ID": "PubMed 30619914"
} | {
"Gene/Insert name": "SaCas9 RuvC gRNA",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"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": "CCTTCATATTTGCATATACGATACAAGGCTGTTAG",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"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": ""
} |
|
pHBS981 TDP43 RRM-GFP G348D | 107835 | To test the effect of sequence on TDP43 droplet properties | Rajat Rohatgi | Rajat Rohatgi TDP43 plasmids (unpublished) | https://media.addgene.org/snapgene-media/v2.0.0/sequences/207991/b46b5066-1f1e-4b15-9998-fc2795ed797e/addgene-plasmid-107835-sequence-207991.gbk | full | LOCUS sequence-207991- 5682 bp DNA circular SYN 26-AUG-2024
DEFINITION To test the effect of sequence on TDP43 droplet properties.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5682)
TITLE Rajat Rohatgi TDP43 plasmids
REFERENCE 2 (bases 1 to 5682)
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..5682
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 30..48
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 30..47
/label=SP6
/note="SP6 promoter, forward primer"
protein_bind 103..127
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
regulatory 146..155
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
protein_bind complement(1660..1684)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
polyA_signal 1729..1863
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1753..1772)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 1807..1826
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind complement(1976..1996)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(1976..1994)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(2015..2031)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2015..2031)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(2028..2050)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 2039..2055
/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(2063..2093)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2108..2129
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(2246..2263)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2417..3005)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2497..2516)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3176..4036)
/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 3799..3818
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(4037..4141)
/gene="bla"
/label=AmpR promoter
rep_origin complement(4167..4622)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4304..4325)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 4516..4535
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 4693..5677
/label=CMV IE94 promoter
/note="enhancer/promoter region of simian cytomegalovirus
major immediate early transcription unit IE94"
ORIGIN
1 ttctgcctgg ggacgtcgga gcaagcttga tttaggtgac actatagaat acaagctact
61 tgttcttttt gcaggatccc atcgattcga attcaaggat caacaagttt gtacaaaaaa
121 gcaggctggc gccggtacca ccggtgccac catgggatcc atgtctgaat atattcgggt
181 aaccgaagat gagaacgatg agcccattga aataccatcg gaagacgatg ggacggtgct
241 gctctccacg gttacagccc agtttccagg ggcgtgtggg cttcgctaca ggaatccagt
301 gtctcagtgt atgagaggtg tccggctggt agaaggaatt ctgcatgccc cagatgctgg
361 ctggggaaat ctggtgtatg ttgtcaacta tccaaaagat aacaaaagaa aaatggatga
421 gacagatgct tcatcagcag tgaaagtgaa aagagcagtc cagaaaacat ccagtaaagg
481 agaggaacta tttacaggta ctgtgcctat caaagtcgag ctggacggcg acgtaaacgg
541 ccacaagttc agcgtgcgtg gcgagggcga gggcgatgcc accgagggca agctgaccct
601 gaagttcatc tgcaccaccg gcaagctgcc ggtgccgtgg ccgaccctcg tgaccaccct
661 gacctacggc gtgcagtgct tcagccgcta cccggaccac atgaagcgtc acgacttctt
721 caagtccgcc atgccggaag gctacgtcca ggagcgcacc atcgagttca aggacgacgg
781 cacctacaag acccgcgcgg aggtgaagtt cgagggcgac accctggtga accgcatcga
841 gctgaagggc aacgacttca aggaggacgg caacatcctg ggtcacaagc tggagtacaa
901 ccacaacagc cacaacgtcc gtatcgaagc cgacaagcag aagaacggca tcaaggctaa
961 cttcaagatc cgccacaacg ttgaggacgg cagccagcag gaagccgacc acaaacagca
1021 gaacaccccg atcggcgacg gcccggtgcg cctgccggac aaccactatc tgagcaccca
1081 gaccacgctg agcaaagacc cgaacgagaa gcgcgatcac atggtcctga aggagttcgt
1141 gacggcagcc ggtatcacta aaggtgaaga tgaaagtgga tccaatgccg aacctaagca
1201 caatagcaat agacagttag aaagaagtgg aagatttggt ggtaatccag gtggctttgg
1261 gaatcagggt ggatttggta atagcagagg gggtggagct ggtttgggaa acaatcaagg
1321 tagtaatatg ggtggtggga tgaactttgg tgcgttcagc attaatccag ccatgatggc
1381 tgccgcccag gcagcactac agagcagttg gggtatgatg ggcatgttag ccagccagca
1441 gaaccagtca gacccatcgg gtaataacca aaaccaaggc aacatgcaga gggagccaaa
1501 ccaggccttc ggttctggaa ataactctta tagtggctct aattctggtg cagcaattgg
1561 ttggggatca gcatccaatg cagggtcggg cagtggtttt aatggaggct ttggctcaag
1621 catggattct aagtcttctg gctggggaat gtagactaga cccagctttc ttgtacaaag
1681 tggttgatcc tctcgagcct ctagaactat agtgagtcgt attacgtaga tccagacatg
1741 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
1801 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
1861 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
1921 ttttaattcg cggccgcggc gccaatgcat tgggcccggt acccagcttt tgttcccttt
1981 agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
2041 gttatccgct cacaattcca cacaacatac gagccgggag cataaagtgt aaagcctggg
2101 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
2161 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
2221 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
2281 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
2341 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
2401 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
2461 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
2521 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
2581 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
2641 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
2701 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
2761 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
2821 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
2881 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
2941 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
3001 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
3061 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
3121 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
3181 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
3241 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
3301 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
3361 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
3421 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
3481 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
3541 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
3601 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
3661 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
3721 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
3781 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
3841 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
3901 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
3961 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
4021 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
4081 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
4141 gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc
4201 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
4261 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
4321 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
4381 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
4441 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
4501 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
4561 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
4621 gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
4681 gctattacgc cagtcgaccg ccaattcaat atggcgtata tggactcatg ccaattcaat
4741 atggtggatc tggacctgtg ccaattcaat atggcgtata tggactcgtg ccaattcaat
4801 atggtggatc tggaccccag ccaattcaat atggcggact tggcaccatg ccaattcaat
4861 atggcggacc tggcactgtg ccaactgggg aggggtctac ttggcacggt gccaagtttg
4921 aggaggggtc ttggccctgt gccaagtccg ccatattgaa ttggcatggt gccaataatg
4981 gcggccatat tggctatatg ccaggatcaa tatataggca atatccaata tggccctatg
5041 ccaatatggc tattggccag gttcaatact atgtattggc cctatgccat atagtattcc
5101 atatatgggt tttcctattg acgtagatag cccctcccaa tgggcggtcc catataccat
5161 atatggggct tcctaatacc gcccatagcc actcccccat tgacgtcaat ggtctctata
5221 tatggtcttt cctattgacg tcatatgggc ggtcctattg acgtatatgg cgcctccccc
5281 attgacgtca attacggtaa atggcccgcc tggctcaatg cccattgacg tcaataggac
5341 cacccaccat tgacgtcaat gggatggctc attgcccatt catatccgtt ctcacgcccc
5401 ctattgacgt caatgacggt aaatggccca cttggcagta catcaatatc tattaatagt
5461 aacttggcaa gtacattact attggaagta cgccagggta cattggcagt actcccattg
5521 acgtcaatgg cggtaaatgg cccgcgatgg ctgccaagta catccccatt gacgtcaatg
5581 gggaggggca atgacgcaaa tgggcgttcc attgacgtaa atgggcggta ggcgtgccta
5641 atgggaggtc tatataagca atgctcgttt agggaaccgc ca
//
| {
"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": "",
"Publication": "",
"DOI": "",
"Authors": "",
"PubMed Link": "",
"PubMed ID": ""
} | {
"Gene/Insert name": "TARDBP mutant",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "G348D mutation in LLPS 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": ""
} |
|
pEPYCα2rKN0010 | 154617 | GoldenBraid level α luciferase reporter | Nicola Patron | Cai et al Nucleic Acids Res. 2020 Aug 28. pii: 5897334. doi: 10.1093/nar/gkaa682. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/299491/10e2623a-ad47-4789-b1dc-632dd03d5dcd/addgene-plasmid-154617-sequence-299491.gbk | full | LOCUS sequence-299491- 7334 bp DNA circular SYN 26-AUG-2024
DEFINITION GoldenBraid level alpha luciferase reporter.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7334)
AUTHORS Cai YM, Kallam K, Tidd H, Gendarini G, Salzman A, Patron NJ
TITLE Rational design of minimal synthetic promoters for plants.
JOURNAL Nucleic Acids Res. 2020 Aug 28. pii: 5897334. doi:
10.1093/nar/gkaa682.
PUBMED 32856047
REFERENCE 2 (bases 1 to 7334)
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. 2020 Aug 28. pii: 5897334. doi: 10.1093/nar/gkaa682."
FEATURES Location/Qualifiers
source 1..7334
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS complement(1220..1849)
/codon_start=1
/product="stability protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/label=pVS1 StaA
/translation="MKVIAVLNQKGGSGKTTIGTHLARALQLAGADVLLVDSDPQGSAR
DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
misc_feature 3150..3174
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
CDS 3283..3795
/codon_start=1
/product="NanoLuc(R) luciferase"
/label=Nluc
/note="human codon-optimized"
/translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
RLCERILA"
CDS 3856..3879
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
terminator 3895..4147
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
misc_feature 4175..4199
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
CDS 4632..5447
/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 4632..5447
/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(4705..4724)
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
rep_origin 5535..6123
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6024..6043
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 6277..6294
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 6309..6449
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(6375..6394)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 6535..6557
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
rep_origin 6793..6987
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
CDS complement(join(7053..7334,1..786))
/codon_start=1
/product="replication protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/label=pVS1 RepA
/translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA
AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR
DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR
VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL
ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA
RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV
MRYRNLIEGEASAGS"
ORIGIN
1 aaccgtcagc acgtccgggt tcttgccttc tgtgatctcg cggtacatcc aatcagctag
61 ctcgatctcg atgtactccg gccgcccggt ttcgctcttt acgatcttgt agcggctaat
121 caaggcttca ccctcggata ccgtcaccag gcggccgttc ttggccttct tcgtacgctg
181 catggcaacg tgcgtggtgt ttaaccgaat gcaggtttct accaggtcgt ctttctgctt
241 tccgccatcg gctcgccggc agaacttgag tacgtccgca acgtgtggac ggaacacgcg
301 gccgggcttg tctcccttcc cttcccggta tcggttcatg gattcggtta gatgggaaac
361 cgccatcagt accaggtcgt aatcccacac actggccatg ccggccggcc ctgcggaaac
421 ctctacgtgc ccgtctggaa gctcgtagcg gatcacctcg ccagctcgtc ggtcacgctt
481 cgacagacgg aaaacggcca cgtccatgat gctgcgacta tcgcgggtgc ccacgtcata
541 gagcatcgga acgaaaaaat ctggttgctc gtcgcccttg ggcggcttcc taatcgacgg
601 cgcaccggct gccggcggtt gccgggattc tttgcggatt cgatcagcgg ccgcttgcca
661 cgattcaccg gggcgtgctt ctgcctcgat gcgttgccgc tgggcggcct gcgcggcctt
721 caacttctcc accaggtcat cacccagcgc cgcgccgatt tgtaccgggc cggatggttt
781 gcgaccgtca cgccgattcc tcgggcttgg gggttccagt gccattgcag ggccggcaga
841 caacccagcc gcttacgcct ggccaaccgc ccgttcctcc acacatgggg cattccacgg
901 cgtcggtgcc tggttgttct tgattttcca tgccgcctcc tttagccgct aaaattcatc
961 tactcattta ttcatttgct catttactct ggtagctgcg cgatctattc agatagcagc
1021 tcggtaatgg tcttgccttg gcgtaccgcg tacatcttca gcttggtgtg atcctccgcc
1081 ggcaactgaa agttgacccg cttcatggct ggcgtgtctg ccaggctggc caacgttgca
1141 gccttgctgc tgcgtgcgct cggacggccg gcacttagcg tgtttgtgct tttgctcatt
1201 ttctctttac ctcattaact caaatgagtt ttgatttaat ttcagcggcc agcgcctgga
1261 cctcgcgggc agcgtcgccc tcgggttctg attcaagaac ggttgtgccg gcggcggcag
1321 tgcctgggta gctcacgcgc tgcgtgatac gggactcaag aatgggcagc tcgtacccgg
1381 ccagcgcctc ggcaacctca ccgccgatgc gcgtgccttt gatcgcccgc gacacgacaa
1441 aggccgcttg tagccttcca tccgtgacct caatgcgctg cttaaccagc tccaccaggt
1501 cggcggtggc ccatatgtcg taagggcttg gctgcaccgg aatcagcacg aagtcggctg
1561 ccttgatcgc ggacacagcc aagtccgccg cctggggcgc tccgtcgatc actacgaagt
1621 cgcgccggcc gatggccttc acgtcgcggt caatcgtcgg gcggtcgatg ccgacaacgg
1681 ttagcggttg atcttcccgc acggccgccc aatcgcgggc actgccctgg ggatcggaat
1741 cgactaacag aacatcggcc ccggcgagtt gcagggcgcg ggctagatgg gttccgatgg
1801 tcgtcttgcc tgacccgcct ttctggttaa gtacagcgat aaccttcatg cgttcccctt
1861 gcgtatttgt ttatttactc atcggatcat atacgcagcg accgcatgac gcaagctgtt
1921 ttactcaaat acacatcacc tttttagacg gcggcgctcg gtttcttcag cggccaagct
1981 ggccggccag gccgccagct tggcatcaga caaaccggcc aggatttcat gcagccgcac
2041 ggttgagagg tgcgcgggcg gctcgaacac gtacccggcc gcgatcatct ccgcctcgat
2101 ctcttcggta atgaaaaacg gttcgtcctg gccgtcctgg tgcggtttca tgcttgttcc
2161 tcttggcgtt cattctcggc ggccgccagg gcgtcggcct cggtcaatgc gtcctcacgg
2221 aaggcaccgc gccgcctggc ctcggtgggc gtcacttcct cgctgcgctc aagtgcgcgg
2281 tacagggtcg agcgatgcac gccaagcagt gcagccgcct ctttcacggt gcggccttcc
2341 tggtcgatca gctcgcgggc gtgcgcgatc tgtgccgggg tgagggtagg gcgggggcca
2401 aacttcacgc ctcgggcctt ggcggcctcg cgcccgctcc gggtgcggtc gatgattagg
2461 gaacgctcga actcggcaat gccggcgaac acggtcaaca ccatgcggcc ggccggcgtg
2521 gtggtgtcgg cccacggctc tgccaggcta cgcaggcccg cgccggcctc ctggatgcgc
2581 tcggcaatgt ccagtaggtc gcgggtgctg cgggccaggc ggtctagcct ggtcactgtc
2641 acaacgtcgc cagggcgtag gtggtcaagc atcctggcca gctccgggcg gtcgcgcctg
2701 gtgccggtga tcttctcgga aaacagcttg gtgcagccgg ccgcgtgcag ttcggcccgt
2761 tggttggtca agtcctggtc gtcggtgctg acgcgggcat agcccagcag gccagcggcg
2821 gcgctcttgt tcatggcgta atgtctccgg ttctagtcgc aagtattcta ctttatgcga
2881 ctaaaacacg cgacaagaaa acgccaggaa aagggcaggg cggcagcctg tcgcgtaact
2941 taggacttgt gcgacatgtc gttttcagat gacggctgca ctgaacgtca gaagccgact
3001 gcactatagc agcggagggg ttggatcaaa gtactttgat cccgagggga accctgtggt
3061 tggcatgcac atacaaatgg acgaacggat aaaccttttc acgccctttt aaatatccga
3121 ttattctaat aaacgctctt ttctcttagg tttacccgcc aatatatcct gtcagatatc
3181 gtctcaagcg gaggttctgt gggttttttg ggtagaagaa gaaacccata gctatataat
3241 gagaaacagg ggcaaatccc cgaaaacaaa taacgcgaag caatggtatt cacactcgaa
3301 gatttcgttg gggactggcg acagacagcc ggctacaacc tggaccaagt ccttgaacag
3361 ggaggtgtgt ccagtttgtt tcagaatctc ggggtgtccg taactccgat ccaaaggatt
3421 gtcctgagcg gtgaaaatgg gctgaagatc gacatccatg tcatcatccc gtatgaaggt
3481 ctgagcggcg accaaatggg ccagatcgaa aaaattttta aggtggtgta ccctgtggat
3541 gatcatcact ttaaggtgat cctgcactat ggcacactgg taatcgacgg ggttacgccg
3601 aacatgatcg actatttcgg acggccgtat gaaggcatcg ccgtgttcga cggcaaaaag
3661 atcactgtaa cagggaccct gtggaacggc aacaaaatta tcgacgagcg cctgatcaac
3721 cccgacggct ccctgctgtt ccgagtaacc atcaacggag tgaccggctg gcggctgtgc
3781 gaacgcattc tggcttcgga ttataaggac catgacggag actataagga ccatgacctc
3841 gacgctgcag cagcggatta taaggacgat gacgataagt gagcttgtca agcagatcgt
3901 tcaaacattt ggcaataaag tttcttaaga ttgaatcctg ttgccggtct tgcgatgatt
3961 atcatataat ttctgttgaa ttacgttaag catgtaataa ttaacatgta atgcatgacg
4021 ttatttatga gatgggtttt tatgattaga gtcccgcaat tatacattta atacgcgata
4081 gaaaacaaaa tatagcgcgc aaactaggat aaattatcgc gcgcggtgtc atctatgtta
4141 ctagatcgac gcttgactga gacgatatcg ttatgtttac accacaatat atcctgccac
4201 cagccagcca acagctcccc gaccggcagc tcggcacaaa atcaccactc gatacaggca
4261 gcccatcagt ccgggacggc gtcagcggga gagccgttgt aaggcggcag actttgctca
4321 tgttaccgat gctattcgga agaacggcaa ctaagctgcc gggtttgaaa cacggatgat
4381 ctcgcggagg gtagcatgtt gattgtaacg atgacagagc gttgctgcct gtgatcaccg
4441 cggtttcaaa atcggctccg tcgatactat gttatacgcc aactttgaaa acaactttga
4501 aaaagctgtt ttctggtatt taaggtttta gaatgcaagg aacagtgaat tggagttcgt
4561 cttgttataa ttagcttctt ggggtatctt taaatactgt agaaaagagg aaggaaataa
4621 taagttctgt tatgagccat attcaacggg aaacgtcttg ctcaaggccg cgattaaatt
4681 ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag
4741 gtgcgacaat ctaccgattg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg
4801 gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg
4861 aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac
4921 tcaccactgc gatcccaggg aaaacagcat tccaggtatt agaagaatat cctgattcag
4981 gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt
5041 gtaattgtcc ttttaacagc gatcgcgtat ttcgactcgc tcaggcgcaa tcacgaatga
5101 ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac
5161 aagtctggaa agaaatgcat aaacttttgc cattctcacc ggattcagtc gtcactcatg
5221 gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg
5281 ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg
5341 gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg
5401 atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaatca cctagaatgc
5461 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
5521 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
5581 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
5641 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
5701 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
5761 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
5821 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
5881 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
5941 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
6001 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
6061 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
6121 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
6181 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
6241 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
6301 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
6361 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc
6421 tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc
6481 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtcaccggga
6541 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ggtgccttga
6601 tgtgggcgcc ggcggtcgag tggcgacggc gcggcttgtc cgcgccctgg tagattgcct
6661 ggccgtaggc cagccatttt tgagcggcca gcggccgcga taggccgacg cgaagcggcg
6721 gggcgtaggg agcgcagcga ccgaagggta ggcgcttttt gcagctcttc ggctgtgcgc
6781 tggccagaca gttatgcaca ggccaggcgg gttttaagag ttttaataag ttttaaagag
6841 ttttaggcgg aaaaatcgcc ttttttctct tttatatcag tcacttacat gtgtgaccgg
6901 ttcccaatgt acggctttgg gttcccaatg tacgggttcc ggttcccaat gtacggcttt
6961 gggttcccaa tgtacgtgct atccacagga aagagtcctt ttcgaccttt ttcccctgct
7021 agggcaattt gccctagcat ctgctccgta cattaggaac cggcggatgc ttcgccctcg
7081 atcaggttgc ggtagcgcat gactaggatc gggccagcct gccccgcctc ctccttcaaa
7141 tcgtactccg gcaggtcatt tgacccgatc agcttgcgca cggtgaaaca gaacttcttg
7201 aactctccgg cgctgccact gcgttcgtag atcgtcttga acaaccatct ggcttctgcc
7261 ttgcctgcgg cgcggcgtgc caggcggtag agaaaacggc cgatgccggg atcgatcaaa
7321 aagtaatcgg ggtg
//
| {
"Vector backbone": "pDGB3_?2R",
"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)": "NEB Stable",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA ; NanoLuc Luciferase Limited Use Label License",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Rational design of minimal synthetic promoters for plants",
"Publication": "Nucleic Acids Res. 2020 Aug 28. pii: 5897334. doi: 10.1093/nar/gkaa682.",
"DOI": "10.1093/nar/gkaa682.",
"Authors": "Cai YM, Kallam K, Tidd H, Gendarini G, Salzman A, Patron NJ /nar/gkaa",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/32856047",
"PubMed ID": "PubMed 32856047"
} | {
"Gene/Insert name": "MinSyn_064 Nanoluc expression cassette [MinSyn_064::LucN::Flag::nosT]",
"Species": "Synthetic",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "BsaI (destroyed during cloning)",
"3′ cloning site": "BsaI (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": ""
} |
|
pY006 (pcDNA3.1-hBpCpf1) | 69978 | Expresses humanized BpCpf1 | Feng Zhang | Zetsche et al Cell. 2015 Sep 23. pii: S0092-8674(15)01200-3. doi: 10.1016/j.cell.2015.09.038. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/125573/1757cf59-6693-4762-b8b1-66dfecab3af3/addgene-plasmid-69978-sequence-125573.gbk | partial | LOCUS sequence-125573- 921 bp DNA linear UNA 26-AUG-2024
DEFINITION Expresses humanized BpCpf1.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE natural DNA sequence
ORGANISM unspecified
REFERENCE 1 (bases 1 to 921)
AUTHORS Zetsche B, Gootenberg JS, Abudayyeh OO, Slaymaker IM, Makarova KS,
Essletzbichler P, Volz SE, Joung J, van der Oost J, Regev A, Koonin
EV, Zhang F
TITLE Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas
System.
JOURNAL Cell. 2015 Sep 23. pii: S0092-8674(15)01200-3. doi:
10.1016/j.cell.2015.09.038.
PUBMED 26422227
REFERENCE 2 (bases 1 to 921)
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. 2015
Sep 23. pii: S0092-8674(15)01200-3. doi:
10.1016/j.cell.2015.09.038."
FEATURES Location/Qualifiers
source 1..921
/mol_type="genomic DNA"
/organism="unspecified"
CDS complement(50..130)
/codon_start=1
/product="three tandem HA epitope tags"
/label=3xHA
/translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
CDS complement(137..184)
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
ORIGIN
1 ctctagactc gagcggccgc cactgtgctg gatatctgca gaattcttag gcatagtcgg
61 ggacatcata tgggtatgca taatcaggca cgtcgtaggg ataagcgtaa tctggaacat
121 cgtatgggta ggatcccttt ttcttttttg cctggccggc ctttttcgtg gccgccggcc
181 ttttcacggc gatatcctgg gcctgcatgt aattcaggaa ggcgtccaga aaatcgccgt
241 cgcctctata ctcctgcttg aacatgaact tcttgccgaa gttgtatgtg gtgttcttgc
301 tcacgtcgat ctctgtgccg gtgatggggc tccggtagat cttctcgccg tccttctttg
361 tgatgatgcc atacagggcg ttggcgaacc acttgtagat cagctccacg gacttgccgc
421 tgatggcggt cttatccaga aacttctcgg ccatattgcc atcgaacacg gccttgtcct
481 ccagcagggc tgggatgatc tgctctctca cgatgtcgtt cacgtacacg tactcgccct
541 tatccttgtc gaaataggag ccatcgccgg cgatcttcac ggtccactcc ttgcggtcca
601 gcagcctctt tgtagggtac tcctccaggt taaaggtgaa cagcatctcg ccatcgtcga
661 ttctgatgtc cttcatcttg gcaaagaact gtctcttaga ctgcattgtg atcagccggg
721 tcttatcgaa cacgtccaca aagccggtat cagggcacag ggtccgggta taggcgttat
781 tcaggaagta gatcacgccg tcctgataag aattgccgtt attgttggac acctggatgg
841 ggttagagat ggagccggcc tcgccctcct tgattgtatt aaaagacagg tcggacattc
901 tggcgcacag ccgggcctca a
//
| {
"Vector backbone": "pcDNA3.1",
"Backbone size w/o insert (bp)": "5354",
"Vector type": "Mammalian Expression, CRISPR",
"Selectable markers": "Neomycin (select with G418)"
} | {
"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": "Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System",
"Publication": "Cell. 2015 Sep 23. pii: S0092-8674(15)01200-3. doi: 10.1016/j.cell.2015.09.038.",
"DOI": "10.1016/j.cell.2015.09.038.",
"Authors": "Zetsche B, Gootenberg JS, Abudayyeh OO, Slaymaker IM, Makarova KS, Essletzbichler P, Volz SE, Joung J, van der Oost J, Regev A, Koonin EV, Zhang F /jcell",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/26422227",
"PubMed ID": "PubMed 26422227"
} | {
"Gene/Insert name": "hBpCpf1",
"Species": "Butyrivibrio_proteoclasticus",
"Insert Size (bp)": "3858",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "CMV",
"Tag / Fusion Protein": "NLS & 3xHA (C terminal on insert)"
} | {
"Cloning method": "Unknown",
"5′ cloning site": "",
"3′ cloning site": "",
"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": ""
} |
|
hGFAP-GFP | 40592 | Medium | Albee Messing | Zhuo et al Dev Biol. 1997 Jul 1;187(1):36-42. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/240470/ab60d8bc-22d0-40d8-8622-2ba588758baf/addgene-plasmid-40592-sequence-240470.gbk | full | LOCUS sequence-240470- 6181 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 6181)
AUTHORS Zhuo L, Sun B, Zhang CL, Fine A, Chiu SY, Messing A
TITLE Live astrocytes visualized by green fluorescent protein in
transgenic mice.
JOURNAL Dev Biol. 1997 Jul 1;187(1):36-42.
PUBMED 9224672
REFERENCE 2 (bases 1 to 6181)
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.";
date: "1997-07-1"; volume: "187(1)"; pages: "36-42"
FEATURES Location/Qualifiers
source 1..6181
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 1..105
/gene="bla"
/label=AmpR promoter
CDS 106..966
/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"
rep_origin 1137..1725
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 1626..1645
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 1879..1896
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 2013..2034
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2049..2079
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2087..2103
/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 2092..2114
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 2111..2127
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 2111..2127
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
regulatory 4382..4391
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 4388..5107
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLM
EFVTAAGITLGMDELYK"
primer_bind complement(4433..4454)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(4694..4713)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind complement(5692..5709)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(5692..5708)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5701..5723)
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind complement(5917..5936)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 6036..6058
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(6096..6114)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
ORIGIN
1 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
61 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
121 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
181 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
241 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
301 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
361 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
421 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
481 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
541 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
601 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
661 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
721 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
781 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
841 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
901 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
961 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
1021 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
1081 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
1141 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
1201 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
1261 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
1321 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
1381 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
1441 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
1501 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
1561 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
1621 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
1681 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
1741 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
1801 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
1861 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
1921 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
1981 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
2041 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
2101 caatttcaca caggaaacag ctatgacatg attacgaatt cgagctcggt accagatctg
2161 agctcccacc tccctctctg tgctgggact cacagaggga gacctcagga ggcagtctgt
2221 ccatcacatg tccaaatgca gagcataccc tgggctgggc gcagtggcgc acaactgtaa
2281 ttccagcact ttgggaggct gatgtggaag gatcacttga gcccagaagt tctagaccag
2341 cctgggcaac atggcaagac cctatctcta caaaaaaagt taaaaaatca gccacgtgtg
2401 gtgacacaca cctgtagtcc cagctattca ggaggctgag gtgaggggat cacttaaggc
2461 tgggaggttg aggctgcagt gagtcgtggt tgcgccactg cactccagcc tgggcaacag
2521 tgagaccctg tctcaaaaga caaaaaaaaa aaaaaaaaaa aaaagaacat atcctggtgt
2581 ggagtagggg acgctgctct gacagaggct cgggggcctg agctggctct gtgagctggg
2641 gaggaggcag acagccaggc cttgtctgca agcagacctg gcagcattgg gctggccgcc
2701 ccccagggcc tcctcttcat gcccagtgaa tgactcacct tggcacagac acaatgttcg
2761 gggtgggcac agtgcctgct tcccgccgca ccccagcccc cctcaaatgc cttccgagaa
2821 gcccattgag cagggggctt gcattgcacc ccagcctgac agcctggcat cttgggataa
2881 aagcagcaca gccccctagg ggctgccctt gctgtgtggc gccaccggcg gtggagaaca
2941 aggctctatt cagcctgtgc ccaggaaagg ggatcagggg atgcccaggc atggacagtg
3001 ggtggcaggg ggggagagga gggctgtctg cttcccagaa gtccaaggac acaaatgggt
3061 gaggggactg ggcagggttc tgaccctgtg ggaccagagt ggagggcgta gatggacctg
3121 aagtctccag ggacaacagg gcccaggtct caggctccta gttgggccca gtggctccag
3181 cgtttccaaa cccatccatc cccagaggtt cttcccatct ctccaggctg atgtgtggga
3241 actcgaggaa ataaatctcc agtgggagac ggaggggtgg ccagggaaac ggggcgctgc
3301 aggaataaag acgagccagc acagccagct catgtgtaac ggctttgtgg agctgtcaag
3361 gcctggtctc tgggagagag gcacagggag gccagacaag gaaggggtga cctggaggga
3421 cagatccagg ggctaaagtc ctgataaggc aagagagtgc cggccccctc ttgccctatc
3481 aggacctcca ctgccacata gaggccatga ttgaccctta gacaaagggc tggtgtccaa
3541 tcccagcccc cagccccaga actccaggga atgaatgggc agagagcagg aatgtgggac
3601 atctgtgttc aagggaagga ctccaggagt ctgctgggaa tgaggcctag taggaaatga
3661 ggtggccctt gagggtacag aacaggttca ttcttcgcca aattcccagc accttgcagg
3721 cacttacagc tgagtgagat aatgcctggg ttatgaaatc aaaaagttgg aaagcaggtc
3781 agaggtcatc tggtacagcc cttccttccc tttttttttt ttttttttgt gagacaaggt
3841 ctctctctgt tgcccaggct ggagtggcgc aaacacagct cactgcagcc tcaacctact
3901 gggctcaagc aatcctccag cctcagcctc ccaaagtgct gggattacaa gcatgagcca
3961 ccccactcag ccctttcctt cctttttaat tgatgcataa taattgtaag tattcatcat
4021 ggtccaacca accctttctt gacccacctt cctagagaga gggtcctctt gcttcagcgg
4081 tcagggcccc agacccatgg tctggctcca ggtaccacct gcctcatgca ggagttggcg
4141 tgcccaggaa gctctgcctc tgggcacagt gacctcagtg gggtgagggg agctctcccc
4201 atagctgggc tgcggcccaa ccccaccccc tcaggctatg ccagggggtg ttgccagggg
4261 cacccgggca tcgccagtct agcccactcc ttcataaagc cctcgcatcc caggagcgag
4321 cagagccaga gcaggttgga gaggagacgc atcacctccg ctgctcgcgg ggaagcttgc
4381 cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
4441 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
4501 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
4561 gcccaccctc gtgaccacct tcacctacgg cgtgcagtgc ttcagccgct accccgacca
4621 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
4681 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
4741 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
4801 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
4861 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
4921 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
4981 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
5041 catggtcctg atggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
5101 caagtaaagc ggccgcgact ctaggatccg cagatcccgg ccagataccg atgctgccgc
5161 agcaaaagca ggagcagatg ccgccgtcgc aggcgaagat gtcgcagacg gaggaggcga
5221 tgctgccggc ggaggaggcg aagtaagtag agggctgggc tgggctgtgg ggggtgtggg
5281 gtgcgggact gggcagtctg ggagtccctc tcaccacttt tcttaccttt ctaggatgct
5341 gccgtcgccg ccgctcatac accataaggt gtaaaaaata ctagatgcac agaatagcaa
5401 gtccatcaaa actcctgcgt gagaatttta ccagacttca agagcatctc gccacatctt
5461 gaaaaatgcc accgtccgat gaaaaacagg agcctgctaa ggaacaatgc cacctgtcaa
5521 taaatgttga aaactcatcc cattcctgcc tcttggtcct tgggcttggg gaggggtgcg
5581 cggatgtggt tagggaacat gactggtcaa atgggaaggg cttcaaaaga attcccaata
5641 ttgactacca agccacctgt acagatctgc ctgcaggcat gcaagcttgg cactggccgt
5701 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
5761 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
5821 acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg tattttctcc ttacgcatct
5881 gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg atgccgcata
5941 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
6001 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
6061 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
6121 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
6181 g
//
| {
"Vector backbone": "pgfa2-lac2",
"Backbone size w/o insert (bp)": "5500",
"Vector type": "Mammalian Expression, Bacterial Expression, Mouse Targeting",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "XL1 Blue",
"Copy number": "Unknown"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Live astrocytes visualized by green fluorescent protein in transgenic mice",
"Publication": "Dev Biol. 1997 Jul 1;187(1):36-42.",
"DOI": "10.1006/dbio.1997.8601",
"Authors": "Zhuo L, Sun B, Zhang CL, Fine A, Chiu SY, Messing A",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/9224672",
"PubMed ID": "PubMed 9224672"
} | {
"Gene/Insert name": "GFP-S65T",
"Species": "H. sapiens (human), M. musculus (mouse), Synthetic; Aequorea victoria (humanized)",
"Insert Size (bp)": "750",
"Mutation": "Construct contains a mutated, humanized hGFP(S65T) cDNA followed by a heterologous mouse intron and polyadenylation signal, all under the control of the gfa2 promoter",
"Entrez Gene": "",
"Promoter": "hGFAP promoter",
"Tag / Fusion Protein": "MP-1 fragment (mouse protamine 1 intron and polyadenylation signal) (C terminal on backbone)"
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "HindIII (not destroyed)",
"3′ cloning site": "XbaI (unknown if destroyed)",
"5′ sequencing primer": "GFAPpro-F (5'-ACTCCTTCATAAAGCCCTCG-3')",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"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-m4134125 | 152235 | Mammalian expression plasmid for SARS-CoV-2 endoRNAse | 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/295247/89b7cdb0-796a-4c82-a3e3-234635d38f17/addgene-plasmid-152235-sequence-295247.gbk | full | LOCUS sequence-295247- 7222 bp DNA circular SYN 26-AUG-2024
DEFINITION Mammalian expression plasmid for SARS-CoV-2 endoRNAse.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7222)
TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for
protein expression and/or purification
REFERENCE 2 (bases 1 to 7222)
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..7222
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 1040..1060
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQS"
CDS 1061..1138
/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"
polyA_signal 1191..1312
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1202..1221)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 1256..1275
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
terminator 1427..1473
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
terminator 1503..1639
/label=rrnG terminator
/note="transcription terminator from the E. coli ribosomal
RNA rrnG operon (Albrechtsen et al., 1991)"
misc_feature 1684..1755
/label=3' beta-globin insulator
/note="insulator downstream of the human beta-globin locus
(Farrell et al., 2002)"
rep_origin complement(1981..2569)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2061..2080)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
promoter 3069..4278
/label=UbC promoter
/note="human ubiquitin C promoter"
primer_bind 4115..4134
/label=hUBCpro-F
/note="Human Ubiquitin C (UbC) promoter, forward primer"
CDS 4282..4881
/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(4282..4301)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 4778..4798
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
polyA_signal 4908..4956
/note="synthetic polyadenylation signal"
CDS 5222..6082
/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(5440..5459)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
terminator 6095..6231
/label=rrnG terminator
/note="transcription terminator from the E. coli ribosomal
RNA rrnG operon (Albrechtsen et al., 1991)"
misc_feature 6376..6447
/label=5' beta-globin insulator
/note="insulator upstream of the human beta-globin locus
(Farrell et al., 2002)"
terminator complement(6452..6498)
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
polyA_signal 6516..6564
/note="synthetic polyadenylation signal"
enhancer 6645..6948
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 6950..7152
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 7103..7123
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
ORIGIN
1 caccatgtta gaaaatgtgg cttttaatgt tgtaaataag ggacactttg atggacaaca
61 gggtgaagta ccagtttcta tcattaataa cactgtttac acaaaagttg atggtgttga
121 tgtagaattg tttgaaaata aaacaacatt acctgttaat gtagcatttg agctttgggc
181 taagcgcaac attaaaccag taccagaggt gaaaatactc aataatttgg gtgtggacat
241 tgctgctaat actgtgatct gggactacaa aagagatgct ccagcacata tatctactat
301 tggtgtttgt tctatgactg acatagccaa gaaaccaact gaaacgattt gtgcaccact
361 cactgtcttt tttgatggta gagttgatgg tcaagtagac ttatttagaa atgcccgtaa
421 tggtgttctt attacagaag gtagtgttaa aggtttacaa ccatctgtag gtcccaaaca
481 agctagtctt aatggagtca cattaattgg agaagccgta aaaacacagt tcaattatta
541 taagaaagtt gatggtgttg tccaacaatt acctgaaact tactttactc agagtagaaa
601 tttacaagaa tttaaaccca ggagtcaaat ggaaattgat ttcttagaat tagctatgga
661 tgaattcatt gaacggtata aattagaagg ctatgccttc gaacatatcg tttatggaga
721 ttttagtcat agtcagttag gtggtttaca tctactgatt ggactagcta aacgttttaa
781 ggaatcacct tttgaattag aagattttat tcctatggac agtacagtta aaaactattt
841 cataacagat gcgcaaacag gttcatctaa gtgtgtgtgt tctgttattg atttattact
901 tgatgatttt gttgaaataa taaaatccca agatttatct gtagtttcta aggttgtcaa
961 agtgactatt gactatacag aaatttcatt tatgctttgg tgtaaagatg gccatgtaga
1021 aacattttac ccaaaattag aaaaccttta ctttcagtcg aagaggcgat ggaaaaagaa
1081 ttttatcgcg gtgagtgcag ccaacagatt caaaaagata tcctcgtctg gtgccctcta
1141 agctagcttg actgactgag atacagcgta ccttcagctc acagacatga taagatacat
1201 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
1261 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
1321 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa
1381 gtaaaacctc tacaaatgtg gtattggccc atctctatcg gtatcgtagc ataacccctt
1441 ggggcctcta aacgggtctt gaggggtttt ttgtgcccct cgggccggat tgctatctac
1501 cggcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat
1561 gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt
1621 accagaaatt tatccttaag gtcgtcagct atcctgcagg cgatctctcg atttcgatca
1681 agacattcct ttaatggtct tttctggaca ccactagggg tcagaagtag ttcatcaaac
1741 tttcttccct ccctaatctc attggttacc ttgggctatc gaaacttaat taaccagtca
1801 agtcagctac ttggcgagat cgacttgtct gggtttcgac tacgctcaga attgcgtcag
1861 tcaagttcga tctggtcctt gctattgcac ccgttctccg attacgagtt tcatttaaat
1921 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
1981 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
2041 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
2101 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
2161 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
2221 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
2281 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
2341 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
2401 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
2461 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
2521 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
2581 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
2641 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
2701 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
2761 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcattta aatttccgaa
2821 ctctccaagg ccctcgtcgg aaaatcttca aacctttcgt ccgatccatc ttgcaggcta
2881 cctctcgaac gaactatcgc aagtctcttg gccggccttg cgccttggct attgcttggc
2941 agcgcctatc gccaggtatt actccaatcc cgaatatccg agatcgggat cacccgagag
3001 aagttcaacc tacatcctca atcccgatct atccgagatc cgaggaatat cgaaatcggg
3061 gcgcgcctgg cctccgcgcc gggttttggc gcctcccgcg ggcgcccccc tcgtcacggc
3121 gagcgctgcc acgtcagacg aagggcgcag gagcgtcctg atccttccgc ccggacgctc
3181 aggacagcgg cccgctgctc ataagactcg gccttagaac cccagtatca gcagaaggac
3241 attttaggac gggacttggg tgactctagg gcactggttt tctttccaga gagcggaaca
3301 ggcgaggaaa agtagtccct tctcggcgat tctgcggagg gatctccgtg gggcggtgaa
3361 cgccgatgat tatataagga cgcgccgggt gtggcacagc tagttccgtc gcagccggga
3421 tttgggtcgc ggttcttgtt tgtggatcgc tgtgatcgtc acttggtgag tagcgggctg
3481 ctgggctggc cggggctttc gtggccgccg ggccgctcgg tgggacggaa gcgtgtggag
3541 agaccgccaa gggctgtagt ctgggtccgc gagcaaggtt gccctgaact gggggttggg
3601 gggagcgcag caaaatggcg gctgttcccg agtcttgaat ggaagacgct tgtgaggcgg
3661 gctgtgaggt cgttgaaaca aggtgggggg catggtgggc ggcaagaacc caaggtcttg
3721 agcccttcgc taatgcggga aagctcttat tcgggtgaga tgggctgggc accatctggg
3781 gaccctgacg tgaagtttgt cactgactgg agaactcggt ttgtcgtctg ttgcgggggc
3841 ggcagttatg gcggtgccgt tgggcagtgc acccgtacct ttgggagcgc gcgccctcgt
3901 cgtgtcgtga cgtcacccgt tctgttggct tataatgcag ggtggggcca cctgccggta
3961 ggtgtgcggt aggcttttct ccgtcgcagg acgcagggtt cgggcctagg gtaggctctc
4021 ctgaatcgac aggcgccgga cctctggtga ggggagggat aagtgaggcg tcagtttctt
4081 tggtcggttt tatgtaccta tcttcttaag tagctgaagc tccggttttg aactatgcgc
4141 tcggggttgg cgagtgtgtt ttgtgaagtt ttttaggcac cttttgaaat gtaatcattt
4201 gggtcaatat gtaattttca gtgttagact tgtaaattgt ccgctaaatt ctggccgttt
4261 ttggcttttt tgttagacaa catgaccgag tacaagccca cggtgcgcct cgccacccgc
4321 gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg
4381 cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc
4441 ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg
4501 gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg
4561 cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga gggcctcctg
4621 gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac
4681 caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc cgccgagcgc
4741 gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg
4801 ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg
4861 acccgcaagc ccggtgccta ggctagtatg taagcctagt cttagataat aaaatcgcta
4921 tccatcgaag atggatgtgt gttggttttt tgtgtgtgta acgctaggcg cgcctggtgt
4981 accgagaacg atcctctcag tgcgagtctc gacgatccat atcgttgctt ggcagtcagc
5041 cagtcggaat ccagcttggg acccaggaag tccaatcgtc agatattgta ctcaagcctg
5101 gtcacggcag cgtaccgatc tgtttaaacc tagatattga tagtctgatc ggtcaacgta
5161 taatcgagtc ctagcttttg caaacatcta tcaagagaca ggatcagcag gaggctttcg
5221 catgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
5281 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
5341 gcgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
5401 cgaagaacgc tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
5461 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
5521 ggttgagtat tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
5581 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
5641 tggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
5701 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
5761 gcctgtagca atggcaacaa ccttgcgtaa actattaact ggcgaactac ttactctagc
5821 ttcccggcaa cagttgatag actggatgga ggcggataaa gttgcaggac cacttctgcg
5881 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
5941 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
6001 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
6061 ctcactgatt aagcattggt aaccgattct aggtgcattg gcgcagaaaa aaatgcctga
6121 tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga tacggcttcc
6181 ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta agatcgttta
6241 aactcgactc tggctctatc gaatctccgt cgtttcgagc ttacgcgaac agccgtggcg
6301 ctcatttgct cgtcgggcat cgaatctcgt cagctatcgt cagcttacct ttttggcagc
6361 gatcgcggct cccgacatct tggaccatta gctccacagg tatcttcttc cctctagtgg
6421 tcataacagc agcttcagct acctctcaat tcaaaaaacc cctcaagacc cgtttagagg
6481 ccccaagggg ttatgctatc aatcgttgcg ttacacacac aaaaaaccaa cacacatcca
6541 tcttcgatgg atagcgattt tattatctaa ctgctgatcg agtgtagcca gatctagtaa
6601 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
6661 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
6721 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
6781 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
6841 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
6901 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatgct gatgcggttt
6961 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
7021 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
7081 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
7141 ataagcagag ctggtttagt gaaccgtcag atcagatctt tgtcgatcct accatccact
7201 cgacacaccc gccagcggcc gc
//
| {
"Vector backbone": "pTwist_CMV_Puro_6",
"Backbone size w/o insert (bp)": "6080",
"Vector type": "Mammalian Expression",
"Selectable markers": "Puromycin"
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "OpenMTA",
"Industry Terms": "OpenMTA"
} | {
"Title": "",
"Publication": "",
"DOI": "",
"Authors": "",
"PubMed Link": "",
"PubMed ID": ""
} | {
"Gene/Insert name": "SARS-CoV-2 endoRNAse",
"Species": "Severe acute respiratory syndrome coronavirus 2",
"Insert Size (bp)": "1031",
"Mutation": "wt",
"Entrez Gene": "ORF1ab",
"Promoter": "CMV",
"Tag / Fusion Protein": "Cleavable TEV;CBP (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": ""
} |
||
p11L2w | 37331 | Low | John Schiller | Kieback et al Virology. 2006 Feb 5;345(1):199-208. Epub 2005 Oct 27. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/46948/cd230051-9dfa-4745-bad6-5b733980c320/addgene-plasmid-37331-sequence-46948.gbk | full | LOCUS sequence-46948-e 5974 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 5974)
AUTHORS Kieback E, Muller M
TITLE Factors influencing subcellular localization of the human
papillomavirus L2 minor structural protein.
JOURNAL Virology. 2006 Feb 5;345(1):199-208. Epub 2005 Oct 27.
PUBMED 16257028
REFERENCE 2 (bases 1 to 5974)
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: "Virology.";
date: "2006-02-5"; volume: "345(1)"; pages: "199-208. Epub 2005 Oct
27"
FEATURES Location/Qualifiers
source 1..5974
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 113..1291
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 344..1282
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 1239..1259
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
misc_feature 2791..3379
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(2844..2864)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(3262..3273)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
polyA_signal 3436..3557
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3447..3466)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 3501..3520
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
rep_origin 3745..4333
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4234..4253
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind complement(4417..4437)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 4422..4751
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 4602..4737
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4664..4683
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter 4799..4846
/label=EM7 promoter
/note="synthetic bacterial promoter "
promoter 4838..4856
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 4866..5660
/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 4866..5660
/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(4920..4939)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 5530..5549
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(5718..5737)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 5762..5809
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
1 ccgctaccgg actcagatct cgagctcaag cttgcaaaga tggataaagt tttaaacaga
61 gaggaatctt tgcagctaat ggaccttgta ggtcttgaaa ggagtgggaa ttggctccgg
121 tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt
181 cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt
241 gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc
301 cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg ccgtgtgtgg
361 ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa ttacttccac
421 ctggctgcag tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt gggagagttc
481 gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct ggcctgggcg
541 ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg ctttcgataa
601 gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct ggcaagatag
661 tcttgtaaat gcgggccaag atctgcacac tggtatttcg gtttttgggg ccgcgggcgg
721 cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc gagcgcggcc
781 accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc ctggcctcgc
841 gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac cagttgcgtg
901 agcggaaaga tggccgcttc ccggccctgc tgcagggagc tcaaaatgga ggacgcggcg
961 ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc cgtcctcagc
1021 cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg attagttctc
1081 gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga tggagtttcc
1141 ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt aattctcctt
1201 ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga cagtggttca
1261 aagttttttt cttccatttc aggtgtcgtg aggaattctc tagagatccc tcgacctcga
1321 gatccatcct gtacggaagt gttacttctg ctctaaaagc tgcggaattg tacccgcggc
1381 cgctctagat gaagcccaga gcccggcgga gaaagcgggc cagcgccacc cagctgtacc
1441 agacctgcaa ggccaccggc acctgccccc ctgacgtgat ccccaaggtg gagcacacca
1501 ccatcgccga ccagatcctg aagtggggca gcctgggcgt gttcttcggc ggcctgggca
1561 tcggcaccgg cgccggcagc ggcggccggg ccggctacat ccccctgggc agcagcccta
1621 agcccgccat caccgggggc cctgccgcca gaccccccgt gctggtggag cctgtggccc
1681 ccagcgaccc cagcatcgtg agtctgatcg aggagagcgc catcatcaac gccggcgccc
1741 ctgaggtggt gcctcctacc cagggcggct tcaccatcac cagcagcgag agcaccaccc
1801 ctgccatcct ggacgtgagc gtgaccaacc acaccaccac cagcgtgttc cagaaccccc
1861 tgttcaccga gcctagcgtg atccagcccc agcctcccgt ggaggccagc ggccacatcc
1921 tgatcagcgc ccctaccatc accagccagc acgtggagga catccccctg gacaccttcg
1981 tggtgagcag cagcgacagc ggccccacca gcagcacccc tctgccccgg gccttcccca
2041 gacctcgggt gggcctgtac agcagagccc tgcagcaggt gcaggtgacc gaccccgcct
2101 tcctgagcac cccccagcgg ctggtgacct acgacaaccc tgtgtacgag ggcgaggacg
2161 tgagcctgca gttcacccac gagagcatcc acaacgcccc cgacgaggcc ttcatggata
2221 tcatcagact gcaccggccc gccatcacca gcagacgggg cctggtgcgc ttcagcagaa
2281 tcggccagcg gggcagcatg tacacccgga gcggccagca catcggcgcc agaattcact
2341 acttccagga catcagccct gtgacccagg ccgccgagga gatcgagctg caccccctgg
2401 tggccgccga gaacgacacc ttcgacatct acgccgagcc cttcgaccct atccccgacc
2461 ccgtgcagca cagcgtgacc cagagctacc tgaccagcac ccctaacacc ctgagccaga
2521 gctggggcaa caccaccgtg cccctgagca tccccagcga ctggttcgtg cagagcggcc
2581 ccgacatcac cttccctacc gctagcatgg gcaccccctt cagccccgtg acccctgccc
2641 tgcccaccgg ccccgtgttc atcaccggca gcgacttcta cctgcaccct acctggtact
2701 tcgcccggcg gagacggaag cggatccccc tgttcttcac cgacgtggcc gcctaagctt
2761 atcgataccg tcgatatcaa gcttatcgat aatcaacctc tggattacaa aatttgtgaa
2821 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
2881 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
2941 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
3001 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
3061 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
3121 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
3181 gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg
3241 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
3301 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
3361 ctttgggccg cctccccgca tcgataccgt cggccgcttc cctttagtga gggttaatgc
3421 ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
3481 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
3541 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
3601 aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatg gaattcttaa
3661 ttaactagcc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
3721 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
3781 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
3841 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
3901 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
3961 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
4021 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
4081 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
4141 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
4201 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
4261 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
4321 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
4381 ttttgctcac atgttcttaa ttaagctgta cactgtggaa tgtgtgtcag ttagggtgtg
4441 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
4501 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
4561 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
4621 ccagttccgc ccattctccg ccccatgact gactaatttt ttttatttat gcagaggccg
4681 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
4741 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat cagcacgtgt
4801 tgacaattaa tcatcggcat agtatatcgg catagtataa tacgactcac tataggaggg
4861 ccaccatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
4921 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
4981 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
5041 aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
5101 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
5161 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
5221 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
5281 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
5341 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc
5401 ccgacggcga ggatctcgtc gtgacacatg gcgatgcctg cttgccgaat atcatggtgg
5461 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
5521 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
5581 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
5641 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgaattc
5701 gctagaggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
5761 acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg
5821 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
5881 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
5941 gcagccaacg tcggggcggc aggccctgcc atag
//
| {
"Vector backbone": "custom",
"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",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Factors influencing subcellular localization of the human papillomavirus L2 minor structural protein",
"Publication": "Virology. 2006 Feb 5;345(1):199-208. Epub 2005 Oct 27.",
"DOI": "10.1016/j.virol.2005.09.047",
"Authors": "Kieback E, Muller M",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/16257028",
"PubMed ID": "PubMed 16257028"
} | {
"Gene/Insert name": "HPV11 L2",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "EF1a",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Unknown",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "NA",
"3′ sequencing primer": "NA"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pT7-EGFP-C1-HseIF4E-F47AI79A_Z | 148136 | Mammalian Expression of HseIF4E-F47AI79A | Elisa Izaurralde | Izaurralde lab plasmids (unpublished) | https://media.addgene.org/snapgene-media/v2.0.0/sequences/390263/9d580a51-1d17-418f-b06c-e7c47544ffb6/addgene-plasmid-148136-sequence-390263.gbk | full | LOCUS sequence-390263- 5419 bp DNA circular SYN 26-AUG-2024
DEFINITION Mammalian Expression of HseIF4E-F47AI79A.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5419)
TITLE Izaurralde lab plasmids
REFERENCE 2 (bases 1 to 5419)
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..5419
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 61..364
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 365..568
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 519..539
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 613..632
/label=T7
/note="T7 promoter, forward primer"
promoter 613..631
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory 659..668
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 665..1381
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 665..1381
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(710..731)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(971..990)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1318..1339
/label=EGFP-C
/note="EGFP, forward primer"
polyA_signal 2207..2328
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2244..2263)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2298..2317
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(2335..2790)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2472..2493)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2684..2703
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2817..2921
/gene="bla"
/label=AmpR promoter
promoter 2923..3280
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
enhancer 2923..3114
/label=SV40 enhancer
/note="enhancer for the SV40 early promoter (Herr, 1993)"
primer_bind complement(2946..2966)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 3131..3266
/label=SV40 ori
/note="SV40 origin of replication"
CDS 3315..4109
/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 3315..4109
/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(3369..3388)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3979..3998
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(4297..4316)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 4341..4388
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4717..5305
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5206..5225
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
541 acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg
601 gcttatcgaa attaatacga ctcactatag ggagacccaa gctctagcgc taccggtcgc
661 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
721 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
781 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
841 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
901 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
961 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
1021 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
1081 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
1141 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
1201 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
1261 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
1321 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
1381 gtccggactc agatctcgag ctcaagcttc gaattctgca atggcgactg tcgaaccgga
1441 aaccacccct actcctaatc ccccgactac agaagaggag aaaacggaat ctaatcagga
1501 ggttgctaac ccagaacact atattaaaca tcccctacag aacagatggg cactctgggc
1561 ttttaaaaat gataaaagca aaacttggca agcaaacctg cggctgatct ccaagtttga
1621 tactgttgaa gacttttggg ctctgtacaa ccatgcccag ttgtctagta atttaatgcc
1681 tggctgtgac tactcacttt ttaaggatgg tattgagcct atgtgggaag atgagaaaaa
1741 caaacgggga ggacgatggc taattacatt gaacaaacag cagagacgaa gtgacctcga
1801 tcgcttttgg ctagagacac ttctgtgcct tattggagaa tcttttgatg actacagtga
1861 tgatgtatgt ggcgctgttg ttaatgttag agctaaaggt gataagatag caatatggac
1921 tactgaatgt gaaaacagag aagctgttac acatataggg agggtataca aggaaaggtt
1981 aggacttcct ccaaagatag tgattggtta tcagtcccac gcagacacag ctactaagag
2041 cggctccacc actaaaaata ggtttgttgt ttaaccggga tccaccggat ctagataact
2101 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca
2161 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
2221 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
2281 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaac gcgtaaattg
2341 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
2401 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
2461 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
2521 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
2581 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
2641 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
2701 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
2761 ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg
2821 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
2881 aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca
2941 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
3001 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
3061 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
3121 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
3181 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
3241 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat
3301 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
3361 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
3421 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
3481 ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
3541 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
3601 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
3661 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
3721 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
3781 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
3841 cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
3901 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
3961 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
4021 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
4081 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
4141 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
4201 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
4261 gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc
4321 ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc
4381 gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag
4441 accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc
4501 gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg
4561 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
4621 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
4681 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
4741 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
4801 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
4861 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
4921 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
4981 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
5041 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
5101 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
5161 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
5221 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
5281 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
5341 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
5401 ccgtattacc gccatgcat
//
| {
"Vector backbone": "Unknown",
"Backbone size w/o insert (bp)": "",
"Vector type": "Mammalian Expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Kanamycin, 50 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "",
"Publication": "",
"DOI": "",
"Authors": "",
"PubMed Link": "",
"PubMed ID": ""
} | {
"Gene/Insert name": "HseIF4E-F47AI79A",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "EIF4E",
"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": ""
} |
|
pLOVE empty vector | 15948 | High | (Empty Backbone) | Miguel Ramalho-Santos | Blelloch et al Cell Stem Cell. 2007 Sep 13. ():. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/110663/501225e1-a634-4c79-9404-bc14d8fbbe7f/addgene-plasmid-15948-sequence-110663.gbk | full | LOCUS sequence-110663- 8263 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 8263)
AUTHORS Blelloch R, Venere M, Yen J, Ramalho-Santos M
TITLE Generation of Induced Pluripotent Stem Cells in the Absence of Drug
Selection.
JOURNAL Cell Stem Cell. 2007 Sep 13. ():.
PUBMED 18371358
REFERENCE 2 (bases 1 to 8263)
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 Stem
Cell. 2007 Sep 13. ():."
FEATURES Location/Qualifiers
source 1..8263
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(56..75)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 247..626
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 628..826
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
LTR 844..1024
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 1071..1196
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1695..1928
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 2113..2157
/codon_start=1
/product="antigenic peptide corresponding to amino acids
655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
et al., 2013)"
/label=gp41 peptide
/note="recognized by the 2H10 single-chain llama nanobody"
/translation="KNEQELLELDKWASL"
misc_feature 2455..2572
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
enhancer 2638..2941
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 2942..3145
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
protein_bind 3147..3165
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 3168..3186
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 3227..3351
/gene="mutant version of attR"
/label=attR1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
promoter 3376..3406
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
CDS 3460..4119
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind complement(3526..3545)
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
CDS 4461..4766
/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 4645..4664
/label=ccdB-fwd
/note="ccdB gene, forward primer"
protein_bind complement(4807..4931)
/gene="mutant version of attR"
/label=attR2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
CDS 4984..5025
/codon_start=1
/product="epitope tag from simian virus 5"
/label=V5 tag
/translation="GKPIPNPLLGLDST"
protein_bind complement(5063..5096)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
misc_feature 5152..5740
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(5205..5225)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(5623..5634)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(5743..5759)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5744..5760)
/label=pBluescriptKS
/note="For pBluescript vector"
LTR 6269..6449
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
rep_origin complement(6511..7099)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6591..6610)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(7270..8130)
/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 7893..7912
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(8131..8235)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gcttaagcgg tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa
61 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
121 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
181 tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata
241 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
301 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
361 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
421 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
481 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
541 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
601 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
661 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
721 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
781 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagcgcg ttttgcctgt
841 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
901 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
961 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
1021 agcagtggcg cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac
1081 gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta
1141 cgccaaaaat tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta
1201 ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga
1261 aaaaatataa attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta
1321 atcctggcct gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat
1381 cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca accctctatt
1441 gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag
1501 agcaaaacaa aagtaagacc accgcacagc aagcggccgg ccgcgctgat cttcagacct
1561 ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa
1621 attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa
1681 agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg
1741 ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag
1801 cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc
1861 tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa
1921 cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg
1981 aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag
2041 tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa
2101 aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg
2161 aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga
2221 ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag
2281 ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc
2341 gaaggaatag aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac
2401 ggatcggcac tgcgtgcgcc aattctgcag acaaatggca gtattcatcc acaattttaa
2461 aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac
2521 agacatacaa actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttcgggttta
2581 ttacagggac agcagagatc cagtttggtt agtaccgggc ccgctctagc agttccgcgt
2641 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
2701 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
2761 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
2821 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
2881 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
2941 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
3001 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
3061 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
3121 gtgggaggtc tatataagca gagctctccc tatcagtgat agagatctcc ctatcagtga
3181 tagagatcgt cgactagtcc agtgtggtgg aattctgcag atatcaacaa gtttgtacaa
3241 aaaagctgaa cgagaaacgt aaaatgatat aaatatcaat atattaaatt agattttgca
3301 taaaaaacag actacataat actgtaaaac acaacatatc cagtcactat ggcggccgca
3361 ttaggcaccc caggctttac actttatgct tccggctcgt ataatgtgtg gattttgagt
3421 taggatccgt cgagattttc aggagctaag gaagctaaaa tggagaaaaa aatcactgga
3481 tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc atttcagtca
3541 gttgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt tttaaagacc
3601 gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc ccgcctgatg
3661 aatgctcatc cggaattccg tatggcaatg aaagacggtg agctggtgat atgggatagt
3721 gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc gctctggagt
3781 gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt ggcgtgttac
3841 ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt cgtctcagcc
3901 aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga caacttcttc
3961 gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct gatgccgctg
4021 gcgattcagg ttcatcatgc cgtctgtgat ggcttccatg tcggcagaat gcttaatgaa
4081 ttacaacagt actgcgatga gtggcagggc ggggcgtaaa gatctggatc cggcttacta
4141 aaagccagat aacagtatgc gtatttgcgc gctgattttt gcggtataag aatatatact
4201 gatatgtata cccgaagtat gtcaaaaaga ggtgtgctat gaagcagcgt attacagtga
4261 cagttgacag cgacagctat cagttgctca aggcatatat gatgtcaata tctccggtct
4321 ggtaagcaca accatgcaga atgaagcccg tcgtctgcgt gccgaacgct ggaaagcgga
4381 aaatcaggaa gggatggctg aggtcgcccg gtttattgaa atgaacggct cttttgctga
4441 cgagaacagg gactggtgaa atgcagttta aggtttacac ctataaaaga gagagccgtt
4501 atcgtctgtt tgtggatgta cagagtgata ttattgacac gcccgggcga cggatggtga
4561 tccccctggc cagtgcacgt ctgctgtcag ataaagtctc ccgtgaactt tacccggtgg
4621 tgcatatcgg ggatgaaagc tggcgcatga tgaccaccga tatggccagt gtgccggtct
4681 ccgttatcgg ggaagaagtg gctgatctca gccaccgcga aaatgacatc aaaaacgcca
4741 ttaacctgat gttctgggga atataaatgt caggctccct tatacacagc cagtctgcag
4801 gtcgaccata gtgactggat atgttgtgtt ttacagtatt atgtagtctg ttttttatgc
4861 aaaatctaat ttaatatatt gatatttata tcattttacg tttctcgttc agctttcttg
4921 tacaaagtgg ttgatatcca gcacagtggc ggccgctcga gtctagaggg cccgcggttc
4981 gaaggtaagc ctatccctaa ccctctcctc ggtctcgatt ctacgcgtac cggttagtaa
5041 tgacaattcg tcgagggacc taataacttc gtatagcata cattatacga agttatacat
5101 gtttaagggt tccggttcca ctaggtacaa ttcgatatca agcttatcga taatcaacct
5161 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
5221 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
5281 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
5341 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
5401 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
5461 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
5521 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt
5581 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
5641 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
5701 cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg tcgacctcga
5761 tcgagaccta gaaaaacatg gagcaatcac aagtagcaat acagcagcta ccaatgctga
5821 ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt tttccagtca cacctcaggt
5881 acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa
5941 ggggggactg gaagggctaa ttcactccca acgaagacaa gatatccttg atctgtggat
6001 ctaccacaca caaggctact tccctgattg gcagaactac acaccagggc cagggatcag
6061 atatccactg acctttggat ggtgctacaa gctagtacca gttgagcaag agaaggtaga
6121 agaagccaat gaaggagaga acacccgctt gttacaccct gtgagcctgc atgggatgga
6181 tgacccggag agagaagtat tagagtggag gtttgacagc cgcctagcat ttcatcacat
6241 ggcccgagag ctgcatccgg actgtactgg gtctctctgg ttagaccaga tctgagcctg
6301 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
6361 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
6421 cttttagtca gtgtggaaaa tctctagcag catgtgagca aaaggccagc aaaaggccag
6481 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
6541 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
6601 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
6661 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
6721 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
6781 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
6841 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
6901 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
6961 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
7021 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
7081 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
7141 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
7201 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
7261 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
7321 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
7381 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
7441 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
7501 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
7561 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
7621 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
7681 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
7741 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
7801 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
7861 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
7921 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
7981 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
8041 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
8101 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
8161 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
8221 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gac
//
| {
"Vector backbone": "pLOVE (for Lentiviral Over-Expression)",
"Backbone size w/o insert (bp)": "",
"Vector type": "Mammalian Expression, Lentiviral",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Chloramphenicol and Ampicillin, 25 & 100 μg/mL",
"Growth Temperature": "30°C",
"Growth Strain(s)": "DB3.1",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Generation of Induced Pluripotent Stem Cells in the Absence of Drug Selection",
"Publication": "Cell Stem Cell. 2007 Sep 13. ():.",
"DOI": "10.1016/j.stem.2007.08.008",
"Authors": "Blelloch R, Venere M, Yen J, Ramalho-Santos M",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/18371358",
"PubMed ID": "PubMed 18371358"
} | {
"Gene/Insert name": "None",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Gateway Cloning",
"5′ cloning site": "attR1 (not destroyed)",
"3′ cloning site": "attR2 (not destroyed)",
"5′ sequencing primer": "n/a",
"3′ sequencing primer": "n/a"
} | {
"Gene/Insert name": "",
"Species": "",
"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": ""
} |
IDP379 | 87006 | plasmid for cell culture expression of SYNseq components | Anthony Zador | Peikon et al Nucleic Acids Res. 2017 Jul 7;45(12):e115. doi: 10.1093/nar/gkx292. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/169737/417f5ae5-931d-4a54-9c17-202e6133e1c6/addgene-plasmid-87006-sequence-169737.gbk | full | LOCUS sequence-169737- 5647 bp DNA circular SYN 26-AUG-2024
DEFINITION plasmid for cell culture expression of SYNseq components.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5647)
AUTHORS Peikon ID, Kebschull JM, Vagin VV, Ravens DI, Sun YC, Brouzes E,
Correa IR Jr, Bressan D, Zador AM
TITLE Using high-throughput barcode sequencing to efficiently map
connectomes.
JOURNAL Nucleic Acids Res. 2017 Jul 7;45(12):e115. doi: 10.1093/nar/gkx292.
PUBMED 28449067
REFERENCE 2 (bases 1 to 5647)
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."; date: "2017-07-7"; volume: "45(12):e115. doi"; pages: "
10.1093/nar/gkx292"
FEATURES Location/Qualifiers
source 1..5647
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 108..1286
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 339..1277
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 1234..1254
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
regulatory 1545..1554
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
primer_bind complement(1681..1699)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 1949..1968
/label=mCherry-F
/note="mCherry, forward primer"
primer_bind 2143..2166
/label=DsRed1-C
/note="DsRed1, forward primer"
polyA_signal 2435..2556
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2472..2491)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2526..2545
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(2563..3018)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2700..2721)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2912..2931
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 3045..3149
/gene="bla"
/label=AmpR promoter
promoter 3151..3508
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
enhancer 3151..3342
/label=SV40 enhancer
/note="enhancer for the SV40 early promoter (Herr, 1993)"
primer_bind complement(3174..3194)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 3359..3494
/label=SV40 ori
/note="SV40 origin of replication"
CDS 3543..4337
/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 3543..4337
/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(3597..3616)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4207..4226
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(4525..4544)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 4569..4616
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4945..5533
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5434..5453
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 tagttattaa tcctgcagga taagctttgc aaagatggat aaagttttaa acagagagga
61 atctttgcag ctaatggacc ttctaggtct tgaaaggagt gggaattggc tccggtgccc
121 gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca
181 attgaaccgg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact
241 ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga
301 acgttctttt tcgcaacggg tttgccgcca gaacacaggt aagtgccgtg tgtggttccc
361 gcgggcctgg cctctttacg ggttatggcc cttgcgtgcc ttgaattact tccacctggc
421 tgcagtacgt gattcttgat cccgagcttc gggttggaag tgggtgggag agttcgaggc
481 cttgcgctta aggagcccct tcgcctcgtg cttgagttga ggcctggcct gggcgctggg
541 gccgccgcgt gcgaatctgg tggcaccttc gcgcctgtct cgctgctttc gataagtctc
601 tagccattta aaatttttga tgacctgctg cgacgctttt tttctggcaa gatagtcttg
661 taaatgcggg ccaagatctg cacactggta tttcggtttt tggggccgcg ggcggcgacg
721 gggcccgtgc gtcccagcgc acatgttcgg cgaggcgggg cctgcgagcg cggccaccga
781 gaatcggacg ggggtagtct caagctggcc ggcctgctct ggtgcctggc ctcgcgccgc
841 cgtgtatcgc cccgccctgg gcggcaaggc tggcccggtc ggcaccagtt gcgtgagcgg
901 aaagatggcc gcttcccggc cctgctgcag ggagctcaaa atggaggacg cggcgctcgg
961 gagagcgggc gggtgagtca cccacacaaa ggaaaagggc ctttccgtcc tcagccgtcg
1021 cttcatgtga ctccacggag taccgggcgc cgtccaggca cctcgattag ttctcgagct
1081 tttggagtac gtcgtcttta ggttgggggg aggggtttta tgcgatggag tttccccaca
1141 ctgagtgggt ggagactgaa gttaggccag cttggcactt gatgtaattc tccttggaat
1201 ttgccctttt tgagtttgga tcttggttca ttctcaagcc tcagacagtg gttcaaagtt
1261 tttttcttcc atttcaggtg tcgtgaggta cgaccaccgc tagcgctggt cggtacggta
1321 acggagagtc cgtcccctct tatcctcggc gttgtgtgtc aaatggcgta gatctggatt
1381 gactctatga cggtatctgc tgatcggtag ggagaccgag aatctatcgg gctaacgcgt
1441 cttataacct ccgaaaatgc tggcaccgtt ctctaagatc ggaagagcgg ttcagcagga
1501 atgccgagac cgatctcgta tgccgtcttc tgcttggcgg ccgcgccacc atggcctcct
1561 ccgaggacgt catcaaggag ttcatgcgct tcaaggtgcg catggagggc tccgtgaacg
1621 gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg
1681 ccaagctgaa ggtgaccaag ggcggccccc tgcccttcgc ctgggacatc ctgtcccctc
1741 agttccagta cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga
1801 agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg
1861 tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc
1921 tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg
1981 aggcctccac cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagatga
2041 ggctgaagct gaaggacggc ggccactacg acgccgaggt caagaccacc tacatggcca
2101 agaagcccgt gcagctgccc ggcgcctaca agaccgacat caagctggac atcacctccc
2161 acaacgagga ctacaccatc gtggaacagt acgagcgcgc cgagggccgc cactcctaat
2221 gtacagccct gaaaaagggc tcgagccctg aaaaagggca attgccctga aaaagggcgt
2281 cgacgccctg aaaaagggcg gtaccgcggg cccggggccg cgactctaga tcataatcag
2341 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
2401 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
2461 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
2521 tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata
2581 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
2641 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
2701 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
2761 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
2821 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
2881 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
2941 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
3001 cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
3061 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
3121 tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt
3181 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
3241 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
3301 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
3361 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
3421 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
3481 cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc
3541 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
3601 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
3661 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
3721 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
3781 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
3841 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
3901 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
3961 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
4021 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
4081 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
4141 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
4201 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
4261 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
4321 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
4381 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
4441 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
4501 cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
4561 cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa
4621 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
4681 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
4741 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat
4801 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
4861 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
4921 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
4981 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
5041 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
5101 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
5161 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
5221 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
5281 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
5341 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
5401 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
5461 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
5521 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
5581 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
5641 catgcat
//
| {
"Vector backbone": "unknown",
"Backbone size w/o insert (bp)": "",
"Vector type": "Mammalian 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 ; Takara Bio Limited Use Label License (formerly Clontech)",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Using high-throughput barcode sequencing to efficiently map connectomes",
"Publication": "Nucleic Acids Res. 2017 Jul 7;45(12):e115. doi: 10.1093/nar/gkx292.",
"DOI": "10.1093/nar/gkx292.",
"Authors": "Peikon ID, Kebschull JM, Vagin VV, Ravens DI, Sun YC, Brouzes E, Correa IR Jr, Bressan D, Zador AM /nar/gkx",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/28449067",
"PubMed ID": "PubMed 28449067"
} | {
"Gene/Insert name": "postLib-mRFP-4xboxB",
"Species": "M. musculus (mouse)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "EF1a",
"Tag / Fusion Protein": "mRFP (C terminal on insert)"
} | {
"Cloning method": "Gibson Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "EF1a-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": ""
} |
|
pAc5.1B-lambdaN-HA-DmNot1_1467-1717-K1537SQ1658A_W | 147869 | Insect Expression of DmNot1_1467-1717-K1537SQ1658A | Elisa Izaurralde | Chen et al Mol Cell. 2014 Jun 5;54(5):737-50. doi: 10.1016/j.molcel.2014.03.034. Epub 2014 Apr 24. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/290316/833c1a41-0fc4-47e0-a3d6-7ab601a3c79e/addgene-plasmid-147869-sequence-290316.gbk | full | LOCUS sequence-290316- 6219 bp DNA circular SYN 26-AUG-2024
DEFINITION Insect Expression of DmNot1_1467-1717-K1537SQ1658A.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6219)
AUTHORS Chen Y, Boland A, Kuzuoglu-Ozturk D, Bawankar P, Loh B, Chang CT,
Weichenrieder O, Izaurralde E
TITLE A DDX6-CNOT1 complex and W-binding pockets in CNOT9 reveal direct
links between miRNA target recognition and silencing.
JOURNAL Mol Cell. 2014 Jun 5;54(5):737-50. doi:
10.1016/j.molcel.2014.03.034. Epub 2014 Apr 24.
PUBMED 24768540
REFERENCE 2 (bases 1 to 6219)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1016/j.molcel.2014.03.034"; journalName: "Mol Cell"; date:
"2014-06-5- 5"; volume: "54"; issue: "5"; pages: "737-50"
FEATURES Location/Qualifiers
source 1..6219
/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 3504..3545
/codon_start=1
/product="epitope tag from simian virus 5"
/label=V5 tag
/translation="GKPIPNPLLGLDST"
CDS 3555..3572
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(3595..3612)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 3645..3779
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3669..3688)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 3723..3742
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
rep_origin complement(4401..4989)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4481..4500)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5160..6020)
/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 5783..5802
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6021..6125)
/gene="bla"
/label=AmpR promoter
primer_bind 6193..6211
/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 agcttaatta agatgcgcta ttcatacgtg gacgtaaacg tgagcaactt
2761 ccagcttatt ggtcagcagt tggtactgcc acccaatact ccgttcctgc acgccaatcc
2821 cggaatcaaa cacatggtcg tcaacgccgt ggagcgcacg ataaccgact ggttgcagcc
2881 tattgtggac cgaagcatcc gcattgcctg tgccaccacc gagcagatca tccgcagcga
2941 cttcgccctg gatgctgacg agaaccgaat gcgcactgcc gctcatcaga tggtacgaaa
3001 ccttgccgcc ggcatggcca tgattaccgg taaggatgag atagcgcgtg ctattagcca
3061 gaatctgcac aaggccttgc tgtcgggtct aaatgggatg cccagcatgg ctgagatcca
3121 agctgctgct atgcagttag ccagcgaaaa tgttgagcta gtttgcgctt tcattcagaa
3181 aacatctgct gaaaaggcag ccgcagagat cgataggcgt ctaagcaccg atttcgagac
3241 cagaaaaatt gctcgtgaag agggcaaccg ctttgtggac gcccagatcc tcacctatgc
3301 acaggagcgt cttccagaag cagtgcgcat caaggtgggc gcggctccag ctacacttta
3361 tgctgtgtac tcggagttcg ctagaagcat acctggcttc cagcagatga gcgatcgcga
3421 tattgccctg tttgtgccca agccgacgga tttgtctcag ccaaatgtgt ttgctaattg
3481 atctagaggg cccgcggttc gaaggtaagc ctatccctaa ccctctcctc ggtctcgatt
3541 ctacgcgtac cggtcatcat caccatcacc attgagttta aacccgctga tcagcctcga
3601 ctgtgccttc taaggcctga gctcgctgat cagcctcgat cgaggatcca gacatgataa
3661 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
3721 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
3781 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt
3841 aaagcaagta aaacctctac aaatgtggta tggctgatta tgatcagtcg accgatgccc
3901 ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc
3961 gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc agcgctctgg
4021 gtcattttcg gcgaggaccg ctttcgctgg agcgcgacga tgatcggcct gtcgcttgcg
4081 gtattcggaa tcttgcacgc cctcgctcaa gccttcgtca ctggtcccgc caccaaacgt
4141 ttcggcgaga agcaggccat tatcgccggc atggcggccg acgcgctggg ctacgtcttg
4201 ctggcgttcg cgacgcgagg ctggatggcc ttccccatta tgattcttct cgcttccggc
4261 ggcatcggga tgcccgcgtt gcaggccatg ctgtccaggc aggtagatga cgaccatcag
4321 ggacagcttc aaggatcgct cgcggctctt accagccagc aaaaggccag gaaccgtaaa
4381 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
4441 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
4501 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
4561 gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt
4621 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
4681 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
4741 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
4801 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
4861 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
4921 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
4981 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
5041 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
5101 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
5161 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
5221 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
5281 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
5341 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
5401 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
5461 gttgttgcca ttgctgcagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
5521 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
5581 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
5641 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
5701 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
5761 tcttgcccgg cgtcaacacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
5821 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
5881 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
5941 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
6001 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
6061 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
6121 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
6181 ttaacctata aaaataggcg tatcacgagg ccctttcgt
//
| {
"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": "A DDX6-CNOT1 complex and W-binding pockets in CNOT9 reveal direct links between miRNA target recognition and silencing",
"Publication": "Mol Cell. 2014 Jun 5;54(5):737-50. doi: 10.1016/j.molcel.2014.03.034. Epub 2014 Apr 24.",
"DOI": "10.1016/j.molcel.2014.03.034.",
"Authors": "Chen Y, Boland A, Kuzuoglu-Ozturk D, Bawankar P, Loh B, Chang CT, Weichenrieder O, Izaurralde E /jmolcel",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/24768540",
"PubMed ID": "PubMed 24768540"
} | {
"Gene/Insert name": "DmNot1_1467-1717-K1537SQ1658A",
"Species": "D. melanogaster (fly)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "Not1",
"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": ""
} |
|
TFORF2854 | 143384 | Lentiviral vector for overexpressing transcription factor ORFs with unique 24-bp barcodes. Barcodes facilitate identification of transcription factors in pooled screens. | Feng Zhang | Joung et al Cell. 2023 Jan 5;186(1):209-229.e26. doi: 10.1016/j.cell.2022.11.026. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/392750/80f25500-e518-4ae1-9cc5-7b60d7af3846/addgene-plasmid-143384-sequence-392750.gbk | full | LOCUS sequence-392750- 9082 bp DNA circular SYN 25-AUG-2024
DEFINITION Lentiviral vector for overexpressing transcription factor ORFs with
unique 24-bp barcodes. Barcodes facilitate identification of
transcription factors in pooled screens..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9082)
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 9082)
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..9082
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 1..1179
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 232..1170
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 1127..1147
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
misc_feature 2020..2608
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(2073..2093)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(2491..2502)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 2680..2913
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 2991..3112
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3028..3047)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3082..3101
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 3152..3287
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(3307..3326)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(3308..3326)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(3336..3353)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(3336..3352)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3345..3367)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 3494..3949
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3581..3600)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3791..3812
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 3975..4079
/gene="bla"
/label=AmpR promoter
CDS 4080..4940
/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(4298..4317)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 5111..5699
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5600..5619
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 5853..5870
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 5987..6008
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 6023..6053
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6061..6077
/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 6066..6088
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 6085..6101
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 6085..6101
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 6120..6140
/label=T3
/note="T3 promoter, forward primer"
promoter 6122..6140
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 6168..6394
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 6395..6575
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 6622..6747
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 7240..7473
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 7658..7702
/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 7933..8143
/label=SV40 promoter
/note="SV40 early promoter"
promoter 7955..8151
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 8002..8137
/label=SV40 ori
/note="SV40 origin of replication"
CDS 8183..8785
/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(8183..8202)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 8679..8699
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
misc_feature 8828..8945
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
ORIGIN
1 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
61 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
121 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
181 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc
241 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt
301 acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
361 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
421 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
481 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
541 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
601 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
661 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
721 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
781 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
841 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
901 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
961 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
1021 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
1081 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
1141 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gctagcaccg ccaccatgag
1201 acagagcggc gcctcccagc ccctgctgat caacatgtac ctgccagatc ccgtcggaga
1261 cggtctcttc aaggacggga agaacccgag ctgggggccg ctgagccccg cggttcagaa
1321 aggcagcgga cagatccagc tgtggcagtt tctgctggag ctgctggctg accgcgcgaa
1381 cgccggctgc atcgcgtggg agggcggtca cggcgagttc aagctcacgg acccggacga
1441 ggtggcgcgg cggtggggcg agcgcaagag caagcccaac atgaactacg acaagctgag
1501 ccgcgccctg cgctactact acgacaagaa catcatgagc aaggtgcatg gcaagcgcta
1561 cgcctaccgc ttcgacttcc agggcctggc gcaggcctgc cagccgccgc ccgcgcacgc
1621 tcatgccgcc gccgcagctg ctgccgccgc cgcggccgcc caggacggcg cgctctacaa
1681 gctgcccgcc ggcctcgccc cgctgccctt ccccggcctc tccaaactca acctcatggc
1741 cgcctcggcc ggggtcgcgc ccgccggctt ctcctactgg ccgggcccgg gccccgccgc
1801 caccgctgcc gccgccaccg ccgcgctcta ccccagtccc agcttgcagc ccccgcccgg
1861 gcccttcggg gccgtggccg cagcctcgca cttggggggc cattaccact agtaatgaac
1921 tagtccgtaa cttgaaagta tttcgatttc ttggctttat atatcttgtg gaaaggacga
1981 tcctgaataa atacaattcc tgctacgcgt taagtcgaca atcaacctct ggattacaaa
2041 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
2101 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
2161 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
2221 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
2281 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
2341 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
2401 gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt
2461 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
2521 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
2581 cggatctccc tttgggccgc ctccccgcgt cgactttaag accaatgact tacaaggcag
2641 ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc
2701 aacgaagaca agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag
2761 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
2821 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
2881 gaccctttta gtcagtgtgg aaaatctcta gcagtacgta tagtagttca tgtcatctta
2941 ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg aacttgttta
3001 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
3061 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
3121 ggctctagct atcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
3181 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
3241 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag ggacgtaccc
3301 aattcgccct atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt
3361 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
3421 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
3481 aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
3541 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
3601 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
3661 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
3721 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
3781 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
3841 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
3901 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt
3961 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
4021 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
4081 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
4141 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
4201 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
4261 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
4321 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
4381 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
4441 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
4501 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
4561 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
4621 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
4681 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
4741 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
4801 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
4861 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
4921 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
4981 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
5041 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
5101 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
5161 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
5221 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
5281 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
5341 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
5401 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
5461 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
5521 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
5581 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
5641 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
5701 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
5761 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
5821 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
5881 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
5941 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
6001 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
6061 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg
6121 caattaaccc tcactaaagg gaacaaaagc tggagctgca agcttaatgt agtcttatgc
6181 aatactcttg tagtcttgca acatggtaac gatgagttag caacatgcct tacaaggaga
6241 gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg tacgatcgtg ccttattagg
6301 aaggcaacag acgggtctga catggattgg acgaaccact gaattgccgc attgcagaga
6361 tattgtattt aagtgcctag ctcgatacat aaacgggtct ctctggttag accagatctg
6421 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
6481 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
6541 cagacccttt tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa
6601 gcgaaaggga aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg
6661 gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag
6721 aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg
6781 gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg
6841 gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc
6901 tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga
6961 tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac
7021 accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag
7081 caagcggccg ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga
7141 attatataaa tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa
7201 gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt
7261 cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag
7321 acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca
7381 acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc
7441 tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact
7501 catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat
7561 ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat
7621 acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga
7681 attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat
7741 aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact
7801 ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc
7861 aaccccgagg ggacccatgc attgcatctc aattagtcag caaccaggtg tggaaagtcc
7921 ccaggctccc cagcaggcag aagtatgcaa agcatgcgtc tcaattagtc agcaaccata
7981 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
8041 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
8101 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctttct
8161 agaggtacca ccggagctta ccatgaccga gtacaagccc acggtgcgcc tcgccacccg
8221 cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac
8281 gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt
8341 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgccgt
8401 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc
8461 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct
8521 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga
8581 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg
8641 cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg
8701 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat
8761 gacccgcaag cccggtgcca gttaacgaat tctagatctt gagacaaatg gcagtattca
8821 tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag
8881 acataatagc aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa
8941 attttcgggt ttattacagg gacagcagag atccactttg gcgccggctc gaggcctgca
9001 ggtgcaaaga tggataaagt tttaaacaga gaggaatctt tgcagctaat ggaccttcta
9061 ggtcttgaaa ggagtgggaa tt
//
| {
"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": "FEV",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "818",
"Mutation": "",
"Entrez Gene": "FEV",
"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": ""
} |
|
pMH5 | 52531 | Low | expresses FLP recombinase in Drosophila | Klaus Foerstemann | Bottcher et al Nucleic Acids Res. 2014 Apr 19. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/81491/475ac21e-126c-40be-b246-fe6f92c52f6d/addgene-plasmid-52531-sequence-81491.gbk | full | LOCUS sequence-81491-e 11335 bp DNA circular SYN 26-AUG-2024
DEFINITION expresses FLP recombinase in Drosophila .
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11335)
AUTHORS Bottcher R, Hollmann M, Merk K, Nitschko V, Obermaier C,
Philippou-Massier J, Wieland I, Gaul U, Forstemann K
TITLE Efficient chromosomal gene modification with CRISPR/cas9 and
PCR-based homologous recombination donors in cultured Drosophila
cells.
JOURNAL Nucleic Acids Res. 2014 Apr 19.
PUBMED 24748663
REFERENCE 2 (bases 1 to 11335)
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. 2014 Apr 19."
FEATURES Location/Qualifiers
source 1..11335
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 2060..3331
/codon_start=1
/product="site-specific recombinase"
/label=FLP
/note="FLP is a site-specific recombinase from
Saccharomyces cerevisiae. Recombination occurs at FRT
sequences."
/translation="MPQFGILCKTPPKVLVRQFVERFERPSGEKIALCAAELTYLCWMI
THNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKKLIPAWEFTI
IPYYGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESIWEITEKILN
SFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGVIIQCLVTET
KTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEYQLLKDNLVR
SYNKALKKNAPYSIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWSDKRASAVAR
TTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQLKGSAEGSIR
YPAWNGIISQEVLDYLSSYINRRI"
polyA_signal 3453..3534
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3490..3509)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3544..3563
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
gene 3576..7704
/gene="white"
/label=mini-white
/note="This modified version of the white gene lacks part
of the first intron."
primer_bind complement(3585..3603)
/label=pCasper-F
/note="Drosophila mini-white gene, reverse primer"
CDS join(4055..4126,4521..4794,4869..5523,5585..5900,6121..6252,
6323..6937)
/codon_start=1
/gene="white"
/product="Drosophila white gene eye color pigment"
/label=mini-white
/note="This is a modified version of the white gene lacking
part of the first intron."
/translation="MGQEDQELLIRGGSKHPSAEHLNNGDSGAASQSCINQGFGQAKNY
GTLLPPSPPEDSGSGSGQLAENLTYAWHNMDIFGAVNQPGSGWRQLVNRTRGLFCNERH
IPAPRKHLLKNVCGVAYPGELLAVMGSSGAGKTTLLNALAFRSPQGIQVSPSGMRLLNG
QPVDAKEMQARCAYVQQDDLFIGSLTAREHLIFQAMVRMPRHLTYRQRVARVDQVIQEL
SLSKCQHTIIGVPGRVKGLSGGERKRLAFASEALTDPPLLICDEPTSGLDSFTAHSVVQ
VLKKLSQKGKTVILTIHQPSSELFELFDKILLMAEGRVAFLGTPSEAVDFFSYVGAQCP
TNYNPADFYVQVLAVVPGREIESRDRIAKICDNFAISKVARDMEQLLATKNLEKPLEQP
ENGYTYKATWFMQFRAVLWRSWLSVLKEPLLVKVRLIQTTMVAILIGLIFLGQQLTQVG
VMNINGAIFLFLTNMTFQNVFATINVFTSELPVFMREARSRLYRCDTYFLGKTIAELPL
FLTVPLVFTAIAYPMIGLRAGVLHFFNCLALVTLVANVSTSFGYLISCASSSTSMALSV
GPPVIIPFLLFGGFFLNSGSVPVYLKWLSYLSWFRYANEGLLINQWADVEPGEISCTSS
NTTCPSSGKVILETLNFSAADLPLDYVGLAILIVSFRVLAYLALRLRARRKE"
misc_feature 7715..8300
/label=P element 5' end
rep_origin complement(8535..9123)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(8615..8634)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(9294..10154)
/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 9917..9936
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(10155..10259)
/gene="bla"
/label=AmpR promoter
primer_bind 10327..10345
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(10383..10405)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 10505..10524
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
protein_bind 10545..10578
/label=attB
/bound_moiety="phage phi-C31 integrase"
/note="minimal attB site for the phi-C31 integrase (Groth
et al., 2000)"
misc_feature 11103..11335
/label=P element 3' end
/note="P element 3' end"
ORIGIN
1 aagcttgggc tgcaggtcga ctctagctag aggatgtcgc cggaacgcag cgacagggat
61 tccaatgtgt ccgtatcctt caggcttttg cccttcagtt ccaggcgcag cgactggcga
121 ttcgcgtgtg gggtctgctt cagggtcttg tggattaggg cgcgcagatc gccgatgggc
181 gtggcgccgg agggcacctt caccttgccg tacggcttgc tgttcttcgc gttcaaaatc
241 tccagctcca ttttgctttc ggtgcgcttg caatcagtac tgtccaaaat cgaaaatcgc
301 cgaaccgtag tgtgaccgtg cggggctctg cgaaaataaa cttttttagg tatatggcca
361 cacacgggga aagcacagtg gattatatgt attaatatat tatgcaggtt ttcattactt
421 atccagatgt aagcccactt aaagcgattt aacaattatt tgccgaaaga gtataaacaa
481 atttcacata aaaatggatt aagaaaagct tgtgtaagat tatgcgcagc gttgccagat
541 agctccattt aaaacacttc aaaaacaata agtttagaaa atatatacat aaatagcagt
601 cgttgccgca acgctcaaca catcacactt ttaaaacacc ctttacctac acagaattac
661 tttttaaatt tccagtcaag ctgcgagttt caaaattata gccggtagag aagacagtgc
721 tatttcaaaa gcaaactaac aagggtctta aatttcaaaa caccaatcct aacaagcctt
781 ggacttttgt aagtttagat caaaggtggc attgcattca atgtcatggt aagaagtagg
841 tcgtctaggt agaaatcctc attcagccgg tcaagtcagt acgagaaagg tctcaatttg
901 aaattgtctt aaaaatattt tattgttttg tactgtggtg agtttaaacg aaaaacacaa
961 aaaaaaagtg atacacagaa atcataaaaa attttaatac aaggtattcg tacgtatcaa
1021 aaacatttcg gcacaatttt ttttctctgt actaaagtgt tacgaacact acggtatttt
1081 ttagtgattt tcaacggaca ccgaaggtat ataaacagcg ttcgcgaacg gtcgccttca
1141 aaaccaattg acatttgcag cagcaagtac aagtagaaag taaagcgcaa tcagcgaaaa
1201 atttatactt aattgttggt gattaaagta caattaaaag aacattctcg aaagtcacaa
1261 gaaacgtaag tttttaactc gctgttacca attagtaata agagcaacaa gacgttgagt
1321 aatttcaaga aaaactgcat ttcaaggtct ttgttcggcc attttttttt tattcaacgc
1381 tctacgtaat tacaaaataa gaaattggca gccacgcatc ttgttttccc aatgaattgg
1441 catcaaaacg caaacaaatc tataaataaa acttgcgtgt tgattttcgc caagatttat
1501 tggcaaattg tgaaattcgc agtgacgcat ttgaaaattc gagaaatcac gaacgcactc
1561 gagcatttgt gtgcatgtta ttagttagtt gtttagttaa ttgaagtatt taccaacgaa
1621 atccacttat ttttagctga aatagagtag gttgcttaaa caaagccacg tctgaaaatt
1681 tcttattgct tgtagttgtg acgtcaccat atacacacaa aataatgtgt atgcatgcat
1741 ttcagctgtg tatatataca tgcacacact cgcaacacga aaacgatgac gaagcaacgg
1801 aacaaaggtt tctcaactac cctttgttcc ctgtttcttc gctttccttt gttccaatat
1861 tcgtagaggg ttaatagggg tttctcaaca aagttggcgt cgataaataa gtttcccatt
1921 tttattcccc agccagaagt tagtttcaat agttttgtaa tttcaacgaa actcatttga
1981 tttcgtacta attttccaca tctctatttt ctgcccgcag aataatccaa actgcaggtc
2041 gactctagct agaggatcca tgccacaatt tggtatatta tgtaaaacac cacctaaggt
2101 gcttgttcgt cagtttgtgg aaaggtttga aagaccttca ggtgagaaaa tagcattatg
2161 tgctgctgaa ctaacctatt tatgttggat gattacacat aacggaacag caatcaagag
2221 agccacattc atgagctata atactatcat aagcaattcg ctgagtttcg atattgtcaa
2281 taaatcactc cagtttaaat acaagacgca aaaagcaaca attctggaag cctcattaaa
2341 gaaattgatt cctgcttggg aatttacaat tattccttac tatggacaaa aacatcaatc
2401 tgatatcact gatattgtaa gtagtttgca attacagttc gaatcatcgg aagaagcaga
2461 taagggaaat agccacagta aaaaaatgct taaagcactt ctaagtgagg gtgaaagcat
2521 ctgggagatc actgagaaaa tactaaattc gtttgagtat acttcgagat ttacaaaaac
2581 aaaaacttta taccaattcc tcttcctagc tactttcatc aattgtggaa gattcagcga
2641 tattaagaac gttgatccga aatcatttaa attagtccaa aataagtatc tgggagtaat
2701 aatccagtgt ttagtgacag agacaaagac aagcgttagt aggcacatat acttctttag
2761 cgcaaggggt aggatcgatc cacttgtata tttggatgaa tttttgagga attctgaacc
2821 agtcctaaaa cgagtaaata ggaccggcaa ttcttcaagc aataaacagg aataccaatt
2881 attaaaagat aacttagtca gatcgtacaa taaagctttg aagaaaaatg cgccttattc
2941 aatctttgct ataaaaaatg gcccaaaatc tcacattgga agacatttga tgacctcatt
3001 tctttcaatg aagggcctaa cggagttgac taatgttgtg ggaaattgga gcgataagcg
3061 tgcttctgcc gtggccagga caacgtatac tcatcagata acagcaatac ctgatcacta
3121 cttcgcacta gtttctcggt actatgcata tgatccaata tcaaaggaaa tgatagcatt
3181 gaaggatgag actaatccaa ttgaggagtg gcagcatata gaacagctaa agggtagtgc
3241 tgaaggaagc atacgatacc ccgcatggaa tgggataata tcacaggagg tactagacta
3301 cctttcatcc tacataaata gacgcatata agcggccgcg actctagatc ataatcagcc
3361 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
3421 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
3481 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
3541 gttgtggttt gtccaaactc atcaatgtat gaattctagt atgtatgtaa gttaataaaa
3601 cccatttttg cggaaagtag ataaaaaaaa catttttttt ttttactgca ctggatatca
3661 ttgaacttat ctgatcagtt ttaaatttac ttcgatccaa gggtatttga tgtaccaggt
3721 tctttcgatt acctctcact caaaatgaca ttccactcaa agtcagcgct gtttgcctcc
3781 ttctctgtcc acagaaatat cgccgtctct ttcgccgctg cgtccgctat ctctttcgcc
3841 accgtttgta gcgttacgta gcgtcaatgt ccgccttcag ttgcattttg tcagcggttt
3901 cgtgacgaag ctccaagcgg tttacgccat caattaaaca caaagtgctg tgccaaaact
3961 cctctcgctt cttatttttg tttgtttttt gagtgattgg ggtggtgatt ggttttgggt
4021 gggtaagcag gggaaagtgt gaaaaatccc ggcaatgggc caagaggatc aggagctatt
4081 aattcgcgga ggcagcaaac acccatctgc cgagcatctg aacaatgtga gtagtacatg
4141 tgcatacatc ttaagttcac ttgatctata ggaactgcga ttgcaacatc aaattgtctg
4201 cggcgtgaga actgcgaccc acaaaaatcc caaaccgcaa tcgcacaaac aaatagtgac
4261 acgaaacaga ttattctggt agctgtgctc gctatataag acaattttta agatcatatc
4321 atgatcaaga catctaaagg cattcatttt cgactacatt cttttttaca aaaaatataa
4381 caaccagata ttttaagctg atcctagatg cacaaaaaat aaataaaagt ataaacctac
4441 ttcgtaggat acttcgtttt gttcggggtt agatgagcat aacgcttgta gttgatattt
4501 gagatcccct atcattgcag ggtgacagcg gagcggcttc gcagagctgc attaaccagg
4561 gcttcgggca ggccaaaaac tacggcacgc tcctgccacc cagtccgccg gaggactccg
4621 gttcagggag cggccaacta gccgagaacc tcacctatgc ctggcacaat atggacatct
4681 ttggggcggt caatcagccg ggctccggat ggcggcagct ggtcaaccgg acacgcggac
4741 tattctgcaa cgagcgacac ataccggcgc ccaggaaaca tttgctcaag aacggtgagt
4801 ttctattcgc agtcggctga tctgtgtgaa atcttaataa agggtccaat taccaatttg
4861 aaactcagtt tgcggcgtgg cctatccggg cgaacttttg gccgtgatgg gcagttccgg
4921 tgccggaaag acgaccctgc tgaatgccct tgcctttcga tcgccgcagg gcatccaagt
4981 atcgccatcc gggatgcgac tgctcaatgg ccaacctgtg gacgccaagg agatgcaggc
5041 caggtgcgcc tatgtccagc aggatgacct ctttatcggc tccctaacgg ccagggaaca
5101 cctgattttc caggccatgg tgcggatgcc acgacatctg acctatcggc agcgagtggc
5161 ccgcgtggat caggtgatcc aggagctttc gctcagcaaa tgtcagcaca cgatcatcgg
5221 tgtgcccggc agggtgaaag gtctgtccgg cggagaaagg aagcgtctgg cattcgcctc
5281 cgaggcacta accgatccgc cgcttctgat ctgcgatgag cccacctccg gactggactc
5341 atttaccgcc cacagcgtcg tccaggtgct gaagaagctg tcgcagaagg gcaagaccgt
5401 catcctgacc attcatcagc cgtcttccga gctgtttgag ctctttgaca agatccttct
5461 gatggccgag ggcagggtag ctttcttggg cactcccagc gaagccgtcg acttcttttc
5521 ctagtgagtt cgatgtgttt attaagggta tctagcatta cattacatct caactcctat
5581 ccagcgtggg tgcccagtgt cctaccaact acaatccggc ggacttttac gtacaggtgt
5641 tggccgttgt gcccggacgg gagatcgagt cccgtgatcg gatcgccaag atatgcgaca
5701 attttgctat tagcaaagta gcccgggata tggagcagtt gttggccacc aaaaatttgg
5761 agaagccact ggagcagccg gagaatgggt acacctacaa ggccacctgg ttcatgcagt
5821 tccgggcggt cctgtggcga tcctggctgt cggtgctcaa ggaaccactc ctcgtaaaag
5881 tgcgacttat tcagacaacg gtgagtggtt ccagtggaaa caaatgatat aacgcttaca
5941 attcttggaa acaaattcgc tagattttag ttagaattgc ctgattccac acccttctta
6001 gtttttttca atgagatgta tagtttatag ttttgcagaa aataaataaa tttcatttaa
6061 ctcgcgaaca tgttgaagat atgaatatta atgagatgcg agtaacattt taatttgcag
6121 atggttgcca tcttgattgg cctcatcttt ttgggccaac aactcacgca agtgggcgtg
6181 atgaatatca acggagccat cttcctcttc ctgaccaaca tgacctttca aaacgtcttt
6241 gccacgataa atgtaagtct tgtttagaat acatttgcat attaataatt tactaacttt
6301 ctaatgaatc gattcgattt aggtgttcac ctcagagctg ccagttttta tgagggaggc
6361 ccgaagtcga ctttatcgct gtgacacata ctttctgggc aaaacgattg ccgaattacc
6421 gctttttctc acagtgccac tggtcttcac ggcgattgcc tatccgatga tcggactgcg
6481 ggccggagtg ctgcacttct tcaactgcct ggcgctggtc actctggtgg ccaatgtgtc
6541 aacgtccttc ggatatctaa tatcctgcgc cagctcctcg acctcgatgg cgctgtctgt
6601 gggtccgccg gttatcatac cattcctgct ctttggcggc ttcttcttga actcgggctc
6661 ggtgccagta tacctcaaat ggttgtcgta cctctcatgg ttccgttacg ccaacgaggg
6721 tctgctgatt aaccaatggg cggacgtgga gccgggcgaa attagctgca catcgtcgaa
6781 caccacgtgc cccagttcgg gcaaggtcat cctggagacg cttaacttct ccgccgccga
6841 tctgccgctg gactacgtgg gtctggccat tctcatcgtg agcttccggg tgctcgcata
6901 tctggctcta agacttcggg cccgacgcaa ggagtagccg acatatatcc gaaataactg
6961 cttgtttttt tttttaccat tattaccatc gtgtttactg tttattgccc cctcaaaaag
7021 ctaatgtaat tatatttgtg ccaataaaaa caagatatga cctatagaat acaagtattt
7081 ccccttcgaa catccccaca agtagacttt ggatttgtct tctaaccaaa agacttacac
7141 acctgcatac cttacatcaa aaactcgttt atcgctacat aaaacaccgg gatatatttt
7201 ttatatacat acttttcaaa tcgcgcgccc tcttcataat tcacctccac cacaccacgt
7261 ttcgtagttg ctctttcgct gtctcccacc cgctctccgc aacacattca ccttttgttc
7321 gacgaccttg gagcgactgt cgttagttcc gcgcgattcg gttcgctcaa atggttccga
7381 gtggttcatt tcgtctcaat agaaattagt aataaatatt tgtatgtaca atttatttgc
7441 tccaatatat ttgtatatat ttccctcaca gctatattta ttctaattta atattatgac
7501 tttttaaggt aattttttgt gacctgttcg gagtgattag cgttacaatt tgaactgaaa
7561 gtgacatcca gtgtttgttc cttgtgtaga tgcatctcaa aaaaatggtg ggcataatag
7621 tgttgtttat atatatcaaa aataacaact ataataataa gaatacattt aatttagaaa
7681 atgcttggat ttcactggaa ctagaattaa ttcggctgct gctctaaacg acgcatttcg
7741 tactccaaag tacgaatttt ttccctcaag ctcttatttt cattaaacaa tgaacaggac
7801 ctaacgcaca gtcacgttat tgtttacata aatgattttt tttactattc aaacttactc
7861 tgtttgtgta ctcccactgg tatagccttc ttttatcttt tctggttcag gctctatcac
7921 tttactaggt acggcatctg cgttgagtcg cctcctttta aatgtctgac cttttgcagg
7981 tgcagccttc cactgcgaat ctttaaagtg ggtatcacaa atttgggagt tttcaccaag
8041 gctgcaccca aggctctgct cccacaattt tctcttaata gcacacttcg gcacgtgaat
8101 taattttact ccagtcacag ctttgcagca aaatttgcaa tatttcattt ttttttattc
8161 cacgtaaggg ttaatgtttt caaaaaaaaa ttcgtccgca cacaaccttt cctctcaaca
8221 agcaaacgtg cactgaattt aagtgtatac ttcggtaagc ttcggctatc gacgggacca
8281 ccttatgtta tttcatcatg ggccagaccc acgtagtcca gcggcagatc ggcggcggag
8341 aagttaagcg tctccaggat gaccttgccc gaactggggc acgtggtgtt cgacgatgtg
8401 cagctaattt cgcccggctc cacgtccgcc cattggttaa tcagcagacc ctcgttggcg
8461 taacggaacc atgagaggta cgacaaccat ttgaggtata ctggcaccga gcccgagttc
8521 aagaagaagg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
8581 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
8641 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
8701 ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg
8761 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
8821 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
8881 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
8941 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
9001 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
9061 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
9121 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
9181 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
9241 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
9301 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
9361 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
9421 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
9481 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
9541 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
9601 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
9661 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
9721 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
9781 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
9841 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
9901 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
9961 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
10021 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
10081 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
10141 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
10201 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
10261 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
10321 tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
10381 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
10441 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
10501 tatgcggcat cagagcagat tgtactgaga gtgcaccata tggggtgcca gggcgtgccc
10561 ttgggctccc cgggcgcgta tatgcggtgt gaaataccgc accgaatcgc gcggaactaa
10621 cgacagtcgc tccaaggtcg tcgaacaaaa ggtgaatgtg ttgcggagag cgggtgggag
10681 acagcgaaag agcaactacg aaacgtggtg tggtggaggt gaattatgaa gagggcgcgc
10741 gatttgaaaa gtatgtatat aaaaaatata tcccggtgtt ttatgtagcg ataaacgagt
10801 ttttgatgta aggtatgcag gtgtgtaagt cttttggtta gaagacaaat ccaaagtcta
10861 cttgtgggga tgttcgaagg ggaaatactt gtattctata ggtcatatct tgtttttatt
10921 ggcacaaata taattacatt agctttttga gggggcaata aacagtaaac acgatggtaa
10981 taatggtaaa aaaaaaaaca agcagttatt tcggatatat gtcggctact ccttgcgtcg
11041 ggcccgaagt cttagagcca gatatgcgag cacccggaag ctcacgatga gaatggccag
11101 accatgatga aataacataa ggtggtcccg tcggcaagag acatccactt aacgtatgct
11161 tgcaataagt gcgagtgaaa ggaatagtat tctgagtgtc gtattgagtc tgagtgagac
11221 agcgatatga ttgttgatta acccttagca tgtccgtggg gtttgaatta actcataata
11281 ttaattagac gaaattattt ttaaagtttt atttttaata atttgcgagt acgca
//
| {
"Vector backbone": "pCASPER2",
"Backbone size w/o insert (bp)": "10063",
"Vector type": "Insect Expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "XL1 Blue",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Efficient chromosomal gene modification with CRISPR/cas9 and PCR-based homologous recombination donors in cultured Drosophila cells",
"Publication": "Nucleic Acids Res. 2014 Apr 19.",
"DOI": "10.1093/nar/gku289",
"Authors": "Bottcher R, Hollmann M, Merk K, Nitschko V, Obermaier C, Philippou-Massier J, Wieland I, Gaul U, Forstemann K",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/24748663",
"PubMed ID": "PubMed 24748663"
} | {
"Gene/Insert name": "FLP recombinase",
"Species": "S. cerevisiae (budding yeast)",
"Insert Size (bp)": "1272",
"Mutation": "",
"Entrez Gene": "flp",
"Promoter": "ubiquitin",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "BamHI (not destroyed)",
"3′ cloning site": "NotI (not destroyed)",
"5′ sequencing primer": "catttgatttcgtactaattttccac",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"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": ""
} |
I-ATGC | 43020 | Ding et al Cell Stem Cell. 2012 Dec 12. pii: S1934-5909(12)00645-5. doi: 10.1016/j.stem.2012.11.011. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/59941/cb21ef27-4ff8-4d9e-b9c5-4bbbbdd1e589/addgene-plasmid-43020-sequence-59941.gbk | partial | LOCUS sequence-59941-e 871 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 871)
AUTHORS Ding Q, Lee YK, Schaefer EA, Peters DT, Veres A, Kim K, Kuperwasser
N, Motola DL, Meissner TB, Hendriks WT, Trevisan M, Gupta RM, Moisan
A, Banks E, Friesen M, Schinzel RT, Xia F, Tang A, Xia Y, Figueroa
E, Wann A, Ahfeldt T, Daheron L, Zhang F, Rubin LL, Peng LF, Chung
RT, Musunuru K, Cowan CA
TITLE A TALEN Genome-Editing System for Generating Human Stem Cell-Based
Disease Models.
JOURNAL Cell Stem Cell. 2012 Dec 12. pii: S1934-5909(12)00645-5. doi:
10.1016/j.stem.2012.11.011.
PUBMED 23246482
REFERENCE 2 (bases 1 to 871)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Cell Stem
Cell. 2012 Dec 12. pii: S1934-5909(12)00645-5. doi:
10.1016/j.stem.2012.11.011."
FEATURES Location/Qualifiers
source 1..871
/mol_type="genomic DNA"
/organism="unspecified"
promoter complement(621..639)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(622..639)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(657..673)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(657..673)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(670..692)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 681..697
/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(705..735)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 750..771
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
ORIGIN
1 ctatagggcg aattgggccc tctagatgca tgctcgagcg gccgccagtg tgatggatat
61 ctgcagaatt cgcccttata tagatgccgt cctagcgcgt ctcctgaccc cagagcaggt
121 cgtggcaatc gcctccaaca ttggcgggaa acaggcactc gagactgtcc agcgcctgct
181 tcccgtgctg tgccaagcgc acggactcac cccagagcag gtcgtggcca ttgcctcgaa
241 tggagggggc aaacaggcgt tggaaaccgt acaacgattg ctgccggtgc tgtgccaagc
301 gcacggcctc accccagagc aggtcgtggc aatcgcgagc aataacggcg gaaaacaggc
361 tttggaaacg gtgcagaggc tccttccagt gctgtgccaa gcgcacggat taaccccaga
421 gcaggtcgtg gcgatcgcaa gccacgacgg aggaaagcaa gccttggaaa cagtacagag
481 gctgttgcct gtgctgtgcc aagcgcacgg cttaagagac gtgggcacag gaaagatact
541 taagggcgaa ttccagcaca ctggcggccg ttactagtgg atccgagctc ggtaccaagc
601 ttgatgcata gcttgagtat tctatagtgt cacctaaata gcttggcgta atcatggtca
661 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
721 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
781 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
841 caacgcgcgg ggagaggcgg tttgcgtatt g
//
| {
"Vector backbone": "pCRII-TOPO",
"Backbone size w/o insert (bp)": "3973",
"Vector type": "Bacterial Cloning",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin and Kanamycin, 100 & 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": "A TALEN Genome-Editing System for Generating Human Stem Cell-Based Disease Models",
"Publication": "Cell Stem Cell. 2012 Dec 12. pii: S1934-5909(12)00645-5. doi: 10.1016/j.stem.2012.11.011.",
"DOI": "10.1016/j.stem.2012.11.011.",
"Authors": "Ding Q, Lee YK, Schaefer EA, Peters DT, Veres A, Kim K, Kuperwasser N, Motola DL, Meissner TB, Hendriks WT, Trevisan M, Gupta RM, Moisan A, Banks E, Friesen M, Schinzel RT, Xia F, Tang A, Xia Y, Figueroa E, Wann A, Ahfeldt T, Daheron L, Zhang F, Rubin LL, Peng LF, Chung RT, Musunuru K, Cowan CA /jstem",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/23246482",
"PubMed ID": "PubMed 23246482"
} | {
"Gene/Insert name": "I-ATGC",
"Species": "Synthetic",
"Insert Size (bp)": "424",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "TOPO Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "T7",
"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": ""
} |
|||
pBlastR-5xQUAS-EBFP2-MiniWhite | 165924 | Blasticidin resistant 5xQUAS EBFP2 Mini-White CDS response vector. Contains an Alpha level GB2.0 entry point for addition of custom assemblies utilizing blue-white bacterial colony screening | Koen Venken | Matinyan et al Cell Rep. 2021 Sep 14;36(11):109700. doi: 10.1016/j.celrep.2021.109700. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/326303/3f66dde2-45ab-40e0-8377-82f4d5e52067/addgene-plasmid-165924-sequence-326303.gbk | full | LOCUS sequence-326303- 9324 bp DNA circular SYN 26-AUG-2024
DEFINITION Blasticidin resistant 5xQUAS EBFP2 Mini-White CDS response vector.
Contains an Alpha level GB2.0 entry point for addition of custom
assemblies utilizing blue-white bacterial colony screening.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9324)
AUTHORS Matinyan N, Karkhanis MS, Gonzalez Y, Jain A, Saltzman A,
Malovannaya A, Sarrion-Perdigones A, Dierick HA, Venken KJT
TITLE Multiplexed drug-based selection and counterselection genetic
manipulations in Drosophila.
JOURNAL Cell Rep. 2021 Sep 14;36(11):109700. doi:
10.1016/j.celrep.2021.109700.
PUBMED 34525356
REFERENCE 2 (bases 1 to 9324)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Cell Rep.";
date: "2021-09-14"; volume: "36(11):109700. doi"; pages: "
10.1016/j.celrep.2021.109700"
FEATURES Location/Qualifiers
source 1..9324
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin complement(950..1538)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1030..1049)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS 2006..2797
/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 2852..3307
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2939..2958)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3149..3170
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 3331..3378
/label=EM7 promoter
/note="synthetic bacterial promoter "
promoter 3432..3450
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 3432..3449
/label=SP6
/note="SP6 promoter, forward primer"
misc_feature 3457..3513
/label=MCS
/note="pUC18/19 multiple cloning site"
primer_bind complement(3515..3534)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(3516..3534)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(3541..3558)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(3541..3557)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3550..3572)
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind complement(3766..3785)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
protein_bind 3842..3911
/label=attB
/bound_moiety="phage phi-C31 integrase"
/note="attB site for the phi-C31 integrase (Groth et al.,
2000)"
primer_bind 4338..4359
/label=pCasper-hs
/note="Drosophila Hsp70 promoter, forward primer"
polyA_signal 5943..6077
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5967..5986)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 6021..6040
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind complement(6165..6183)
/label=pCasper-F
/note="Drosophila mini-white gene, reverse primer"
CDS join(6635..6706,7108..7381,7456..8110,8172..8487,8691..8822,
8893..9324,1..183)
/codon_start=1
/gene="white"
/product="Drosophila white gene eye color pigment"
/label=mini-white
/note="This is a modified version of the white gene lacking
part of the first intron."
/translation="MGQEDQELLIRGGSKHPSAEHLNNGDSGAASQSCINQGFGQAKNY
GTLRPPSPPEDSGSGSGQLAENLTYAWHNMDIFGAVNQPGSGWRQLVNRTRGLFCNERH
IPAPRKHLLKNVCGVAYPGELLAVMGSSGAGKTTLLNALAFRSPQGIQVSPSGMRLLNG
QPVDAKEMQARCAYVQQDDLFIGSLTAREHLIFQAMVRMPRHLTYRQRVARVDQVIQEL
SLSKCQHTIIGVPGRVKGLSGGERKRLAFASEALTDPPLLICDEPTSGLDSFTAHSVVQ
VLKKLSQKGKTVILTIHQPSSELFELFDKILLMAEGRVAFLGTPSEAVDFFSYVGAQCP
TNYNPADFYVQVLAVVPGREIESRDRIAKICDNFAISKVARDMEQLLATKNLEKPLEQP
ENGYTYKATWFMQFRAVLWRSWLSVLKEPLLVKVRLIQTTMVAILIGLIFLGQQLTQVG
VMNINGAIFLFLTNMTFQNVFATINVFTSELPVFMREARSRLYRCDTYFLGKTIAELPL
FLTVPLVFTAIAYPMIGLRAGVLHFFNCLALVTLVANVSTSFGYLISCASSSTSMALSV
GPPVIIPFLLFGGFFLNSGSVPVYLKWLSYLSWFRYANEGLLINQWADVEPGEISCTSS
NTTCPSSGKVILETLNFSAADLPLDYVGLAILIVSFRVLAYLALRLRARRKE"
ORIGIN
1 ggcgaaatta gctgcacatc gtcgaacacc acgtgcccca gttcgggcaa ggtcatcctg
61 gagacactta acttctccgc cgccgatctg ccgctggact acgtgggtct ggccattctc
121 atcgtgagct tccgggtgct cgcatatctg gctctaagac ttcgggcccg acgcaaggag
181 tagccgacat atatccgaaa taactgcttg tttttttttt ttaccattat taccatcgtg
241 tttactgttt attgccccct caaaaagcta atgtaattat atttgtgcca ataaaaacaa
301 gatatgacct atagaataca agtatttccc cttcgaacat ccccacaagt agactttgga
361 tttgtcttct aaccaaaaga cttacacacc tgcatacctt acatcaaaaa ctcgtttatc
421 gctacataaa acaccgggat atatttttta tatacatact tttcaaatcg cgcgccctct
481 tcataattca cctccaccac accacgtttc gtagttgctc tttcgctgtc tcccacccgc
541 tctccgcaac acattcacct tttgttcgac gaccttggag cgactgtcgt tagttccgcg
601 cgattcggtt cgctcaaatg gttccgagtg gttcatttcg gctcaataga aattagtaat
661 aaatatttgt atgtacaatt tatttgctcc aatatatttg tatatatttc cctcacagct
721 atatttattc taatttaata ttatgacttt ttaaggtaat tttttgtgac ctgttcggag
781 tgattagcgt tacaatttga actgaaagtg acatccagtg tttgttcctt gtgtagatgc
841 atcgcttgac acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
901 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
961 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
1021 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
1081 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
1141 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
1201 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
1261 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
1321 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
1381 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
1441 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
1501 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
1561 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
1621 caaaaaggat cttcacctag atccttttgt gaccagccag gacagaaatg cctcgacttc
1681 gctgctaccc aaggttgccg ggtgacgcac accgtggaaa cggatgaagg cacgaaccca
1741 gtggacataa gcctgttcgg ttcgtaagct gtaatgcaag tagcgtatgc gctcacgcaa
1801 ctggtccaga accttgaccg aacgcagcgg tggtaacggc gcagtggcgg ttttcatggc
1861 ttgttatgac tgtttttttg gggtacagtc tatgcctcgg gcatccaagc agcaagcgcg
1921 ttacgccgtg ggtcgatgtt tgatgttatg gagcagcaac gatgttacgc agcagggcag
1981 tcgccctaaa acaaagttaa acattatgag ggaagcggtg atcgccgaag tatcgactca
2041 actatcagag gtagttggcg tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca
2101 tttgtacggc tccgcagtgg atggcggcct gaagccacac agtgatattg atttgctggt
2161 tacggtgacc gtaaggcttg atgaaacaac gcggcgagct ttgatcaacg accttttgga
2221 aacttcggct tcccctggag agagcgagat tctccgcgct gtagaagtca ccattgttgt
2281 gcacgacgac atcattccgt ggcgttatcc agctaagcgc gaactgcaat ttggagaatg
2341 gcagcgcaat gacattcttg caggtatctt cgagccagcc acgatcgaca ttgatctggc
2401 tatcttgctg acaaaagcaa gagaacatag cgttgccttg gtaggtccag cggcggagga
2461 actctttgat ccggttcctg aacaggatct atttgaggcg ctaaatgaaa ccttaacgct
2521 atggaactcg ccgcccgact gggctggcga tgagcgaaat gtagtgctta cgttgtcccg
2581 catttggtac agcgcagtaa ccggcaaaat cgcgccgaag gatgtagctg ccgactgggc
2641 aatggagcgc ctgccggccc agtatcagcc cgtcatactt gaagctagac aggcttatct
2701 tggacaagaa gaagatcgtt tggcctcgcg cgcagatcag ttggaagaat ttgtccacta
2761 cgtgaaaggc gagatcacca aggtagtcgg caaataatca ctagaccaat gttacacata
2821 tatactttag attgatttaa aacttcatgt tacgcgccct gtagcggcgc attaagcgcg
2881 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
2941 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
3001 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
3061 cttgattagg gtgatggttc acgtagtggg ccagcgccct gatagacggt ttttcgccct
3121 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
3181 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
3241 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
3301 acaattttaa cggatcagcg agagacccct gttgacaatt aatcatcggc atagtatatc
3361 ggcatagtat aatacgacaa ggtgaggaac taaacccagg aggcagatca tgaccatgat
3421 tacgccaagc tatttaggtg acactataga atactcaagc ttgcatgcct gcaggtcgac
3481 tctagaggat ccccgggtac cgagctcgaa ttcgccctat agtgagtcgt attacaattc
3541 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
3601 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
3661 cccttcccaa cagttgcgca gcctgaatgg cgaatgacgc ctgatgcggt attttctcct
3721 tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga
3781 tgccgcatag ggtctctctc cggatcccag ggaggaattg gtcgacgatg taggtcactg
3841 tctcgaagcc gcggtgcggg tgccagggcg tgcccttggg ctccccgggc gcgtactcca
3901 cctcacccat ctggtccatc atgatgcgct gtcaggagct agaatcccaa aacaaactgg
3961 ttattgtggt aggtcatttg tttggcagaa agaaaactcg agaaatttct ctggccgtta
4021 ttcgttattc tctcttttct ttttgggtcg ctccctctct gcactaatgc tctctcactc
4081 tgtcacacag taaacggcat actgctctcg ttggttcgag agagcgcgcc tcgaatgttc
4141 gcgaaaagag cgccggagta taaatagagg cgcttcgtct acggagcgac aattcaattc
4201 aaacaagcaa agtgaacacg tcgctaagcg aaagctaagc aaataaacaa gcgcagctga
4261 acaagctaaa caatctgcag taaagtgcaa gttaaagtga atcaattaaa agtaaccagc
4321 aaccaagtaa atcaactgca actactgaaa tctgccaaga agtaattatt gaatacaaga
4381 agagaacact caatactcga atccatgtgg gagtttattc ttgacacaga tatttatgat
4441 ataataactg agtaagctta acataaggag gaaaacaaaa tgaaaacctt caacatcagc
4501 cagcaggacc tggaactggt ggaggtggct accgagaaga ttacgatgct ctacgaggat
4561 aacaagcacc acgtgggagc tgccattcgc accaagacgg gcgagattat cagtgccgtg
4621 cacattgagg cctacatcgg acgcgtgacc gtgtgtgccg aggccattgc cattggcagt
4681 gccgtgagca acggccagaa ggattttgat accattgtgg ccgtgcgtca cccatactcc
4741 gatgaggtgg accgtagcat ccgtgtcgtg tccccgtgtg gaatgtgccg tgagctgatt
4801 agcgattacg cccccgattg cttcgtgctg attgagatga acggcaagct ggtcaagacc
4861 accattgagg aactgattcc cctgaagtat acccgtaact aagcttggca ataaaaagac
4921 agaataaaac gcacgggtgt tgggtcgttt gttccgctgt caggaggggt aatcgcttat
4981 cctcggataa acaattatcc tcacgggtaa tcgcttatcc gggtaatcgc ttatcctcgg
5041 gtaatcgctt atcctttccc gagctcgccc ggggatcgag cgcagcggta taaaagggcg
5101 cggggtggct gagagcatca gttgtgaatg aatgttcgag ccgagcagac gtgccgctgc
5161 cttcgttaat atcctttgaa taagccaact ttgaatcaca agacgcatac caaaccaaat
5221 ggtgtccaag ggcgaggagc tgttcaccgg cgtggtgccc atcctggtgg agctggatgg
5281 cgacgtgaac ggccacaagt tcagcgtgcg cggcgagggc gagggcgacg ccaccaacgg
5341 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
5401 ggtgaccacc ctgagccacg gcgtgcagtg cttcgcccgc taccccgatc acatgaagca
5461 gcacgatttc ttcaagagcg ccatgcccga gggctacgtg caggagcgca ccattttctt
5521 caatgacgac ggcacctaca agacccgcgc cgaggtgaag ttcgagggcg ataccctggt
5581 gaaccgcatc gagctgaagg gcgtggattt caaggaggat ggcaacatcc tgggccacaa
5641 gctggagtac aacttcaaca gccacaacat ctacatcatg gccgtgaagc agaagaacgg
5701 catcaaggtc aacttcaaga tccgccacaa cgtggaggat ggctcagtgc agctggccga
5761 tcactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg atagccacta
5821 cctgagcacc cagagcgtgc tgtccaagga tcccaacgag aagcgcgatc acatggtgct
5881 gctggagttc cgcaccgccg ccggcatcac cctgggcatg gatgagctgt acaagtaagc
5941 ttgatccaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
6001 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
6061 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
6121 aggtgtggga ggttttttcg ctgtcaggag gaattctagt atgtatgtaa gttaataaaa
6181 cccatttttg cggaaagtag ataaaaaaaa catttttttt ttttactgca ctggatatca
6241 ttgaacttat ctgatcagtt ttaaatttac ttcgatccaa gggtatttga tgtaccaggt
6301 tctttcgatt acctctcact caaaatgaca ttccactcaa agtcagcgct gtttgcctcc
6361 ttctctgtcc acagaaatat cgccgtcact ttcgccgctg cgtccgctat ctctttcgcc
6421 accgtttgta gcgttacgta gcgtcaatgt ccgccttcag ttgcattttg tcagcggttt
6481 cgtgacgaag ctccaagcgg tttacgccat caattaaaca caaagtgctg tgccaaaact
6541 cctctcgctt cttatttttg tttgtttttt gagtgattgg ggtggtgatt ggttttgggt
6601 gggtaagcag gggaaagtgt gaaaaatccc ggcaatgggc caagaggatc aggagctatt
6661 aattcgcgga ggcagcaaac acccatctgc cgagcatctg aacaatgtga gtagtacatg
6721 tgcatacatc ttaagttcac ttgatctata ggaactgcga ttgcaacatc aaattgtctg
6781 cggcgtgaga actgcgaccc acaaaaatcc caaaccgcaa ttgcacaaac aaatagtgac
6841 acgaaacaga ttattctggt agctgttctc gctatataag acaatttttg agatcatatc
6901 atgatcaaga catctaaagg cattcatttt cgactatatt cttttttaca aaaaatataa
6961 caaccagata ttttaagctg atcctagatg cacaaaaaat aaataaaagt ataaacctac
7021 ttcgtaggat acttcggggt actttttgtt cggggttaga tgagcataac gcttgtagtt
7081 gatatttgag atcccctatc attgcagggt gacagcggag cggcttcgca gagctgcatt
7141 aaccagggct tcgggcaggc caaaaactac ggcacgctcc ggccacccag tccgccggag
7201 gactccggtt cagggagcgg ccaactagcc gagaacctca cctatgcctg gcacaatatg
7261 gacatctttg gggcggtcaa tcagccgggc tccggatggc ggcagctggt caaccggaca
7321 cgcggactat tctgcaacga gcgacacata ccggcgccca ggaaacattt gctcaagaac
7381 ggtgagtttc tattcgcagt cggctgatct gtgtgaaatc ttaataaagg gtccaattac
7441 caatttgaaa ctcagtttgc ggcgtggcct atccgggcga acttttggcc gtgatgggca
7501 gttccggtgc cggaaagacg accctgctga atgcccttgc ctttcgatcg ccgcagggca
7561 tccaagtatc gccatccggg atgcgactgc tcaatggcca acctgtggac gccaaggaga
7621 tgcaggccag gtgcgcctat gtccagcagg atgacctctt tatcggctcc ctaacggcca
7681 gggaacacct gattttccaa gccatggtgc ggatgccacg acatctgacc tatcggcagc
7741 gagtggcccg cgtggatcag gtgatccagg agctttcgct cagcaaatgt cagcacacga
7801 tcatcggtgt gcccggcagg gtgaaaggtc tgtccggcgg agaaaggaag cgtctggcat
7861 tcgcctccga ggctctaacc gatccgccgc ttctgatctg cgatgagccc acctccggac
7921 tggactcctt taccgcccac agcgtcgtcc aggtgctgaa gaagctgtcg cagaagggca
7981 agaccgtcat cctgaccatt catcagccgt cttccgagct gtttgagctc tttgacaaga
8041 tccttctgat ggccgagggc agggtagctt tcttgggcac tcccagcgaa gccgtcgact
8101 tcttttccta gtgagttcga tgtgtttatt aagggtatct agtattacat aacatctcaa
8161 ctcctatcca gcgtgggtgc ccagtgtcct accaactaca atccggcgga cttttacgta
8221 caggtgttgg ccgttgtgcc cggacgggag atcgagtccc gtgatcggat cgccaagata
8281 tgcgacaatt ttgccattag caaagtagcc cgggatatgg agcagttgtt ggccaccaaa
8341 aatctggaga agccactgga gcagccggag aatgggtaca cctacaaggc cacctggttc
8401 atgcagttcc gggcggtcct gtggcgatcc tggctgtcgg tgctcaagga accactcctc
8461 gtaaaagtgc gacttattca gacaacggtg agtggttcca gtggaaacaa atgatataac
8521 gcttacaatt cttggaaaca aattcgctag attttagata gaattgcctg attccacacc
8581 cttcttagtt tttttcaatg agatgtatag tttatagttt tgcagaagat aaataaattt
8641 catttaactc gcgaatatta atgagatgcg agtaacattt taatttgcag atggttgcca
8701 tcttgattgg cctcatcttt ttgggccaac aactcacgca agtgggtgtg atgaatatca
8761 acggagccat cttcctcttc ctgaccaaca tgacctttca aaacgtcttt gccacgataa
8821 atgtaagtca tgtttagaat acatttgcat ttcaataatt tactaacttt ctaatgaatc
8881 gattcgattt aggtgttcac ctcagagctg ccagttttta tgagggaggc ccgaagtcga
8941 ctttatcgct gtgacacata ctttctgggc aaaacgattg ccgaattgcc gctttttctc
9001 acagtgccac tggtcttcac ggcgattgcc tatccgatga tcggactgcg ggccggagtg
9061 ctgcacttct tcaactgcct ggcgctggtc actctggtgg ccaatgtgtc aacgtccttc
9121 ggatatctaa tatcctgcgc cagctcctcg acctcgatgg cgctgtctgt gggtccgccg
9181 gttatcatac cattcctgct ctttggcggc ttcttcttga actcgggctc ggtgccagta
9241 tacctcaaat ggttgtcgta cctctcatgg ttccgttacg ccaacgaggg tctgctgatt
9301 aaccaatggg cggacgtgga gccg
//
| {
"Vector backbone": "pColE1_AlphaOmega",
"Backbone size w/o insert (bp)": "2967",
"Vector type": "Synthetic Biology",
"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": "Multiplexed drug-based selection and counterselection genetic manipulations in Drosophila",
"Publication": "Cell Rep. 2021 Sep 14;36(11):109700. doi: 10.1016/j.celrep.2021.109700.",
"DOI": "10.1016/j.celrep.2021.109700.",
"Authors": "Matinyan N, Karkhanis MS, Gonzalez Y, Jain A, Saltzman A, Malovannaya A, Sarrion-Perdigones A, Dierick HA, Venken KJT /jcelrep",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/34525356",
"PubMed ID": "PubMed 34525356"
} | {
"Gene/Insert name": "Selectable QUAS Response Vector",
"Species": "Synthetic",
"Insert Size (bp)": "6357",
"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": ""
} |
|
pFA6a-SRS:hphMX4 | 134441 | PCR-based C-terminal SRS tag | Robert Tomko | Nemec et al Yeast. 2020 May 13. doi: 10.1002/yea.3471. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/271056/9a1d5286-e91f-4106-8b96-5dbecced7e1d/addgene-plasmid-134441-sequence-271056.gbk | full | LOCUS sequence-271056- 4413 bp DNA circular SYN 26-AUG-2024
DEFINITION PCR-based C-terminal SRS tag.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4413)
AUTHORS Nemec AA, Tomko RJ Jr
TITLE A suite of PCR-based peptide tagging plasmids to permit
epitope-targeted enzymatic functionalization of yeast proteins.
JOURNAL Yeast. 2020 May 13. doi: 10.1002/yea.3471.
PUBMED 32401365
REFERENCE 2 (bases 1 to 4413)
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. 2020
May 13. doi: 10.1002/yea.3471."
FEATURES Location/Qualifiers
source 1..4413
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 237..1265
/codon_start=1
/product="hygromycin B phosphotransferase"
/label=HygR
/note="confers resistance to hygromycin"
/translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
KE"
CDS 237..1265
/codon_start=1
/gene="aph(4)-Ia"
/product="aminoglycoside phosphotransferase from E. coli"
/label=HygR
/note="confers resistance to hygromycin"
/translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
KE"
terminator 1271..1468
/label=TEF terminator
/note="Ashbya gossypii TEF terminator"
primer_bind complement(1575..1594)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(1576..1594)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(1681..1698)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(1852..2440)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1932..1951)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(2611..3471)
/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 3234..3253
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(3472..3576)
/gene="bla"
/label=AmpR promoter
primer_bind 3644..3662
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(3700..3722)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 3822..3841
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
promoter 3922..3940
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 3922..3939
/label=SP6
/note="SP6 promoter, forward primer"
terminator 4044..4231
/gene="S. cerevisiae ADH1"
/label=ADH1 terminator
/note="transcription terminator for the S. cerevisiae
alcohol dehydrogenase 1 (ADH1) gene"
primer_bind complement(4099..4123)
/label=pHybLex-R
/note="ADH terminator, reverse primer"
gene join(4306..4413,1..1468)
/label=hphMX6
/note="yeast selectable marker conferring hygromycin
resistance"
promoter join(4306..4413,1..236)
/label=TEF promoter
/note="Ashbya gossypii TEF promoter"
ORIGIN
1 tttgcatcca tacattttga tggccgcacg gcgcgaagca aaaattacgg ctcctcgctg
61 cggacctgcg agcagggaaa cgctcccctc acagacgcgt tgaattgtcc ccacgccgcg
121 cccctgtaga gaaatataaa aggttaggat ttgccactga ggttcttctt tcatatactt
181 ccttttaaaa tcttgctagg atacagttct cacatcacat ccgaacataa acaaccatgg
241 gtaaaaagcc tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa aagttcgaca
301 gcgtctccga cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg
361 taggagggcg tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc
421 gttatgttta tcggcacttt gcatcggccg cgctcccgat tccggaagtg cttgacattg
481 gggaattcag cgagagcctg acctattgca tctcccgccg tgcacagggt gtcacgttgc
541 aagacctgcc tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag gccatggatg
601 cgatcgctgc ggccgatctt agccagacga gcgggttcgg cccattcgga ccgcaaggaa
661 tcggtcaata cactacatgg cgtgatttca tatgcgcgat tgctgatccc catgtgtatc
721 actggcaaac tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc
781 tgatgctttg ggccgaggac tgccccgaag tccggcacct cgtgcacgcg gatttcggct
841 ccaacaatgt cctgacggac aatggccgca taacagcggt cattgactgg agcgaggcga
901 tgttcgggga ttcccaatac gaggtcgcca acatcttctt ctggaggccg tggttggctt
961 gtatggagca gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca ggatcgccgc
1021 ggctccgggc gtatatgctc cgcattggtc ttgaccaact ctatcagagc ttggttgacg
1081 gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag
1141 ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg accgatggct
1201 gtgtagaagt actcgccgat agtggaaacc gacgccccag cactcgtccg agggcaaagg
1261 aataatcagt actgacaata aaaagattct tgttttcaag aacttgtcat ttgtatagtt
1321 tttttatatt gtagttgttc tattttaatc aaatgttagc gtgatttata ttttttttcg
1381 cctcgacatc atctgcccag atgcgaagtt aagtgcgcag aaagtaatat catgcgtcaa
1441 tcgtatgtga atgctggtcg ctatactgct gtcgattcga tactaacgcc gccatccagt
1501 gtcgaaaacg agctcgaatt catcgatgat atcagatcca ctagtggcct atgcggccgc
1561 ggatctgccg gtctccctat agtgagtcgt attaatttcg ataagccagg ttaacctgca
1621 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
1681 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
1741 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
1801 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
1861 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
1921 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
1981 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
2041 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
2101 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
2161 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
2221 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
2281 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
2341 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
2401 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
2461 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
2521 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
2581 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
2641 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
2701 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
2761 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
2821 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
2881 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
2941 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
3001 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
3061 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
3121 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
3181 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
3241 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
3301 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
3361 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
3421 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
3481 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
3541 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
3601 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
3661 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
3721 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
3781 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
3841 actgagagtg caccatatgg acatattgtc gttagaacgc ggctacaatt aatacataac
3901 cttatgtatc atacacatac gatttaggtg acactataga acgcggccgc cagctgaagc
3961 ttcgtacgct gcaggtcgac ggatccccgg gttaattaac tgcccgaaac cggtggtggc
4021 taaggcgcgc cacttctaaa taagcgaatt tcttatgatt tatgattttt attattaaat
4081 aagttataaa aaaaataagt gtatacaaat tttaaagtga ctcttaggtt ttaaaacgaa
4141 aattcttatt cttgagtaac tctttcctgt aggtcaggtt gctttctcag gtatagtatg
4201 aggtcgctct tattgaccac acctctaccg gcagatccgc tagggataac agggtaatat
4261 agatctgttt agcttgcctc gtccccgccg ggtcacccgg ccagcgacat ggaggcccag
4321 aataccctcc ttgacagtct tgacgtgcgc agctcagggg catgatgtga ctgtcgcccg
4381 tacatttagc ccatacatcc ccatgtataa tca
//
| {
"Vector backbone": "pFA6a-hphMX4",
"Backbone size w/o insert (bp)": "",
"Vector type": "PCR-based tagging in yeast",
"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": "Industry MTA"
} | {
"Title": "A suite of PCR-based peptide tagging plasmids to permit epitope-targeted enzymatic functionalization of yeast proteins",
"Publication": "Yeast. 2020 May 13. doi: 10.1002/yea.3471.",
"DOI": "10.1002/yea.3471.",
"Authors": "Nemec AA, Tomko RJ Jr /yea",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/32401365",
"PubMed ID": "PubMed 32401365"
} | {
"Gene/Insert name": "SRS",
"Species": "Synthetic",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "PacI (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": ""
} |
|
pZC04 (pFA6a-PSc-6xGly-PrtA-His3MX) | 44066 | (Empty Backbone) | Tim Formosa | Formosa lab yeast 3' tagging constructs (unpublished) | https://media.addgene.org/snapgene-media/v2.0.0/sequences/61687/b40a09c0-163a-47db-a58e-e3cfe6063709/addgene-plasmid-44066-sequence-61687.gbk | full | LOCUS sequence-61687-e 4522 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 4522)
TITLE Formosa lab yeast 3' tagging constructs
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 4522)
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..4522
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter complement(199..303)
/gene="bla"
/label=AmpR promoter
primer_bind 371..389
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(427..449)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 549..568
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
promoter 649..667
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 649..666
/label=SP6
/note="SP6 promoter, forward primer"
CDS 728..751
/codon_start=1
/product="recognition and cleavage site for human
rhinovirus 3C and PreScission proteases"
/label=HRV 3C site
/translation="LEVLFQGP"
CDS 809..982
/codon_start=1
/product="IgG-binding unit of Staphylococcus aureus protein
A"
/label=ProtA
/translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
AEAKKLNDAQAPK"
CDS 983..1156
/codon_start=1
/product="IgG-binding unit of Staphylococcus aureus protein
A"
/label=ProtA
/translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
AEAKKLNGAQAPK"
terminator 1215..1402
/gene="S. cerevisiae ADH1"
/label=ADH1 terminator
/note="transcription terminator for the S. cerevisiae
alcohol dehydrogenase 1 (ADH1) gene"
primer_bind complement(1270..1294)
/label=pHybLex-R
/note="ADH terminator, reverse primer"
promoter 1477..1820
/label=TEF promoter
/note="Ashbya gossypii TEF promoter"
CDS 1821..2519
/codon_start=1
/product="S. kluyveri imidazoleglycerol phosphate
dehydratase"
/label=k-HIS3
/translation="MAEPAQKKQKQTVQERKAFISRITNETKIQIAISLNGGYIQIKDS
ILPAKKDDDVASQATQSQVIDIHTGVGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHT
TEDCGIALGQAFKEAMGAVRGVKRFGTGFAPLDEALSRAVVDLSNRPFAVIDLGLKREM
IGDLSTEMIPHFLESFAEAARITLHVDCLRGFNDHHRSESAFKALAVAIREAISSNGTN
DVPSTKGVLM"
terminator 2523..2720
/label=TEF terminator
/note="Ashbya gossypii TEF terminator"
primer_bind complement(2824..2843)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(2825..2843)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(2930..2947)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3101..3689)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3181..3200)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(join(3860..4522,1..198))
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 4483..4502
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
ORIGIN
1 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
61 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
121 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
181 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
241 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
301 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
361 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
421 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
481 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
541 taactatgcg gcatcagagc agattgtact gagagtgcac catatggaca tattgtcgtt
601 agaacgcggc tacaattaat acataacctt atgtatcata cacatacgat ttaggtgaca
661 ctatagaacg cggccgccag ctgaagcttc gtacgctgca ggtcgacgga tccccgggtt
721 aattaacttg gaagtattgt tccaaggtcc aggtggaggc ggtggaggct ctagaaaaac
781 cgcggctctt gcgcaacacg atgaagccgt ggacaacaaa ttcaacaaag aacaacaaaa
841 cgcgttctat gagatcttac atttacctaa cttaaacgaa gaacaacgaa acgccttcat
901 ccaaagttta aaagatgacc caagccaaag cgctaacctt ttagcagaag ctaaaaagct
961 aaatgatgct caggcgccga aagtagacaa caaattcaac aaagaacaac aaaacgcgtt
1021 ctatgagatc ttacatttac ctaacttaaa cgaagaacaa cgaaacgcct tcatccaaag
1081 tttaaaagat gacccaagcc aaagcgctaa ccttttagca gaagctaaaa agctaaatgg
1141 tgctcaggcg ccgaaagtag acgcgaattc cgcggggaag tcaacctgac tcgaggcgcg
1201 ccacttctaa ataagcgaat ttcttatgat ttatgatttt tattattaaa taagttataa
1261 aaaaaataag tgtatacaaa ttttaaagtg actcttaggt tttaaaacga aaattcttat
1321 tcttgagtaa ctctttcctg taggtcaggt tgctttctca ggtatagtat gaggtcgctc
1381 ttattgacca cacctctacc ggcagatccg ctagggataa cagggtaata tagatctgtt
1441 tagcttgcct cgtccccgcc gggtcacccg gccagcgaca tggaggccca gaataccctc
1501 cttgacagtc ttgacgtgcg cagctcaggg gcatgatgtg actgtcgccc gtacatttag
1561 cccatacatc cccatgtata atcatttgca tccatacatt ttgatggccg cacggcgcga
1621 agcaaaaatt acggctcctc gctgcagacc tgcgagcagg gaaacgctcc cctcacagac
1681 gcgttgaatt gtccccacgc cgcgcccctg tagagaaata taaaaggtta ggatttgcca
1741 ctgaggttct tctttcatat acttcctttt aaaatcttgc taggatacag ttctcacatc
1801 acatccgaac ataaacaacc atggcagaac cagcccaaaa aaagcaaaaa caaactgttc
1861 aggagcgcaa ggcgtttatc tcccgtatca ctaatgaaac taaaattcaa atcgctattt
1921 cgctgaatgg tggttatatt caaataaaag attcgattct tcctgcaaag aaggatgacg
1981 atgtagcttc ccaagctact cagtcacagg tcatcgatat tcacacaggt gttggctttt
2041 tggatcatat gatccatgcg ttggcaaaac actctggttg gtctcttatt gttgaatgta
2101 tcggtgacct gcacattgac gatcaccata ctaccgaaga ttgcggtatc gcattagggc
2161 aagcgttcaa agaagcaatg ggtgctgtcc gtggtgtaaa aagattcggt actgggttcg
2221 caccattgga tgaggcgcta tcacgtgccg tagtcgattt atctaataga ccatttgctg
2281 taatcgacct tggattgaag agagagatga ttggtgattt atccactgaa atgattccac
2341 actttttgga aagtttcgcg gaggcggcca gaattacttt gcatgttgat tgtctgagag
2401 gtttcaacga tcaccacaga agtgagagtg cgttcaaggc tttggctgtt gccataagag
2461 aagctatttc tagcaatggc accaatgacg ttccctcaac caaaggtgtt ttgatgtgaa
2521 gtactgacaa taaaaagatt cttgttttca agaacttgtc atttgtatag tttttttata
2581 ttgtagttgt tctattttaa tcaaatgtta gcgtgattta tatttttttt cgcctcgaca
2641 tcatctgccc agatgcgaag ttaagtgcgc agaaagtaat atcatgcgtc aatcgtatgt
2701 gaatgctggt cgctatactg ctgtcgattc gatactaacg ccgccatcca gtttaaacga
2761 gctcgaattc atcgatgata tcagatccac tagtggccta tgcggccgcg gatctgccgg
2821 tctccctata gtgagtcgta ttaatttcga taagccaggt taacctgcat taatgaatcg
2881 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
2941 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
3001 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
3061 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
3121 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
3181 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
3241 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct
3301 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
3361 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
3421 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
3481 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
3541 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
3601 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
3661 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
3721 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
3781 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
3841 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
3901 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
3961 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
4021 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
4081 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
4141 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
4201 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
4261 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
4321 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
4381 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
4441 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
4501 gcagaacttt aaaagtgctc at
//
| {
"Vector backbone": "pFA6a-6xGLY-3xFLAG-HIS3MX6 (Addgene plasmid #20753)",
"Backbone size w/o insert (bp)": "",
"Vector type": "PCR-based yeast C-terminal tagging",
"Selectable markers": "HIS3"
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "Unknown"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "",
"Publication": "",
"DOI": "",
"Authors": "",
"PubMed Link": "",
"PubMed ID": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Ligation Independent Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "Sp6",
"3′ sequencing primer": "T7"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pCAG-WIPI4 | 42528 | Low | James Hurley | Baskaran et al Mol Cell. 2012 Aug 10;47(3):339-48. doi: 10.1016/j.molcel.2012.05.027. Epub 2012 Jun 14. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/67741/babcd94a-ef27-4b7f-b7e6-7444a43362a9/addgene-plasmid-42528-sequence-67741.gbk | partial | LOCUS sequence-67741-e 916 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 916)
AUTHORS Baskaran S, Ragusa MJ, Boura E, Hurley JH
TITLE Two-site recognition of phosphatidylinositol 3-phosphate by PROPPINs
in autophagy.
JOURNAL Mol Cell. 2012 Aug 10;47(3):339-48. doi:
10.1016/j.molcel.2012.05.027. Epub 2012 Jun 14.
PUBMED 22704557
REFERENCE 2 (bases 1 to 916)
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.molcel.2012.05.027"; journalName: "Mol Cell"; date:
"2012-08-10- 10"; volume: "47"; issue: "3"; pages: "339-48"
FEATURES Location/Qualifiers
source 1..916
/mol_type="genomic DNA"
/organism="unspecified"
CDS 27..47
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQG"
ORIGIN
1 tcggatgatt cgatatccca acgaccgaaa acctgtattt tcagggtggt accatgactc
61 aacagccact tcgaggagtg accagcctgc gtttcaacca agaccaaagc tgcttttgct
121 gcgccatgga gacaggtgtg cgcatctaca acgtggagcc cttgatggag aaggggcatc
181 tggaccacga gcaggtgggc agcatgggct tggtggagat gctgcaccgc tccaaccttc
241 tggccttggt gggcggtggt agtagtccca agttctcaga gatctcagtg ctgatctggg
301 acgatgcccg ggagggcaag gactccaagg agaagctggt gctggagttc accttcacca
361 agccagtgct ttctgtgcgc atgcgccatg acaagatcgt gatcgtgctg aagaaccgca
421 tctatgtgta ctccttcccc gacaatcccc gaaagctgtt tgagtttgat acccgggaca
481 accccaaggg gctctgtgac ctctgcccca gcctggagaa gcaactgcta gtgttcccgg
541 gacacaagtg tgggagtctg caacttgtgg acctggcgag cacaaagcct ggcacctcgt
601 ctgctccatt cacgatcaat gcacatcaga gtgacatagc ctgtgtgtct ctaaaccagc
661 caggcactgt agtggcctca gcctcccaga agggtaccct tattcgcctc tttgacacac
721 aatccaagga gaaactggtg gagctgcgcc gaggcactga ccctgccacc ctctactgca
781 ttaacttcag ccacgactcc tccttcctct gcgcttccag tgataagggt actgtccata
841 tctttgctct caaggatacc cgcctcaacc gccgctccgc gctggctcgc gtgggcaagg
901 tggggcctat gattgg
//
| {
"Vector backbone": "pCAG",
"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": "Two-site recognition of phosphatidylinositol 3-phosphate by PROPPINs in autophagy",
"Publication": "Mol Cell. 2012 Aug 10;47(3):339-48. doi: 10.1016/j.molcel.2012.05.027. Epub 2012 Jun 14.",
"DOI": "10.1016/j.molcel.2012.05.027.",
"Authors": "Baskaran S, Ragusa MJ, Boura E, Hurley JH /jmolcel",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/22704557",
"PubMed ID": "PubMed 22704557"
} | {
"Gene/Insert name": "WIPI4",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "1083",
"Mutation": "",
"Entrez Gene": "WDR45",
"Promoter": "",
"Tag / Fusion Protein": "GST-TEV (N terminal on insert)"
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "ClaI (unknown if destroyed)",
"3′ cloning site": "XhoI (unknown if destroyed)",
"5′ sequencing primer": "unknown",
"3′ sequencing primer": "unknown"
} | {
"Gene/Insert name": "",
"Species": "",
"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": ""
} |
|
MVK gRNA (BRDN0001145420) | 77407 | 3rd generation lentiviral gRNA plasmid targeting human MVK | Doench et al Nat Biotechnol. 2016 Jan 18. doi: 10.1038/nbt.3437. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/189028/f07a4fbc-5c2f-4930-b53c-0e71c1ad9b17/addgene-plasmid-77407-sequence-189028.gbk | full | LOCUS sequence-189028- 8323 bp DNA circular SYN 26-AUG-2024
DEFINITION 3rd generation lentiviral gRNA plasmid targeting human MVK.
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 cgcaaagtcc ctcgcaatac cagttttaga 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": "MVK",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "MVK",
"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": ""
} |
||
MAST1 | 39065 | Bacterial expression for structure determination; may not be full ORF | Nicola Burgess-Brown | SGC+year (unpublished) | https://media.addgene.org/snapgene-media/v2.0.0/sequences/53255/e2cb80ee-69d2-4394-b681-b39a30bb1963/addgene-plasmid-39065-sequence-53255.gbk | partial | LOCUS sequence-53255-e 993 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 993)
TITLE SGC+year
JOURNAL unpublished
REFERENCE 2 (bases 1 to 993)
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..993
/mol_type="genomic DNA"
/organism="unspecified"
CDS 48..65
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 90..110
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQS"
CDS 470..487
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(540..558)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 554..601
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
primer_bind complement(725..744)
/label=F1ori-R
/note="F1 origin, reverse primer"
ORIGIN
1 attccctcta gaatattttg tttactttag aaggagatat acatatgcac catcatcatc
61 atcattcttc tggtgtagat ctgggtaccg agaacctgta cttccaatcc atgcgtagcc
121 cgattaccat tcagcgtagc ggcaaaaaat atggttttac cctgcgtgcg attcgcgtgt
181 atatgggcga taccgatgtt tatagcgttc atcatattgt ttggcatgtg gaagaaggtg
241 gtccggcgca ggaagcgggc ctgtgcgcgg gcgatctgat tacccatgtg aatggtgaac
301 cggtgcatgg catggttcat ccggaagtgg ttgaactgat tctgaaaagc ggtaataaag
361 tggcggtgac caccaccccg tttgaaaaca cccagagcct ggtttgacag taaaggtgga
421 tacggatccg aattcgagct ccgtcgacaa gcttgcggcc gcactcgagc accaccacca
481 ccaccactga gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac
541 cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
601 gctgaaagga ggaactatat ccggattggc gaatgggacg cgccctgtag cggcgcatta
661 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
721 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
781 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
841 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
901 tcgccctttt gacgttggga gtccacgttc tttaatagtg gaactcttgt tccaaactgg
961 aacaacactc aacccctatc tcgggtctat ttc
//
| {
"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)": "Mach1",
"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": "MAST1",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "388",
"Mutation": "codon-optimized",
"Entrez Gene": "MAST1",
"Promoter": "",
"Tag / Fusion Protein": "His Tev (N terminal on insert)"
} | {
"Cloning method": "Ligation Independent Cloning",
"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": ""
} |
|
EF1A_Hygro_P2A_RUNX1_P68R_Barcode | 170931 | Overexpression of RUNX1 P68R mutant protein. | Prashant Mali | Ozturk et al Cell Rep. 2024 Jul 4;43(7):114436. doi: 10.1016/j.celrep.2024.114436. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/336433/38d7261e-6017-4fbb-b43b-8bc11d23271a/addgene-plasmid-170931-sequence-336433.gbk | full | LOCUS sequence-336433- 10865 bp DNA circular SYN 26-AUG-2024
DEFINITION Overexpression of RUNX1 P68R mutant protein..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10865)
AUTHORS Ozturk K, Panwala R, Sheen J, Ford K, Jayne N, Portell A, Zhang DE,
Hutter S, Haferlach T, Ideker T, Mali P, Carter H
TITLE Interface-guided phenotyping of coding variants in the transcription
factor RUNX1.
JOURNAL Cell Rep. 2024 Jul 4;43(7):114436. doi:
10.1016/j.celrep.2024.114436.
PUBMED 38968069
REFERENCE 2 (bases 1 to 10865)
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: "2024-07-4"; volume: "43(7):114436. doi"; pages: "
10.1016/j.celrep.2024.114436"
FEATURES Location/Qualifiers
source 1..10865
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(13..33)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 18..347
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 198..333
/label=SV40 ori
/note="SV40 origin of replication"
promoter 395..442
/label=EM7 promoter
/note="synthetic bacterial promoter "
CDS 461..835
/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 965..1086
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1002..1021)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 1056..1075
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(1135..1151)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1135..1151)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(1148..1170)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 1159..1175
/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(1183..1213)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 1228..1249
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(1366..1383)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(1537..2125)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1617..1636)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(2296..3156)
/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 2919..2938
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(3157..3261)
/gene="bla"
/label=AmpR promoter
primer_bind complement(3336..3355)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 3527..3906
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 3908..4106
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 4061..4081
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
LTR 4124..4304
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 4351..4476
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 4969..5202
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 5387..5431
/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 5729..5846
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 5982..7160
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 6213..7151
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 7108..7128
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
primer_bind 7177..7196
/label=T7
/note="T7 promoter, forward primer"
promoter 7177..7195
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 7259..8281
/codon_start=1
/product="hygromycin B phosphotransferase"
/label=HygR
/note="confers resistance to hygromycin"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
E"
CDS 7259..8281
/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 8291..8347
/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"
protein_bind complement(9901..9934)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
LTR 9959..10139
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
primer_bind complement(10165..10182)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 10171..10395
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin join(10441..10865,1..4)
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(10528..10547)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 10738..10759
/label=F1ori-F
/note="F1 origin, forward primer"
ORIGIN
1 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
61 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
121 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
181 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
241 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
301 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
361 cttgtatatc cattttcgga tctgatcagc acgtgttgac aattaatcat cggcatagta
421 tatcggcata gtataatacg acaaggtgag gaactaaacc atggccaagt tgaccagtgc
481 cgttccggtg ctcaccgcgc gcgacgtcgc cggagcggtc gagttctgga ccgaccggct
541 cgggttctcc cgggacttcg tggaggacga cttcgccggt gtggtccggg acgacgtgac
601 cctgttcatc agcgcggtcc aggaccaggt ggtgccggac aacaccctgg cctgggtgtg
661 ggtgcgcggc ctggacgagc tgtacgccga gtggtcggag gtcgtgtcca cgaacttccg
721 ggacgcctcc gggccggcca tgaccgagat cggcgagcag ccgtgggggc gggagttcgc
781 cctgcgcgac ccggccggca actgcgtgca cttcgtggcc gaggagcagg actgacacgt
841 gctacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
901 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
961 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
1021 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
1081 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
1141 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
1201 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
1261 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
1321 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
1381 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
1441 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
1501 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
1561 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
1621 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
1681 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
1741 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
1801 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
1861 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
1921 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
1981 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
2041 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
2101 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
2161 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
2221 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
2281 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
2341 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
2401 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
2461 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
2521 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
2581 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
2641 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
2701 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
2761 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
2821 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
2881 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
2941 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
3001 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
3061 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
3121 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
3181 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
3241 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
3301 gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga tgccgcatag
3361 ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa
3421 atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt
3481 aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca ttgattattg
3541 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
3601 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
3661 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
3721 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
3781 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
3841 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
3901 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
3961 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
4021 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
4081 gtacggtggg aggtctatat aagcagcgcg ttttgcctgt actgggtctc tctggttaga
4141 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
4201 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
4261 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg
4321 gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa
4381 gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc
4441 ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga
4501 tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat
4561 atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca
4621 tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa
4681 gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag
4741 ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc
4801 accgcacagc aagcggccgc tgatcttcag acctggagga ggagatatga gggacaattg
4861 gagaagtgaa ttatataaat ataaagtagt aaaaattgaa ccattaggag tagcacccac
4921 caaggcaaag agaagagtgg tgcagagaga aaaaagagca gtgggaatag gagctttgtt
4981 ccttgggttc ttgggagcag caggaagcac tatgggcgca gcgtcaatga cgctgacggt
5041 acaggccaga caattattgt ctggtatagt gcagcagcag aacaatttgc tgagggctat
5101 tgaggcgcaa cagcatctgt tgcaactcac agtctggggc atcaagcagc tccaggcaag
5161 aatcctggct gtggaaagat acctaaagga tcaacagctc ctggggattt ggggttgctc
5221 tggaaaactc atttgcacca ctgctgtgcc ttggaatgct agttggagta ataaatctct
5281 ggaacagatt tggaatcaca cgacctggat ggagtgggac agagaaatta acaattacac
5341 aagcttaata cactccttaa ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga
5401 attattggaa ttagataaat gggcaagttt gtggaattgg tttaacataa caaattggct
5461 gtggtatata aaattattca taatgatagt aggaggcttg gtaggtttaa gaatagtttt
5521 tgctgtactt tctatagtga atagagttag gcagggatat tcaccattat cgtttcagac
5581 ccacctccca accccgaggg gacccgacag gcccgaagga atagaagaag aaggtggaga
5641 gagagacaga gacagatcca ttcgattagt gaacggatcg gcactgcgtg cgccaattct
5701 gcagacaaat ggcagtattc atccacaatt ttaaaagaaa aggggggatt ggggggtaca
5761 gtgcagggga aagaatagta gacataatag caacagacat acaaactaaa gaattacaaa
5821 aacaaattac aaaaattcaa aattttcggg tttattacag ggacagcaga gatccagttt
5881 ggttaattaa tcgataagct ttgcaaagat ggataaagtt ttaaacagag aggaatcttt
5941 gcagctaatg gaccttctag gtcttgaaag gagtgggaat tggctccggt gcccgtcagt
6001 gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa
6061 ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc
6121 gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc
6181 tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt tcccgcgggc
6241 ctggcctctt tacgggttat ggcccttgcg tgccttgaat tacttccacc tggctgcagt
6301 acgtgattct tgatcccgag cttcgggttg gaagtgggtg ggagagttcg aggccttgcg
6361 cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg gcctgggcgc tggggccgcc
6421 gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc tttcgataag tctctagcca
6481 tttaaaattt ttgatgacct gctgcgacgc tttttttctg gcaagatagt cttgtaaatg
6541 cgggccaaga tctgcacact ggtatttcgg tttttggggc cgcgggcggc gacggggccc
6601 gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg agcgcggcca ccgagaatcg
6661 gacgggggta gtctcaagct ggccggcctg ctctggtgcc tggcctcgcg ccgccgtgta
6721 tcgccccgcc ctgggcggca aggctggccc ggtcggcacc agttgcgtga gcggaaagat
6781 ggccgcttcc cggccctgct gcagggagct caaaatggag gacgcggcgc tcgggagagc
6841 gggcgggtga gtcacccaca caaaggaaaa gggcctttcc gtcctcagcc gtcgcttcat
6901 gtgactccac ggagtaccgg gcgccgtcca ggcacctcga ttagttctcg agcttttgga
6961 gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat ggagtttccc cacactgagt
7021 gggtggagac tgaagttagg ccagcttggc acttgatgta attctccttg gaatttgccc
7081 tttttgagtt tggatcttgg ttcattctca agcctcagac agtggttcaa agtttttttc
7141 ttccatttca ggtgtcgtga ggaattagct tggtactaat acgactcact atagggagac
7201 ccaagctggc taggtaagct tgatatcgaa ttcctgcagc ccgggggatc ccgccaccat
7261 gaaaaagcct gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag
7321 cgtctccgac ctgatgcagc tctcggaggg cgaagaatct cgtgctttca gcttcgatgt
7381 aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg
7441 ttatgtttat cggcactttg catcggccgc gctcccgatt ccggaagtgc ttgacattgg
7501 ggaattcagc gagagcctga cctattgcat ctcccgccgt gcacagggtg tcacgttgca
7561 agacctgcct gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg ccatggatgc
7621 gatcgctgcg gccgatctta gccagacgag cgggttcggc ccattcggac cgcaaggaat
7681 cggtcaatac actacatggc gtgatttcat atgcgcgatt gctgatcccc atgtgtatca
7741 ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc tcgatgagct
7801 gatgctttgg gccgaggact gccccgaagt ccggcacctc gtgcacgcgg atttcggctc
7861 caacaatgtc ctgacggaca atggccgcat aacagcggtc attgactgga gcgaggcgat
7921 gttcggggat tcccaatacg aggtcgccaa catcttcttc tggaggccgt ggttggcttg
7981 tatggagcag cagacgcgct acttcgagcg gaggcatccg gagcttgcag gatcgccgcg
8041 gctccgggcg tatatgctcc gcattggtct tgaccaactc tatcagagct tggttgacgg
8101 caatttcgat gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc
8161 cgggactgtc gggcgtacac aaatcgcccg cagaagcgcg gccgtctgga ccgatggctg
8221 tgtagaagta ctcgccgata gtggaaaccg acgccccagc actcgtccga gggcaaagga
8281 aggatccggc gcaacaaact tctctctgct gaaacaagcc ggagatgtcg aagagaatcc
8341 tggaccgatg cgtatccccg tagatgccag cacgagccgc cgcttcacgc cgccttccac
8401 cgcgctgagc ccaggcaaga tgagcgaggc gttgccgctg ggcgccccgg acgccggcgc
8461 tgccctggcc ggcaagctga ggagcggcga ccgcagcatg gtggaggtgc tggccgacca
8521 cccgggcgag ctggtgcgca ccgacagccg caacttcctc tgctccgtgc tgcctacgca
8581 ctggcgctgc aacaagaccc tgcccatcgc tttcaaggtg gtggccctag gggatgttcc
8641 agatggcact ctggtcactg tgatggctgg caatgatgaa aactactcgg ctgagctgag
8701 aaatgctacc gcagccatga agaaccaggt tgcaagattt aatgacctca ggtttgtcgg
8761 tcgaagtgga agagggaaaa gcttcactct gaccatcact gtcttcacaa acccaccgca
8821 agtcgccacc taccacagag ccatcaaaat cacagtggat gggccccgag aacctcgaag
8881 acatcggcag aaactagatg atcagaccaa gcccgggagc ttgtcctttt ccgagcggct
8941 cagtgaactg gagcagctgc ggcgcacagc catgagggtc agcccacacc acccagcccc
9001 cacgcccaac cctcgtgcct ccctgaacca ctccactgcc tttaaccctc agcctcagag
9061 tcagatgcag gatacaaggc agatccaacc atccccaccg tggtcctacg atcagtccta
9121 ccaatacctg ggatccattg cctctccttc tgtgcaccca gcaacgccca tttcacctgg
9181 acgtgccagc ggcatgacaa ccctctctgc agaactttcc agtcgactct caacggcacc
9241 cgacctgaca gcgttcagcg acccgcgcca gttccccgcg ctgccctcca tctccgaccc
9301 ccgcatgcac tatccaggcg ccttcaccta ctccccgacg ccggtcacct cgggcatcgg
9361 catcggaatg tcggccatgg gctcggccac gcgctaccac acctacctgc cgccgcctta
9421 ccccggctcg tcgcaagcgc agggaggccc gttccaagcc agctcgccct cctaccacct
9481 gtactacggc gcctcggccg gctcctacca gttctccatg gtgggcggcg agcgctcgcc
9541 gccgcgcatc ctgccgccct gcaccaacgc ctccaccggc tccgcgctgc tcaaccccag
9601 cctcccgaac cagagcgacg tggtggaggc cgagggcagc cacagcaact cccccaccaa
9661 catggcgccc tccgcgcgcc tggaggaggc cgtgtggagg ccctactgac tagctaagtt
9721 agaactattt cctggctgtt acgcgtgatg gctgggtcta ctgacgggct tattaaacag
9781 tgaaccgccc gccccacgac ccgcagcgcc cgaccgaaag gagcgcacga ccccatgcat
9841 cgaacaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatatc
9901 ataacttcgt atagcataca ttatacgaag ttatcgccta gctggtccgg actgtactgg
9961 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
10021 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
10081 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
10141 ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
10201 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
10261 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
10321 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
10381 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
10441 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
10501 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
10561 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
10621 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
10681 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
10741 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
10801 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
10861 acgcg
//
| {
"Vector backbone": "EF1a_mCherry_P2A_Hygro_Barcode",
"Backbone size w/o insert (bp)": "9508",
"Vector type": "Mammalian Expression, Lentiviral",
"Selectable markers": "Hygromycin"
} | {
"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": "Interface-guided phenotyping of coding variants in the transcription factor RUNX1",
"Publication": "Cell Rep. 2024 Jul 4;43(7):114436. doi: 10.1016/j.celrep.2024.114436.",
"DOI": "10.1016/j.celrep.2024.114436.",
"Authors": "Ozturk K, Panwala R, Sheen J, Ford K, Jayne N, Portell A, Zhang DE, Hutter S, Haferlach T, Ideker T, Mali P, Carter H /jcelrep",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/38968069",
"PubMed ID": "PubMed 38968069"
} | {
"Gene/Insert name": "RUNX1",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "1359",
"Mutation": "P68R",
"Entrez Gene": "RUNX1",
"Promoter": "Ef1A",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Gibson Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "CTGTGTAGAAGTACTCGCCGATAGTG",
"3′ sequencing primer": "TCTTGTCTTCGTTGGGAGTG"
} | {
"Gene/Insert name": "",
"Species": "",
"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": ""
} |
|
YFP(1-158)-beta-2 in pcDNAI/Amp | 54469 | An amino-terminal YFP fragment was fused to Gbeta-2. When co-expressed with a carboxyl-terminal YFP or CFP fragment fused to a Ggamma subunit, a fluorescent signal is produced. | Catherine Berlot | Hynes et al J Biol Chem. 2004 Jul 16;279(29):30279-86. Epub 2004 May 10. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/114883/568f4bd7-88b9-4540-9801-59d61df1bcdf/addgene-plasmid-54469-sequence-114883.gbk | full | LOCUS sequence-114883- 6283 bp DNA circular SYN 26-AUG-2024
DEFINITION An amino-terminal YFP fragment was fused to Gbeta-2. When
co-expressed with a carboxyl-terminal YFP or CFP fragment fused to a
Ggamma subunit, a fluorescent signal is produced..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6283)
AUTHORS Hynes TR, Tang L, Mervine SM, Sabo JL, Yost EA, Devreotes PN, Berlot
CH
TITLE Visualization of G protein betagamma dimers using bimolecular
fluorescence complementation demonstrates roles for both beta and
gamma in subcellular targeting.
JOURNAL J Biol Chem. 2004 Jul 16;279(29):30279-86. Epub 2004 May 10.
PUBMED 15136579
REFERENCE 2 (bases 1 to 6283)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J Biol
Chem."; date: "2004-07-16"; volume: "279(29)"; pages: "30279-86.
Epub 2004 May 10"
FEATURES Location/Qualifiers
source 1..6283
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 471..490
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
rep_origin complement(603..1061)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(740..761)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 955..974
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind complement(1186..1204)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 1272..1376
/gene="bla"
/label=AmpR promoter
CDS 1377..2237
/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(1595..1614)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
enhancer 2310..2689
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 2690..2893
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 2844..2864
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 2938..2957
/label=T7
/note="T7 promoter, forward primer"
promoter 2938..2956
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory 2988..2997
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 2994..3458
/codon_start=1
/product="improved N-terminal fragment of mVenus for use in
bimolecular fluorescence complementation (BiFC) (Kodama and
Hu, 2010)"
/label=VN155(I152L)
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMA"
primer_bind complement(3039..3060)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(3300..3319)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
promoter complement(4555..4573)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(4556..4573)
/label=SP6
/note="SP6 promoter, forward primer"
intron 4657..4722
/label=small t intron
/note="SV40 (simian virus 40) small t antigen intron"
CDS 4852..4872
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
polyA_signal 5144..5278
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5181..5200)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 5235..5254
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 6124..6259
/label=SV40 ori
/note="SV40 origin of replication"
ORIGIN
1 ggcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
61 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
121 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
181 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
241 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
301 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
361 ctacagcgtg agcattgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
421 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
481 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
541 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcaagcta gcttctagct
601 agaaattgta aacgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
661 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagcccga
721 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
781 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc cgcccactac gtgaaccatc
841 acccaaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
901 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
961 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
1021 caccacaccc gccgcgctta atgcgccgct acagggcgcg tactatggtt gctttgacga
1081 gaccgtataa cgtgctttcc tcgttggaat cagagcggga gctaaacagg aggccgatta
1141 aagggatttt agacaggaac ggtacgccag ctggattacc aaagggcctc gtgatacgcc
1201 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
1261 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
1321 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
1381 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
1441 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
1501 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
1561 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg
1621 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
1681 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
1741 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
1801 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
1861 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
1921 cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
1981 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
2041 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
2101 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
2161 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
2221 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
2281 aacttcattt ttaatttctc tagcgcgttg acattgatta ttgactagtt attaatagta
2341 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
2401 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
2461 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggactattt
2521 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
2581 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
2641 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
2701 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
2761 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
2821 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
2881 tataagcaga gctctctggc taactagaga acccactgct tactggctta tcgaaattaa
2941 tacgactcac tatagggaga cccaagcttg gtaccgagct cggatccgcc accatggtga
3001 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
3061 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
3121 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
3181 ccaccttcgg ctacggcctg atgtgcttcg cccgctaccc cgaccacatg aagcagcacg
3241 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
3301 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
3361 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
3421 agtacaacta caacagccac aacgtctata tcatggccga caagcagaga tccatcgcaa
3481 ccatgagtga gctggagcaa ctgagacagg aggccgagca gctccggaac cagatccggg
3541 atgcccgaaa agcatgtggg gactcaacac tgacccagat cacagctggg ctggacccag
3601 tggggagaat ccagatgagg acccggagga ccctccgtgg gcacctggca aagatttatg
3661 ccatgcactg ggggaccgac tcaaggctgc tggtcagcgc ctcccaggat gggaagctca
3721 tcatctggga cagctacacc accaacaagg tccacgccat cccgctgcgc tcctcctggg
3781 taatgacctg tgcctacgcg ccctccggaa actttgtggc ctgtgggggg ttggacaaca
3841 tctgctccat ctacagcctc aagacccgcg agggcaacgt cagggtcagc cgggagctgc
3901 ctggccacac tgggtacctg tcgtgttgcc gcttcctgga tgacaaccaa atcatcacca
3961 gctctgggga taccacctgt gccctgtggg acattgagac aggccagcag acagtgggtt
4021 ttgctggaca cagtggggat gtgatgtccc tgtccctggc ccccaatggc cgcacgtttg
4081 tgtcaggcgc ctgtgatgcc tctatcaagc tgtgggacgt gcgggattcc atgtgccgac
4141 agaccttcat cggccatgaa tccgacatca atgcagtggc tttcttcccc aacggctacg
4201 ccttcaccac cggctctgac gacgccacgt gccgcctctt cgacctgcgg gccgatcagg
4261 agctcctcat gtactcccat gacaacatca tctgtggcat cacctctgtt gccttctcgc
4321 gcagcggacg gctgctgctc gctggctacg acgacttcaa ctgcaacatc tgggatgcca
4381 tgaagggcga ccgtgcagga gtcctcgctg gccacgacaa ccgcgtgagc tgcctcgggg
4441 tcaccgacga tggcatggct gtggccacgg gctcctggga ctccttcctc aagatttgga
4501 actaaagatc tatcacactg gcggccgctc gagcatgcat ctagagggcc ctattctata
4561 gtgtcaccta aatgctagag gatctttgtg aaggaacctt acttctgtgg tgtgacataa
4621 ttggacaaac tacctacaga gatttaaagc tctaaggtaa atataaaatt tttaagtgta
4681 taatgtgtta aactactgat tctaattgtt tgtgtatttt agattccaac ctatggaact
4741 gatgaatggg agcagtggtg gaatgccttt aatgaggaaa acctgttttg ctcagaagaa
4801 atgccatcta gtgatgatga ggctactgct gactctcaac attctactcc tccaaaaaag
4861 aagagaaagg tagaagaccc caaggacttt ccttcagaat tgctaagttt tttgagtcat
4921 gctgtgttta gtaatagaac tcttgcttgc tttgctattt acaccacaaa ggaaaaagct
4981 gcactgctat acaagaaaat tatggaaaaa tatttgatgt atagtgcctt gactagagat
5041 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
5101 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
5161 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
5221 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatcat
5281 cccgccatgg tatcaacgcc atatttctat ttacagtagg gacctcttcg ttgtgtaggt
5341 accgctgtat tcctagggaa atagtagagg caccttgaac tgtctgcatc agccatatag
5401 cccccgctgt tcgacttaca aacacaggca cagtactgac aaacccatac acctcctctg
5461 aaatacccat agttgctagg gctgtctccg aactcattac accctccaaa gtcagagctg
5521 taatttcgcc atcaagggca gcgagggctt ctccagataa aatagcttct gccgagagtc
5581 ccgtaagggt agacacttca gctaatccct cgatgaggtc tactagaata gtcagtgcgg
5641 ctcccatttt gaaaattcac ttacttgatc agcttcagaa gatggcggag ggcctccaac
5701 acagtaattt tcctcccgac tcttaaaata gaaaatgtca agtcagttaa gcaggaagtg
5761 gactaactga cgcagctggc cgtgcgacat cctcttttaa ttagttgcta ggcaacgccc
5821 tccagagggc gtgtggtttt gcaagaggaa gcaaaagcct ctccacccag gcctagaatg
5881 tttccaccca atcattacta tgacaacagc tgtttttttt agtattaagc agaggccggg
5941 gacccctggg cccgcttact ctggagaaaa agaagagagg cattgtagag gcttccagag
6001 gcaacttgtc aaaacaggac tgcttctatt tctgtcacac tgtctggccc tgtcacaagg
6061 tccagcacct ccataccccc tttaataagc agtttgggaa cgggtgcggg tcttactccg
6121 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
6181 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
6241 ggaggctttt ttggaggcct aggcttttgc aaaaagctaa ttc
//
| {
"Vector backbone": "pcDNAI/Amp",
"Backbone size w/o insert (bp)": "4800",
"Vector type": "Mammalian Expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH10B",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Visualization of G protein betagamma dimers using bimolecular fluorescence complementation demonstrates roles for both beta and gamma in subcellular targeting",
"Publication": "J Biol Chem. 2004 Jul 16;279(29):30279-86. Epub 2004 May 10.",
"DOI": "10.1074/jbc.M401432200",
"Authors": "Hynes TR, Tang L, Mervine SM, Sabo JL, Yost EA, Devreotes PN, Berlot CH",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/15136579",
"PubMed ID": "PubMed 15136579"
} | {
"Gene/Insert name": "YFP(1-158)/beta-2",
"Species": "H. sapiens (human); Aequorea victoria",
"Insert Size (bp)": "1483",
"Mutation": "YFP (1-158) includes a substitution of Met for Gln-69. Gbeta-2 was amplified via PCR, which added an N-terminal linker sequence (RSIAT), a 5' BamHI site, and a 3' BglII site. Cloning into the BglII site of YFP(1-158) destroyed the BamHI site.",
"Entrez Gene": "GNB2",
"Promoter": "CMV",
"Tag / Fusion Protein": "YFP(1-158) was fused to the amino terminus of Gbeta-2. (N terminal on insert)"
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "BamHI (not destroyed)",
"3′ cloning site": "BglII (not destroyed)",
"5′ sequencing primer": "T7",
"3′ sequencing primer": "SP6"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pCN-LYS21-GFP | 45335 | Brendan Cormack | Zordan et al G3 (Bethesda). 2013 Oct 3;3(10):1675-86. doi: 10.1534/g3.113.006908. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/63482/d5a6caa7-2d00-4b18-bf12-ebb4ef5ba91e/addgene-plasmid-45335-sequence-63482.gbk | full | LOCUS sequence-63482-e 6856 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 6856)
AUTHORS Zordan RE, Ren Y, Pan SJ, Rotondo G, De Las Penas A, Iluore J,
Cormack BP
TITLE Expression plasmids for use in Candida glabrata.
JOURNAL G3 (Bethesda). 2013 Oct 3;3(10):1675-86. doi: 10.1534/g3.113.006908.
PUBMED 23934995
REFERENCE 2 (bases 1 to 6856)
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: "10.1534/g3.113";
journalName: "G3 (Bethesda)"; date: "2013-10-3- 3"; volume: "3";
issue: "10"; pages: "1675-86"
FEATURES Location/Qualifiers
source 1..6856
/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 180..585
/gene="S. cerevisiae TEF1"
/label=TEF1 promoter
/note="promoter for EF-1-alpha"
CDS 599..1171
/codon_start=1
/gene="Streptomyces noursei nat1"
/product="nourseothricin acetyltransferase"
/label=NrsR
/note="confers resistance to nourseothricin"
/translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
RLTVEDIEVAPEHRGHGVGRALMGLATEFARELGAGHLWLEVTNVNAPAIHAYRRMGFT
LCGLDTALYDGTASDGEQALYMSMPCP"
terminator 1178..1425
/gene="S. cerevisiae CYC1"
/label=CYC1 terminator
/note="transcription terminator for CYC1"
primer_bind complement(1194..1212)
/label=CYC1
/note="CYC1 transcription termination signal, reverse
primer"
primer_bind 1610..1632
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 1624..1641
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 1625..1641
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
CDS 2847..3563
/codon_start=1
/product="yeast-enhanced green fluorescent protein"
/label=yeGFP
/note="codons optimized for translation in S. cerevisiae
and C. albicans"
/translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
FVTAAGITHGMDELYK"
primer_bind complement(4037..4053)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4037..4053)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4050..4072)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4061..4077
/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(4085..4115)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4130..4151
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4268..4285)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4439..5027)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4519..4538)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5198..6058)
/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 5821..5840
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6059..6163)
/gene="bla"
/label=AmpR promoter
primer_bind 6829..6847
/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 accataccat agcttcaaaa tgtttctact ccttttttac tcttccagat tttctcggac
241 tccgcgcatc gccgtaccac ttcaaaacac ccaagcacag catactaaat ttcccctctt
301 tcttcctcta gggtgtcgtt aattacccgt actaaaggtt tggaaaagaa aaaagagacc
361 gcctcgtttc tttttcttcg tcgaaaaagg caataaaaat ttttatcacg tttctttttc
421 ttgaaaattt ttttttgatt tttttctctt tcgatgacct cccattgata tttaagttaa
481 taaacggtct tcaatttctc aagtttcagt ttcatttttc ttgttctatt acaacttttt
541 ttacttcttg ctcattagaa agaaagcata gcaatctaat ctaagttttg gatctaaaat
601 gggcaccact cttgacgaca cggcttaccg ataccgcacc agtgtcccgg gggacgccga
661 ggccatcgag gcactggatg ggtccttcac caccgacacc gtcttccgcg tcaccgccac
721 cggggacggc ttcaccctgc gggaggtgcc ggtggacccg cccctgacca aggtgttccc
781 cgacgacgaa tcggacgacg aatcggacga cggggaggac ggcgaccctg actcccggac
841 gttcgtcgcg tacggggacg acggcgacct ggcgggcttc gtggtcgtct cgtactccgg
901 ctggaaccgc cggctgaccg tcgaggacat cgaggtcgcc ccggagcacc gggggcacgg
961 ggtcgggcgc gcgttgatgg ggctcgcgac ggagttcgcc cgcgagctgg gcgccgggca
1021 cctctggctg gaggtcacca acgtcaacgc accggcgatc cacgcgtacc ggcggatggg
1081 gttcaccctc tgcggcctgg acaccgccct gtacgacggc accgcctcgg acggcgagca
1141 ggcgctctac atgagcatgc cctgccccta agtcgagtca tgtaattagt tatgtcacgc
1201 ttacattcac gccctccccc cacatccgct ctaaccgaaa aggaaggagt tagacaacct
1261 gaagtctagg tccctattta tttttttata gttatgttag tattaagaac gttatttata
1321 tttcaaattt ttcttttttt tctgtacaga cgcgtgtacg catgtaacat tatactgaaa
1381 accttgcttg agaaggtttt gggacgctcg aaggctttaa tttgcggccg tatgcggtgt
1441 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc gccattcagg
1501 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg
1561 aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
1621 cgttgtaaaa cgacggccag tgaattcgag ctcagaaagc gaagaagata tatctagttt
1681 ttctccgcac tcttacccat tctcattttt tttaactcag cattttacta tagatttttt
1741 ttctttctta taatcaagta catagaatat atgcacatag aaacaaaaaa gataggcaag
1801 ttattattca gacgcattat cataaaaaac tttatttatc tgttaacgaa acaaaattta
1861 gcgctatttc tcaaaggatt gtagcagcat catgagaaaa atatcgtcaa tcggagcgaa
1921 gtttattacg tctacacagg taacaagggt atctgtgtat tgaacctcgg attgaaccgg
1981 catattctca cacaaatgag aagcttgaaa tccccacgga tcctgccaca cccccaacaa
2041 ttaaagtgac taagaaacaa ctctagaccc ttgttactta tttcactcaa acttgtacgt
2101 gccctgctaa cacagtttac agataggagt ccatatgact gcaattccac acacctcgaa
2161 tggctcttat cctctcccct ctccaataga agagacttga gctaagaaaa aatttacatc
2221 ggaattttta cagctgtgtt ggtttcatca ctactttgca agcctagttt ttaccacttc
2281 ctcatatggt tatgactctt ttcctcatct ttttggtgct ttcatttctt tttgcaacgg
2341 tttataattt ttaccttcgc aattttctga tttttttttg caaaaatatt ttcacaattt
2401 acaaaatgca ctaaaagtcc gtagagaagt gggatcagtt attattcctt cacttactaa
2461 catcgataca tttcgaatca gctatttcat agtggtttat gctattctaa tcacacattt
2521 tgttacaaca cttccttcaa tacacgattg tagttgttgt acgtatccat atagtcgact
2581 catttgcgtc gccttctctt gcttttagca acagtcgttc tagtttttgc aagcttctga
2641 ctaattccta aaggcaagta taaatacatg ccgtttcttg ctgaaaccat atgttatctc
2701 gtatctatgt ttcattacac atttttgtaa cagctatttc aactggtgaa atactacaag
2761 taaaaatata ttctacctaa aatatttttg ctgaacaaag aatattaaag actagaacta
2821 gtggatcccc cgggctgcag gaattcatgt ctaaaggtga agaattattc actggtgttg
2881 tcccaatttt ggttgaatta gatggtgatg ttaatggtca caaattttct gtctccggtg
2941 aaggtgaagg tgatgctact tacggtaaat tgaccttaaa atttatttgt actactggta
3001 aattgccggt tccatggcca accttagtca ctactttcgg ttatggtgtt caatgttttg
3061 ctagataccc agatcatatg aaacaacatg actttttcaa gtctgccatg ccagaaggtt
3121 atgttcaaga aagaactatt tttttcaaag atgacggtaa ctacaagacc agagctgaag
3181 tcaagtttga aggtgatacc ttagttaata gaatcgaatt aaaaggtatt gattttaaag
3241 aagatggtaa cattttaggt cacaaattgg aatacaacta taactctcac aatgtttaca
3301 tcatggctga caaacaaaag aatggtatca aagttaactt caaaattaga cacaacattg
3361 aagatggttc tgttcaatta gctgaccatt atcaacaaaa tactccaatt ggtgatggtc
3421 cagtcttgtt accagacaac cattacttat ccactcaatc tgccttatcc aaagatccaa
3481 acgaaaagag agaccacatg gtcttgttag aatttgttac tgctgctggt attacccatg
3541 gtatggatga attgtacaaa taactgcagg tcgacctcga gaacacaacc cacagctacc
3601 accatcaaca atatttatat atataacgta cacatagaaa tcacacaaac agagtattta
3661 ttcttaacta catgaactac catcagaccg tctgggccac tatataatgt gccattcata
3721 aacgtgatca ctttacgtag caggcaaccc caggtgaaaa tttttcagcg agctgccaga
3781 ttgtcaggtg aaaaactgaa aaaaacttct gggcgatgag cttgtggcgg gaaattaagt
3841 atataaagca gttagtttct tctgcttctt gtggttctgg gtattcttgt aatatttgga
3901 agaatagaaa atcaagacta ctgcctttct tttcatatta tagaggaaga aatacgcacg
3961 aacacgatat agaggtaaag gtacccgggg atcctctaga gtcgacctgc aggcatgcaa
4021 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
4081 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
4141 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
4201 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
4261 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
4321 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
4381 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
4441 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
4501 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
4561 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
4621 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
4681 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
4741 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
4801 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
4861 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
4921 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
4981 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
5041 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
5101 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
5161 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
5221 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
5281 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
5341 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
5401 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
5461 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
5521 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
5581 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
5641 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
5701 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
5761 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
5821 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
5881 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
5941 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
6001 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
6061 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
6121 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
6181 tgccacctga cgtcgggatc tctaatgatg caatttttgt tttgtacact aagattctat
6241 ttcggaactc ttacccatgt cattatttac tttattttca ttattgtaat tttttaagct
6301 tttattatga attatttaaa tacaattctt tttgatcatg tgacagacat cactgaaaga
6361 tgtgctctgc gttattagtc agattgaaat atacattttc atgtagatat gatgttctta
6421 ctttaaatgt tgttccattt gctagcttaa atagacgata ttataaattt aagtattcac
6481 taaaaatctg acccattaat gttagaatta gtattatttt ttgtgatatt acatttttat
6541 attttggatt tcaaccaatg ctctgcgata tgattcaaca gaataatatg tacaagctac
6601 tttaattcat atatactaat agattcacag atccagtacg actgcaagaa tttacgtttt
6661 gtttgaaatg gtagtagcta cactaaatta acaatgatcg attaatccac atccacagtg
6721 tgtaatcatc ctaatgaaaa caagaataac tacaaaaatg aatataaact ttttgcatgt
6781 tagtctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
6841 acgaggccct ttcgtc
//
| {
"Vector backbone": "pBM16",
"Backbone size w/o insert (bp)": "",
"Vector type": "C. glabrata expression",
"Selectable markers": "nourseothricin (NAT)"
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH10B",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Expression plasmids for use in Candida glabrata",
"Publication": "G3 (Bethesda). 2013 Oct 3;3(10):1675-86. doi: 10.1534/g3.113.006908.",
"DOI": "10.1534/g3.113.006908.",
"Authors": "Zordan RE, Ren Y, Pan SJ, Rotondo G, De Las Penas A, Iluore J, Cormack BP /g",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/23934995",
"PubMed ID": "PubMed 23934995"
} | {
"Gene/Insert name": "yeGFP",
"Species": "Synthetic",
"Insert Size (bp)": "717",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "LYS21",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "EcoRI (unknown if destroyed)",
"3′ cloning site": "SalI (unknown if destroyed)",
"5′ sequencing primer": "M13F",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"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": ""
} |
||
pDYSCKO | 120263 | Empty backbone for cloning any gRNA to be used with the canavanine co-selection system | Christian Landry | Despres et al G3 (Bethesda). 2018 Oct 3;8(10):3163-3171. doi: 10.1534/g3.118.200461. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/233690/5f78bd0f-a4e8-4bf3-b31c-aa7e60e7dd9e/addgene-plasmid-120263-sequence-233690.gbk | full | LOCUS sequence-233690- 6459 bp DNA circular SYN 26-AUG-2024
DEFINITION Empty backbone for cloning any gRNA to be used with the canavanine
co-selection system.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6459)
AUTHORS Despres PC, Dube AK, Nielly-Thibault L, Yachie N, Landry CR
TITLE Double Selection Enhances the Efficiency of Target-AID and
Cas9-Based Genome Editing in Yeast.
JOURNAL G3 (Bethesda). 2018 Oct 3;8(10):3163-3171. doi:
10.1534/g3.118.200461.
PUBMED 30097473
REFERENCE 2 (bases 1 to 6459)
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: "10.1534/g3.118";
journalName: "G3 (Bethesda)"; date: "2018-10-3- 3"; volume: "8";
issue: "10"; pages: "3163-3171"
FEATURES Location/Qualifiers
source 1..6459
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin 69..1411
/label=2u ori
/note="yeast 2u plasmid origin of replication"
protein_bind 401..448
/label=FRT
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
promoter 1438..1542
/gene="bla"
/label=AmpR promoter
CDS 1543..2403
/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(1761..1780)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 2574..3161
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3062..3081
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 3315..3332
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 3449..3470
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 3485..3515
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3523..3539
/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 3528..3550
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 3547..3563
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 3547..3563
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 3582..3602
/label=T3
/note="T3 promoter, forward primer"
promoter 3584..3602
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 3620..3888
/label=SNR52 promoter
/note="promoter for the S. cerevisiae small nucleolar RNA
gene SNR52"
misc_RNA 3909..3983
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
terminator 3988..4007
/label=SUP4 terminator
/note="transcription terminator for the S. cerevisiae SUP4
tRNA gene"
terminator 4013..4260
/gene="S. cerevisiae CYC1"
/label=CYC1 terminator
/note="transcription terminator for CYC1"
terminator complement(4266..4285)
/label=SUP4 terminator
/note="transcription terminator for the S. cerevisiae SUP4
tRNA gene"
misc_RNA complement(4290..4364)
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
promoter complement(4384..4652)
/label=SNR52 promoter
/note="promoter for the S. cerevisiae small nucleolar RNA
gene SNR52"
rep_origin 4661..5116
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4748..4767)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4958..4979
/label=F1ori-F
/note="F1 origin, forward primer"
CDS complement(5247..6050)
/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(6051..6266)
/gene="S. cerevisiae URA3"
/label=URA3 promoter
primer_bind complement(6290..6309)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 6409..6431
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
ORIGIN
1 gacgaaaggg cctcggcatg cgcctatttt tataggttaa tgtcatgata ataatggttt
61 cttagtatga tccaatatca aaggaaatga tagcattgaa ggatgagact aatccaattg
121 aggagtggca gcatatagaa cagctaaagg gtagtgctga aggaagcata cgataccccg
181 catggaatgg gataatatca caggaggtac tagactacct ttcatcctac ataaatagac
241 gcatataagt acgcatttaa gcataaacac gcactatgcc gttcttctca tgtatatata
301 tatacaggca acacgcagat ataggtgcga cgtgaacagt gagctgtatg tgcgcagctc
361 gcgttgcatt ttcggaagcg ctcgttttcg gaaacgcttt gaagttccta ttccgaagtt
421 cctattctct agaaagtata ggaacttcag agcgcttttg aaaaccaaaa gcgctctgaa
481 gacgcacttt caaaaaacca aaaacgcacc ggactgtaac gagctactaa aatattgcga
541 ataccgcttc cacaaacatt gctcaaaagt atctctttgc tatatatctc tgtgctatat
601 ccctatataa cctacccatc cacctttcgc tccttgaact tgcatctaaa ctcgacctct
661 acatttttta tgtttatctc tagtattact ctttagacaa aaaaattgta gtaagaacta
721 ttcatagagt gaatcgaaaa caatacgaaa atgtaaacat ttcctatacg tagtatatag
781 agacaaaata gaagaaaccg ttcataattt tctgaccaat gaagaatcat caacgctatc
841 actttctgtt cacaaagtat gcgcaatcca catcggtata gaatataatc ggggatgcct
901 ttatcttgaa aaaatgcacc cgcagcttcg ctagtaatca gtaaacgcgg gaagtggagt
961 caggcttttt ttatggaaga gaaaatagac accaaagtag ccttcttcta accttaacgg
1021 acctacagtg caaaaagtta tcaagagact gcattataga gcgcacaaag gagaaaaaaa
1081 gtaatctaag atgctttgtt agaaaaatag cgctctcggg atgcattttt gtagaacaaa
1141 aaagaagtat agattctttg ttggtaaaat agcgctctcg cgttgcattt ctgttctgta
1201 aaaatgcagc tcagattctt tgtttgaaaa attagcgctc tcgcgttgca tttttgtttt
1261 acaaaaatga agcacagatt cttcgttggt aaaatagcgc tttcgcgttg catttctgtt
1321 ctgtaaaaat gcagctcaga ttctttgttt gaaaaattag cgctctcgcg ttgcattttt
1381 gttctacaaa atgaagcaca gatgcttcgt tcaggtggca cttttcgggg aaatgtgcgc
1441 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
1501 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
1561 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
1621 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
1681 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
1741 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
1801 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
1861 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
1921 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
1981 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
2041 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
2101 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
2161 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
2221 tggtttattg ctgataaatc tggagccggt gagcgtgggt cccgcggtat cattgcagca
2281 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
2341 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
2401 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
2461 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
2521 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
2581 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa accaccgcta ccagcggtgg
2641 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
2701 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
2761 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
2821 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
2881 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
2941 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
3001 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
3061 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
3121 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
3181 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
3241 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
3301 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
3361 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
3421 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
3481 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
3541 ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac cctcactaaa
3601 gggaacaaaa gctggagctt ctttgaaaag ataatgtatg attatgcttt cactcatatt
3661 tatacagaaa cttgatgttt tctttcgagt atatacaagg tgattacatg tacgtttgaa
3721 gtacaactct agattttgta gtgccctctt gggctagcgg taaaggtgcg cattttttca
3781 caccctacaa tgttctgttc aaaagatttt ggtcaaacgc tgtagaagtg aaagttggtg
3841 cgcatgtttc ggcgttcgaa acttctccgc agtgaaagat aaatgatcat aacggaatcc
3901 aactgggcgt tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt
3961 gaaaaagtgg caccgagtcg gtggtgcttt ttttgttttt tatgtcttcg agtcatgtaa
4021 ttagttatgt cacgcttacg ttcacgccct ccccccacat ccgctctaac cgaaaaggaa
4081 ggagttagac aacctgaagt ctaggtccct atttattttt ttatagttat gttagtatta
4141 agaacgttat ttatatttca aatttttctt ttttttctgt acagacgcgt gtacgcatgt
4201 aacattatac tgaaaacctt gcttgagaag gttttgggac gctcgaaggc tttaatttgc
4261 ctcgaagaca taaaaaacaa aaaaagcacc accgactcgg tgccactttt tcaagttgat
4321 aacggactag ccttatttta acttgctatt tctagctcta aaactgagac cagatctggt
4381 ctcgatcatt tatctttcac tgcggagaag tttcgaacgc cgaaacatgc gcaccaactt
4441 tcacttctac agcgtttgac caaaatcttt tgaacagaac attgtagggt gtgaaaaaat
4501 gcgcaccttt accgctagcc caagagggca ctacaaaatc tagagttgta cttcaaacgt
4561 acatgtaatc accttgtata tactcgaaag aaaacatcaa gtttctgtat aaatatgagt
4621 gaaagcataa tcatacatta tcttttcaaa gaatggcgcg acgcgccctg tagcggcgca
4681 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
4741 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
4801 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
4861 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
4921 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
4981 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
5041 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
5101 ttaacgctta caatttcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
5161 accgcatagg gtaataactg atataattaa attgaagctc taatttgtga gtttagtata
5221 catgcattta cttataatac agttttttag ttttgctggc cgcatcttct caaatatgct
5281 tcccagcctg cttttctgta acgttcaccc tctaccttag catcccttcc ctttgcaaat
5341 agtcctcttc caacaataat aatgtcagat cctgtagata ccacatcatc cacggttcta
5401 tactgttgac ccaatgcgtc tcccttgtca tctaaaccca caccgggtgt cataatcaac
5461 caatcgtaac cttcatctct tccacccatg tctctttgag caataaagcc gataacaaaa
5521 tctttgtcgc tcttcgcaat gtcaacagta cccttagtat attctccagt agatagggag
5581 cccttgcatg acaattctgc taacatcaaa aggcctctag gttcctttgt tacttcttct
5641 gccgcctgct tcaaaccgct aacaatacct gggcccacca caccgtgtgc attcgtaatg
5701 tctgcccatt ctgctattct gtatacaccc gcagagtact gcaatttgac tgtattacca
5761 atgtcagcaa attttctgtc ttcgaagagt aaaaaattgt acttggcgga taatgccttt
5821 agcggcttaa ctgtgccctc catggaaaaa tcagtcaaga tatccacatg tgtttttagt
5881 aaacaaattt tgggacctaa tgcttcaact aactccagta attccttggt ggtacgaaca
5941 tccaatgaag cacacaagtt tgtttgcttt tcgtgcatga tattaaatag cttggcagca
6001 acaggactag gatgagtagc agcacgttcc ttatatgtag ctttcgacat gatttatctt
6061 cgtttcctgc aggtttttgt tctgtgcagt tgggttaaga atactgggca atttcatgtt
6121 tcttcaacac tacatatgcg tatatatacc aatctaagtc tgtgctcctt ccttcgttct
6181 tccttctgtt cggagattac cgaatcaaaa aaatttcaag gaaaccgaaa tcaaaaaaaa
6241 gaataaaaaa aaaatgatga attgaaaagg tggtatggtg cactctcagt acaatctgct
6301 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
6361 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
6421 tgtgtcagag gttttcaccg tcatcaccga aacgcgcga
//
| {
"Vector backbone": "p426",
"Backbone size w/o insert (bp)": "5500",
"Vector type": "CRISPR, Synthetic Biology",
"Selectable markers": "URA3 ; Canavanine (after mutagenesis)"
} | {
"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": "Double Selection Enhances the Efficiency of Target-AID and Cas9-Based Genome Editing in Yeast",
"Publication": "G3 (Bethesda). 2018 Oct 3;8(10):3163-3171. doi: 10.1534/g3.118.200461.",
"DOI": "10.1534/g3.118.200461.",
"Authors": "Despres PC, Dube AK, Nielly-Thibault L, Yachie N, Landry CR /g",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/30097473",
"PubMed ID": "PubMed 30097473"
} | {
"Gene/Insert name": "DYSCKO cassette",
"Species": "S. cerevisiae (budding yeast)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "SNR52",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Gibson Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "GCGCCGCTACAGGGCGCGT",
"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": ""
} |
|
pENTR223-BXRF1 | 195789 | Gateway Entry Clone of Epstein-Barr virus BXRF1 | Eric Johannsen | Calderwood et al Proc Natl Acad Sci U S A. 2007 May 1;104(18):7606-11. doi: 10.1073/pnas.0702332104. Epub 2007 Apr 19. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/385735/bf361c6e-e186-437a-a5ab-a9b9b22db374/addgene-plasmid-195789-sequence-385735.gbk | full | LOCUS sequence-385735- 3531 bp DNA circular SYN 27-AUG-2024
DEFINITION Gateway Entry Clone of Epstein-Barr virus BXRF1.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3531)
AUTHORS Calderwood MA, Venkatesan K, Xing L, Chase MR, Vazquez A, Holthaus
AM, Ewence AE, Li N, Hirozane-Kishikawa T, Hill DE, Vidal M, Kieff
E, Johannsen E
TITLE Epstein-Barr virus and virus human protein interaction maps.
JOURNAL Proc Natl Acad Sci U S A. 2007 May 1;104(18):7606-11. doi:
10.1073/pnas.0702332104. Epub 2007 Apr 19.
PUBMED 17446270
REFERENCE 2 (bases 1 to 3531)
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:
"10.1073/pnas.0702332104"; journalName: "Proc Natl Acad Sci U S A";
date: "2007-05-1- 1"; volume: "104"; issue: "18"; pages: "7606-11"
FEATURES Location/Qualifiers
source 1..3531
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 92..109
/label=L4440
/note="L4440 vector, forward primer"
terminator complement(268..295)
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator complement(387..473)
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
primer_bind 522..544
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 536..553
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 537..553
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 569..668
/gene="mutant version of attL"
/label=attL1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
protein_bind complement(1410..1509)
/gene="mutant version of attL"
/label=attL2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
primer_bind complement(1526..1545)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(1527..1545)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(1550..1566)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1550..1566)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
CDS 1997..2788
/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 2881..3469
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3370..3389
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
661 agttggcatg gatcccaccc ggggtctgtg tgccctctcc acacacgacc tggcaaaatt
721 tcacagtctt cccccggcta gaaaggcggc aggtaagcga gcgcaccttc ggtgttactc
781 caagctgctc tctcttaaga gctgggagca actggcctct tttttgtctc tgcccccggg
841 acccacgttt acagacttta gactattttt cgaagtcacc ctgggtcgga gaatcgcaga
901 ttgcgttgtg gtagctctgc agccttatcc ccggtgttat attgtagaat ttaagacggc
961 catgagcaac acggccaacc cgcaaagcgt tactcgcaag gcacagaggc tagagggcac
1021 cgcccagttg tgtgactgtg ccaattttct tcgcacgtcc tgcccccccg tgctgggcag
1081 tcagggcctg gaagtcttgg cggcgttggt atttaaaaac cagcgatccc tgagaacgct
1141 ccaggtagag tttccagccc tgggccaaaa gaccctcccc acctccacca ccggcctgct
1201 aaacctcctc tcccgctggc aggatggcgc tctccgggca cgtcttgata gaccccgccc
1261 gactgcccag ggacacaggc cccgaactca tgtgggcccc aagccttcgc aactcactgc
1321 gcgtgtcccc cgaagcgctc gagctggcag agcgggaggc cgaaagggcc aggtcggagc
1381 ggtgggacag gtgtgcccag gtgctcaaaa accaactttc ttgtacaaag ttggcattat
1441 aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc aaaataaaat
1501 cattatttgc catccagctg atatccccta tagtgagtcg tattacatgg tcatagctgt
1561 ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt tacattgcac aagataaaaa
1621 tatatcatca tgcctcctct agaccagcca ggacagaaat gcctcgactt cgctgctacc
1681 caaggttgcc gggtgacgca caccgtggaa acggatgaag gcacgaaccc agtggacata
1741 agcctgttcg gttcgtaagc tgtaatgcaa gtagcgtatg cgctcacgca actggtccag
1801 aaccttgacc gaacgcagcg gtggtaacgg cgcagtggcg gttttcatgg cttgttatga
1861 ctgttttttt ggggtacagt ctatgcctcg ggcatccaag cagcaagcgc gttacgccgt
1921 gggtcgatgt ttgatgttat ggagcagcaa cgatgttacg cagcagggca gtcgccctaa
1981 aacaaagtta aacattatga gggaagcggt gatcgccgaa gtatcgactc aactatcaga
2041 ggtagttggc gtcatcgagc gccatctcga accgacgttg ctggccgtac atttgtacgg
2101 ctccgcagtg gatggcggcc tgaagccaca cagtgatatt gatttgctgg ttacggtgac
2161 cgtaaggctt gatgaaacaa cgcggcgagc tttgatcaac gaccttttgg aaacttcggc
2221 ttcccctgga gagagcgaga ttctccgcgc tgtagaagtc accattgttg tgcacgacga
2281 catcattccg tggcgttatc cagctaagcg cgaactgcaa tttggagaat ggcagcgcaa
2341 tgacattctt gcaggtatct tcgagccagc cacgatcgac attgatctgg ctatcttgct
2401 gacaaaagca agagaacata gcgttgcctt ggtaggtcca gcggcggagg aactctttga
2461 tccggttcct gaacaggatc tatttgaggc gctaaatgaa accttaacgc tatggaactc
2521 gccgcccgac tgggctggcg atgagcgaaa tgtagtgctt acgttgtccc gcatttggta
2581 cagcgcagta accggcaaaa tcgcgccgaa ggatgtcgct gccgactggg caatggagcg
2641 cctgccggcc cagtatcagc ccgtcatact tgaagctaga caggcttatc ttggacaaga
2701 agaagatcgc ttggcctcgc gcgcagatca gttggaagaa tttgtccact acgtgaaagg
2761 cgagatcacc aaggtagtcg gcaaataacc ctcgagccac ccatgaccaa aatcccttaa
2821 cgtgagttac gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
2881 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
2941 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
3001 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
3061 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
3121 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
3181 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
3241 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
3301 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
3361 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
3421 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
3481 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt t
//
| {
"Vector backbone": "pENTR223.1",
"Backbone size w/o insert (bp)": "2875",
"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": "Epstein-Barr virus and virus human protein interaction maps",
"Publication": "Proc Natl Acad Sci U S A. 2007 May 1;104(18):7606-11. doi: 10.1073/pnas.0702332104. Epub 2007 Apr 19.",
"DOI": "10.1073/pnas.0702332104.",
"Authors": "Calderwood MA, Venkatesan K, Xing L, Chase MR, Vazquez A, Holthaus AM, Ewence AE, Li N, Hirozane-Kishikawa T, Hill DE, Vidal M, Kieff E, Johannsen E /pnas",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/17446270",
"PubMed ID": "PubMed 17446270"
} | {
"Gene/Insert name": "BXRF1",
"Species": "Epstein-Barr virus",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "BXRF1",
"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": ""
} |
|
pLKO.1-TSC2 | 15478 | High | Do-Hyung Kim | Vander Haar et al Nat Cell Biol. 2007 Mar . 9(3):316-23. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/153462/64710466-36fb-4acf-a86b-fafef9cad24b/addgene-plasmid-15478-sequence-153462.gbk | full | LOCUS sequence-153462- 7085 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 7085)
AUTHORS Vander Haar E, Lee SI, Bandhakavi S, Griffin TJ, Kim DH
TITLE Insulin signalling to mTOR mediated by the Akt/PKB substrate PRAS40.
JOURNAL Nat Cell Biol. 2007 Mar . 9(3):316-23.
PUBMED 17277771
REFERENCE 2 (bases 1 to 7085)
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 Cell
Biol. 2007 Mar . 9(3):316-23."
FEATURES Location/Qualifiers
source 1..7085
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 426..444
/label=hPGK-F
/note="Human PGK promoter, forward primer"
CDS 529..1128
/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(529..548)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 1025..1045
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
LTR 1256..1489
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 1561..1682
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1598..1617)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 1652..1671
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 1722..1857
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(1877..1896)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(1878..1896)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(1906..1923)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(1906..1922)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1915..1937)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 2064..2519
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2151..2170)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2361..2382
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 2545..2649
/gene="bla"
/label=AmpR promoter
CDS 2650..3510
/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(2868..2887)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 3681..4269
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4170..4189
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 4423..4440
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 4557..4578
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 4593..4623
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4631..4647
/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 4636..4658
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 4655..4671
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 4655..4671
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 4690..4710
/label=T3
/note="T3 promoter, forward primer"
promoter 4692..4710
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 4738..4964
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 4965..5145
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 5192..5317
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 5810..6043
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 6228..6272
/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 6570..6818
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA
(Domitrovich & Kunkel, 2003)"
primer_bind 6570..6590
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 6741..6760
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
misc_feature 6916..7033
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter join(7082..7085,1..507)
/label=hPGK promoter
/note="human phosphoglycerate kinase 1 promoter"
ORIGIN
1 ttggggttgc gccttttcca aggcagccct gggtttgcgc agggacgcgg ctgctctggg
61 cgtggttccg ggaaacgcag cggcgccgac cctgggtctc gcacattctt cacgtccgtt
121 cgcagcgtca cccggatctt cgccgctacc cttgtgggcc ccccggcgac gcttcctgct
181 ccgcccctaa gtcgggaagg ttccttgcgg ttcgcggcgt gccggacgtg acaaacggaa
241 gccgcacgtc tcactagtac cctcgcagac ggacagcgcc agggagcaat ggcagcgcgc
301 cgaccgcgat gggctgtggc caatagcggc tgctcagcag ggcgcgccga gagcagcggc
361 cgggaagggg cggtgcggga ggcggggtgt ggggcggtag tgtgggccct gttcctgccc
421 gcgcggtgtt ccgcattctg caagcctccg gagcgcacgt cggcagtcgg ctccctcgtt
481 gaccgaatca ccgacctctc tccccagggg gatccaccgg agcttaccat gaccgagtac
541 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc
601 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag
661 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg
721 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg
781 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc
841 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc
901 ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg
961 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg
1021 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg
1081 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgacg cccgccccac
1141 gacccgcagc gcccgaccga aaggagcgca cgaccccatg catcggtacc tttaagacca
1201 atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa
1261 gggctaattc actcccaacg aagacaagat ctgctttttg cttgtactgg gtctctctgg
1321 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
1381 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
1441 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tagtagttca
1501 tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg
1561 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
1621 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
1681 tatcatgtct ggctctagct atcccgcccc taactccgcc catcccgccc ctaactccgc
1741 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
1801 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
1861 ggacgtaccc aattcgccct atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt
1921 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
1981 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
2041 gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
2101 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
2161 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
2221 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
2281 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
2341 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
2401 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
2461 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta
2521 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
2581 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
2641 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
2701 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
2761 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
2821 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
2881 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
2941 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
3001 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
3061 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
3121 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
3181 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
3241 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
3301 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
3361 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
3421 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
3481 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
3541 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
3601 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
3661 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
3721 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
3781 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
3841 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
3901 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
3961 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
4021 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
4081 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
4141 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
4201 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
4261 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
4321 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
4381 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
4441 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
4501 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
4561 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
4621 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
4681 gccaagcgcg caattaaccc tcactaaagg gaacaaaagc tggagctgca agcttaatgt
4741 agtcttatgc aatactcttg tagtcttgca acatggtaac gatgagttag caacatgcct
4801 tacaaggaga gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg tacgatcgtg
4861 ccttattagg aaggcaacag acgggtctga catggattgg acgaaccact gaattgccgc
4921 attgcagaga tattgtattt aagtgcctag ctcgatacat aaacgggtct ctctggttag
4981 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
5041 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
5101 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag
5161 ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc ggcttgctga
5221 agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag
5281 cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag
5341 atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca
5401 tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac
5461 atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga
5521 agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga
5581 gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac
5641 caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg agggacaatt
5701 ggagaagtga attatataaa tataaagtag taaaaattga accattagga gtagcaccca
5761 ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt
5821 tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg
5881 tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta
5941 ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa
6001 gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct
6061 ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc
6121 tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca
6181 caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag
6241 aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc
6301 tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt
6361 ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga
6421 cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa gaaggtggag
6481 agagagacag agacagatcc attcgattag tgaacggatc tcgacggtat cgatcacgag
6541 actagcctcg agcggccgcc cccttcaccg agggcctatt tcccatgatt ccttcatatt
6601 tgcatatacg atacaaggct gttagagaga taattggaat taatttgact gtaaacacaa
6661 agatattagt acaaaatacg tgacgtagaa agtaataatt tcttgggtag tttgcagttt
6721 taaaattatg ttttaaaatg gactatcata tgcttaccgt aacttgaaag tatttcgatt
6781 tcttggcttt atatatcttg tggaaaggac gaaacaccgg tcactggcct tggacggtat
6841 tgctcgagca ataccgtcca aggccagtgt ttttgaattc tcgacctcga gacaaatggc
6901 agtattcatc cacaatttta aaagaaaagg ggggattggg gggtacagtg caggggaaag
6961 aatagtagac ataatagcaa cagacataca aactaaagaa ttacaaaaac aaattacaaa
7021 aattcaaaat tttcgggttt attacaggga cagcagagat ccactttggc cgcggctcga
7081 ggggg
//
| {
"Vector backbone": "pLKO.1",
"Backbone size w/o insert (bp)": "7000",
"Vector type": "Mammalian Expression, Lentiviral, RNAi",
"Selectable markers": "Puromycin"
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "Stbl3",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Insulin signalling to mTOR mediated by the Akt/PKB substrate PRAS40",
"Publication": "Nat Cell Biol. 2007 Mar . 9(3):316-23.",
"DOI": "10.1038/ncb1547",
"Authors": "Vander Haar E, Lee SI, Bandhakavi S, Griffin TJ, Kim DH",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/17277771",
"PubMed ID": "PubMed 17277771"
} | {
"Gene/Insert name": "TSC2 shRNA",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "59",
"Mutation": "",
"Entrez Gene": "TSC2",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "AgeI (not destroyed)",
"3′ cloning site": "EcoRI (not destroyed)",
"5′ sequencing primer": "LKO.1 5'",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
H1-Mos-6U | 120531 | dsRNA expression | Petr Svoboda | Petr Svoboda lab unpublished (unpublished) | https://media.addgene.org/snapgene-media/v2.0.0/sequences/234010/7744ed4b-5b7e-456c-a041-de4f92760f17/addgene-plasmid-120531-sequence-234010.gbk | full | LOCUS sequence-234010- 2852 bp DNA circular SYN 26-AUG-2024
DEFINITION dsRNA expression .
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2852)
TITLE Petr Svoboda lab unpublished
REFERENCE 2 (bases 1 to 2852)
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..2852
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 83..331
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA
(Domitrovich & Kunkel, 2003)"
primer_bind 83..103
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 254..273
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
primer_bind complement(689..708)
/label=H1
/note="Human H1 promoter, forward primer"
primer_bind complement(854..872)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 940..1044
/gene="bla"
/label=AmpR promoter
CDS 1045..1905
/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(1263..1282)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 2076..2664
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2565..2584
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 2818..2835
/label=L4440
/note="L4440 vector, forward primer"
ORIGIN
1 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
61 gtttcccgac tggaaagcaa ttgagggcct atttcccatg attccttcat atttgcatat
121 acgatacaag gctgttagag agataattgg aattaatttg actgtaaaca caaagatatt
181 agtacaaaat acgtgacgta gaaagtaata atttcttggg tagtttgcag ttttaaaatt
241 atgttttaaa atggactatc atatgcttac cgtaacttga aagtatttcg atttcttggc
301 tttatatatc ttgtggaaag gacgaaacac cgggaaaaag cttgcaccaa ggacctacgt
361 gcatcccagc ggagtttctg ggctgaactg aacattgcaa gactacgcca cgacaacata
421 gttcgggttg tggctgccag cacgcgcacg cccgaagact ccaacagcct aggtaccata
481 atcatggagt ttgggggcaa cgtgactcta caccaagtca tctacggtgc cacccgctca
541 ccggagcctc tcagctgcag agaacaaccg agtttgggga agtgcctcaa gtattcccta
601 gatgttgtta gcggccgctt tttccctcta tcactgatag ggaacttata agattcccaa
661 atccaaagac atttcacgtt tatggtgatt tcccagaaca catagcgaca tgcaaatatt
721 gcagggcgcc actcccctgt ccctcacagc catcttcctg ccagggcgca cgcgcgctgg
781 gtgttcccgc ctagtgacac tgggcccgcg attccttgga gcgggttgat gacgtcagcg
841 ttctgcaggg gcggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
901 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
961 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
1021 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
1081 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
1141 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
1201 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
1261 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
1321 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
1381 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
1441 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
1501 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
1561 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
1621 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
1681 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
1741 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
1801 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
1861 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
1921 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
1981 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
2041 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
2101 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
2161 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
2221 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
2281 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
2341 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
2401 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
2461 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
2521 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
2581 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
2641 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
2701 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
2761 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
2821 gagtcagtga gcgaggaagc ggaagagcgc cc
//
| {
"Vector backbone": "pJET1.2",
"Backbone size w/o insert (bp)": "",
"Vector type": "RNAi",
"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": "Mos",
"Species": "M. musculus (mouse)",
"Insert Size (bp)": "",
"Mutation": "A>G (see comments)",
"Entrez Gene": "Mos",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "Unknown (unknown if destroyed)",
"3′ cloning site": "Unknown (unknown if destroyed)",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
integrin alpha9 W999A pcDNA1 neo | 13589 | Dean Sheppard | Young et al Mol Biol Cell. 2001 Oct . 12(10):3214-25. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/7708/b4d42d50-ea03-43bd-a1a5-18b66c728ed7/addgene-plasmid-13589-sequence-7708.gbk | full | LOCUS sequence-7708-ex 10098 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 10098)
AUTHORS Young BA, Taooka Y, Liu S, Askins KJ, Yokosaki Y, Thomas SM,
Sheppard D
TITLE The cytoplasmic domain of the integrin alpha9 subunit requires the
adaptor protein paxillin to inhibit cell spreading but promotes cell
migration in a paxillin-independent manner.
JOURNAL Mol Biol Cell. 2001 Oct . 12(10):3214-25.
PUBMED 11598204
REFERENCE 2 (bases 1 to 10098)
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 Biol
Cell. 2001 Oct . 12(10):3214-25."
FEATURES Location/Qualifiers
source 1..10098
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 471..490
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
rep_origin complement(603..1061)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(740..761)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 955..974
/label=F1ori-R
/note="F1 origin, reverse primer"
tRNA 1204..1383
/product="E. coli amber-suppressor tyrosine tRNA"
/label=supF
enhancer 1542..1921
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 1922..2125
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 2076..2096
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 2170..2189
/label=T7
/note="T7 promoter, forward primer"
promoter 2170..2188
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(5382..5398)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5382..5398)
/label=pBluescriptSK
/note="For pBluescript vector"
promoter complement(5434..5452)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(5435..5452)
/label=SP6
/note="SP6 promoter, forward primer"
intron 5536..5601
/label=small t intron
/note="SV40 (simian virus 40) small t antigen intron"
CDS 5731..5751
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
polyA_signal 6023..6157
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(6185..6204)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
promoter 6481..6743
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
CDS 7103..7897
/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(7157..7176)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 7767..7786
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
intron 8318..8383
/label=small t intron
/note="SV40 (simian virus 40) small t antigen intron"
CDS 8513..8533
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
polyA_signal 8958..9092
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
rep_origin 9939..10074
/label=SV40 ori
/note="SV40 origin of replication"
ORIGIN
1 ggcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
61 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
121 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
181 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
241 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
301 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
361 ctacagcgtg agcattgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
421 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
481 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
541 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcaagcta gcttctagct
601 agaaattgta aacgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
661 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagcccga
721 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
781 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc cgcccactac gtgaaccatc
841 acccaaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
901 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
961 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
1021 caccacaccc gccgcgctta atgcgccgct acagggcgcg tactatggtt gctttgacga
1081 gaccgtataa cgtgctttcc tcgttggaat cagagcggga gctaaacagg aggccgatta
1141 aagggatttt agacaggaac ggtacgccag ctggattacc gcggtctttc tcaacgtaac
1201 actttacagc ggcgcgtcat ttgatatgat gcgccccgct tcccgataag ggagcaggcc
1261 agtaaaagca ttacccgtgg tggggttccc gagcggccaa agggagcaga ctctaaatct
1321 gccgtcatcg acttcgaagg ttcgaatcct tcccccacca ccatcacttt caaaagtccg
1381 aaagaatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gtaaaattta
1441 agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg
1501 ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag
1561 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
1621 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
1681 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
1741 ggtggactat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
1801 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
1861 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
1921 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
1981 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
2041 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
2101 gtgggaggtc tatataagca gagctctctg gctaactaga gaacccactg cttactggct
2161 tatcgaaatt aatacgactc actataggga gacccaagct tagagcgggc gctatgggga
2221 gccggacgcc agagtcccct ctccatgccg tgcagctgcg ctggggcccc cggcgccgac
2281 ccccgctgct gccgctgcta ttgctgctcg tgccgccgcc accgcgggtc gggggctaca
2341 acctggatcc gcagcgcccc gtgcacttcc aaggccccgc tgactcgttc ttcggctacg
2401 cagttctgga gcatttccac gacaacacgc gctgggtcct tgtgggcgca ccaaaggcag
2461 attccaaata cagcccttca gtgaagtctc ctggggctgt gtttaagtgc cgtgttcaca
2521 ccaaccctga ccggagatgc accgaactgg acatggctcg agggaagaat cggggcacgt
2581 cctgcggaaa gacctgccgg gaagaccgcg atgatgagtg gatgggggtg agcctggccc
2641 gacagcccaa ggctgatggc cgtgtgttgg cctgtgctca tcgctggaag aacatctact
2701 atgaagccga ccacatccta ccccatggct tctgctacat catcccctcc aacctccagg
2761 ccaaaggcag gacgctgatc ccttgctatg aagagtataa gaagaagtac ggagaggaac
2821 acggctcctg ccaggctggg atagcgggct tcttcaccga ggagctggtg gtgatgggtg
2881 ctccagggtc attttattgg gctggaacca tcaaagtgct gaaccttacg gacaacacct
2941 atttaaaact gaacgacgaa gtgatcatga acaggcggta cacctacctg ggctacgcag
3001 tgaccgctgg ccacttctct cacccgtcca ccattgatgt ggtaggaggt gccccacagg
3061 acaaaggcat cggcaaggtt tatattttca gagctgaccg aagatcaggc accttaatta
3121 agatctttca agcatcaggt aaaaagatgg gctcttactt cggctcctcc ttgtgcgcag
3181 ttgacctgaa tggggacggc ctctctgacc tgctggtggg ggcccccatg ttttctgaga
3241 tcagggatga gggacaggtc actgtctaca tcaacagagg aaatggagcc ctcgaggagc
3301 agctggctct gactggggat ggtgcctaca atgcccactt tggagagagc attgccagcc
3361 tggacgatct ggacaatgat gggttcccag atgtggccat tggtgcaccc aaggaggatg
3421 acttcgcagg ggcggtctat atctatcatg gtgatgccgg tgggatagtc cctcagtact
3481 caatgaaact gtctgggcag aagataaatc cagtgctccg gatgtttggt cagtccatat
3541 cgggaggcat tgatatggat ggaaatggct atcctgatgt cactgttgga gccttcatgt
3601 ccgacagcgt ggttcttctc agagcaaggc ctgtcattac ggtggatgtc tccatcttcc
3661 tcccgggctc catcaacatc acagcgcctc agtgtcacga cggacagcag cctgtgaact
3721 gcctgaacgt caccacctgc ttcagcttcc atggcaaaca cgttccagaa gagattggcc
3781 tgaattatgt tctgatggct gacgtggcca aaaaggagaa gggccagatg cccagggtct
3841 actttgtgct gctgggagag accatgggtc aggtcacaga gaagctgcag ctgacttaca
3901 tggaggagac gtgtcgtcac tatgtggccc atgtgaagcg gagggtgcag gacgtcatca
3961 gcccgatcgt gtttgaagca gcctacagcc tcagtgagca tgtgactgga gaggaggaga
4021 gggaactgcc gcctctgaca ccagttctcc gctggaaaaa gggacaaaag attgcccaaa
4081 agaatcagac tgtttttgaa aggaattgcc gttcagagga ctgtgccgca gacctgcagc
4141 ttcagggtaa actgctgctc tccagtatgg atgagaaaac cctgtatcta gctttggggg
4201 ctgtgaagaa catctcccta aacatctcta tctccaacct cggagatgat gcctatgatg
4261 ccaacgtgtc cttcaatgtt tcccgggagc tcttcttcat caacatgtgg cagaaggagg
4321 agatgggcat ctcctgtgag ctgctggaat cggacttcct caaatgcagc gtgggatttc
4381 ctttcatgag gtcaaagtca aagtatgaat tcagcgtgat ctttgataca agccacctgt
4441 ctggggaaga ggaagttctc agcttcattg ttactgctca gagtggcaac acggagcgct
4501 ctgaatccct gcatgacaac accctcgtgc tgatggtgcc actgatgcac gaggtggaca
4561 cgtccatcac cggaatcatg tctccaacct cctttgtata tggcgagtcc gtggacgcag
4621 ccaacttcat tcagctggat gacctggagt gtcactttca gcccatcaat atcacccttc
4681 aggtctacaa cactggccca agcacccttc cagggtcatc tgtcagcatc tctttcccta
4741 atcgactctc atctggtggt gcagagatgt ttcatgtcca ggaaatggtg gtgggccaag
4801 agaagggaaa ctgctctttc cagaaaaacc caactccctg catcatccct caagaacaag
4861 aaaatatctt ccacacaata tttgcttttt tcacaaagtc tggaagaaaa gtcttggact
4921 gtgaaaaacc aggaatttct tgcctaacag cacactgtaa ctttagtgct cttgctaaag
4981 aagaaagtcg tactatagac atttacatgc tgctgaacac agaaatactg aaaaaggaca
5041 gttcgtctgt catccagttc atgtcccgcg ccaaggtgaa ggtggatcct gccctaaggg
5101 tggtggaaat agctcatggg aacccagaag aggtgacggt ggtcttcgag gccctgcaca
5161 atctggagcc ccgtggctac gtcgtggggt ggatcatcgc catcagtttg ttggtgggaa
5221 tcctcatctt cctgctgctg gccgtgctgc tctggaagat gggcttcttt cgccgaaggt
5281 acaaagaaat tatcgaagct gagaagaacc ggaaagagaa tgaagacagt gcggactggg
5341 tccagaaaaa ccagtgagct gccacaccag tcacatgacc tgatccacta gttctagagc
5401 ggccgcctcg agcatgcatc tagagggccc tattctatag tgtcacctaa atgctagagg
5461 atctttgtga aggaacctta cttctgtggt gtgacataat tggacaaact acctacagag
5521 atttaaagct ctaaggtaaa tataaaattt ttaagtgtat aatgtgttaa actactgatt
5581 ctaattgttt gtgtatttta gattccaacc tatggaactg atgaatggga gcagtggtgg
5641 aatgccttta atgaggaaaa cctgttttgc tcagaagaaa tgccatctag tgatgatgag
5701 gctactgctg actctcaaca ttctactcct ccaaaaaaga agagaaaggt agaagacccc
5761 aaggactttc cttcagaatt gctaagtttt ttgagtcatg ctgtgtttag taatagaact
5821 cttgcttgct ttgctattta caccacaaag gaaaaagctg cactgctata caagaaaatt
5881 atggaaaaat atttgatgta tagtgccttg actagagatc ataatcagcc ataccacatt
5941 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
6001 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
6061 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
6121 gtccaaactc atcaatgtat cttatcatgt ctggatctcc cgatccccta tggtgcactc
6181 tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt
6241 tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg
6301 acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg
6361 ccagatatac gcgtatctga ggggactagg gtgtgtttag gcgaaaagcg gggcttcggt
6421 tgtacgcggt taggagtccc ctcaggatat agtagtttcg cttttgcata gggaggggga
6481 aatgtagtct tatgcaatac acttgtagtc ttgcaacatg gtaacgatga gttagcaaca
6541 tgccttacaa ggagagaaaa agcaccgtgc atgccgattg gtggaagtaa ggtggtacga
6601 tcgtgcctta ttaggaaggc aacagacagg tctgacatgg attggacgaa ccactgaatt
6661 ccgcattgca gagataattg tatttaagtg cctagctcga tacaataaac gccatttgac
6721 cattcaccac attggtgtgc acctcctaag ctagctttgc cgcaagcact cagggcgcaa
6781 gggctgctaa aggaagcgga acacgtagaa agccagtccg cagaaacggt gctgaccccg
6841 gatgaatgtc agctactggg ctatctggac aagggaaaac gcaagcgcaa agagaaagca
6901 ggtagcttgc agtgggctta catggcgata gctagactgg gcggttttat ggacagcaag
6961 cgaaccggaa ttgccagctg gggcgccctc tggtaaggtt gggaagccct gcaaagtaaa
7021 ctggatggct ttcttgccgc caaggatctg atggcgcagg ggatcaagat ctgatcaaga
7081 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
7141 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
7201 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
7261 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
7321 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
7381 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
7441 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
7501 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
7561 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
7621 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
7681 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
7741 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
7801 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
7861 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
7921 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
7981 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
8041 gatctcatgc tggagttctt cgcccacccc gggctcgatc ccctcgcgag ttggttcagc
8101 tgctgcctga ggctggacga cctcgcggag ttctaccggc agtgcaaatc cgtcggcatc
8161 caggaaacca gcagcggcta tccgcgcatc catgcccccg aactgcagga gtggggaggc
8221 acgatggccg ctttggtccc ggatctttgt gaaggaacct tacttctgtg gtgtgacata
8281 attggacaaa ctacctacag agatttaaag ctctaaggta aatataaaat ttttaagtgt
8341 ataatgtgtt aaactactga ttctaattgt ttgtgtattt tagattccaa cctatggaac
8401 tgatgaatgg gagcagtggt ggaatgcctt taatgaggaa aacctgtttt gctcagaaga
8461 aatgccatct agtgatgatg aggctactgc tgactctcaa cattctactc ctccaaaaaa
8521 gaagagaaag gtagaagacc ccaaggactt tccttcagaa ttgctaagtt ttttgagtca
8581 tgctgtgttt agtaatagaa ctcttgcttg ctttgctatt tacaccacaa aggaaaaagc
8641 tgcactgcta tacaagaaaa ttatggaaaa atattctgta acctttataa gtaggcataa
8701 cagttataat cataacatac tgttttttct tactccacac aggcatagag tgtctgctat
8761 taataactat gctcaaaaat tgtgtacctt tagcttttta atttgtaaag gggttaataa
8821 ggaatatttg atgtatagtg ccttgactag agatcataat cagccatacc acatttgtag
8881 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
8941 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
9001 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
9061 aactcatcaa tgtatcttat catgtctgga tcgatcccgc catggtatca acgccatatt
9121 tctatttaca gtagggacct cttcgttgtg taggtaccgc tgtattccta gggaaatagt
9181 agaggcacct tgaactgtct gcatcagcca tatagccccc gctgttcgac ttacaaacac
9241 aggcacagta ctgacaaacc catacacctc ctctgaaata cccatagttg ctagggctgt
9301 ctccgaactc attacaccct ccaaagtcag agctgtaatt tcgccatcaa gggcagcgag
9361 ggcttctcca gataaaatag cttctgccga gagtcccgta agggtagaca cttcagctaa
9421 tccctcgatg aggtctacta gaatagtcag tgcggctccc attttgaaaa ttcacttact
9481 tgatcagctt cagaagatgg cggagggcct ccaacacagt aattttcctc ccgactctta
9541 aaatagaaaa tgtcaagtca gttaagcagg aagtggacta actgacgcag ctggccgtgc
9601 gacatcctct tttaattagt tgctaggcaa cgccctccag agggcgtgtg gttttgcaag
9661 aggaagcaaa agcctctcca cccaggccta gaatgtttcc acccaatcat tactatgaca
9721 acagctgttt tttttagtat taagcagagg ccggggaccc ctgggcccgc ttactctgga
9781 gaaaaagaag agaggcattg tagaggcttc cagaggcaac ttgtcaaaac aggactgctt
9841 ctatttctgt cacactgtct ggccctgtca caaggtccag cacctccata ccccctttaa
9901 taagcagttt gggaacgggt gcgggtctta ctccgcccat cccgccccta actccgccca
9961 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
10021 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
10081 tttgcaaaaa gctaattc
//
| {
"Vector backbone": "pcDNAI neo",
"Backbone size w/o insert (bp)": "7000",
"Vector type": "Mammalian Expression",
"Selectable markers": "Neomycin (select with G418)"
} | {
"Bacterial Resistance(s)": "Ampicillin and Tetracycline, 100 & 10 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "Top10/P3",
"Copy number": "Unknown"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "The cytoplasmic domain of the integrin alpha9 subunit requires the adaptor protein paxillin to inhibit cell spreading but promotes cell migration in a paxillin-independent manner",
"Publication": "Mol Biol Cell. 2001 Oct . 12(10):3214-25.",
"DOI": "10.1091/mbc.12.10.3214",
"Authors": "Young BA, Taooka Y, Liu S, Askins KJ, Yokosaki Y, Thomas SM, Sheppard D",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/11598204",
"PubMed ID": "PubMed 11598204"
} | {
"Gene/Insert name": "integrin alpha 9 W999A",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "3200",
"Mutation": "W999A",
"Entrez Gene": "ITGA9",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "HindIII (not destroyed)",
"3′ cloning site": "NotI (not destroyed)",
"5′ sequencing primer": "T7",
"3′ sequencing primer": "SP6"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
||
pEMS1435 | 29230 | Expression of the reporter gene under the control of the MiniPromoter in this plasmid has not been tested | Elizabeth Simpson | Portales-Casamar et al Proc Natl Acad Sci U S A. 2010 Sep 21. 107(38):16589-94. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/13645/b87155dc-4b3e-4229-984c-152d7b58d32e/addgene-plasmid-29230-sequence-13645.gbk | full | LOCUS sequence-13645-e 17760 bp DNA circular SYN 26-AUG-2024
DEFINITION Expression of the reporter gene under the control of the
MiniPromoter in this plasmid has not been tested.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 17760)
AUTHORS Portales-Casamar E, Swanson DJ, Liu L, de Leeuw CN, Banks KG, Ho Sui
SJ, Fulton DL, Ali J, Amirabbasi M, Arenillas DJ, Babyak N, Black
SF, Bonaguro RJ, Brauer E, Candido TR, Castellarin M, Chen J, Chen
Y, Cheng JC, Chopra V, Docking TR, Dreolini L, D'Souza CA, Flynn EK,
Glenn R, Hatakka K, Hearty TG, Imanian B, Jiang S, Khorasan-zadeh S,
Komljenovic I, Laprise S, Liao NY, Lim JS, Lithwick S, Liu F, Liu J,
Lu M, McConechy M, McLeod AJ, Milisavljevic M, Mis J, O'Connor K,
Palma B, Palmquist DL, Schmouth JF, Swanson MI, Tam B, Ticoll A,
Turner JL, Varhol R, Vermeulen J, Watkins RF, Wilson G, Wong BK,
Wong SH, Wong TY, Yang GS, Ypsilanti AR, Jones SJ, Holt RA,
Goldowitz D, Wasserman WW, Simpson EM
TITLE A regulatory toolbox of MiniPromoters to drive selective expression
in the brain.
JOURNAL Proc Natl Acad Sci U S A. 2010 Sep 21. 107(38):16589-94.
PUBMED 20807748
REFERENCE 2 (bases 1 to 17760)
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. 2010 Sep 21. 107(38):16589-94."
FEATURES Location/Qualifiers
source 1..17760
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 49..67
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
regulatory 1475..1484
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 1481..2197
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 1481..2197
/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(1526..1547)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1787..1806)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 2134..2155
/label=EGFP-C
/note="EGFP, forward primer"
CDS 2210..2227
/codon_start=1
/product="tetracysteine peptide that binds biarsenical
labeling reagents"
/label=tetracysteine tag
/translation="CCPGCC"
CDS 2240..2260
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 2264..2284
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 2288..2308
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
protein_bind 2315..2348
/label=FRT (minimal)
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="supports FLP-mediated excision but not integration
(Turan and Bode, 2011)"
polyA_signal 2471..2552
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2508..2527)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 10215..10236
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 10399..10503
/gene="bla"
/label=AmpR promoter
CDS 10504..11364
/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(15571..15593)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
misc_feature 15679..15819
/label=bom
/note="basis of mobility region from pBR322"
primer_bind 15734..15753
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(15834..15851)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(16005..16593)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(16085..16104)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(16764..17624)
/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"
promoter complement(17625..17729)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 cggccgcacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
61 acgaggccct ttcgtcttca agaattaatt ccgatcatat tcaataaccc ttaatataac
121 ttcgtataat gtatgcgaat tcgagctcct aggccggcca cgcgtggcgc gccaaatacc
181 acaagatgca tagaaggaag tgcaaagaaa ggagaaaaga gagacagaga tatactcaca
241 gcaaggtgca gagacatgga gaccctataa gagatggaaa tggagagcca cagaggggga
301 actgcgggca ctaagagaga catggagaga actagaaaga tagagacgtg aacacagaca
361 cagagaccca gagccaaaaa gagacagaaa tggggtcata ggtggacaga aatgcacaga
421 gagactgtga gcccccacag acacaaacgc tgagaggggc agacatgcag agacacccac
481 agaaatgcag agacaaaacc acccggagac acatgccgag aaacacactg aggggctcag
541 aaagcgacag atgcagagag agcccaagac agacccagca aggggccccg agaaatgcac
601 agaaacacaa agaccgagac caccagagac agaggtacgg agaggcgggg agggtgagga
661 agcctgctct ccagggctgg cttgaaccta gaactcaagg tcatggcttg gctgggaagc
721 gggaggggga taccaaaaaa gatcagcgtt agatggaggt ggggagggag ggtaggtttc
781 accctaattg acaccccacc ctccccctcc ccctccccac acctgcttcc cggctttagt
841 gcctgttgtc accaggatgg aatcctggcg gcagacaaag gggtgggggt gggaggagga
901 gacaaaaaga tggacagaga cagaggagga gactaagaag tagagacaga cacacagaga
961 cacacagatg gagagccaca gagacaacag aaatgggggt ccagaaagag acctagatag
1021 aaagagggaa cagagagaag aaaacaggaa acattcctta tagcccatgt gtcattagta
1081 ggtcctccaa ggacctactg tgtgccgggc atggggacgt caaagcacag ggagctataa
1141 gggaggtagt tggagcacca cgggagcctg gggtgtgggg tgggagcagg ggcggggctc
1201 ctgggccatg cagatggggt tggcatgggg ttggggaggg gcccacggag gcttctggac
1261 tctgagtgtg gagccagggg cagcagggct gtctctgaac tcccaggctt gtgtttgagc
1321 aaaggaattc tctccccctc cctctcccct cggggcgggc ccgagcctcc agctataaac
1381 tccccggagc ttcagtgccc tcagcaaggc tcagcctcaa gattcacagc aggcgcgccg
1441 aagttcctat tcttcaaaag gtataggaac ttccgccacc atggtgagca agggcgagga
1501 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
1561 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
1621 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
1681 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
1741 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
1801 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
1861 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
1921 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa
1981 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
2041 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc
2101 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
2161 cgccgggatc actctcggca tggacgagct gtacaagtta tttctgaact gctgcccggg
2221 ctgctgcatg gaaccggatc caaaaaagaa gagaaaggta gatccaaaaa agaagagaaa
2281 ggtagatcca aaaaagaaga gaaaggtaat ataagaagtt cctattctct agaaagtata
2341 ggaacttcag cggccgcgac tctagatcat aatcagccat accacatttg tagaggtttt
2401 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
2461 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
2521 aaatttcaca aataaagcat ttttttcact gcgaattctc gtaaaactct tcatggcagt
2581 agttattatt ctctctctct ctctttttct tttttcttga gacaggatat ttctctgttg
2641 cccaggctgg agtgcagtgg cacagtcttg gctcactgca gcctggacct cctgggctca
2701 agccatcctc ccacctcagc ctcccaagta gctggggcta caggcacatg gccaccaggc
2761 cagataattt ttcatttttg tagagactga gtctcaccat gttacccagg tttattattc
2821 tcatttttta gatgaagaga ctgaggtcca gagaagctca atgacttgcc tagttttaca
2881 aatctcctgc catcacatac ccctcagcgt ccttaataag agggaggcca ccaactatgt
2941 gctgggcact gtggtggatg ctggagctat agggttgagt atataagaaa tggtgttgct
3001 ggagcaactg ttgcttgctt acctgaccta tctgagaatt aattagcagg ggaacatatt
3061 tttgttttca gattcaatat aagaacttgt gtgggcaaaa ataaagatca gtagtaataa
3121 cagtagttcc catttgctga ctgtactgtc ctaagtgcat atatatatac atacacacac
3181 gcatacctat actcctctaa tactcaaaat gatcctgttt atgtattgtt aatatgctca
3241 ttttattttt aaatttttat ttatttttat ttttatttat ttttgagacg gagtctcatt
3301 ctgtcgcgga ggctgaagtg cagtggtgcg atctcagctc agtgcgacct ccgcctcccg
3361 ggttcaagtg attctcctgc ctcagctccg gactagctgg gattacaggc gcccgcctcc
3421 acgcccagct aatttttgta tttttagtag agatggggtt tcgccatgtt ggccaggctg
3481 gtctcgtact cctgaccttg agtgatccac ctgcctcggc ctcccaaagt gctgggatta
3541 caggcatgag ccaccgcgcc gggctaatat gctcatttta gtgaggcaaa aatagaggct
3601 cagagtctga tttgtacaaa actacagagc agttaagtgt cctctcagat gtgtaccctg
3661 atctgggtga ctctaggact ctaggtctca actgttacaa ccagttaagg gtttggggaa
3721 gcactgggcc aagagtcagg aaaatggaag ccacaggtag tgcaaggtct tgggaatggg
3781 acgtctggtc caaggattca cgcgatgact ggaacccgaa gagccggggc ccggtttacg
3841 gccgccatga agcaacgcgc gccggtaggt ttgggaatca gggagccctc tgaataggag
3901 actgagttgg gagggaaagg ggcttcgctg ggggagcctc ggcttcttct gggagaaaat
3961 tcccacggct acctagtgag cctgcaaact ggtaggcgcc ggcgtaggcg cgcgggcggg
4021 gccgggggcg gggcctgcgg ggcgtggcgg ggcgggcaga gggcggggcc tgcttctcct
4081 cagcttcagg cggctgcgac gagccctcag gcgaacctct cggctttccc gcgcggcgcc
4141 gcctcttgct gcgcctccgc ctcctcctct gctccgccac cggcttcctc ctcctgagca
4201 gtcagcccgc gcgccggccg gctccgttat ggcgacccgc agccctggcg tcgtggtgag
4261 cagctcggcc tgccggccct ggccggttca ggcccacgcg gcaggtggcg gccgggccct
4321 gaggcgcggg atcgatccat gacacccccc cgccccccat atctcactat aattcactgt
4381 caaatggctt ttatgatttg gagtttgggg atacttttaa agacagggtc ttggtagctc
4441 ctagatggct tgggattcac ttagtaaccc ttgctggcct aaaattcagg aacttctacc
4501 tcatcttccc aagtactggg ttagtaggga tgtccagcat atttagcttg ctttaatgat
4561 tttgctgtac ttttttctgt gtgtacaggt gcatgtatat ttataaaact aacataatat
4621 attttacccc caaagtacca taaccctaat taatgctatt aaatgttttg tatgctgggc
4681 tctgggcata ctaatataac caatattata caatgcaaaa caattttgga atatcttctg
4741 taaaattatc ttattagtaa gttgtaacca tctcagactt ctaagtttga gtttgtaaag
4801 gatcagtttt ttttggtgac ctacacacac acacacacac acacacacac acacacacac
4861 aggggggtgg attctgagaa tttagagaat ttagatggtt aataccagtt tttattttta
4921 ttctgtgtga tatttattta ttaagattta tttatttatt acatgtaggt acactgtatc
4981 tgtcttcaga cacaccagaa gagggcatca gatctcgtta cagatggttg tgagtcacca
5041 tgtggttgct gggatttgaa ctcaggacct ctggaagaac agtcagtgct cttaaccgtt
5101 gagccatctc accagccccg tgatatttta tttaacttac tagaatatac tgtgttaaaa
5161 catattttag gggattgggg agatggccca ttctgtaaag tgtctgccat gtcatcttca
5221 ggacctgcgt ttgtaacccc agcatggaga agacagagac agggagaccc ctgggcttcc
5281 tggcaagcta gactagcaaa tttggttaac taggttcaat gagagactca gtctcaaaac
5341 ataagaagtg aattttttag acccaatttc aatctctgaa ctccatacgt atgcacatgt
5401 acctgcatat aaatatatat acacacccat aaacacatgc acacacaaaa aatggttcaa
5461 aagttcttaa aatgtataag agtagaaagt gtagagccgg gagtggtggc gcacgccttt
5521 aatcccagca cttgggagca gaggcaggtg gatttctgag ttcgaggcca gcctggtcta
5581 cagagtgagt tccaggacag ccagagctat acagagaaat cctgcctcaa aaaaccaaaa
5641 ataaagtgta tatactaatc tttcatgcct aaaatattta tttttgccac atttttcata
5701 cctctttaat atttttcttt ttctctttgt gcttcaaagg aaattttgtg tcatgtttta
5761 cttttcattc ttgactctat tcagcagtaa gacgcagcat ggctttctta gagaagtata
5821 atctattaca aaacggtaac atttaactgc tctacatacc ctcttgtagc taaattcctc
5881 tgttaaacta agacttatta tctaaacgta attaaatgtt gatatcttat aatgctccac
5941 tttgaaacag ctgtctttct tttgcagatt agcgatgatg aaccaggtta tgacctagat
6001 ttgttttgta tacctaatca ttatgccgag gatttggaaa aagtgtttat tcctcatgga
6061 ctgattatgg acaggttagt aagacctcga ttgaagttat ttttagaggg tgtgtgtgtg
6121 gtgtgtgtgt gtgtgtgtgt gtgtgtgtgt gtgtgtgtgt aagagagaga ggagaggcat
6181 gtgtgatata tatgtgaagt gtattgcatg tacatggaat ccagagaagg acattaggtg
6241 tcctactctg ttgctctctt attcccttga atcagtgtct cttgctgaac ctggggcata
6301 catactgatt tggctaggct gcctggctag caagacccag ggatctgctt gtctctgccc
6361 cccagtgctg acgttacaag caggtgcaac cacatctggt ttttatatgg gtactggggg
6421 atctgaactt agggcctgat gcgtgcccaa gtgctctcac ccgccaagcc acctctccag
6481 ccaaagggtt ttttttcttg tattaattct aacttagcaa ctgttgagta gttgatgttt
6541 gaatataagc taagccaact tattagaata aatgttttct attttttact tgtatgtgtg
6601 agtgcctgca tgtatgtatg tgaagcatat gtattccagg atgtcagaat tgtgatggat
6661 tccctggaac tggagttaca gaccattgtg aaccactgtg tatgggtgct gggaactgaa
6721 ctttaggtcc tctgcaaaaa cagtaaggat cttaactgat aaactatctt tctagcccta
6781 aggctttcta cttttaaagt tcgtttctct ttctctctct ctctctctct ctctctctct
6841 ctctccctct ccctccctcc tcctcctcct cctctttccc ccctctcgcc tcctctttct
6901 ctctcccctc cctctctctc tctcacacac acacacagac agagagagga gagagaaagg
6961 aagccttact aattaattac ctagttttag acataactag tgggaatcca tagtaagtaa
7021 atcaacattc ttgaagagag gttagaagaa ttggctctcg ttgtccatcg aggctgtata
7081 agattattct tctgttttct cagcttacca tacatatttt ctacgtagac aataccaatt
7141 caaatacact gtacgtaaag tatatgcgca cacacacaca cacacacaca cacacacaca
7201 cacaagttac atattgtcat atagtactaa aatttatgga ctttttcata ttaatacatc
7261 tagagctatc atgtttttta tcttttaaaa attactttac ggggctacag agatgtctca
7321 gcagttagga gcactgactg ctcttccaga ggtcctgagt tcaattccca gcaaccacat
7381 ggtggctcac aaccatctgt aatgggatct gatccactct tatggtgtgt ctgaagacag
7441 tgacagtgta ctcatataca tgaaataaat ctttaaaaaa ttactttacc atcagggtat
7501 atatatatat atatatgtgt gtgtgtgtgt gtgtgtgtat gaagtatttg ctatttgaga
7561 gggtctcact gtttagctca gtctggcttg gaacgttctg tatagcccag actggcctca
7621 actcagcagt ccccctgaaa ggctctcata tgtggagatt acaggtatgt gctactgtgc
7681 ccagccctac atgttacttt gtttttgaaa tagggtatca aatagcttag actggcttct
7741 aagtcagtgt gtagctaagg atggtctgag cttctgaacc tgctacttca actcctggtg
7801 tgccatcatg gccagtttgc acagttgtga gagtagaact caggacctcc tatgtgttac
7861 tagacatgtt ctctaccagc tgagctacat cccaattcgc ctacatttac ttttattttt
7921 taaaagattt atttatgtat acagcactct gcctgcatgt acatctgcat gccagaagag
7981 ggcaccagat ctcatcatac atagttgtga gccaccatgt ggttgctggg gattgaactc
8041 agggcctctg gaagagcagt cagtgctctt aacctctgag ccatctctgt agccccctac
8101 atttatttta aagattatag gctattctat tcatagatgt gccataactg atttaaccag
8161 atcttcatta ttttgtactt agtagagtat attagaggga atgaatctta aatacacatt
8221 ttgacacttg ctcagcttta gacaagtaaa aatcactggt caaggaaatg gtgcttagtg
8281 gatgcttctg aactgtcaaa ttgtcctcca acaagttaca tcattttata cttccaacgt
8341 tatgtgagag ttcatttttc tttaggctct tctagtattt tatatttgag aaataaaatt
8401 taaccaaaat ttgccaatct agtaagcttg tttttatttg tatctttaaa ttttatgata
8461 gatgttatag tgtactctcc tctcccctcc cctcccctct cctcctcagc aggatcttgt
8521 tatgtagccc taagggatct aatacttact ctgtagtcca ggctgtcctt cagctcatga
8581 gacacaccat catgcttggc atgtttcttg ttctgaaagg tatctggttt taattttggt
8641 tgagctggtg tgttggtgta tatctgtaat ctcagcacag gcaagtagag acaaggatta
8701 tgattacagt ccagcctggg ctatttagag ggattctctt taatctttcc tcatgcccca
8761 aaatcttacc tttggtatat gaaaaatagt ctccacttct gcaaaatatt gctttatgaa
8821 gtaagaatta attcccttca tagagacaag gaatgtgtcc tgtaaaagtt taatgtgtaa
8881 gaagtatttg ttataaaaga taaatattca gaatcttctt tttaattcct gattttattt
8941 ctataggact gaaagacttg ctcgagatgt catgaaggag atgggaggcc atcacattgt
9001 ggccctctgt gtgctcaagg ggggctataa gttctttgct gacctgctgg attacattaa
9061 agcactgaat agaaatagtg atagatccat tcctatgact gtagatttta tcagactgaa
9121 gagctactgt gtaagtataa ttaacttata attaaaaaaa tagggccatt ctagttttat
9181 ctatattttt tttaaacttg tgcaaactat gctacataat caacttactt gattatgtaa
9241 tatatatagt aatagttgga attactatta ttacatattt atttttctgt gctggggtag
9301 aacccaggga actaagtact gtaataaccg agcttcattc atcctagtca tttttaattt
9361 tttttttttt ttttgttttt tcgagacaag gtttctctgt gtagccctgg ctgtcctgga
9421 actcactctg tagaccaggc tggcctcgaa ctcagaaatc tgcctgcctc tgcctcccag
9481 tgctgagatt aatgacgtgc accaccactg cctggcattt ttttaaagtt ttaatttgag
9541 accctttctc actaacttgc tcacactatc ctcgaactca ttacgtgggc tggtctagaa
9601 atttcagtca acttgcctca gcttctcaag tagctgggat tacaggcagg tgctaccact
9661 tctggccatg atcattgttg ttgttgctat tattattatt attatttcac tgatacttca
9721 tatcacaaca cagaatagta actgtagttc atgaccaaca aaaaagttgt acagtttctc
9781 tgttgtctta ttttgatctg gattttgatt tgtttcatgg cactgggcag gctaatctca
9841 aactgcatgg ctcaaatagt ccttctgtct taatctccca tgttctggaa gtataggtat
9901 cactacacct agctgtattt tcatatttta aaaaaaacgc ctaagaattg ctattgaatg
9961 aggcggagag atggctcagc agtcacaagt acttgtagag gactggggtt tggttactag
10021 cacctcaaag atggcttaca attatctgta gctccaattc cagaggatct gtgtggtaaa
10081 gctgcatgta tatgggcatg caggtaaaac tgctaaacat ataaaaatag ctcagtctta
10141 agagaattgg tcgacgtaag ctcattcgag tcgacttttt cgccctttga cgttggagtc
10201 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
10261 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
10321 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc
10381 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
10441 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
10501 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
10561 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
10621 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
10681 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
10741 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
10801 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
10861 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
10921 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
10981 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
11041 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
11101 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
11161 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
11221 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
11281 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
11341 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
11401 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
11461 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
11521 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
11581 aaaccaccgc taccaggtcg agtagggata acagggtaat ggatcctcgt tgggcacctg
11641 cactcatgta cacaggccca catatagatg tacacacgca tatcattcaa aatagaaatt
11701 atttatacaa gtagtgcgtt ggagagctgg ctcaggagtt gttaatgctt gctacccttg
11761 caaaggaccc aagttcaatt tccagcacct gtgttgggca tctcacaacc acatgtaaat
11821 ctagtttcag gggatttcac ccctcttctg aactcttcag gcacttatac ccacaaatgg
11881 acatctccac atagagacac atacttttaa aaaagtaatc tagggtctag agaaatggtt
11941 atagaccacg gactgggatc agttcccaga agccacatcc tgtggctcac aattgcctgt
12001 gaatcaagtt ctagatctgg tggcctcgtc tagcctgcag tcatcaacaa acacatgcac
12061 acatccatac acacataatt taaaagaaag cgaaagtaac ctataatgct aaacaaagca
12121 cattctcagt gtagactgca gtggtgatga accttgaaaa tatattaata gaagccagac
12181 acgtaagtca caaatggtat aactcaattt atatgaaatt cccagagagt ccaaagttaa
12241 aaaaaacaaa aaggcagatt attggttgtc agtagctaca gagatgggac attgaagata
12301 ttaataggta tagggttgag gattcatggc ctttccctct tccatgttag catgtctatt
12361 gttgttaacc ttattcaact catgttaggg caatcatatt gatgagactt aatgagtata
12421 gcttgacatt cctagaagac agagtctcac atcagactct ctgatcctct gactcttaga
12481 atctttcaac accctcttct gaaatgatct ctgagccttt ggttgtgact gaattatgag
12541 aaagaatttt atgttgaatg aattttactt cagcattttt acaaaactga agctggggat
12601 ggcgtttcat gcgtgcagtc cctaccattt gcctataagc ctggaggagg cagaagtaga
12661 attaggcttt tcagggacat cttcagctgc atcatgagct tgaggacaac ttgagctaca
12721 taagattccc atccccaccc ccacacacac acacacatct agtatgtacc tagtatgcag
12781 tcttttcata tgtttaaaat gtagaaaggt atatgtgaag tctccctttt ccccacaact
12841 tagtccctcc aaagtattgc atcagtttcc ttgtacatga ttttatgcat atataagata
12901 cactatttct ctctctctct ctctccttcc ctccttccct ccaacctctg ggtatgagtt
12961 gagggttatg tatgtttcaa gttgtaataa gaattgccac tcaaacattt catcccattt
13021 taagttaagt ttgtctgatt ttccacttta gaagggcagc catgatggct catgcctata
13081 atcccatcct agcacttgta agcctaatac ataagactgg aggattgtca tcagcttgag
13141 gctagcctgg tctacctaat gagttccagc cagccaagag tagaatataa gaccctgtct
13201 ccaaaaaatg taccaaaggg gctggagggg taactcagtg gttaagaaca gtagctgctc
13261 ttccagagga cctgggttca attcccagca cccacatggc agcttgcaac tatctgtaac
13321 tgtccagctt tgggatatcc gacaccatta taccaattac ataaaatgaa aaaattgtaa
13381 aatgtgctaa aatgagtggg gagggggagc atggtgtggt tctgtgggct tgggattttg
13441 ccatgaatcc atgactatat ctcagcaata tgtggattat ttctggaaac ctgtggggca
13501 tgggtgtatg ggaagagggg agagagagag agcccgcatc cagccagagt tcctgtgctc
13561 tgggcaggcg gacgtgggag gactgctgga cgatttccac ttggccctgg gtgggcatct
13621 ggctgtgtga agccactgac cccactcagc ggggggtaga taaggggcat ttcccatacc
13681 aggttctcag actccagcat tccacgctgg atacagcaga agagctgaga acagaatagg
13741 ggccccaggg agacactctc gtcccagaga taaagaaggc agagggagag agattcctac
13801 gcaggcgaga ggccttagtc tgggccttga tgaacagaga cggtctatgg ttttagagct
13861 ttattgtaga aaggcagggg gaaagagaga aggtagaaag agagagacca gccatggcca
13921 agaggagaga aggggggaaa ggagagagag aagaaaggct agagagtaag aaaggttaaa
13981 gagacagaag taaaagctta gagtaagaga gagagagaga gcaagaggga gagtaagaaa
14041 agtaaaagct taagagagcg aggaggggcc agcccctcta atagtgggct tgttatcttg
14101 ttgttgctag gtaactgtga agcgtctagc ctgaaggcca gaagcttggg acattgtcta
14161 tgtgactgaa agccatgagc ctctactgtg ggggctgtgg aggtggtaac ttggacagga
14221 gccaggggtt ccaggagaca tgagggaatg ccttctgtcc catgtaggta ggaattacca
14281 ccaccggctt tcaccgtagt tcactggaga ctgtttgtac atagcccatt gccccacaga
14341 aaccaggcac tcactgtttc atttgcataa gaattagaag gctttatttt ggctgggatt
14401 ggtgacttgc cccagaaatt ccagcttttg ggaagccgaa tcctggagaa ccaggagttc
14461 aaggtcatcc tcaggtacct catttgttca agaccagcct gtgctaccat gagaccttgt
14521 ccctactccc ctcaacccca aaacacgctg ttttcatttt ataggttgta ctaaccatgg
14581 aggatggtca tttgcctttc tcttgatttc agaagcgtag cgcgggcttt gcaccatgat
14641 ctagtgtttc cagataggct tgcctctcca cagccaggct tatcctgctt cagtcccatg
14701 tttggcaagg gcaacatatg actgaagaac ttgcatagcc ttatggtgta tcatgactct
14761 tagccccagt ttataactgt ctaacagctg gtggcttttg tgaaaaagga aagcatcatt
14821 tgaactttta acttttcagc tgtttgtttt gactttgcat aaatactgag actagctagg
14881 aaatgagctt ttgtttcatt tgctaaaaat gatttttttt tcatgaggca atttcaagca
14941 atttgaattt agcaatctct acttgtctga aaagaagtta ttaattaaaa aaaaacaaac
15001 aaacaggaac ctattatgct ggctagtcac atgaaagaat ccaaatgaac acagtcaagg
15061 ctgaatcaac acatgaaatt attttctaat tccccaaatt acatttatgc taattagtat
15121 gtagtaaaac attttaattg tttccaatgt taaacattac agaaataatt ttgatgtttt
15181 cctaataacc cagcctttga tcatgtttgc atttatttga ttgttgtcag tttattaaat
15241 aattaagtaa gagcaacaag ataaaatgac ttcatgagag tccttttctg taacttagaa
15301 cgtcagtagt cataggaact gcggtcgttc ccttaccttt cagaagtagg ctaaagagtt
15361 gaacgcaaag gtggtttttc gaatcttcct gatccttaag cccccactag gtttatattc
15421 tatatttgat atcggattcg aacccctcga gcttaggatc cggaaccctt aatataacta
15481 agttattagg tccctcgaag aggttcacta gtactggcca ttgcggccgc ttcgaggctg
15541 cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
15601 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
15661 tgttggcggg tgtcggggcg cagccatgac ccagtcacgt agcgatagcg gagtgtatac
15721 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa
15781 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc
15841 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
15901 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
15961 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
16021 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
16081 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
16141 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
16201 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
16261 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
16321 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
16381 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
16441 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
16501 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
16561 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
16621 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
16681 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
16741 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
16801 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
16861 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
16921 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
16981 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
17041 tcgccagtta atagtttgcg caacgttgtt gccattgctg caggcatcgt ggtgtcacgc
17101 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
17161 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
17221 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
17281 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
17341 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca
17401 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
17461 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
17521 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
17581 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
17641 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
17701 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtg
//
| {
"Vector backbone": "pEMS1306",
"Backbone size w/o insert (bp)": "16454",
"Vector type": "Pleiades Promoter Project [sic, Pleaides Plieades]",
"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 regulatory toolbox of MiniPromoters to drive selective expression in the brain",
"Publication": "Proc Natl Acad Sci U S A. 2010 Sep 21. 107(38):16589-94.",
"DOI": "10.1073/pnas.1009158107",
"Authors": "Portales-Casamar E, Swanson DJ, Liu L, de Leeuw CN, Banks KG, Ho Sui SJ, Fulton DL, Ali J, Amirabbasi M, Arenillas DJ, Babyak N, Black SF, Bonaguro RJ, Brauer E, Candido TR, Castellarin M, Chen J, Chen Y, Cheng JC, Chopra V, Docking TR, Dreolini L, D'Souza CA, Flynn EK, Glenn R, Hatakka K, Hearty TG, Imanian B, Jiang S, Khorasan-zadeh S, Komljenovic I, Laprise S, Liao NY, Lim JS, Lithwick S, Liu F, Liu J, Lu M, McConechy M, McLeod AJ, Milisavljevic M, Mis J, O'Connor K, Palma B, Palmquist DL, Schmouth JF, Swanson MI, Tam B, Ticoll A, Turner JL, Varhol R, Vermeulen J, Watkins RF, Wilson G, Wong BK, Wong SH, Wong TY, Yang GS, Ypsilanti AR, Jones SJ, Holt RA, Goldowitz D, Wasserman WW, Simpson EM",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/20807748",
"PubMed ID": "PubMed 20807748"
} | {
"Gene/Insert name": "Ple231",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "1258",
"Mutation": "",
"Entrez Gene": "TNNT1",
"Promoter": "",
"Tag / Fusion Protein": "EGFP-NLS (C terminal on backbone)"
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "FseI (unknown if destroyed)",
"3′ cloning site": "AscI (unknown if destroyed)",
"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": ""
} |
|
HOXB8 (murine) HIS-tag pET | 8540 | Low | Corey Largman | Shen et al Nucleic Acids Res 1996 Mar 1;24(5):898-906. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/17353/15708b12-e65a-482e-9fcc-bf05eb869963/addgene-plasmid-8540-sequence-17353.gbk | partial | LOCUS sequence-17353-e 885 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 885)
AUTHORS Shen WF, Chang CP, Rozenfeld S, Sauvageau G, Humphries RK, Lu M,
Lawrence HJ, Cleary ML, Largman C
TITLE Hox homeodomain proteins exhibit selective complex stabilities with
Pbx and DNA.
JOURNAL Nucleic Acids Res 1996 Mar 1;24(5):898-906.
PUBMED 8600458
REFERENCE 2 (bases 1 to 885)
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"; date: "1996-03-1"; volume: "24(5)"; pages: "898-906"
FEATURES Location/Qualifiers
source 1..885
/mol_type="genomic DNA"
/organism="unspecified"
CDS 94..126
/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 aatattttgt ttactttaag aaggagatat accatgggca gcagccatta tcatcatcat
61 cacagcagcg gcctggtgcc gcgcggctgc catatggcta gcatgactgg tggacagcaa
121 atgggtcggg atccgaattc gagctcttat ttcgtcaact cactgttctc caaatacaaa
181 accggggagt ccctgcgccc caattattat gactgcggct tcgcccagga cctgggcggc
241 cgacccaccg tggtgtacgg tcccagcagc ggcggcagct tccagcaccc ttcgcaaatc
301 caggagttct accacgggcc atcgtcgctg tccacagctc cctaccagca gaacccgtgc
361 gccgtggcgt gccacggcga ccccggcaac ttctacggct acgaccctct gcagcgccag
421 agcctgttcg gtgcgcagga tccagacctg gtgcagtacg cagactgcaa gctcgcggca
481 gccagcggcc tgggcgagga ggccgagggg tctgagcaga gcccgtcgcc cacacagctc
541 tttccctgga tgcgccctca agccgccgga cgcaggcgag gccgccagac ctacagtcgc
601 taccagaccc tggagctgga gaaggagttc ctatttaatc cctatctgac tcgcaagcgg
661 aggatcgagg tatcgcacgc gctgggactg acagagagac aggtcaaaat ctggttccag
721 aatcggagaa tgaagtggaa aaaggagaac aacaaagaca agtttcccag cagtaaatgc
781 gagcaggagg agctggagaa agagaagctg gagcgggcac cagagaccgc cgagcagggc
841 gatgcgcaga aaggtgacaa gaagtaggct ccagctggga ctgct
//
| {
"Vector backbone": "pET28b",
"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": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Hox homeodomain proteins exhibit selective complex stabilities with Pbx and DNA",
"Publication": "Nucleic Acids Res 1996 Mar 1;24(5):898-906.",
"DOI": "10.1093/nar/24.5.898",
"Authors": "Shen WF, Chang CP, Rozenfeld S, Sauvageau G, Humphries RK, Lu M, Lawrence HJ, Cleary ML, Largman C",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/8600458",
"PubMed ID": "PubMed 8600458"
} | {
"Gene/Insert name": "HOXB8",
"Species": "M. musculus (mouse)",
"Insert Size (bp)": "828",
"Mutation": "starts at amino acid #2 (Ser)",
"Entrez Gene": "Hoxb8",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "SacI (not destroyed)",
"3′ cloning site": "SacI (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": ""
} |
|
pBHRSF184 | 89482 | Low | Express asparaginyl endopeptidase 1 from O. affinis (OaAEP1) | Hideo Iwai | Mikula et al FEBS Lett. 2017 Mar 30. doi: 10.1002/1873-3468.12640. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/180027/65c767da-1445-4c9e-ad0d-c95e1d1d9fd7/addgene-plasmid-89482-sequence-180027.gbk | full | LOCUS sequence-180027- 5245 bp DNA circular SYN 25-AUG-2024
DEFINITION Express asparaginyl endopeptidase 1 from O. affinis (OaAEP1).
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5245)
AUTHORS Mikula KM, Tascon I, Tommila JJ, Iwai H
TITLE Segmental isotopic labeling of a single-domain globular protein
without any refolding step by an asparaginyl endopeptidase.
JOURNAL FEBS Lett. 2017 Mar 30. doi: 10.1002/1873-3468.12640.
PUBMED 28369872
REFERENCE 2 (bases 1 to 5245)
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: "FEBS Lett.
2017 Mar 30. doi: 10.1002/1873-3468.12640."
FEATURES Location/Qualifiers
source 1..5245
/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)."
CDS 100..117
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 124..414
/codon_start=1
/gene="S. cerevisiae SMT3 (truncated)"
/product="cleavable ubiquitin-like protein tag"
/label=SUMO
/translation="SDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPLR
RLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIGG"
CDS 1799..1843
/codon_start=1
/product="affinity and epitope tag derived from pancreatic
ribonuclease A"
/label=S-Tag
/translation="KETAAAKFERQHMDS"
primer_bind complement(1881..1899)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 1895..1942
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(2172..2987)
/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(2172..2987)
/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 2895..2914
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
promoter complement(2988..3079)
/gene="bla"
/label=AmpR promoter
rep_origin complement(3095..3844)
/direction=LEFT
/label=RSF ori
/note="Plasmids containing the RSF 1030 origin of
replication can be propagated in E. coli cells that contain
additional plasmids with compatible origins."
primer_bind complement(3105..3122)
/label=L4440
/note="L4440 vector, forward primer"
primer_bind complement(3345..3364)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4040..5122)
/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 5084..5103
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(5123..5200)
/gene="lacI"
/label=lacI promoter
ORIGIN
1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctgta gaaataattt
61 tgtttaactt taataaggag atataccatg ggcagcagcc atcatcatca tcatcacagc
121 actagtgact cagaagtcaa tcaagaagct aagccagagg tcaagccaga agtcaagcct
181 gagactcaca tcaatttaaa ggtgtccgat ggatcttcag aaatcttctt taagatcaaa
241 aagaccactc ctttaagaag gctgatggaa gcgttcgcta aaagacaggg taaggaaatg
301 gactccttaa gattcttgta cgacggtatt agaattcaag ctgatcagac ccctgaagat
361 ttggacatgg aggataacga tattattgag gctcacagag aacagattgg tggatccgct
421 cgcgatggtg attatctgca tctgccgagc gaagtgagcc gtttctttcg cccgcaggaa
481 acaaacgatg atcatggtga agatagcgtg ggtacccgct gggcggtgct gattgcgggc
541 tctaaaggct atgcgaacta tcgccatcag gctggcgtgt gccatgcgta tcagattctg
601 aaacgcggcg gcctgaaaga tgaaaacatt gttgtgttca tgtatgatga tattgcgtat
661 aacgaaagca atccacgtcc gggcgtgatt attaacagcc cgcatggtag cgatgtttat
721 gctggcgtgc cgaaagatta tacaggcgaa gaagtgaacg cgaaaaactt tctggcggcg
781 attctgggca acaaaagcgc gattaccggc ggcagcggca aagtggtgga tagcggtccg
841 aacgaccata tttttattta ttataccgat catggcgctg cgggcgtgat tggcatgccg
901 agcaaaccgt atctgtatgc agatgaactg aacgatgcgc tgaaaaagaa acatgcgagc
961 ggtacatata aaagcctggt gttttatctg gaagcgtgcg aaagcggcag catgtttgaa
1021 ggtattttgc cggaagattt gaacatttat gcgctgacca gcaccaacac caccgaaagc
1081 agctggtgct attattgtcc ggcgcaggaa aacccgccac cgccggaata taacgtgtgc
1141 ctgggcgatc tgtttagcgt ggcgtggctg gaagatagcg atgtgcagaa ttcctggtat
1201 gaaaccctga accagcagta tcatcatgtg gataaacgca ttagccatgc gagccatgcg
1261 acccagtatg gcaacctgaa attgggcgaa gaaggcctgt ttgtgtatat gggcagcaac
1321 ccggcgaacg ataactatac ctctctggat ggcaacgcgc tgaccccgtc atcgattgtg
1381 gtgaaccagc gcgatgcgga tctgctgcat ctgtgggaaa aattccgcaa agcgccggaa
1441 ggcagcgcgc gcaaagaaga agcgcagacc cagattttta aagcgatgag ccatcgcgtg
1501 catattgata gcagcattaa actgattggc aaactgctgt ttggcattga aaaatgcacc
1561 gaaattctga acgcggttcg tccggcaggt caaccgctgg tggatgattg ggcgtgcctg
1621 cgtagcctgg tgggcacctt tgaaacccat tgcggcagcc tgagcgaata tggcatgcgc
1681 catacccgca ccattgcgaa catttgcaac gcgggcatta gcgaagaaca gatggcggaa
1741 gcggcgagcc aggcgtgcgc gagcattccg taagcttgcg gccgcactcg agtctggtaa
1801 agaaaccgct gctgcgaaat ttgaacgcca gcacatggac tcgtctacta gcgcagctta
1861 attaacctag gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa
1921 acgggtcttg aggggttttt tgctgaaacc tcaggcattt gagaagcaca cggtcacact
1981 gcttccggta gtcaataaac cggtaaacca gcaatagaca taagcggcta tttaacgacc
2041 ctgccctgaa ccgacgacaa gctgacgacc gggtctccgc aagtggcact tttcggggaa
2101 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
2161 tgaattaatt cttagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca
2221 ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg
2281 aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca
2341 tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca
2401 tgagtgacga ctgaatccgg tgagaatggc aaaagtttat gcatttcttt ccagacttgt
2461 tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc
2521 attcgtgatt gcgcctgagc gagacgaaat acgcggtcgc tgttaaaagg acaattacaa
2581 acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct
2641 gaatcaggat attcttctaa tacctggaat gctgttttcc cggggatcgc agtggtgagt
2701 aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc
2761 gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca
2821 tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat tgtcgcacct
2881 gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa
2941 tttaatcgcg gcctagagca agacgtttcc cgttgaatat ggctcatact cttccttttt
3001 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
3061 atttagaaaa ataaacaaat aggcatgcag cgctcttccg cttcctcgct cactgactcg
3121 ctacgctcgg tcgttcgact gcggcgagcg gtgtcagctc actcaaaagc ggtaatacgg
3181 ttatccacag aatcagggga taaagccgga aagaacatgt gagcaaaaag caaagcaccg
3241 gaagaagcca acgccgcagg cgtttttcca taggctccgc ccccctgacg agcatcacaa
3301 aaatcgacgc tcaagccaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
3361 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
3421 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gttggtatct
3481 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
3541 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
3601 atcgccactg gcagcagcca ttggtaactg atttagagga ctttgtcttg aagttatgca
3661 cctgttaagg ctaaactgaa agaacagatt ttggtgagtg cggtcctcca acccacttac
3721 cttggttcaa agagttggta gctcagcgaa ccttgagaaa accaccgttg gtagcggtgg
3781 tttttcttta tttatgagat gatgaatcaa tcggtctatc aagtcaacga acagctattc
3841 cgttactcta gatttcagtg caatttatct cttcaaatgt agcacctgaa gtcagcccca
3901 tacgatataa gttgtaattc tcatgttagt catgccccgc gcccaccgga aggagctgac
3961 tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag tgagctaact
4021 tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
4081 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gccagggtgg
4141 tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag
4201 agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg
4261 tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga
4321 tgtccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgccatct
4381 gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt tgcatggttt
4441 gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc tgaatttgat
4501 tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca gaacttaatg
4561 ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc acgcccagtc
4621 gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca gagacatcaa
4681 gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc tggtcatcca
4741 gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc accgccgctt
4801 tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca cccagttgat
4861 cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc agactggagg
4921 tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg cggttgggaa
4981 tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca gaaacgtggc
5041 tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac tctgcgacat
5101 cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc gggcgctatc
5161 atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg acgctctccc
5221 ttatgcgact cctgcattag gaaat
//
| {
"Vector backbone": "pHYRSF53",
"Backbone size w/o insert (bp)": "4540",
"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": "Segmental isotopic labeling of a single-domain globular protein without any refolding step by an asparaginyl endopeptidase",
"Publication": "FEBS Lett. 2017 Mar 30. doi: 10.1002/1873-3468.12640.",
"DOI": "10.1002/1873-3468.12640.",
"Authors": "Mikula KM, Tascon I, Tommila JJ, Iwai H /-",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/28369872",
"PubMed ID": "PubMed 28369872"
} | {
"Gene/Insert name": "Asparaginyl endopeptidase from Oldenlandia affinis",
"Species": "Synthetic",
"Insert Size (bp)": "1353",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "T7",
"Tag / Fusion Protein": "Smt3 (N terminal on backbone)"
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "BamHI (not destroyed)",
"3′ cloning site": "HindIII (not destroyed)",
"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": ""
} |
pTALYM16-Telo1 | 47882 | Low | TALE-mRuby2 against telomere sequence | Maria-Elena Torres-Padilla | Miyanari et al Nat Struct Mol Biol. 2013 Oct 6. doi: 10.1038/nsmb.2680. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/68421/48dd618b-393d-4dc3-acc6-f5685aa6d537/addgene-plasmid-47882-sequence-68421.gbk | full | LOCUS sequence-68421-e 7999 bp DNA circular SYN 26-AUG-2024
DEFINITION TALE-mRuby2 against telomere sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7999)
AUTHORS Miyanari Y, Ziegler-Birling C, Torres-Padilla ME
TITLE Live visualization of chromatin dynamics with fluorescent TALEs.
JOURNAL Nat Struct Mol Biol. 2013 Oct 6. doi: 10.1038/nsmb.2680.
PUBMED 24096363
REFERENCE 2 (bases 1 to 7999)
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 Struct
Mol Biol. 2013 Oct 6. doi: 10.1038/nsmb.2680."
FEATURES Location/Qualifiers
source 1..7999
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 12..42
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 50..66
/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 55..77
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 74..90
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 74..90
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
enhancer 110..489
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 491..768
/label=chicken beta-actin promoter
intron 769..1777
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
primer_bind 1701..1723
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 1790..1809
/label=T7
/note="T7 promoter, forward primer"
promoter 1790..1808
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
5'UTR 1833..1875
/label=Xenopus globin 5'-UTR
/note="translational enhancer from the 5'-UTR of the major
beta-globin gene of Xenopus laevis"
CDS 4089..4799
/codon_start=1
/product="monomeric red fluorescent protein derived from
mRuby, with improved photophysical properties (Lam et al.,
2012)"
/label=mRuby2
/translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
AKFAGLGGGMDELYK"
CDS 4999..5028
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 5044..5061
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(5084..5101)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 5090..5314
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(5324..5341)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(5324..5340)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5333..5355)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 5482..5937
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5569..5588)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 5779..5800
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 5963..6067
/gene="bla"
/label=AmpR promoter
CDS 6068..6928
/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(6286..6305)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 7099..7687
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 7588..7607
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 7841..7858
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 7975..7996
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
ORIGIN
1 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
61 taacaatttc acacaggaaa cagctatgac catgaggcgc gccggattcg acattgatta
121 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
181 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
241 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
301 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
361 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
421 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
481 attaccatgg tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc
541 ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg
601 gggggggggg ggcgcgcgcc aggcggggcg gggcggggcg aggggcgggg cggggcgagg
661 cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg
721 aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg agtcgctgcg
781 ttgccttcgc cccgtgcccc gctccgcgcc gcctcgcgcc gcccgccccg gctctgactg
841 accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag
901 cgcttggttt aatgacggct cgtttctttt ctgtggctgc gtgaaagcct taaagggctc
961 cgggagggcc ctttgtgcgg gggggagcgg ctcggggggt gcgtgcgtgt gtgtgtgcgt
1021 ggggagcgcc gcgtgcggcc cgcgctgccc ggcggctgtg agcgctgcgg gcgcggcgcg
1081 gggctttgtg cgctccgcgt gtgcgcgagg ggagcgcggc cgggggcggt gccccgcggt
1141 gcgggggggc tgcgagggga acaaaggctg cgtgcggggt gtgtgcgtgg gggggtgagc
1201 agggggtgtg ggcgcggcgg tcgggctgta acccccccct gcacccccct ccccgagttg
1261 ctgagcacgg cccggcttcg ggtgcggggc tccgtgcggg gcgtggcgcg gggctcgccg
1321 tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg ggcggggccg cctcgggccg
1381 gggagggctc gggggagggg cgcggcggcc ccggagcgcc ggcggctgtc gaggcgcggc
1441 gagccgcagc cattgccttt tatggtaatc gtgcgagagg gcgcagggac ttcctttgtc
1501 ccaaatctgg cggagccgaa atctgggagg cgccgccgca ccccctctag cgggcgcggg
1561 cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga gggccttcgt gcgtcgccgc
1621 gccgccgtcc ccttctccat ctccagcctc ggggctgccg cagggggacg gctgccttcg
1681 ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct ctagagcctc
1741 tgctaaccat gttcatgcct tcttcttttt cctacagatc cttaattaat aatacgactc
1801 actataggga acaaaagctg gagctaagct tgcttgttct ttttgcagaa gctcagaata
1861 aacgctcaac tttggcagcc gccaccatga gatctggcaa gagggctgag gacgatgaag
1921 atgacgacgt ggacaccaag aagcagaaga ccgtcgacca ggtggatcta cgcacgctcg
1981 gctacagtca gcagcagcaa gagaagatca aaccgaaggt gcgttcgaca gtggcgcagc
2041 accacgaggc actggtgggc catgggttta cacacgcgca catcgttgcg ctcagccaac
2101 acccggcagc gttagggacc gtcgctgtca cgtatcagca cataatcacg gcgttgccag
2161 aggcgacaca cgaagacatc gttggcgtcg gcaaacagtg gtccggcgca cgcgccctgg
2221 aggccttgct cacggatgcg ggggagttga gaggtccgcc gttacagttg gacacaggcc
2281 aacttgtgaa gattgcaaaa cgtggcggcg tgaccgcaat ggaggcagtg catgcatcgc
2341 gcaatgcact gacgggtgcc cccctgaacc tgaccccgga ccaagtggtg gctatcgcca
2401 gcaacattgg cggcaagcaa gcgctcgaaa cggtgcagcg gctgttgccg gtgctgtgcc
2461 aggaccatgg cctgaccccg gaccaagtgg tggctatcgc cagcaaccac ggcggcaagc
2521 aagcgctcga aacggtgcag cggctgttgc cggtgctgtg ccaggaccat ggcctgaccc
2581 cggaccaagt ggtggctatc gccagcaacc acggcggcaa gcaagcgctc gaaacggtgc
2641 agcggctgtt gccggtgctg tgccaggacc atggcctgac cccggaccaa gtggtggcta
2701 tcgccagcaa ccacggcggc aagcaagcgc tcgaaacggt gcagcggctg ttgccggtgc
2761 tgtgccagga ccatggcctg accccggacc aagtggtggc tatcgccagc aacggtggcg
2821 gcaagcaagc gctcgaaacg gtgcagcggc tgttgccggt gctgtgccag gaccatggcc
2881 tgaccccgga ccaagtggtg gctatcgcca gcaacggtgg cggcaagcaa gcgctcgaaa
2941 cggtgcagcg gctgttgccg gtgctgtgcc aggaccatgg cctgaccccg gaccaagtgg
3001 tggctatcgc cagcaacatt ggcggcaagc aagcgctcga aacggtgcag cggctgttgc
3061 cggtgctgtg ccaggaccat ggcctgaccc cggaccaagt ggtggctatc gccagcaacc
3121 acggcggcaa gcaagcgctc gaaacggtgc agcggctgtt gccggtgctg tgccaggacc
3181 atggcctgac cccggaccaa gtggtggcta tcgccagcaa ccacggcggc aagcaagcgc
3241 tcgaaacggt gcagcggctg ttgccggtgc tgtgccagga ccatggcctg accccggacc
3301 aagtggtggc tatcgccagc aaccacggcg gcaagcaagc gctcgaaacg gtgcagcggc
3361 tgttgccggt gctgtgccag gaccatggcc tgaccccgga ccaagtggtg gctatcgcca
3421 gcaacggtgg cggcaagcaa gcgctcgaaa cggtgcagcg gctgttgccg gtgctgtgcc
3481 aggaccatgg cctgaccccg gaccaagtgg tggctatcgc cagcaacggt ggcggcaagc
3541 aagcgctcga aacggtgcag cggctgttgc cggtgctgtg ccaggaccat ggcctgaccc
3601 cggaccaagt ggtggctatc gccagcaaca ttggcggcaa gcaagcgctc gaaacggtgc
3661 agcggctgtt gccggtgctg tgccaggacc atggcctgac cccggaccaa gtggtggcta
3721 tcgccagcaa ccacggcggc aagcaagcgc tcgaaacggt gcagcggctg ttgccggtgc
3781 tgtgccagga ccatggcctg accccggacc aagtggtggc tatcgccagc aaccacggcg
3841 gcaagcaagc gctcgaaagc attgtggccc agctgagccg gcctgatccg gcgttggccg
3901 cgttgaccaa cgaccacctc gtcgccttgg cctgcctcgg cggacgtcct gccatggatg
3961 cagtgaaaaa gggattgccg cacgcgccgg aattgatcag aagagtcaat cgccgtattg
4021 gcgaacgcac gtcccatcgc gttgcgggat ccactggagg cggaggtggc tccggcaccg
4081 cctcctccat ggtgtctaag ggcgaagagc tgatcaagga aaatatgcgt atgaaggtgg
4141 tcatggaagg ttcggtcaac ggccaccaat tcaaatgcac aggtgaagga gaaggcaatc
4201 cgtacatggg aactcaaacc atgaggatca aagtcatcga gggaggaccc ctgccatttg
4261 cctttgacat tcttgccacg tcgttcatgt atggcagccg tacttttatc aagtacccga
4321 aaggcattcc tgatttcttt aaacagtcct ttcctgaggg ttttacttgg gaaagagtta
4381 cgagatacga agatggtgga gtcgtcaccg tcatgcagga caccagcctt gaggatggct
4441 gtctcgttta ccacgtccaa gtcagagggg taaactttcc ctccaatggt cccgtgatgc
4501 agaagaagac caagggttgg gagcctaata cagagatgat gtatccagca gatggtggtc
4561 tgaggggata cactcatatg gcactgaaag ttgatggtgg tggccatctg tcttgctctt
4621 tcgtaacaac ttacaggtca aaaaagaccg tcgggaacat caagatgccc ggtatccatg
4681 ccgttgatca ccgcctggaa aggttagagg aaagtgacaa tgaaatgttc gtagtacaac
4741 gcgaacacgc agttgccaag ttcgccgggc ttggtggtgg gatggacgag ctgtacaagt
4801 aagaattctg tacatatggg caccaaagaa cctgtaaacg ttatcttttt aaattgaatg
4861 tgcacaaata aaagtttgga aaagaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
4921 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaatacgcat catacgctgt ggcgtacctc
4981 gagtctagag ggcccttcga acaaaaactc atctcagaag aggatctgaa tatgcatacc
5041 ggtcatcatc accatcacca ttgagtttaa acccgctgat cagcctcgac tgtgccttct
5101 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
5161 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
5221 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
5281 agcaggcatg ctggggatgc ggtgggctct atgggcggcc gccactggcc gtcgttttac
5341 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
5401 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
5461 gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
5521 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
5581 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
5641 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
5701 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
5761 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
5821 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
5881 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg
5941 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
6001 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
6061 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
6121 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
6181 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
6241 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
6301 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
6361 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
6421 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
6481 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
6541 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
6601 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
6661 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
6721 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
6781 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
6841 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
6901 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
6961 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
7021 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
7081 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
7141 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
7201 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
7261 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
7321 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
7381 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
7441 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
7501 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
7561 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
7621 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
7681 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
7741 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
7801 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
7861 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
7921 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
7981 gagttagctc actcattag
//
| {
"Vector backbone": "pTALYM16",
"Backbone size w/o insert (bp)": "",
"Vector type": "Mammalian Expression, TALEN",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "SURE",
"Copy number": "Low Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Live visualization of chromatin dynamics with fluorescent TALEs",
"Publication": "Nat Struct Mol Biol. 2013 Oct 6. doi: 10.1038/nsmb.2680.",
"DOI": "10.1038/nsmb.2680.",
"Authors": "Miyanari Y, Ziegler-Birling C, Torres-Padilla ME /nsmb",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/24096363",
"PubMed ID": "PubMed 24096363"
} | {
"Gene/Insert name": "TALE-mRuby2",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "CAG",
"Tag / Fusion Protein": "mRuby2 (C terminal on backbone)"
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "BsmBI (destroyed during cloning)",
"3′ cloning site": "BsmBI (destroyed during cloning)",
"5′ sequencing primer": "catcgcgcaatgcactgac",
"3′ sequencing primer": "ggcgacgaggtggtcgttgg"
} | {
"Gene/Insert name": "",
"Species": "",
"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": ""
} |
pSWAP_Entry_T1 | 184863 | pSwap plasmid part containing the T1 sgRNA module for the Swap and Drop recombination system. | Patrick Sobetzko | Teufel et al Nat Commun. 2022 Jun 14;13(1):3430. doi: 10.1038/s41467-022-30843-1. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/363180/3e5dadf1-596c-4302-a254-2ef18c2de4ad/addgene-plasmid-184863-sequence-363180.gbk | full | LOCUS sequence-363180- 6328 bp DNA circular SYN 25-AUG-2024
DEFINITION pSwap plasmid part containing the T1 sgRNA module for the Swap and
Drop recombination system..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6328)
AUTHORS Teufel M, Klein CA, Mager M, Sobetzko P
TITLE A multifunctional system for genome editing and large-scale
interspecies gene transfer.
JOURNAL Nat Commun. 2022 Jun 14;13(1):3430. doi: 10.1038/s41467-022-30843-1.
PUBMED 35701417
REFERENCE 2 (bases 1 to 6328)
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: "2022-06-14"; volume: "13(1):3430. doi"; pages: "
10.1038/s41467-022-30843-1"
FEATURES Location/Qualifiers
source 1..6328
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind 24..42
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 126..147
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 162..192
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 200..216
/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 205..227
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 224..240
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 224..240
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
misc_feature 253..309
/label=MCS
/note="pUC18/19 multiple cloning site"
primer_bind complement(310..327)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(310..326)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(319..341)
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind complement(535..554)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
CDS 901..1206
/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 1085..1104
/label=ccdB-fwd
/note="ccdB gene, forward primer"
misc_RNA 1314..1389
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
primer_bind complement(4516..4534)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 4602..4706
/gene="bla"
/label=AmpR promoter
CDS 4707..5501
/codon_start=1
/gene="aphA-3"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin"
/translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
rep_origin complement(5561..6149)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5641..5660)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
protein_bind join(6327..6328,1..17)
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
1 cctatcagtg atagagattg acatccctat cagtgataga gatactgagc actgagaccc
61 agtggtatgg gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
121 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
181 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
241 catgattacg ccaagcttgc atgcctgcag gtcgactcta gaggatcccc gggtaccgag
301 ctcgaattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
361 acttaatcgc cttgcaggac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
421 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgcc tgatgcggta
481 ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc tcagtacaat
541 ctgctctgat gccgcatagt cgcgtggatc cggcttacta aaagccagat aacagtatgc
601 gtatttgcgc gctgattttt gcggtataag aatatatact gatatgtata cccgaagtat
661 gtcaaaaaga ggtatgctat gaagcagcgt attacagtga cagttgacag cgacagctat
721 cagttgctca aggcatatat gatgtcaata tctccggtct ggtaagcaca accatgcaga
781 atgaagcccg tcgtctgcgt gccgaacgct ggaaagcgga aaatcaggaa gggatggctg
841 aggtcgcccg gtttattgaa atgaacggct cttttgctga cgagaacagg ggctggtgaa
901 atgcagttta aggtttacac ctataaaaga gagagccgtt atcgtctgtt tgtggatgta
961 cagagtgata ttattgacac gcccgggcga cggatggtga tccccctggc cagtgcacgt
1021 ctgctgtcag ataaagtctc ccgtgaactt tacccggtgg tgcatatcgg ggatgaaagc
1081 tggcgcatga tgaccaccga tatggccagt gtgcctgtct ccgttatcgg ggaagaagtg
1141 gctgatctca gccaccgcga aaatgacatc aaaaacgcca ttaacctgat gttctgggga
1201 atataattaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
1261 ctgctcccgg catccgctta cagacaagct gtgacccact caagtgggtc tcagttttag
1321 agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg
1381 agtcggtgct ttttttacgg tcatgtaaaa aaaaaccccg cccctgacag ggcggggttt
1441 tttttcagtc aagcctctct tgacatcttt ccccgagata aatcggtaag ttgctgatta
1501 aaaaactcca acatttctat cggcgccatg ccgatagcat cgctctccag gctgcgcgcg
1561 tactgcgcca gcgccaggca tctctgcgcc agcatgtcgc agcccaggct gtagtcgctg
1621 ctgacccggc tgctgaccgg ctcgggcacg ggcggtccag catggcggcc aggcaggccg
1681 gacggcatgt tcatcagggc cgccgacagg gggcgcaaca ggcctgtcat gatgtcgatg
1741 gacgcgttca tcaggcgcga gcggcgcagg ctgcccagcg gctgctgcgc aaaatggtgc
1801 gccatcatca ggaaggtcag ttcatagcag ccctgataca gctgcatcag cgcgcgcgcc
1861 ttctgatcgg tcaccacgct gcagtccgcg cgcgcttcca gcacgggatt cttcagcgcc
1921 ggcagcgccg gttcgaacgg cttgtcgcag gccgaaaacc tgccggcgat ggcgcgcagc
1981 cgctggaagt gcgaggaggc gaaatgcggc gaatcgacgg ccacgccttc gccctgttcc
2041 gtgacgaaat cgatggcgaa caacgcgctg tcgcggtcgg atacttccag ctggtagccg
2101 ggataggcgc ggttggtcag ttctttcagg aacaggtggt gctcgccgcc gcgcttgccg
2161 ccgccgcttt cgaacaggcc gggcagctcg gccagggcct ggcggatcgc gatatagaag
2221 gtggcgatgg atttgcccgg cgtgggaatg tagtcgggcc attcaaaacg cacgaagcgg
2281 gccagcacgt gttcggaaaa tggttcgaac gcaaattccg tgtccaggcc gaaacgctgg
2341 cgcgcctgct ggcccagcag cggggtaccg ctgtaaaacg gttcgccaag cgccatcaat
2401 acattgttca ccaataagta gtgaatcatt tcttcatggg cgatttccag cagcgcgccg
2461 cgcgtgccgc tgttgcggcg ccgatcctcg gcgccacacg ctaactccag ttccgcggga
2521 agccagcgac cagactgcac caacgcggca ccttgcgcat aattcgggat gctatacgca
2581 gcgtacagat actgcagcat caggcttaat tccaggtcaa tcgccttctt cagctcatct
2641 atcagttcag ctttgccgcc aatcgcatga ggcgccgctg gttccggagc cgccgctttt
2701 gcggccgcgg cggcttctac ttgggtcaga tatttcagaa acaagcggga tttaggcagc
2761 gacagttcgc gagtgctggg catgtagtag cttttgtccc gattttgcgg gtcacacatc
2821 tgccacatca ggcgtgaata cgtttcgcat ttgcactgat cagccaggct gaacacctta
2881 tcggacataa aggggtaaac caattcgtag taactcatca catggcggta caagaaggca
2941 tagtccactt gttctgctgg aacgtcgtct aagtcccagt catccggcag aacccgcacg
3001 ccaagatact gtttcgcttt gccagttcca aggacaatgc gcgttgctcc cggtttacga
3061 cctgtgaggg tcagcgtggt atagccgtgg cctaacggac tcggttgctg ctcgacggct
3121 aagaggacgc catcttccgc ttcggccagt gacggaggtg cagccagttc accacggaaa
3181 cgcgattgaa ccgtgatggt ctgcggaaac tgctcgtgtt tgttgcgatt cggtgcctcc
3241 aggtacagct ggttagaatc gctttggagg acccaatcgg cttcatccca ctgtgcctgg
3301 gctgaaccga gcgaaagaga acctggtgca gctcccgcat gctgtaatgg aacatcaaac
3361 acgccatgat gacgccaata gtcgcgatac aactgttccg gaatacgggc gagtacggtc
3421 cctgccccat cgtgcagcag cagatcgccc aatgcttgtt taccacccag cgcatgtgtc
3481 ggatgctgct cggatacagc cccggcatca cgggtagtaa acgggattgc ggtaggcatg
3541 ttgaaactca cgcgatccgc atgcactttc accagcaccg gccccaggct gccctggcgc
3601 ggctgcagca ggcggccggc cggataggtg gccagctcgc cgcgccgcca caggccgatg
3661 ctgccggcca ggtcgtagaa cacgggcgaa tcgggttttt gcggcgtcga catattgaac
3721 aggcagtatt gcaccgtcag gcccagcacc tcgtcgtcgg ccagggcttg ctgcaaggca
3781 tgcatactgg ccggcagcgg caggtccgga ttgaacagga aatgcgggtc ctgcttggcc
3841 acggaaaact ggaacagcct ggagcggccg aattcctcgt ccaggaaatg cccgctgcgt
3901 tccgtgatgt ggccgctgcc gagccagcgt accgagtgcg cctgcgcgat gtcggccgtg
3961 aacagcgtgg gcgtattggg cgtggccagc ttgtcgctca aggtgaactg gcccgcgtag
4021 atcagcgtgg tgtcgggctg cgccgggttg ttgtcgatcc accttgcgcg gttgaacgtc
4081 gtgcgcaggt actcgttgta gtggccccac aggcccagct tggcgcccac cagcgcgtcc
4141 tgggtatcga cctcgccatc acgcaactgg acgcccgtga tccgcgcgtt ttcccacgag
4201 aaatggttgt tcccgcaaaa attatagcct gtcgcctggc tgaagatgcc gtccggatcg
4261 ggcttgccct gtgcgttgaa gcggggggcg agctgtttca ggtgcgcatg gaattcggct
4321 ggcggccggc tcaggtcgac agcctcgccc gccatcgata ccgcattcgt ggcgatatcg
4381 atgttgccgt gcgtattgcg attccccgtc ggcacattgg cgcgggcaaa accccgaaaa
4441 tgcaggcggg ggaagtcaag taggctcatt acgactctcc tctgtaaaag cgtccttgtc
4501 ttccgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
4561 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
4621 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
4681 ttcaataata ttgaaaaagg aagagtatgg ctaaaatgag aatatcaccg gaattgaaaa
4741 aactgatcga aaaataccgc tgcgtaaaag atacggaagg aatgtctcct gctaaggtat
4801 ataagctggt gggagaaaat gaaaacctat atttaaaaat gacggacagc cggtataaag
4861 ggaccaccta tgatgtggaa cgggaaaagg acatgatgct atggctggaa ggaaagctgc
4921 ctgttccaaa ggtcctgcac tttgaacggc atgatggctg gagcaatctg ctaatgagtg
4981 aggccgatgg cgtcctttgc tcggaagagt atgaagatga acaaagccct gaaaagatta
5041 tcgagctgta tgcggagtgc atcaggctct ttcactccat cgacatatcg gattgtccct
5101 atacgaatag cttagacagc cgcttagccg aattggatta cttactgaat aacgatctgg
5161 ccgatgtgga ttgcgaaaac tgggaagagg acactccatt taaagatccg cgcgagctgt
5221 atgatttttt aaagacggaa aagcccgaag aggaacttgt cttttcccac ggcgacctgg
5281 gagacagcaa catctttgtg aaagatggca aagtaagtgg ctttattgat cttgggagaa
5341 gcggcagggc ggacaagtgg tatgacattg ccttctgcgt ccggtcgctc agggaggata
5401 tcggggaaga acagtatgtc gagctatttt ttgacttact ggggatcaag cctgattggg
5461 agaaaataaa atattatatt ttactggatg aattgtttta gctgtcagac caagtttact
5521 ccggggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
5581 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
5641 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
5701 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
5761 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
5821 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
5881 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
5941 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
6001 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
6061 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
6121 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
6181 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
6241 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
6301 agtaaacttg gtctggcaga agaccatc
//
| {
"Vector backbone": "Entry vector",
"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)": "ccdB Survival",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Industry MTA"
} | {
"Title": "A multifunctional system for genome editing and large-scale interspecies gene transfer",
"Publication": "Nat Commun. 2022 Jun 14;13(1):3430. doi: 10.1038/s41467-022-30843-1.",
"DOI": "10.1038/s41467-022-30843-1.",
"Authors": "Teufel M, Klein CA, Mager M, Sobetzko P /s---",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/35701417",
"PubMed ID": "PubMed 35701417"
} | {
"Gene/Insert name": "lacZalpha,ccdB",
"Species": "E. coli, lambda phage",
"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": ""
} |
|
TFORF0462 | 142632 | Lentiviral vector for overexpressing transcription factor ORFs with unique 24-bp barcodes. Barcodes facilitate identification of transcription factors in pooled screens. | Feng Zhang | Joung et al Cell. 2023 Jan 5;186(1):209-229.e26. doi: 10.1016/j.cell.2022.11.026. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/397730/2ef43c54-ec3e-4321-b56d-19c8c27aa475/addgene-plasmid-142632-sequence-397730.gbk | full | LOCUS sequence-397730- 9475 bp DNA circular SYN 26-AUG-2024
DEFINITION Lentiviral vector for overexpressing transcription factor ORFs with
unique 24-bp barcodes. Barcodes facilitate identification of
transcription factors in pooled screens..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9475)
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 9475)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1016/j.cell.2022.11"; journalName: "Cell"; date: "2023-01-5- 5";
volume: "186"; issue: "1"; pages: "209-229.e26"
FEATURES Location/Qualifiers
source 1..9475
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 5..231
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 232..412
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 459..584
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1077..1310
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1495..1539
/codon_start=1
/product="antigenic peptide corresponding to amino acids
655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
et al., 2013)"
/label=gp41 peptide
/note="recognized by the 2H10 single-chain llama nanobody"
/translation="KNEQELLELDKWASL"
promoter 1770..1980
/label=SV40 promoter
/note="SV40 early promoter"
promoter 1792..1988
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 1839..1974
/label=SV40 ori
/note="SV40 origin of replication"
CDS 2020..2622
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGAS"
primer_bind complement(2020..2039)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 2516..2536
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
misc_feature 2665..2782
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2920..4098
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 3151..4089
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 4046..4066
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
misc_feature 5332..5920
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(5385..5405)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(5803..5814)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 5992..6225
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 6303..6424
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(6340..6359)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 6394..6413
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 6464..6599
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(6619..6638)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(6620..6638)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(6648..6665)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(6648..6664)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6657..6679)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 6806..7261
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(6893..6912)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 7103..7124
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 7287..7391
/gene="bla"
/label=AmpR promoter
CDS 7392..8252
/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(7610..7629)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 8423..9011
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 8912..8931
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 9165..9182
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 9299..9320
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 9335..9365
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 9373..9389
/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 9378..9400
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 9397..9413
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 9397..9413
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 9432..9452
/label=T3
/note="T3 promoter, forward primer"
promoter 9434..9452
/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 caccatgttc tgcacctcag gagggaattc
4141 agcctcagtg atgtccatga ggcctgtccc aggctctcta tcttctctgc tacatctcca
4201 caacagacag agacagccca tgccagcctc catgcctggg accctgccca accctacaat
4261 gccaggatct tccgccgtct tgatgccaat ggagcgacaa atgtcagtga actccagcat
4321 catggggatg caaggtccaa atctcagcaa cccctgtgct tctccccagg tccagccaat
4381 gcattcagaa gccaaaatga ggttgaaggc tgcattgact caccaccctg ctgccatgtc
4441 aaatgggaac atgaacacca tgggacacat gatggagatg atgggctccc ggcaggacca
4501 gacgccacac catcacatgc actcgcaccc gcatcagcac cagacactgc caccccatca
4561 cccttaccca caccagcacc agcacccagc acaccatcct caccctcaac cccatcacca
4621 gcagaaccat ccacatcacc actcccattc ccaccttcat gcacacccag cacatcacca
4681 gacctcgcca catccgcccc tgcacaccgg caaccaagca caggtttcac cagcaacaca
4741 acagatgcag ccaacccaga caatacagcc accccagccc acaggggggc gccggcgaag
4801 ggtggtagac gaggatccgg acgagaggcg gcggaaattt ctggaacgga accgggcagc
4861 tgccacccgc tgcagacaga agaggaaggt ctgggtgatg tcattggaaa agaaagcaga
4921 agaactcacc cagacaaaca tgcagcttca gaatgaagtg tctatgttga aaaatgaggt
4981 ggcccagctg aaacagttgt tgttaacaca taaagactgc ccaataacag ccatgcagaa
5041 agaatcacaa ggatatctaa gtccagagag tagccctcct gctagtcctg tcccagcttg
5101 ctcccagcaa caagtcatcc agcataatac catcactact tcctcatcgg tcagcgaggt
5161 ggtaggaagc tccaccctca gccagctcac cactcacaga acagacctga atccgattct
5221 ttagtaatga actagtccgt aacttgaaag tatttcgatt tcttggcttt atatatcttg
5281 tggaaaggac gaggcgggag ggttgctgca cttacgacgc gttaagtcga caatcaacct
5341 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
5401 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
5461 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
5521 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
5581 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
5641 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
5701 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt
5761 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
5821 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
5881 cctcagacga gtcggatctc cctttgggcc gcctccccgc gtcgacttta agaccaatga
5941 cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc
6001 taattcactc ccaacgaaga caagatctgc tttttgcttg tactgggtct ctctggttag
6061 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
6121 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
6181 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtacg tatagtagtt
6241 catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga
6301 ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
6361 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
6421 cttatcatgt ctggctctag ctatcccgcc cctaactccg cccatcccgc ccctaactcc
6481 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
6541 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
6601 agggacgtac ccaattcgcc ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt
6661 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
6721 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
6781 ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt
6841 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
6901 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
6961 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
7021 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
7081 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
7141 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
7201 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
7261 taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
7321 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
7381 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
7441 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
7501 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
7561 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
7621 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
7681 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
7741 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
7801 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
7861 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
7921 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
7981 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
8041 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
8101 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
8161 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
8221 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
8281 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
8341 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
8401 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
8461 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
8521 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
8581 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
8641 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
8701 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
8761 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
8821 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
8881 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
8941 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
9001 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
9061 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
9121 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
9181 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
9241 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
9301 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
9361 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
9421 acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctg caagc
//
| {
"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": "CREB5",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "1211",
"Mutation": "",
"Entrez Gene": "CREB5",
"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": ""
} |
|
pAc5.1B-EGFP-DmGW182-delM1_N | 146974 | Insect Expression of DmGW182-delM1 | Elisa Izaurralde | Izaurralde lab plasmids (unpublished) | https://media.addgene.org/snapgene-media/v2.0.0/sequences/289421/c701d3e9-a603-4035-ba5a-bc8a5f447ce8/addgene-plasmid-146974-sequence-289421.gbk | full | LOCUS sequence-289421- 10017 bp DNA circular SYN 26-AUG-2024
DEFINITION Insect Expression of DmGW182-delM1.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10017)
TITLE Izaurralde lab plasmids
REFERENCE 2 (bases 1 to 10017)
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..10017
/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 7302..7343
/codon_start=1
/product="epitope tag from simian virus 5"
/label=V5 tag
/translation="GKPIPNPLLGLDST"
CDS 7353..7370
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(7393..7410)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 7443..7577
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(7467..7486)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 7521..7540
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
rep_origin complement(8199..8787)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(8279..8298)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(8958..9818)
/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 9581..9600
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(9819..9923)
/gene="bla"
/label=AmpR promoter
primer_bind 9991..10009
/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 ttgaattctg cagatatcca gcacagtggc ggccgcatgc gtgaagccct
3361 tttttcccaa gatggctggg gctgtcagca tgttaaccag gatactaatt gggaagttcc
3421 cagttcgcca gaaccagcca ataaggatgc acccggtcca ccaatgtgga agccaagcat
3481 taacaatggt actgatcttt gggagtccaa tttgagaaac ggaggtcagc cggccgcaca
3541 gcaagttcca aagccgtcgt ggggtcatac accatcctct aacttaggtg gaacatgggg
3601 tgaggacgac gatggcgccg atagtagtag tgtgtggact ggaggagctg ttagcaacgc
3661 gggatccgga gctgcagtgg gagtaaacca agccggagtt aatgtcggtc caggcggtgt
3721 tgtttcgtct ggcggacctc agtggggaca aggtgtcgtt ggcgtcggac ttggatcaac
3781 tggaggtaac gggtcaagca atataactgg atcgtctgga gtcgcaacag gtagtagcgg
3841 aaactccagc aacgctggta acggttgggg agaccctcgt gaaatacgcc ctttgggagt
3901 tggtggctcc atggatattc gaaatgttga acatcgcggc ggtaacggtt ctggagcaac
3961 ttcgagcgat ccacgagaca ttcgcatgat cgatccgcgt gaccctattc gaggagatcc
4021 ccgtggaata tctggtcgtc ttaatgggac ctctgaaatg tggggtcatc atccacaaat
4081 gtcccataac cagttgcaag gtatcaacaa aatggttggt caaagtgtag caactgccag
4141 caccagtgtc ggaacatctg gctcgggcat cggtcctgga ggtcccggtc ctagtacagt
4201 atcaggcaat atcccaacac agtgggggcc tgctcaaccg gtaagcgttg gtgtaagtgg
4261 tcccaaagac atgtcaaaac agataagtgg atgggaggaa ccatcaccac cgcctcagcg
4321 tcgcagtatt cctaactacg atgatggtac atcgttgtgg ggtcagcaaa ctcgtgttcc
4381 cgctgcaagc ggtcactgga aagacatgac tgattcgata ggtcgtagta gtcatctcat
4441 gcgtggccaa agccaaacgg gaggtatagg aatagccggc gttggaaata gcaatgttcc
4501 agtgggagcc aatccaagta atcctataag cagtgtagtt ggacctcaag cccggattcc
4561 atctgtgggc ggcgtacaac acaaaccaga cggcggcgct atgtgggtgc attccggcaa
4621 tgtaggtggc agaaataatg ttgctgctgt tactacttgg ggagatgaca ctcatagcgt
4681 taatgtcggc gctcccagca gtggcagtgt atccagcaac aattgggttg atgacaagtc
4741 caactcaacc ttggcacaaa actcttggag cgacccggcc cctgttggag ttagttgggg
4801 caataagcaa agcaaaccgc caagcaatag tgcttcatca ggttggagca ctgctgcggg
4861 cgtggtggat ggggttgatc taggatctga gtggaacacg cacgggggga ttattggaaa
4921 atctcagcaa caacaaaaac tagcgggact taacgtggga atggtgaacg taattaacgc
4981 ggagatcatt aagcaaagca agcaatacag gatccttgtc gagaacggct ttaaaaagga
5041 agatgtagag cgggcattag tgattgctaa tatgaacatc gaagaggcag ccgatatgct
5101 ccgtgccaac tcatccctat caatggatgg ttggcgtcga catgatgagt cccttggatc
5161 ttatgccgac cacaatagtt caacaagcag cggtggattt gctggtcgtt acccggtcaa
5221 cagtggacaa ccttcaatgt cctttcctca taataacctt atgaataaca tgggaggtac
5281 cgctgttact ggaggtaaca acaatacaaa catgacagct ttacaggtgc aaaagtattt
5341 aaatcaaggg caacatggtg tcgctgttgg accgcaagcc gttggtaatt cttcagcagt
5401 atctgtcgga tttggtcaga acacgtctaa cgcagcagtg gcaggagcag cctctgcaaa
5461 tatagcagca aatacaaaca accaaccgtc tggtcagcaa attcgcatgc taggccagca
5521 aattcagttg gccattcata gtggtttcat atctagtcag atattgactc aaccgctaac
5581 tcaaacaacc cttaaccttt taaaccaact tcttagcaat attaagcatc tccaggctgc
5641 acagcaatcc cttacccgcg ggggaaatgt caatccaatg gcagtgaatg tggctatatc
5701 taaatacaag cagcaaatcc agaatttaca gaaccagata agtgcacaac aggctgtgta
5761 tgtaaaacag caaaatatgc aaccaacttc acaacaacaa cagccccaac aacagcaact
5821 tccttctgtt catctaagta actcaggcaa cgactattta agaggtcacg atgcaataaa
5881 taatttgcaa agcaactttt ctgagctcaa tattaataag ccaagtggat ataacgagaa
5941 taaagactgg tctgttgctc agccaacttc agcagcaact gcttacactg atctggtcca
6001 agagtttgag ccaggcaagc catggaaggg ttcacagatc aaaagcatag aagatgatcc
6061 cagcattaca ccaggaagcg ttgctagatc tccattgtct attaattcga cgccaaaaga
6121 tgctgacata tttgccaata ccggtaaaaa ttcaccgact gatttaccgc cactaagttt
6181 atcgtcgtct acatggagtt ttaatccaaa ccaaaattat ccgagtcaca gttggtctga
6241 caatagtcaa caatgtaccg ccacttcgga gctttggaca agcccgctaa ataaatcatc
6301 gtctcgaggt cccccgccag gattgactgc caattcaaat aaatctgcaa atagtaatgc
6361 gtcaacgcca acaactatta ccggaggtgc gaatggatgg ttacagcctc gaagtggcgg
6421 tgttcaaacc acaaacacta attggacagg tggtaacacc acttggggct ccagttggtt
6481 gcttttgaaa aatctaacag cacagattga tggtcctact ttgcgtacac tgtgtatgca
6541 gcatgggccc cttgtcagct ttcacccgta tttgaaccaa ggaattgcct tatgtaaata
6601 tactactcgt gaggaggcga acaaggcgca aatggcgtta aacaactgtg tcctcgccaa
6661 caccacaata tttgctgaat ctcccagcga gaacgaggtg caaagcatta tgcagcactt
6721 accacaaact ccttcctcta caagctctag tggaactagt ggtggcaacg tcggaggcgt
6781 cggcacttca gccaataatg caaacagtgg ttctgcagct tgtctgtccg gaaacaatag
6841 cggcaacgga aacggcagcg cgagcggcgc cggcagcggc aacaatggca acagtagctg
6901 caacaacagt gccgccgggg ggggcagcag cagcaacaac acgattacca ctgtagcaaa
6961 ttcgaatctt gttggttcta gtggctctgt ctcaaattcc tctggcgtta ctgctaactc
7021 tagtactgtt tctgtagtta gttgtacagc gagtgggaat tccataaatg gggcaggtac
7081 tgcaaacagt tctggttcaa agagtagtgc aaacaattta gctagcggcc agtctagcgc
7141 ttctaactta actaatagca ccaattcaac atggcgacaa actagccaaa accaagctct
7201 tcaaagtcaa agcaggccat caggcagaga agctgacttt gattatatat ctctcgttta
7261 ttccattgtt gatgattaat ctagagggcc cgcggttcga aggtaagcct atccctaacc
7321 ctctcctcgg tctcgattct acgcgtaccg gtcatcatca ccatcaccat tgagtttaaa
7381 cccgctgatc agcctcgact gtgccttcta aggcctgagc tcgctgatca gcctcgatcg
7441 aggatccaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
7501 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
7561 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
7621 aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatg gctgattatg
7681 atcagtcgac cgatgccctt gagagccttc aacccagtca gctccttccg gtgggcgcgg
7741 ggcatgacta tcgtcgccgc acttatgact gtcttcttta tcatgcaact cgtaggacag
7801 gtgccggcag cgctctgggt cattttcggc gaggaccgct ttcgctggag cgcgacgatg
7861 atcggcctgt cgcttgcggt attcggaatc ttgcacgccc tcgctcaagc cttcgtcact
7921 ggtcccgcca ccaaacgttt cggcgagaag caggccatta tcgccggcat ggcggccgac
7981 gcgctgggct acgtcttgct ggcgttcgcg acgcgaggct ggatggcctt ccccattatg
8041 attcttctcg cttccggcgg catcgggatg cccgcgttgc aggccatgct gtccaggcag
8101 gtagatgacg accatcaggg acagcttcaa ggatcgctcg cggctcttac cagccagcaa
8161 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
8221 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
8281 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
8341 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca
8401 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
8461 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
8521 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
8581 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
8641 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
8701 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
8761 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
8821 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
8881 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
8941 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
9001 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
9061 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
9121 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
9181 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
9241 gttaatagtt tgcgcaacgt tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg
9301 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
9361 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
9421 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
9481 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
9541 atgcggcgac cgagttgctc ttgcccggcg tcaacacggg ataataccgc gccacatagc
9601 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
9661 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
9721 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
9781 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
9841 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
9901 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
9961 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgt
//
| {
"Vector backbone": "Unknown",
"Backbone size w/o insert (bp)": "",
"Vector type": "Insect Expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "Low Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "",
"Publication": "",
"DOI": "",
"Authors": "",
"PubMed Link": "",
"PubMed ID": ""
} | {
"Gene/Insert name": "DmGW182-delM1",
"Species": "D. melanogaster (fly)",
"Insert Size (bp)": "",
"Mutation": "Two non silent mutations V704A and N799S and one silent mutation G2295A compared to the sequence given by NM_166780 varA",
"Entrez Gene": "gw",
"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": ""
} |
|
pDUAL CD81 (GFP) | 86980 | Lentiviral expression construct encoding CD81 and GFP from separate promoters | Joe Grove | Grove lab plasmids (unpublished) | https://media.addgene.org/snapgene-media/v2.0.0/sequences/173290/392d6b3b-f22b-45b0-b801-709ca7258473/addgene-plasmid-86980-sequence-173290.gbk | full | LOCUS sequence-173290- 10799 bp DNA circular SYN 25-AUG-2024
DEFINITION Lentiviral expression construct encoding CD81 and GFP from separate
promoters.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10799)
TITLE Grove lab plasmids
REFERENCE 2 (bases 1 to 10799)
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..10799
/mol_type="other DNA"
/organism="synthetic DNA construct"
LTR 16..649
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 696..821
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1318..1551
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1736..1780
/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 2065..2182
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2335..2742
/label=SFFV promoter
/note="spleen focus-forming virus long terminal repeat
(LTR) promoter"
regulatory 2753..2762
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
primer_bind complement(3565..3585)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(3983..3994)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
promoter 4030..4540
/label=hPGK promoter
/note="human phosphoglycerate kinase 1 promoter"
primer_bind 4459..4477
/label=hPGK-F
/note="Human PGK promoter, forward primer"
regulatory 4555..4564
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 4561..5280
/codon_start=1
/product="Emerald GFP"
/label=EmGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 4561..5280
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(4606..4627)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(4867..4886)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 5214..5235
/label=EGFP-C
/note="EGFP, forward primer"
LTR 5376..5609
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
promoter complement(5634..5652)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(5635..5652)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(5942..5961)
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind complement(6175..6194)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 6294..6316
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(6354..6372)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 6440..6544
/gene="bla"
/label=AmpR promoter
CDS 6545..7405
/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(6763..6782)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 7576..8164
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 8065..8084
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 8318..8335
/label=L4440
/note="L4440 vector, forward primer"
primer_bind complement(8405..8425)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 8410..8739
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 8590..8725
/label=SV40 ori
/note="SV40 origin of replication"
intron 9873..9938
/label=small t intron
/note="SV40 (simian virus 40) small t antigen intron"
CDS 10068..10088
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
polyA_signal 10513..10647
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(10550..10569)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 10604..10623
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
ORIGIN
1 tacaaactta gtagttggaa gggctaattc actcccaaag aagacaagat atccttgatc
61 tgtggatcta ccacacacaa ggctacttcc ctgattagca gaactacaca ccagggccag
121 gggtcagata tccactgacc tttggatggt gctacaagct agtaccagtt gagccagata
181 aggtagaaga ggccaataaa ggagagaaca ccagcttgtt acaccctgtg agcctgcatg
241 ggatggatga cccggagaga gaagtgttag agtggaggtt tgacagccgc ctagcatttc
301 atcacgtggc ccgagagctg catccggagt acttcaagaa ctgctgatat cgagcttgct
361 acaagggact ttccgctggg gactttccag ggaggcgtgg cctgggcggg actggggagt
421 ggcgagccct cagatcctgc atataagcag ctgctttttg cctgtactgg gtctctctgg
481 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
541 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
601 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga
661 acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg
721 ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga
781 ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa
841 ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa
901 aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag
961 aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat
1021 cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga
1081 tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta
1141 agaccaccgc acagcaagcg gccggccgct gatcttcaga cctggaggag gagatatgag
1201 ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt
1261 agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg
1321 agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag cgtcaatgac
1381 gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct
1441 gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct
1501 ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg
1561 gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa
1621 taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca gagaaattaa
1681 caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa
1741 tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac
1801 aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag
1861 aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc
1921 gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa tagaagaaga
1981 aggtggagag agagacagag acagatccat tcgattagtg aacggatctc gacggtatcg
2041 ccaaatggca gtattcatcc acaattttaa aagaaaaggg gggattgggg ggtacagtgc
2101 aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat tacaaaaaca
2161 aattacaaaa attcaaaatt ttcgggttta ttacagggac agcagagatc cagtttggat
2221 cgataagctt gatatcgaat tcctgcagcc ccgataaaat aaaagatttt atttagtctc
2281 cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg gcaagctagc tgcagtaacg
2341 ccattttgca aggcatggaa aaataccaaa ccaagaatag agaagttcag atcaagggcg
2401 ggtacatgaa aatagctaac gttgggccaa acaggatatc tgcggtgagc agtttcggcc
2461 ccggcccggg gccaagaaca gatggtcacc gcagtttcgg ccccggcccg aggccaagaa
2521 cagatggtcc ccagatatgg cccaaccctc agcagtttct taagacccat cagatgtttc
2581 caggctcccc caaggacctg aaatgaccct gcgccttatt tgaattaacc aatcagcctg
2641 cttctcgctt ctgttcgcgc gcttctgctt cccgagctct ataaaagagc tcacaacccc
2701 tcactcggcg cgccagtcct ccgacagact gagtcgcccg gggggatccg ccgccaccat
2761 gggagtggag ggctgcacca agtgcatcaa gtacctgctc ttcgtcttca atttcgtctt
2821 ctggctggct ggaggcgtga tcctgggtgt ggccctgtgg ctccgccatg acccgcagac
2881 caccaacctc ctgtatctgg agctgggaga caagcccgcg cccaacacct tctatgtagg
2941 catctacatc ctcatcgctg tgggcgctgt catgatgttc gttggcttcc tgggctgcta
3001 cggggccatc caggaatccc agtgcctgct ggggacgttc ttcacctgcc tggtcatcct
3061 gtttgcctgt gaggtggccg ccggcatctg gggctttgtc aacaaggacc agatcgccaa
3121 ggatgtgaag cagttctatg accaggccct acagcaggcc gtggtggatg atgacgccaa
3181 caacgccaag gctgtggtga agaccttcca cgagacgctt gactgctgtg gctccagcac
3241 actgactgct ttgaccacct cagtgctcaa gaacaatttg tgtccctcgg gcagcaacat
3301 catcagcaac ctcttcaagg aggactgcca ccagaagatc gatgacctct tctccgggaa
3361 gctgtacctc atcggcattg ctgccatcgt ggtcgctgtg atcatgatct tcgagatgat
3421 cctgagcatg gtgctgtgct gtggcatccg gaacagctcc gtgtactgag tcgacctgca
3481 ggcatgcaag cttgatatca agcttatcga taatcaacct ctggattaca aaatttgtga
3541 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
3601 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
3661 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
3721 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
3781 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
3841 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
3901 ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg
3961 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggtcg
4021 aattcccacg gggttggggt tgcgcctttt ccaaggcagc cctgggtttg cgcagggacg
4081 cggctgctct gggcgtggtt ccgggaaacg cagcggcgcc gaccctgggt ctcgcacatt
4141 cttcacgtcc gttcgcagcg tcacccggat cttcgccgct acccttgtgg gccccccggc
4201 gacgcttcct gctccgcccc taagtcggga aggttccttg cggttcgcgg cgtgccggac
4261 gtgacaaacg gaagccgcac gtctcactag taccctcgca gacggacagc gccagggagc
4321 aatggcagcg cgccgaccgc gatgggctgt ggccaatagt ggctgctcag cagggcgcgc
4381 cgagagcagc ggccgggaag gggcggtgcg ggaggcgggg tgtggggcgg tagtgtgggc
4441 cctgttcctg cccgcacggt gttccgcatt ctgcaagcct ccggagcgca cgtcggcagt
4501 cggctccctc gttgaccgaa tcaccgacct ctctccccag gacgcgtacc ggtcgccacc
4561 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
4621 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
4681 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
4741 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
4801 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
4861 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
4921 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
4981 aagctggagt acaactacaa cagccacaag gtctatatca ccgccgacaa gcagaagaac
5041 ggcatcaagg tgaacttcaa gacccgccac aacatcgagg acggcagcgt gcagctcgcc
5101 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
5161 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
5221 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa
5281 actcgaggga attaattcga gctcggtacc tttaagacca atgacttaca aggcagctgt
5341 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg
5401 aagacaagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg
5461 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
5521 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
5581 cttttagtca gtgtggaaaa tctctagcag catctagaat taattccgtg tattctatag
5641 tgtcacctaa atcgtatgtg tatgatacat aaggttatgt attaattgta gccgcgttct
5701 aacgacaata tgtacaagcc taattgtgta gcatctggct tactgaagca gaccctatca
5761 tctctctcgt aaactgccgt cagagtcggt ttggttggac gaaccttctg agtttctggt
5821 aacgccgtcc cgcacccgga aatggtcagc gaaccaatca gcagggtcat cgctagccag
5881 atcctctacg ccggacgcat cgtggccggc atcaccggcg ccacaggtgc ggttgctggc
5941 gcctatatcg ccgacatcac cgatggggaa gatcgggctc gccacttcgg gctcatgagc
6001 gcttgtttcg gcgtgggtat ggtggcaggc cccgtggccg ggggactgtt gggcgccatc
6061 tccttgcatg caccattcct tgcggcggcg gtgctcaacg gcctcaacct actactgggc
6121 tgcttcctaa tgcaggagtc gcataaggga gagcgtcgaa tggtgcactc tcagtacaat
6181 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
6241 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
6301 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
6361 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
6421 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
6481 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
6541 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
6601 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
6661 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
6721 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
6781 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
6841 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
6901 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
6961 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
7021 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
7081 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
7141 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
7201 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
7261 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
7321 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
7381 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
7441 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
7501 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
7561 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
7621 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
7681 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
7741 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
7801 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
7861 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
7921 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
7981 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
8041 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
8101 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
8161 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
8221 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
8281 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
8341 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
8401 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
8461 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
8521 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
8581 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
8641 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
8701 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agcttggaca caagacaggc
8761 ttgcgagata tgtttgagaa taccacttta tcccgcgtca gggagaggca gtgcgtaaaa
8821 agacgcggac tcatgtgaaa tactggtttt tagtgcgcca gatctctata atctcgcgca
8881 acctattttc ccctcgaaca ctttttaagc cgtagataaa caggctggga cacttcacat
8941 gagcgaaaaa tacatcgtca cctgggacat gttgcagatc catgcacgta aactcgcaag
9001 ccgactgatg ccttctgaac aatggaaagg cattattgcc gtaagccgtg gcggtctgta
9061 ccgggtgcgt tactggcgcg tgaactgggt attcgtcatg tcgataccgt ttgtatttcc
9121 agctacgatc acgacaacca gcgcgagctt aaagtgctga aacgcgcaga aggcgatggc
9181 gaaggcttca tcgttattga tgacctggtg gataccggtg gtactgcggt tgcgattcgt
9241 gaaatgtatc caaaagcgca ctttgtcacc atcttcgcaa aaccggctgg tcgtccgctg
9301 gttgatgact atgttgttga tatcccgcaa gatacctgga ttgaacagcc gtgggatatg
9361 ggcgtcgtat tcgtcccgcc aatctccggt cgctaatctt ttcaacgcct ggcactgccg
9421 ggcgttgttc tttttaactt caggcgggtt acaatagttt ccagtaagta ttctggaggc
9481 tgcatccatg acacaggcaa acctgagcga aaccctgttc aaaccccgct ttaaacatcc
9541 tgaaacctcg acgctagtcc gccgctttaa tcacggcgca caaccgcctg tgcagtcggc
9601 ccttgatggt aaaaccatcc ctcactggta tcgcatgatt aaccgtctga tgtggatctg
9661 gcgcggcatt gacccacgcg aaatcctcga cgtccaggca cgtattgtga tgagcgatgc
9721 cgaacgtacc gacgatgatt tatacgatac ggtgattggc taccgtggcg gcaactggat
9781 ttatgagtgg gccccggatc tttgtgaagg aaccttactt ctgtggtgtg acataattgg
9841 acaaactacc tacagagatt taaagctcta aggtaaatat aaaattttta agtgtataat
9901 gtgttaaact actgattcta attgtttgtg tattttagat tccaacctat ggaactgatg
9961 aatgggagca gtggtggaat gcctttaatg aggaaaacct gttttgctca gaagaaatgc
10021 catctagtga tgatgaggct actgctgact ctcaacattc tactcctcca aaaaagaaga
10081 gaaaggtaga agaccccaag gactttcctt cagaattgct aagttttttg agtcatgctg
10141 tgtttagtaa tagaactctt gcttgctttg ctatttacac cacaaaggaa aaagctgcac
10201 tgctatacaa gaaaattatg gaaaaatatt ctgtaacctt tataagtagg cataacagtt
10261 ataatcataa catactgttt tttcttactc cacacaggca tagagtgtct gctattaata
10321 actatgctca aaaattgtgt acctttagct ttttaatttg taaaggggtt aataaggaat
10381 atttgatgta tagtgccttg actagagatc ataatcagcc ataccacatt tgtagaggtt
10441 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca
10501 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
10561 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
10621 atcaatgtat cttatcatgt ctggatcaac tggataactc aagctaacca aaatcatccc
10681 aaacttccca ccccataccc tattaccact gccaattacc tgtggtttca tttactctaa
10741 acctgtgatt cctctgaatt attttcattt taaagaaatt gtatttgtta aatatgtac
//
| {
"Vector backbone": "pDUAL GFP",
"Backbone size w/o insert (bp)": "",
"Vector type": "Mammalian Expression, Lentiviral",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "NEB Stable",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "",
"Publication": "",
"DOI": "",
"Authors": "",
"PubMed Link": "",
"PubMed ID": ""
} | {
"Gene/Insert name": "CD81",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "CD81",
"Promoter": "SFFV/hPGK",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "unknown (unknown if destroyed)",
"3′ cloning site": "unknown (unknown if destroyed)",
"5′ sequencing primer": "SFFVpro-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": ""
} |
|
pGW045 AAV-CRISPRa base vector pAAV_OE-U6sgbbSapI(MS2)-EF1a-MPH-WPRE | 192161 | (Empty Backbone)AAV-CRISPRa base vector | Sidi Chen | Wang et al Nat Immunol. 2019 Nov;20(11):1494-1505. doi: 10.1038/s41590-019-0500-4. Epub 2019 Oct 14. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/375163/a98ddf3c-96c9-4c8a-87c5-3ad274070c13/addgene-plasmid-192161-sequence-375163.gbk | full | LOCUS sequence-375163- 6717 bp DNA circular SYN 26-AUG-2024
DEFINITION AAV-CRISPRa base vector .
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6717)
AUTHORS Wang G, Chow RD, Bai Z, Zhu L, Errami Y, Dai X, Dong MB, Ye L, Zhang
X, Renauer PA, Park JJ, Shen L, Ye H, Fuchs CS, Chen S
TITLE Multiplexed activation of endogenous genes by CRISPRa elicits potent
antitumor immunity.
JOURNAL Nat Immunol. 2019 Nov;20(11):1494-1505. doi:
10.1038/s41590-019-0500-4. Epub 2019 Oct 14.
PUBMED 31611701
REFERENCE 2 (bases 1 to 6717)
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/s41590-019-0500-4"; journalName: "Nat Immunol"; date:
"2019-11"; volume: "20"; issue: "11"; pages: "1494-1505"
FEATURES Location/Qualifiers
source 1..6717
/mol_type="other DNA"
/organism="synthetic DNA construct"
repeat_region 1..130
/label=AAV2 ITR (alternate)
/note="Functional equivalent of wild-type AAV2 ITR"
promoter 152..400
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA
(Domitrovich & Kunkel, 2003)"
primer_bind 152..172
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 323..342
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
misc_RNA 436..454
/label=MS2 stem loop
/note="stem loop that binds the bacteriophage MS2 coat
protein"
misc_RNA 506..524
/label=MS2 stem loop
/note="stem loop that binds the bacteriophage MS2 coat
protein"
promoter 714..1892
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 945..1883
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 1840..1860
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
regulatory 1899..1908
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 1908..2294
/codon_start=1
/product="bacteriophage MS2 coat protein"
/label=MS2-N55K
/note="binds with high affinity to a specific stem-loop
structure in the viral RNA (Lim et al., 1994)"
/translation="ASNFTQFVLVDNGGTGDVTVAPSNFANGVAEWISSNSRSQAYKVT
CSVRQSSAQKRKYTIKVEVPKVATQTVGGVELPVAAWRSYLNMELTIPIFATNSDCELI
VKAMQGLLKDGNPIPSAIAANSGIY"
CDS 2349..2369
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 2385..2927
/codon_start=1
/product="C-terminal portion of the p65 subunit of mouse
NF-kappa-B"
/label=p65
/translation="PSGQISNQALALAPSSAPVLAQTMVPSSAMVPLAQPPAPAPVLTP
GPPQSLSAPVPKSTQAGEGTLSEALLHLQFDADEDLGALLGNSTDPGVFTDLASVDNSE
FQQLLNQGVSMSHSTAEPMLMEYPEAITRLVTGSQRPPDPAPTPLGTSGLPNGLSGDED
FSSIADMDFSALLSQISS"
CDS 2952..3323
/codon_start=1
/product="C-terminal activation domain from the human heat
shock transcription factor HSF1"
/label=HSF1
/translation="GFSVDTSALLDLFSPSVTVPDMSLPDLDSSLASIQELLSPQEPPR
PPEAENSSPDSGKQLVHYTAQPLFLLDPGSVDTGSNDLPVLFELGEGSYFSEGDGFAED
PTISLLTGSEPPKAKDPTVS"
misc_feature 3348..3936
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(3401..3421)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(3819..3830)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
repeat_region 3980..4120
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 3980..4109
/label=AAV2 ITR
rep_origin 4195..4650
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4282..4301)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4492..4513
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(4667..4686)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 4786..4808
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(4846..4864)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 4932..5036
/gene="bla"
/label=AmpR promoter
CDS 5037..5897
/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(5255..5274)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 6068..6656
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6557..6576
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
121 aggggttcct gcggccgcac gcgttctaga agagggccta tttcccatga ttccttcata
181 tttgcatata cgatacaagg ctgttagaga gataattgga attaatttga ctgtaaacac
241 aaagatatta gtacaaaata cgtgacgtag aaagtaataa tttcttgggt agtttgcagt
301 tttaaaatta tgttttaaaa tggactatca tatgcttacc gtaacttgaa agtatttcga
361 tttcttggct ttatatatct tgtggaaagg acgaaacacc ggaagagcga gctcttctgt
421 tttagagcta ggccaacatg aggatcaccc atgtctgcag ggcctagcaa gttaaaataa
481 ggctagtccg ttatcaactt ggccaacatg aggatcaccc atgtctgcag ggccaagtgg
541 caccgagtcg gtgctttttt ggatccaagc ttggcgtaac tagatcttga gacaaatggg
601 acagcagaga tccagtttgg ttaattagct agctgcaaag atggataaag ttttaaacag
661 agaggaatct ttgcagctaa tggaccttct aggtcttgaa aggagtggga attggctccg
721 gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg gggggagggg
781 tcggcaattg aaccggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg
841 tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg
901 ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acaggtaagt gccgtgtgtg
961 gttcccgcgg gcctggcctc tttacgggtt atggcccttg cgtgccttga attacttcca
1021 cctggctgca gtacgtgatt cttgatcccg agcttcgggt tggaagtggg tgggagagtt
1081 cgaggccttg cgcttaagga gccccttcgc ctcgtgcttg agttgaggcc tggcctgggc
1141 gctggggccg ccgcgtgcga atctggtggc accttcgcgc ctgtctcgct gctttcgata
1201 agtctctagc catttaaaat ttttgatgac ctgctgcgac gctttttttc tggcaagata
1261 gtcttgtaaa tgcgggccaa gatctgcaca ctggtatttc ggtttttggg gccgcgggcg
1321 gcgacggggc ccgtgcgtcc cagcgcacat gttcggcgag gcggggcctg cgagcgcggc
1381 caccgagaat cggacggggg tagtctcaag ctggccggcc tgctctggtg cctggcctcg
1441 cgccgccgtg tatcgccccg ccctgggcgg caaggctggc ccggtcggca ccagttgcgt
1501 gagcggaaag atggccgctt cccggccctg ctgcagggag ctcaaaatga aggacgcggc
1561 gctcgggaga gcgggcgggt gagtcaccca cacaaaggaa aagggccttt ccgtcctcag
1621 ccgtcgcttc atgtgactcc acggagtacc gggcgccgtc caggcacctc gattagttct
1681 cgagcttttg gagtacgtcg tctttaggtt ggggggaggg gttttatgcg atggagtttc
1741 cccacactga gtgggtggag actgaagtta ggccagcttg gcacttgatg taattctcct
1801 tggaatttgc cctttttgag tttggatctt ggttcattct caagcctcag acagtggttc
1861 aaagtttttt tcttccattt caggtgtcgt gacgtacggc caccatggct tcaaacttta
1921 ctcagttcgt gctcgtggac aatggtggga caggggatgt gacagtggct ccttctaatt
1981 tcgctaatgg ggtggcagag tggatcagct ccaactcacg gagccaggcc tacaaggtga
2041 catgcagcgt caggcagtct agtgcccaga agagaaagta taccatcaag gtggaggtcc
2101 ccaaagtggc tacccagaca gtgggcggag tcgaactgcc tgtcgccgct tggaggtcct
2161 acctgaacat ggagctcact atcccaattt tcgctaccaa ttctgactgt gaactcatcg
2221 tgaaggcaat gcaggggctc ctcaaagacg gtaatcctat cccttccgcc atcgccgcta
2281 actcaggtat ctacagcgct ggaggaggtg gaagcggagg aggaggaagc ggaggaggag
2341 gtagcggacc taagaaaaag aggaaggtgg cggccgctgg atccccttca gggcagatca
2401 gcaaccaggc cctggctctg gcccctagct ccgctccagt gctggcccag actatggtgc
2461 cctctagtgc tatggtgcct ctggcccagc cacctgctcc agcccctgtg ctgaccccag
2521 gaccacccca gtcactgagc gctccagtgc ccaagtctac acaggccggc gaggggactc
2581 tgagtgaagc tctgctgcac ctgcagttcg acgctgatga ggacctggga gctctgctgg
2641 ggaacagcac cgatcccgga gtgttcacag atctggcctc cgtggacaac tctgagtttc
2701 agcagctgct gaatcagggc gtgtccatgt ctcatagtac agccgaacca atgctgatgg
2761 agtaccccga agccattacc cggctggtga ccggcagcca gcggcccccc gaccccgctc
2821 caactcccct gggaaccagc ggcctgccta atgggctgtc cggagatgaa gacttctcaa
2881 gcatcgctga tatggacttt agtgccctgc tgtcacagat ttcctctagt gggcagggag
2941 gaggtggaag cggcttcagc gtggacacca gtgccctgct ggacctgttc agcccctcgg
3001 tgaccgtgcc cgacatgagc ctgcctgacc ttgacagcag cctggccagt atccaagagc
3061 tcctgtctcc ccaggagccc cccaggcctc ccgaggcaga gaacagcagc ccggattcag
3121 ggaagcagct ggtgcactac acagcgcagc cgctgttcct gctggacccc ggctccgtgg
3181 acaccgggag caacgacctg ccggtgctgt ttgagctggg agagggctcc tacttctccg
3241 aaggggacgg cttcgccgag gaccccacca tctccctgct gacaggctcg gagcctccca
3301 aagccaagga ccccactgtc tcctaagaat tcacgcgtta agtcgacaat caacctctgg
3361 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
3421 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
3481 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
3541 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
3601 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
3661 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
3721 attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca
3781 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
3841 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
3901 agacgagtcg gatctccctt tgggccgcct ccccgcgtcg actttaagac caatgaccac
3961 gtgcggaccg agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc
4021 gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg
4081 gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt
4141 ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc
4201 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
4261 ttgccagcgc cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
4321 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
4381 tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc
4441 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
4501 tgttccaaac tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga
4561 ttttgccgat ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
4621 attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg
4681 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
4741 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
4801 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
4861 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
4921 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
4981 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
5041 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
5101 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
5161 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
5221 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
5281 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
5341 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
5401 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
5461 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
5521 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
5581 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
5641 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
5701 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
5761 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
5821 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
5881 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
5941 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
6001 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
6061 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
6121 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
6181 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
6241 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
6301 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
6361 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
6421 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
6481 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
6541 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
6601 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
6661 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgt
//
| {
"Vector backbone": "pAAV",
"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": "Multiplexed activation of endogenous genes by CRISPRa elicits potent antitumor immunity",
"Publication": "Nat Immunol. 2019 Nov;20(11):1494-1505. doi: 10.1038/s41590-019-0500-4. Epub 2019 Oct 14.",
"DOI": "10.1038/s41590-019-0500-4.",
"Authors": "Wang G, Chow RD, Bai Z, Zhu L, Errami Y, Dai X, Dong MB, Ye L, Zhang X, Renauer PA, Park JJ, Shen L, Ye H, Fuchs CS, Chen S /s---",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/31611701",
"PubMed ID": "PubMed 31611701"
} | {
"Gene/Insert name": "None",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "NA",
"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": ""
} |
|
pAAV-STAT3 | 195575 | Expresses STAT3 | Kevin Park | Luo et al Cell Rep. 2016 Apr 12;15(2):398-410. doi: 10.1016/j.celrep.2016.03.029. Epub 2016 Mar 31. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/384024/df6d7db3-e21b-45e4-8db7-53fb85cdf704/addgene-plasmid-195575-sequence-384024.gbk | full | LOCUS sequence-384024- 6903 bp DNA circular SYN 26-AUG-2024
DEFINITION Expresses STAT3.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6903)
AUTHORS Luo X, Ribeiro M, Bray ER, Lee DH, Yungher BJ, Mehta ST, Thakor KA,
Diaz F, Lee JK, Moraes CT, Bixby JL, Lemmon VP, Park KK
TITLE Enhanced Transcriptional Activity and Mitochondrial Localization of
STAT3 Co-induce Axon Regrowth in the Adult Central Nervous System.
JOURNAL Cell Rep. 2016 Apr 12;15(2):398-410. doi:
10.1016/j.celrep.2016.03.029. Epub 2016 Mar 31.
PUBMED 27050520
REFERENCE 2 (bases 1 to 6903)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1016/j.celrep.2016.03.029"; journalName: "Cell Rep"; date:
"2016-04-12- 12"; volume: "15"; issue: "2"; pages: "398-410"
FEATURES Location/Qualifiers
source 1..6903
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(104..123)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 917..1505
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 1406..1425
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
repeat_region 1567..1696
/label=AAV2 ITR (alternate)
/note="Functional equivalent of wild-type AAV2 ITR"
enhancer 1772..2075
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 2076..2279
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 2230..2250
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 2276..2300
/label=LNCX
/note="Human CMV promoter, forward primer"
intron 2428..2806
/label=chimeric intron
/note="fusion between human cytomegalovirus intron A and
human beta-globin intron 2"
polyA_signal 5216..5692
/label=hGH poly(A) signal
/note="human growth hormone polyadenylation signal"
primer_bind complement(5415..5434)
/label=hGH-PA-R
/note="Human growth hormone terminator, reverse primer"
repeat_region 5732..5872
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 5732..5861
/label=AAV2 ITR
rep_origin 5947..6402
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(6034..6053)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 6244..6265
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(6419..6438)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 6538..6560
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(6598..6616)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 6684..6788
/gene="bla"
/label=AmpR promoter
CDS join(6789..6903,1..746)
/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"
ORIGIN
1 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
61 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
121 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
181 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
241 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
301 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
361 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
421 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
481 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
541 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
601 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
661 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
721 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
781 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
841 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
901 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
961 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
1021 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
1081 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
1141 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
1201 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
1261 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
1321 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
1381 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
1441 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
1501 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
1561 acatgtcctg caggcagctg cgcgctcgct cgctcactga ggccgcccgg gcgtcgggcg
1621 acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc
1681 atcactaggg gttcctgcgg ccgcacgcgt ctagttatta atagtaatca attacggggt
1741 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
1801 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
1861 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
1921 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
1981 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
2041 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca
2101 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
2161 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
2221 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc
2281 gtttagtgaa ccgtcagatc gcctggagac gccatccacg ctgttttgac ctccatagaa
2341 gacaccggga ccgatccagc ctccgcggat tcgaatcccg gccgggaacg gtgcattgga
2401 acgcggattc cccgtgccaa gagtgacgta agtaccgcct atagagtcta taggcccaca
2461 aaaaatgctt tcttctttta atatactttt ttgtttatct tatttctaat actttcccta
2521 atctctttct ttcagggcaa taatgataca atgtatcatg cctctttgca ccattctaaa
2581 gaataacagt gataatttct gggttaaggc aatagcaata tttctgcata taaatatttc
2641 tgcatataaa ttgtaactga tgtaagaggt ttcatattgc taatagcagc tacaatccag
2701 ctaccattct gcttttattt tatggttggg ataaggctgg attattctga gtccaagcta
2761 ggcccttttg ctaatcatgt tcatacctct tatcttcctc ccacagctcc tgggcaacgt
2821 gctggtctgt gtgctggccc atcactttgg caaagaattg ggattcgaac atcgattgaa
2881 ttcccaccat ggctcagtgg aaccagctgc agcagctgga cacacgctac ctggagcagc
2941 tgcaccagct gtacagcgac agcttcccca tggagctgcg gcagttcctg gcaccttgga
3001 ttgagagtca agactgggca tatgcagcca gcaaagagtc acatgccacg ttggtgtttc
3061 ataatctctt gggtgaaatt gaccagcaat atagccgatt cctgcaagag tccaatgtcc
3121 tctatcagca caaccttcga agaatcaagc agtttctgca gagcaggtat cttgagaagc
3181 caatggaaat tgcccggatc gtggcccgat gcctgtggga agagtctcgc ctcctccaga
3241 cggcagccac ggcagcccag caagggggcc aggccaacca cccaacagcc gccgtagtga
3301 cagagaagca gcagatgttg gagcagcatc ttcaggatgt ccggaagcga gtgcaggatc
3361 tagaacagaa aatgaaggtg gtggagaacc tccaggacga ctttgatttc aactacaaaa
3421 ccctcaagag ccaaggagac atgcaggatc tgaatggaaa caaccagtct gtgaccagac
3481 agaagatgca gcagctggaa cagatgctca cagccctgga ccagatgcgg agaagcattg
3541 tgagtgagct ggcggggctc ttgtcagcaa tggagtacgt gcagaagaca ctgactgatg
3601 aagagctggc tgactggaag aggcggcagc agatcgcgtg catcggaggc cctcccaaca
3661 tctgcctgga ccgtctggaa aactggataa cttcattagc agaatctcaa cttcagaccc
3721 gccaacaaat taagaaactg gaggagctgc agcagaaagt gtcctacaag ggcgacccta
3781 tcgtgcagca ccggcccatg ctggaggaga ggatcgtgga gctgttcaga aacttaatga
3841 agagtgcctt cgtggtggag cggcagccct gcatgcccat gcacccggac cggcccttag
3901 tcatcaagac tggtgtccag tttaccacga aagtcaggtt gctggtcaaa tttcctgagt
3961 tgaattatca gcttaaaatt aaagtgtgca ttgataaaga ctctggggat gttgctgccc
4021 tcagagggtc tcggaaattt aacattctgg gcacgaacac aaaagtgatg aacatggagg
4081 agtctaacaa cggcagcctg tctgcagagt tcaagcacct gacccttagg gagcagagat
4141 gtgggaatgg aggccgtgcc aattgtgatg cctccttgat cgtgactgag gagctgcacc
4201 tgatcacctt cgagactgag gtgtaccacc aaggcctcaa gattgaccta gagacccact
4261 ccttgccagt tgtggtgatc tccaacatct gtcagatgcc aaatgcttgg gcatcaatcc
4321 tgtggtataa catgctgacc aataacccca agaacgtgaa cttcttcact aagccgccaa
4381 ttggaacctg ggaccaagtg gccgaggtgc tcagctggca gttctcgtcc accaccaagc
4441 gggggctgag catcgagcag ctgacaacgc tggctgagaa gctcctaggg cctggtgtga
4501 actactcagg gtgtcagatc acatgggcta aattctgcaa agaaaacatg gctggcaagg
4561 gcctctcctt ctgggtctgg ctagacaata tcatcgacct tgtgaaaaag tatatcttgg
4621 ccctttggaa tgaagggtac atcatgggtt tcatcagcag ggagcgggag cgggccatcc
4681 taagcacaaa gcccccgggc accttcctac tgcgcttcag cgagagcagc aaagaaggag
4741 gggtcacttt cacttgggtg gaaaaggaca tcagtggcaa gacccagatc cagtctgtag
4801 agccatacac caagcagcag ctgaacaaca tgtcatttgc tgaaatcatc atgggctata
4861 agatcatgga tgcgaccaac atcctggtgt ctccacttgt ctacctctac cccgacattc
4921 ccaaggagga ggcatttgga aagtactgta ggcccgagag ccaggagcac cccgaagccg
4981 acccaggtag tgctgccccg tacctgaaga ccaagttcat ctgtgtgaca ccaacgacct
5041 gcagcaatac cattgacctg ccgatgtccc cccgcacttt agattcattg atgcagtttg
5101 gaaataacgg tgaaggtgct gagccctcag caggagggca gtttgagtcg ctcacgtttg
5161 acatggatct gacctcggag tgtgctacct cccccatgta gctcgagaga tctacgggtg
5221 gcatccctgt gacccctccc cagtgcctct cctggccctg gaagttgcca ctccagtgcc
5281 caccagcctt gtcctaataa aattaagttg catcattttg tctgactagg tgtccttcta
5341 taatattatg gggtggaggg gggtggtatg gagcaagggg caagttggga agacaacctg
5401 tagggcctgc ggggtctatt gggaaccaag ctggagtgca gtggcacaat cttggctcac
5461 tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct cagcctcccg agttgttggg
5521 attccaggca tgcatgacca ggctcagcta atttttgttt ttttggtaga gacggggttt
5581 caccatattg gccaggctgg tctccaactc ctaatctcag gtgatctacc caccttggcc
5641 tcccaaattg ctgggattac aggcgtgaac cactgctccc ttccctgtcc ttctgatttt
5701 gtaggtaacc acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact
5761 ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg
5821 ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg
5881 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac
5941 catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
6001 tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc
6061 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
6121 gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta
6181 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
6241 atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg
6301 atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa
6361 aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta
6421 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
6481 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
6541 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc
6601 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
6661 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
6721 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
6781 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
6841 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
6901 tgg
//
| {
"Vector backbone": "pAAV-MCS",
"Backbone size w/o insert (bp)": "4650",
"Vector type": "Mammalian Expression, AAV",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "NEB Stable",
"Copy number": "Unknown"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Enhanced Transcriptional Activity and Mitochondrial Localization of STAT3 Co-induce Axon Regrowth in the Adult Central Nervous System",
"Publication": "Cell Rep. 2016 Apr 12;15(2):398-410. doi: 10.1016/j.celrep.2016.03.029. Epub 2016 Mar 31.",
"DOI": "10.1016/j.celrep.2016.03.029.",
"Authors": "Luo X, Ribeiro M, Bray ER, Lee DH, Yungher BJ, Mehta ST, Thakor KA, Diaz F, Lee JK, Moraes CT, Bixby JL, Lemmon VP, Park KK /jcelrep",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/27050520",
"PubMed ID": "PubMed 27050520"
} | {
"Gene/Insert name": "Signal transducer and activator of transcription 3",
"Species": "M. musculus (mouse)",
"Insert Size (bp)": "2310",
"Mutation": "",
"Entrez Gene": "Stat3",
"Promoter": "CMV enhancer and promoter",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "EcoRI (not destroyed)",
"3′ cloning site": "XhoI (not destroyed)",
"5′ sequencing primer": "CGCAAATGGGCGGTAGGCGTG",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pCAG-B19N | 59924 | Low | Expression vector for Rabies virus SAD B19 nucleoprotein gene | Ian Wickersham | Genetic Neuroengineering Group Plasmids (unpublished) | https://media.addgene.org/snapgene-media/v2.0.0/sequences/94984/40123d70-312d-4355-b3ab-a457ab224942/addgene-plasmid-59924-sequence-94984.gbk | full | LOCUS sequence-94984-e 6148 bp DNA circular SYN 26-AUG-2024
DEFINITION Expression vector for Rabies virus SAD B19 nucleoprotein gene.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6148)
TITLE Genetic Neuroengineering Group Plasmids
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 6148)
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..6148
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 4..383
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 386..661
/label=chicken beta-actin promoter
intron 662..1670
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
primer_bind 1594..1616
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 1678..1697
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
regulatory 1728..1737
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
primer_bind complement(3172..3191)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_signal 3237..3292
/label=beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(3291..3310)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
primer_bind complement(3651..3667)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3651..3667)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3664..3686)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3675..3691
/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(3699..3729)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3744..3765
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 3824..4019
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 3870..4005
/label=SV40 ori
/note="SV40 origin of replication"
polyA_signal 4025..4159
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4062..4081)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4116..4135
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4227..4244)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4398..4986)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4478..4497)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5157..6017)
/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 5780..5799
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6018..6122)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct
1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
1381 gacttccttt gtcccaaatc tggcggagcc gaaatctggg aggcgccgcc gcaccccctc
1441 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
1501 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg
1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
1621 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
1681 acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attcgccgcc accatggatg
1741 ccgacaagat tgtattcaaa gtcaataatc aggtggtctc tttgaagcct gagattatcg
1801 tggatcaata tgagtacaag taccctgcca tcaaagattt gaaaaagccc tgtataaccc
1861 taggaaaggc tcccgattta aataaagcat acaagtcagt tttgtcaggc atgagcgccg
1921 ccaaacttaa tcctgacgat gtatgttcct atttggcagc ggcaatgcag ttttttgagg
1981 ggacatgtcc ggaagactgg accagctatg gaattgtgat tgcacgaaaa ggagataaga
2041 tcaccccagg ttctctggtg gagataaaac gtactgatgt agaagggaat tgggctctga
2101 caggaggcat ggaactgaca agagacccca ctgtccctga gcatgcgtcc ttagtcggtc
2161 ttctcttgag tctgtatagg ttgagcaaaa tatccgggca aaacactggt aactataaga
2221 caaacattgc agacaggata gagcagattt ttgagacagc cccttttgtt aaaatcgtgg
2281 aacaccatac tctaatgaca actcacaaaa tgtgtgctaa ttggagtact ataccaaact
2341 tcagattttt ggccggaacc tatgacatgt ttttctcccg gattgagcat ctatattcag
2401 caatcagagt gggcacagtt gtcactgctt atgaagactg ttcaggactg gtatcattta
2461 ctgggttcat aaaacaaatc aatctcaccg ctagagaggc aatactatat ttcttccaca
2521 agaactttga ggaagagata agaagaatgt ttgagccagg gcaggagaca gctgttcctc
2581 actcttattt catccacttc cgttcactag gcttgagtgg gaaatctcct tattcatcaa
2641 atgctgttgg tcacgtgttc aatctcattc actttgtagg atgctatatg ggtcaagtca
2701 gatccctaaa tgcaacggtt attgctgcat gtgctcctca tgaaatgtct gttctagggg
2761 gctatctggg agaggaattc ttcgggaaag ggacatttga aagaagattc ttcagagatg
2821 agaaagaact tcaagaatac gaggcggctg aactgacaaa gactgacgta gcactggcag
2881 atgatggaac tgtcaactct gacgacgagg actacttttc aggtgaaacc agaagtccgg
2941 aggctgttta tactcgaatc atgatgaatg gaggtcgact aaagagatct cacatacgga
3001 gatatgtctc agtcagttcc aatcatcaag cccgtccaaa ctcattcgcc gagtttctaa
3061 acaagacata ttcgagtgac tcataagcgg ccgcactcct caggtgcagg ctgcctatca
3121 gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa taccactgag atctttttcc
3181 ctctgccaaa aattatgggg acatcatgaa gccccttgag catctgactt ctggctaata
3241 aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct cactcggaag
3301 gacatatggg agggcaaatc atttaaaaca tcagaatgag tatttggttt agagtttggc
3361 aacatatgcc atatgctggc tgccatgaac aaaggtggct ataaagaggt catcagtata
3421 tgaaacagcc ccctgctgtc cattccttat tccatagaaa agccttgact tgaggttaga
3481 ttttttttat attttgtttt gtgttatttt tttctttaac atccctaaaa ttttccttac
3541 atgttttact agccagattt ttcctcctct cctgactact cccagtcata gctgtccctc
3601 ttctcttatg aagatccctc gacctgcagc ccaagcttgg cgtaatcatg gtcatagctg
3661 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
3721 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
3781 ctgcccgctt tccagtcggg aaacctgtcg tgccagcgga tccgcatctc aattagtcag
3841 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
3901 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
3961 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
4021 agctaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
4081 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
4141 atcttatcat gtctggatcc gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
4201 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
4261 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
4321 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
4381 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
4441 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
4501 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
4561 tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg
4621 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
4681 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
4741 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
4801 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
4861 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
4921 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
4981 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
5041 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
5101 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
5161 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
5221 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
5281 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
5341 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
5401 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
5461 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
5521 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
5581 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
5641 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
5701 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
5761 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
5821 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
5881 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
5941 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
6001 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
6061 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
6121 cgcacatttc cccgaaaagt gccacctg
//
| {
"Vector backbone": "pCAGEN",
"Backbone size w/o insert (bp)": "4795",
"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": "",
"Publication": "",
"DOI": "",
"Authors": "",
"PubMed Link": "",
"PubMed ID": ""
} | {
"Gene/Insert name": "Rabies virus nucleoprotein",
"Species": "Rabies virus",
"Insert Size (bp)": "1353",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "CAG",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Unknown",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "CTCTAGAGCCTCTGCTAACc",
"3′ sequencing primer": "CAGAAGTCAGATGCTCAAG"
} | {
"Gene/Insert name": "",
"Species": "",
"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": ""
} |
pCPH2A.L-GS3 | 204756 | Expression of Cas9 and human H2A.L | Yuichiro Miyaoka | Kato-Inui et al PLoS One. 2024 May 13;19(5):e0288578. doi: 10.1371/journal.pone.0288578. eCollection 2024. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/407626/213be6b6-c933-4314-9070-2c04946d345a/addgene-plasmid-204756-sequence-407626.gbk | full | LOCUS sequence-407626- 9177 bp DNA circular SYN 27-AUG-2024
DEFINITION Expression of Cas9 and human H2A.L.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9177)
AUTHORS Kato-Inui T, Takahashi G, Ono T, Miyaoka Y
TITLE Fusion of histone variants to Cas9 suppresses non-homologous end
joining.
JOURNAL PLoS One. 2024 May 13;19(5):e0288578. doi:
10.1371/journal.pone.0288578. eCollection 2024.
PUBMED 38739603
REFERENCE 2 (bases 1 to 9177)
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: "PLoS One.";
date: "2024-05-13"; volume: "19(5):e0288578. doi"; pages: "
10.1371/journal.pone.0288578. eCollection 2024"
FEATURES Location/Qualifiers
source 1..9177
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 12..297
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer;
contains an 18-bp deletion relative to the standard CMV
enhancer"
promoter 299..576
/label=chicken beta-actin promoter
intron 577..804
/label=hybrid intron
/note="hybrid between chicken beta-actin (CBA) and minute
virus of mice (MMV) introns (Gray et al., 2011)"
regulatory 816..825
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 825..890
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
CDS 897..917
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 1374..5474
/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 1374..5474
/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 5475..5522
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
CDS 5538..5591
/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 5592..6191
/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(5592..5611)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 6088..6108
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
primer_bind complement(6219..6236)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 6225..6432
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
repeat_region 6441..6581
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 6441..6570
/label=AAV2 ITR
rep_origin 6656..7111
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(6743..6762)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 6953..6974
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(7128..7147)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 7247..7269
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(7307..7325)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 7393..7497
/gene="bla"
/label=AmpR promoter
CDS 7498..8358
/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(7716..7735)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 8529..9117
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 9018..9037
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 ctagaggtac ccgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
61 ccccgcccat tgacgtcaat agtaacgcca atagggactt tccattgacg tcaatgggtg
121 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
181 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attgtgccca gtacatgacc
241 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtc
301 gaggtgagcc ccacgttctg cttcactctc cccatctccc ccccctcccc acccccaatt
361 ttgtatttat ttatttttta attattttgt gcagcgatgg gggcgggggg gggggggggg
421 cgcgcgccag gcggggcggg gcggggcgag gggcggggcg gggcgaggcg gagaggtgcg
481 gcggcagcca atcagagcgg cgcgctccga aagtttcctt ttatggcgag gcggcggcgg
541 cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgcg ctgccttcgc
601 cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta
661 ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta gctgagcaag
721 aggtaagggt ttaagggatg gttggttggt ggggtattaa tgtttaatta cctggagcac
781 ctgcctgaaa tcactttttt tcaggttgga ccggtgccac catggactat aaggaccacg
841 acggagacta caaggatcat gatattgatt acaaagacga tgacgataag atggccccaa
901 agaagaagcg gaaggtcggg atccacggag tccctgcagc ctcgggacgc ggcaagcagg
961 gaggcaaagc tcgcgccaaa gccaagagcc gctcttctcg tgccggtctc cagttccccg
1021 tgggccgagt gcaccgactg ctccgcaagg gcaactatgc tgagcgggtc ggggccggcg
1081 cgccggtgta cctggcggcg gtgctggagt acctgactgc cgagatcctg gagctggcgg
1141 gcaacgccgc ccgcgacaac aagaagaccc gcattatccc gcgccacttg cagctggcca
1201 tccgcaacga cgaggagctc aacaagctgc tgggcagagt aaccatcgct cagggtggtg
1261 tcctgcccaa catccaggct gtgctactgc ccaagaagac cgagagtcac cacaaggcca
1321 aaggcaaagg tggcggtgga tccggtggcg gtggaagtgg tggcggtgga tctgacaaga
1381 agtacagcat cggcctggac atcggcacca actctgtggg ctgggccgtg atcaccgacg
1441 agtacaaggt gcccagcaag aaattcaagg tgctgggcaa caccgaccgg cacagcatca
1501 agaagaacct gatcggagcc ctgctgttcg acagcggcga aacagccgag gccacccggc
1561 tgaagagaac cgccagaaga agatacacca gacggaagaa ccggatctgc tatctgcaag
1621 agatcttcag caacgagatg gccaaggtgg acgacagctt cttccacaga ctggaagagt
1681 ccttcctggt ggaagaggat aagaagcacg agcggcaccc catcttcggc aacatcgtgg
1741 acgaggtggc ctaccacgag aagtacccca ccatctacca cctgagaaag aaactggtgg
1801 acagcaccga caaggccgac ctgcggctga tctatctggc cctggcccac atgatcaagt
1861 tccggggcca cttcctgatc gagggcgacc tgaaccccga caacagcgac gtggacaagc
1921 tgttcatcca gctggtgcag acctacaacc agctgttcga ggaaaacccc atcaacgcca
1981 gcggcgtgga cgccaaggcc atcctgtctg ccagactgag caagagcaga cggctggaaa
2041 atctgatcgc ccagctgccc ggcgagaaga agaatggcct gttcggaaac ctgattgccc
2101 tgagcctggg cctgaccccc aacttcaaga gcaacttcga cctggccgag gatgccaaac
2161 tgcagctgag caaggacacc tacgacgacg acctggacaa cctgctggcc cagatcggcg
2221 accagtacgc cgacctgttt ctggccgcca agaacctgtc cgacgccatc ctgctgagcg
2281 acatcctgag agtgaacacc gagatcacca aggcccccct gagcgcctct atgatcaaga
2341 gatacgacga gcaccaccag gacctgaccc tgctgaaagc tctcgtgcgg cagcagctgc
2401 ctgagaagta caaagagatt ttcttcgacc agagcaagaa cggctacgcc ggctacattg
2461 acggcggagc cagccaggaa gagttctaca agttcatcaa gcccatcctg gaaaagatgg
2521 acggcaccga ggaactgctc gtgaagctga acagagagga cctgctgcgg aagcagcgga
2581 ccttcgacaa cggcagcatc ccccaccaga tccacctggg agagctgcac gccattctgc
2641 ggcggcagga agatttttac ccattcctga aggacaaccg ggaaaagatc gagaagatcc
2701 tgaccttccg catcccctac tacgtgggcc ctctggccag gggaaacagc agattcgcct
2761 ggatgaccag aaagagcgag gaaaccatca ccccctggaa cttcgaggaa gtggtggaca
2821 agggcgcttc cgcccagagc ttcatcgagc ggatgaccaa cttcgataag aacctgccca
2881 acgagaaggt gctgcccaag cacagcctgc tgtacgagta cttcaccgtg tataacgagc
2941 tgaccaaagt gaaatacgtg accgagggaa tgagaaagcc cgccttcctg agcggcgagc
3001 agaaaaaggc catcgtggac ctgctgttca agaccaaccg gaaagtgacc gtgaagcagc
3061 tgaaagagga ctacttcaag aaaatcgagt gcttcgactc cgtggaaatc tccggcgtgg
3121 aagatcggtt caacgcctcc ctgggcacat accacgatct gctgaaaatt atcaaggaca
3181 aggacttcct ggacaatgag gaaaacgagg acattctgga agatatcgtg ctgaccctga
3241 cactgtttga ggacagagag atgatcgagg aacggctgaa aacctatgcc cacctgttcg
3301 acgacaaagt gatgaagcag ctgaagcggc ggagatacac cggctggggc aggctgagcc
3361 ggaagctgat caacggcatc cgggacaagc agtccggcaa gacaatcctg gatttcctga
3421 agtccgacgg cttcgccaac agaaacttca tgcagctgat ccacgacgac agcctgacct
3481 ttaaagagga catccagaaa gcccaggtgt ccggccaggg cgatagcctg cacgagcaca
3541 ttgccaatct ggccggcagc cccgccatta agaagggcat cctgcagaca gtgaaggtgg
3601 tggacgagct cgtgaaagtg atgggccggc acaagcccga gaacatcgtg atcgaaatgg
3661 ccagagagaa ccagaccacc cagaagggac agaagaacag ccgcgagaga atgaagcgga
3721 tcgaagaggg catcaaagag ctgggcagcc agatcctgaa agaacacccc gtggaaaaca
3781 cccagctgca gaacgagaag ctgtacctgt actacctgca gaatgggcgg gatatgtacg
3841 tggaccagga actggacatc aaccggctgt ccgactacga tgtggaccat atcgtgcctc
3901 agagctttct gaaggacgac tccatcgaca acaaggtgct gaccagaagc gacaagaacc
3961 ggggcaagag cgacaacgtg ccctccgaag aggtcgtgaa gaagatgaag aactactggc
4021 ggcagctgct gaacgccaag ctgattaccc agagaaagtt cgacaatctg accaaggccg
4081 agagaggcgg cctgagcgaa ctggataagg ccggcttcat caagagacag ctggtggaaa
4141 cccggcagat cacaaagcac gtggcacaga tcctggactc ccggatgaac actaagtacg
4201 acgagaatga caagctgatc cgggaagtga aagtgatcac cctgaagtcc aagctggtgt
4261 ccgatttccg gaaggatttc cagttttaca aagtgcgcga gatcaacaac taccaccacg
4321 cccacgacgc ctacctgaac gccgtcgtgg gaaccgccct gatcaaaaag taccctaagc
4381 tggaaagcga gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag atgatcgcca
4441 agagcgagca ggaaatcggc aaggctaccg ccaagtactt cttctacagc aacatcatga
4501 actttttcaa gaccgagatt accctggcca acggcgagat ccggaagcgg cctctgatcg
4561 agacaaacgg cgaaaccggg gagatcgtgt gggataaggg ccgggatttt gccaccgtgc
4621 ggaaagtgct gagcatgccc caagtgaata tcgtgaaaaa gaccgaggtg cagacaggcg
4681 gcttcagcaa agagtctatc ctgcccaaga ggaacagcga taagctgatc gccagaaaga
4741 aggactggga ccctaagaag tacggcggct tcgacagccc caccgtggcc tattctgtgc
4801 tggtggtggc caaagtggaa aagggcaagt ccaagaaact gaagagtgtg aaagagctgc
4861 tggggatcac catcatggaa agaagcagct tcgagaagaa tcccatcgac tttctggaag
4921 ccaagggcta caaagaagtg aaaaaggacc tgatcatcaa gctgcctaag tactccctgt
4981 tcgagctgga aaacggccgg aagagaatgc tggcctctgc cggcgaactg cagaagggaa
5041 acgaactggc cctgccctcc aaatatgtga acttcctgta cctggccagc cactatgaga
5101 agctgaaggg ctcccccgag gataatgagc agaaacagct gtttgtggaa cagcacaagc
5161 actacctgga cgagatcatc gagcagatca gcgagttctc caagagagtg atcctggccg
5221 acgctaatct ggacaaagtg ctgtccgcct acaacaagca ccgggataag cccatcagag
5281 agcaggccga gaatatcatc cacctgttta ccctgaccaa tctgggagcc cctgccgcct
5341 tcaagtactt tgacaccacc atcgaccgga agaggtacac cagcaccaaa gaggtgctgg
5401 acgccaccct gatccaccag agcatcaccg gcctgtacga gacacggatc gacctgtctc
5461 agctgggagg cgacaaaagg ccggcggcca cgaaaaaggc cggccaggca aaaaagaaaa
5521 aggaattcgg cagtggagag ggcagaggaa gtctgctaac atgcggtgac gtcgaggaga
5581 atcctggccc aatgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc
5641 ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg
5701 tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg
5761 tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga
5821 ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg
5881 agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc
5941 ggcccaagga gcccgcgtgg ttcctggcca ccgtcggagt ctcgcccgac caccagggca
6001 agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc
6061 ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca
6121 ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc
6181 ccggtgcctg agaattctaa ctagagctcg ctgatcagcc tcgactgtgc cttctagttg
6241 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
6301 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
6361 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag agaatagcag
6421 gcatgctggg gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg
6481 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg
6541 ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt
6601 tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg
6661 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
6721 cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
6781 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
6841 ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg
6901 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
6961 ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga tttataaggg
7021 attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
7081 aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct
7141 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
7201 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
7261 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
7321 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
7381 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
7441 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
7501 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
7561 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
7621 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
7681 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
7741 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
7801 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
7861 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
7921 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
7981 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
8041 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
8101 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
8161 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tggaagccgc
8221 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
8281 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
8341 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
8401 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
8461 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
8521 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
8581 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
8641 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
8701 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
8761 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
8821 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
8881 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
8941 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
9001 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
9061 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
9121 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatg
//
| {
"Vector backbone": "pSpCas9(BB)-2A-Puro(PX459)V2.0(#62988)",
"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": "Fusion of histone variants to Cas9 suppresses non-homologous end joining",
"Publication": "PLoS One. 2024 May 13;19(5):e0288578. doi: 10.1371/journal.pone.0288578. eCollection 2024.",
"DOI": "10.1371/journal.pone.0288578.",
"Authors": "Kato-Inui T, Takahashi G, Ono T, Miyaoka Y /journalpone",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/38739603",
"PubMed ID": "PubMed 38739603"
} | {
"Gene/Insert name": "H2AL3",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "H2AC6",
"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": ""
} |
|
CLXSN 5myc-CDC6-mut | 109333 | express tagged Cdc6 mutant | Jean Cook | Matson et al Elife. 2017 Nov 17;6. doi: 10.7554/eLife.30473. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/213795/34556b8a-f9c6-4b7e-a222-5606ea755cb8/addgene-plasmid-109333-sequence-213795.gbk | full | LOCUS sequence-213795- 9401 bp DNA circular SYN 25-AUG-2024
DEFINITION express tagged Cdc6 mutant.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9401)
AUTHORS Matson JP, Dumitru R, Coryell P, Baxley RM, Chen W, Twaroski K,
Webber BR, Tolar J, Bielinsky AK, Purvis JE, Cook JG
TITLE Rapid DNA replication origin licensing protects stem cell
pluripotency.
JOURNAL Elife. 2017 Nov 17;6. doi: 10.7554/eLife.30473.
PUBMED 29148972
REFERENCE 2 (bases 1 to 9401)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Elife.";
date: "2017-11-17"; volume: "6. doi"; pages: " 10.7554/eLife.30473"
FEATURES Location/Qualifiers
source 1..9401
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 7..386
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 387..590
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 541..561
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
LTR 591..766
/label=5' LTR (truncated)
/note="truncated long terminal repeat from Moloney murine
sarcoma virus"
misc_feature 829..1028
/label=MMLV Psi
/note="packaging signal of Moloney murine leukemia virus
(MMLV)"
CDS 1229..1645
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
primer_bind 1575..1597
/label=pLXSN 5'
/note="Murine stem cell virus, forward primer. Also called
MSCV"
primer_bind 1613..1629
/label=pBABE 5'
/note="Psi packaging signal, forward primer for pBABE
vectors"
primer_bind 1707..1726
/label=T7
/note="T7 promoter, forward primer"
promoter 1707..1725
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 1793..1822
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 1826..1855
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 1865..1894
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 1898..1927
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
primer_bind complement(3639..3659)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 3644..3973
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3824..3959
/label=SV40 ori
/note="SV40 origin of replication"
CDS 4033..4827
/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(4087..4106)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4697..4716
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
LTR 4887..5480
/note="long terminal repeat from Moloney murine leukemia
virus"
intron 5825..5890
/label=small t intron
/note="SV40 (simian virus 40) small t antigen intron"
CDS 6020..6040
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
polyA_signal 6312..6446
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(6349..6368)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 6403..6422
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 6904..7038
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(7103..7120)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(7274..7862)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(7354..7373)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(8033..8893)
/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 8656..8675
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(8894..8998)
/gene="bla"
/label=AmpR promoter
primer_bind 9066..9084
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind 9244..9263
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
61 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
121 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
181 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
241 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
301 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
361 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
421 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
481 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
541 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct caataaaaga
601 gcccacaacc cctcactcgg cgcgccagtc ttccgataga ctgcgtcgcc cgggtacccg
661 tattcccaat aaagcctctt gctgtttgca tccgaatcgt ggtctcgctg ttccttggga
721 gggtctcctc tgagtgattg actacccacg acgggggtct ttcatttggg ggctcgtccg
781 ggatttggag acccctgccc agggaccacc gacccaccac cgggaggtaa gctggccagc
841 aacttatctg tgtctgtccg attgtctagt gtctatgttt gatgttatgc gcctgcgtct
901 gtactagtta gctaactagc tctgtatctg gcggacccgt ggtggaactg acgagttctg
961 aacacccggc cgcaaccctg ggagacgtcc cagggacttt gggggccgtt tttgtggccc
1021 gacctgagga agggagtcga tgtggaatcc gaccccgtca ggatatgtgg ttctggtagg
1081 agacgagaac ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg tttggaaccg
1141 aagccgcgcg tcttgtctgc tgcagcgctg cagcatcgtt ctgtgttgtc tctgtctgac
1201 tgtgtttctg tatttgtctg aaaattaggg ccagactgtt accactccct taagtttgac
1261 cttaggtcac tggaaagatg tcgagcggat cgctcacaac cagtcggtag atgtcaagaa
1321 gagacgttgg gttaccttct gctctgcaga atggccaacc tttaacgtcg gatggccgcg
1381 agacggcacc tttaaccgag acctcatcac ccaggttaag atcaaggtct tttcacctgg
1441 cccgcatgga cacccagacc aggtccccta catcgtgacc tgggaagcct tggcttttga
1501 cccccctccc tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc ttcctccatc
1561 cgccccgtct ctcccccttg aacctcctcg ttcgaccccg cctcgatcct ccctttatcc
1621 agccctcact ccttctctag gcgccggaat tcgttaactc gactctggct aactagagaa
1681 cccactgctt actggcttat cgaaattaat acgactcact atagggagac ccaagcttgg
1741 taccaccatg gaacagaagc taatctctga ggaagttttg ctggggatct tggagcagaa
1801 attgataagc gaggaagacc ttggagaaca aaagctgatt tcggaagagg atctggggat
1861 cttggagcag aaattgataa gcgaggaaga ccttggagaa caaaagctga tttcggaaga
1921 ggatctgggg atccgaatgc ctcaaacccg atcccaggca caggctacaa tcagttttcc
1981 aaaaaggaag ctgtctcggg cattgaacaa agctaaaaac tccagtgatg ccaaactaga
2041 accaacaaat gtccaaaccg taacctgttc tcctcgtgta aaagccctgc ctctcagccc
2101 cgccaaacgt gccggcgatg acaacctatg caacactccc catttacctc cttgttctcc
2161 accaaagcaa ggcaaggcgg ccgcaggtcc ccctcactca catacactta agggacgaag
2221 attggtattt gacaatcagc tgacaattaa gtctcctagc aaaagagaac tagccaaagt
2281 tcaccaaaac aaaatacttt cttcagttag aaaaagtcaa gagatcacaa caaattctga
2341 gcagagatgt ccactgaaga aagaatctgc atgtgtgaga ctattcaagc aagaaggcac
2401 ttgctaccag caagcaaagc tggtcctgaa cacagctgtc ccagatcggc tgcctgccag
2461 ggaaagggag atggatgtca tcaggaattt cttgagggaa cacatctgtg ggaaaaaagc
2521 tggaagcctt tacctttctg gtgctcctgg aactggaaaa actgcctgct taagccggat
2581 tctgcaagac ctcaagaagg aactgaaagg ctttaaaact atcatgctga attgcatgtc
2641 cttgaggact gcccaggctg tattcccagc tattgctcag gagatttgtc aggaagaggt
2701 atccaggcca gctgggaagg acatgatgag gaaattggaa aaacatatga ctgcagagaa
2761 gggccccatg attgtgttgg tattggacga gatggatcaa ctggacagca aaggccagga
2821 tgtattgtac acgctatttg aatggccatg gctaagcaat tctcacttgg tgctgattgg
2881 tattgctaat accctggatc tcacagatag aattctacct aggcttcaag ctagagaaaa
2941 atgtaagcca cagctgttga acttcccacc ttataccaga aatcagatag tcactatttt
3001 gcaagatcga cttaatcagg tatctagaga tcaggttctg gacaatgctg cagttcaatt
3061 ctgtgcccgc aaagtctctg ctgtttcagg agatgttcgc aaagcactgg atgtttgcag
3121 gagagctatt gaaattgtag agtcagatgt caaaagccag actattctca aaccactgtc
3181 tgaatgtaaa tcaccttctg agcctctgat tcccaagagg gttggtctta ttcacatatc
3241 ccaagtcatc tcagaagttg atggtaacag gatgaccctg agccaagaag gagcacaaga
3301 ttccttccct cttcagcaga agatcttggt ttgctctttg atgctcttga tcaggcagtt
3361 gaaaatcaaa gaggtcactc tggggaagtt atatgaagcc tacagtaaag tctgtcgcaa
3421 acagcaggtg gcggctgtgg accagtcaga gtgtttgtca ctttcagggc tcttggaagc
3481 caggggcatt ttaggattaa agagaaacaa ggaaacccgt ttgacaaagg tgtttttcaa
3541 gattgaagag aaagaaatag aacatgctct gaaagataaa gctttaattg gaaatatctt
3601 agctactgga ttgccttaag cgtcgaggat ccggctgtgg aatgtgtgtc agttagggtg
3661 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
3721 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
3781 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
3841 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
3901 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
3961 aggcttttgc aaaaagcttg ggctgcaggt cgaggcggat ctgatcaaga gacaggatga
4021 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
4081 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
4141 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
4201 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
4261 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
4321 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
4381 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
4441 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
4501 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
4561 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
4621 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
4681 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
4741 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
4801 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgataaa ataaaagatt
4861 ttatttagtc tccagaaaaa ggggggaatg aaagacccca cctgtaggtt tggcaagcta
4921 gcttaagtaa cgccattttg caaggcatgg aaaaatacat aactgagaat agagaagttc
4981 agatcaaggt caggaacaga tggaacagct gaatatgggc caaacaggat atctgtggta
5041 agcagttcct gccccggctc agggccaaga acagatggaa cagctgaata tgggccaaac
5101 aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggtccccag
5161 atgcggtcca gccctcagca gtttctagag aaccatcaga tgtttccagg gtgccccaag
5221 gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt
5281 tcgcgcgctt ctgctccccg agctcaataa aagagcccac aacccctcac tcggggcgcc
5341 agtcctccga ttgactgagt cgcccgggta cccgtgtatc caataaaccc tcttgcagtt
5401 gcatccgact tgtggtctcg ctgttccttg ggagggtctc ctctgagtga ttgactaccc
5461 gtcagcgggg gtctttcatt tgggggctcg tccgggatcg ggagacccct gcccagggac
5521 caccgaccca ccaccgggag gtaagctggc tgcctcgcgc gtttcggtga tgacggtgaa
5581 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
5641 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg
5701 acccagtcac gtagcgatag cggagtgtat ctagctaggt agctagagga tctttgtgaa
5761 ggaaccttac ttctgtggtg tgacataatt ggacaaacta cctacagaga tttaaagctc
5821 taaggtaaat ataaaatttt taagtgtata atgtgttaaa ctactgattc taattgtttg
5881 tgtattttag attccaacct atggaactga tgaatgggag cagtggtgga atgcctttaa
5941 tgaggaaaac ctgttttgct cagaagaaat gccatctagt gatgatgagg ctactgctga
6001 ctctcaacat tctactcctc caaaaaagaa gagaaaggta gaagacccca aggactttcc
6061 ttcagaattg ctaagttttt tgagtcatgc tgtgtttagt aatagaactc ttgcttgctt
6121 tgctatttac accacaaagg aaaaagctgc actgctatac aagaaaatta tggaaaaata
6181 tttgatgtat agtgccttga ctagagatca taatcagcca taccacattt gtagaggttt
6241 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
6301 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
6361 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
6421 tcaatgtatc ttatcatgtc tggatcagct tcagaagatg gcggagggcc tccaacacag
6481 taattttcct cccgactctt aaaatagaaa atgtcaagtc agttaagcag gaagtgacta
6541 actgaccgca gctggccgtg cgacatcctc ttttaattag ttgctaggca actgccctcc
6601 agagggcagt gtggttttgc aagaggaagc aaaaagcctc tccacccagg cctagaatgt
6661 ttccacccaa tcattactat gacaacagct gtttttttta gtattaagca gaggccgggg
6721 gcccctggcc tccgcttact ctggagaaaa agaagagagg cattgtagag gcttccagag
6781 gcaacttgtc aaaacaggac tgcttctatt tctgtcacac tgtctggccc tgtcacaagg
6841 tccagcacct ccataccccc tttaataagc agtttgggaa cgggtgcggg tcttactccg
6901 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
6961 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
7021 ggaggctttt ttggaggctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
7081 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
7141 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
7201 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
7261 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
7321 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
7381 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
7441 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
7501 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
7561 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
7621 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
7681 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
7741 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
7801 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
7861 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
7921 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
7981 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
8041 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
8101 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
8161 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
8221 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
8281 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
8341 ccattgctgc aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
8401 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
8461 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
8521 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
8581 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
8641 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
8701 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
8761 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
8821 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
8881 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
8941 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
9001 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
9061 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
9121 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
9181 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
9241 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca
9301 gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt
9361 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca g
//
| {
"Vector backbone": "pCLXSN",
"Backbone size w/o insert (bp)": "",
"Vector type": "Mammalian Expression, Retroviral",
"Selectable markers": "Neomycin (select with G418)"
} | {
"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": "Rapid DNA replication origin licensing protects stem cell pluripotency",
"Publication": "Elife. 2017 Nov 17;6. doi: 10.7554/eLife.30473.",
"DOI": "10.7554/eLife.30473.",
"Authors": "Matson JP, Dumitru R, Coryell P, Baxley RM, Chen W, Twaroski K, Webber BR, Tolar J, Bielinsky AK, Purvis JE, Cook JG /eLife",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/29148972",
"PubMed ID": "PubMed 29148972"
} | {
"Gene/Insert name": "CDC6",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "R56A, L59A, K81A, E82A, N83A",
"Entrez Gene": "CDC6",
"Promoter": "",
"Tag / Fusion Protein": "5x Myc (N terminal on insert)"
} | {
"Cloning method": "Unknown",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "psi packaging",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"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": ""
} |
|
pBAD-bARGSer-AxeTxe | 192473 | Second-generation bacterial acoustic reporter gene derived from Serratia | Mikhail Shapiro | Hurt et al Nat Biotechnol. 2023 Jan 2. doi: 10.1038/s41587-022-01581-y. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/376957/35f1298a-c32f-4566-9d83-8487148c8757/addgene-plasmid-192473-sequence-376957.gbk | full | LOCUS sequence-376957- 23202 bp DNA circular SYN 25-AUG-2024
DEFINITION Second-generation bacterial acoustic reporter gene derived from
Serratia.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 23202)
AUTHORS Hurt RC, Buss MT, Duan M, Wong K, You MY, Sawyer DP, Swift MB, Dutka
P, Barturen-Larrea P, Mittelstein DR, Jin Z, Abedi MH, Farhadi A,
Deshpande R, Shapiro MG
TITLE Genomically mined acoustic reporter genes for real-time in vivo
monitoring of tumors and tumor-homing bacteria.
JOURNAL Nat Biotechnol. 2023 Jan 2. doi: 10.1038/s41587-022-01581-y.
PUBMED 36593411
REFERENCE 2 (bases 1 to 23202)
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. 2023 Jan 2. doi: 10.1038/s41587-022-01581-y."
FEATURES Location/Qualifiers
source 1..23202
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin 1..546
/direction=RIGHT
/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 447..466
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 663..680
/label=L4440
/note="L4440 vector, forward primer"
promoter 1072..1174
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
CDS 1175..1834
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWCEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind complement(1241..1260)
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
rep_origin complement(2390..2845)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2527..2548)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2739..2758
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter complement(3303..3394)
/gene="bla"
/label=AmpR promoter
terminator complement(3413..3440)
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator complement(3532..3618)
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
primer_bind 3771..3788
/label=pBAD Reverse
/note="For vectors with E. coli araBAD promoter, reverse
primer"
CDS 4895..4906
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
CDS complement(11533..11544)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
promoter complement(20531..20815)
/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 complement(20589..20608)
/label=pBAD Forward
/note="For vectors with E. coli araBAD promoter, forward
primer"
CDS 20842..21720
/codon_start=1
/gene="araC"
/product="L-arabinose regulatory protein"
/label=araC
/translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
ILNLTIRGQGVVKNQGREFVCRPGDILLFPQGEIHHYGRHPEAREWYHQWVYFRPRAYW
HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
EKVNDVAVKLS"
primer_bind complement(21900..21919)
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
ORIGIN
1 ttgagatcgt tttggtctgc gcgtaatctc ttgctctgaa aacgaaaaaa ccgccttgca
61 gggcggtttt tcgaaggttc tctgagctac caactctttg aaccgaggta actggcttgg
121 aggagcgcag tcaccaaaac ttgtcctttc agtttagcct taaccggcgc atgacttcaa
181 gactaactcc tctaaatcaa ttaccagtgg ctgctgccag tggtgctttt gcatgtcttt
241 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcggac tgaacggggg
301 gttcgtgcat acagtccagc ttggagcgaa ctgcctaccc ggaactgagt gtcaggcgtg
361 gaatgagaca aacgcggcca taacagcgga atgacaccgg taaaccgaaa ggcaggaaca
421 ggagagcgca cgagggagcc gccaggggga aacgcctggt atctttatag tcctgtcggg
481 tttcgccacc actgatttga gcgtcagatt tcgtgatgct tgtcaggggg gcggagccta
541 tggaaaaacg gctttgccgc ggccctctca cttccctgtt aagtatcttc ctggcatctt
601 ccaggaaatc tccgccccgt tcgtaagcca tttccgctcg ccgcagtcga acgaccgagc
661 gtagcgagtc agtgagcgag gaagcggaat atatcctgta tcacatattc tgctgacgca
721 ccggtgcagc cttttttctc ctgccacatg aagcacttca ctgacaccct catcagtgcc
781 aacatagtaa gccagtatac actccgctag cgctgatgtc cggcggtgct tttgccgtta
841 cgcaccaccc cgtcagtagc tgaacaggag ggacagctga tagaaacaga agccactgga
901 gcacctcaaa aacaccatca tacactaaat cagtaagttg gcagcatcac ccgacgcact
961 ttgcgccgaa taaatacctg tgacggaaga tcacttcgca gaataaataa atcctggtgt
1021 ccctgttgat accgggaagc cctgggccaa cttttggcga aaatgagacg ttgatcggca
1081 cgtaagaggt tccaactttc accataatga aataagatca ctaccgggcg tattttttga
1141 gttatcgaga ttttcaggag ctaaggaagc taaaatggag aaaaaaatca ctggatatac
1201 caccgttgat atatcccaat ggcatcgtaa agaacatttt gaggcatttc agtcagttgc
1261 tcaatgtacc tataaccaga ccgttcagct ggatattacg gcctttttaa agaccgtaaa
1321 gaaaaataag cacaagtttt atccggcctt tattcacatt cttgcccgcc tgatgaatgc
1381 tcatccggaa ttccgtatgg caatgaaaga cggtgagctg gtgatatggg atagtgttca
1441 cccttgttac accgttttcc atgagcaaac tgaaacgttt tcatcgctct ggtgtgaata
1501 ccacgacgat ttccggcagt ttctacacat atattcgcaa gatgtggcgt gttacggtga
1561 aaacctggcc tatttcccta aagggtttat tgagaatatg tttttcgtct cagccaatcc
1621 ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat atggacaact tcttcgcccc
1681 cgttttcacc atgggcaaat attatacgca aggcgacaag gtgctgatgc cgctggcgat
1741 tcaggttcat catgccgttt gtgatggctt ccatgtcggc agaatgctta atgaattaca
1801 acagtactgc gatgagtggc agggcggggc gtaatttttt taaggcagtt attggtgccc
1861 ttaaacgcct ggttgctacg cctgaataag tgataataag cggatgaatg gcagaaattc
1921 gaaagcaaat tcgacccggt cgtcggttca gggcagggtc gttaaatagc cgcttatgtc
1981 tattgctggt ttaccggttt attgactacc ggaagcagtg tgaccgtgtg cttctcaaat
2041 gcctgactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
2101 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
2161 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
2221 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
2281 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
2341 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattgtaaac
2401 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
2461 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agcccgagat agggttgagt
2521 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
2581 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccaa atcaagtttt
2641 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
2701 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
2761 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
2821 cttaatgcgc cgctacaggg cgcgtaaatc aatctaaagt atatatgagt aaacttggtc
2881 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
2941 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
3001 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
3061 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
3121 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
3181 gctgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
3241 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
3301 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
3361 tacatatttg aatgtattta gaaaaataaa caaaagagtt tgtagaaacg caaaaaggcc
3421 atccgtcagg atggccttct gcttaatttg atgcctggca gtttatggcg ggcgtcctgc
3481 ccgccaccct ccgggccgtt gcttcgcaac gttcaaatcc gctcccggcg gatttgtcct
3541 actcaggaga gcgttcaccg acaaacaaca gataaaacga aaggcccagt ctttcgactg
3601 agcctttcgt tttatttgat gcctggcagt tccctactct cgcatgggga gaccccacac
3661 taccatcggc gctacggcgt ttcacttctg agttcggcat ggggtcaggt gggaccaccg
3721 cgctactgcc gccaggcaaa ttctgtttta tcagaccgct tctgcgttct gatttaatct
3781 gtatcaggct gaaaatcttc tctcaatgtg acggaagtcg cctatcaagt agggtttaat
3841 agtacaagct attttattaa tcagtttcga aaagaaatga agatgactcc gctcatgtat
3901 cgaaaaaaga agctgcacat ctcatcacat cacgctaacg ataaaacacc atttgcacct
3961 taaaacgagc actatcgtca taatgcccgt tttaaggtgc agcgtgtcag taattccccc
4021 cgaagaagca gagcgttgat taagctagcg acgcaactac tcgctcggct gtgatagtgt
4081 ctgtaatgtt ttcagcagct ctttattggt aaaaggcttc gccagaaaac cgcagataat
4141 gccttcaacg atagcatggc gaataaccgg atcatcgtaa tagtgataac cggaaaccag
4201 cactacacgc ggtcttcgct ctagcaaggt actggctcga cgcgccactt ccagaccgtc
4261 catatccggt aacttggcat ctaaaaacac aaaggaaaaa ttgccggtca tcagcttaac
4321 gcaagcatct tccccactgt aagcggtaat gacactcatg ccttgtgatt gaataatatg
4381 ggaaatggcc caacacatgt caggttcatc atcaactatc agaatgggtg cgttcatcat
4441 tttagagtat aggtaaggta acaataatag tggaaccttc acccagacaa ctctctgcct
4501 gtatggaacc aaggtgctgt tggataatgc tgtaacaaat ggacaaaccc aaaccagtac
4561 cttggcctac gggtgaagac gtgtggaacg gatcgaaaat tttatccaac tcgccggcgg
4621 gaatgcctac gccagtatca gaaatacgca caataacccg gtcttcaaag gtctctaccg
4681 caatacttaa cacaccgcct tccggcatcg cattaatggc attaagaaaa atattcatga
4741 acacctgttg cagtaagccc gacacacctg acaccaatat atttcctttt tgcaacggca
4801 acaggatctg aattttctgc actttggcct ggttggcgat caggtcgata gtttctttca
4861 gcaccgtgat cagattgacc tctaccattt cggtatcgag ggaagaacgg gcaaaacgga
4921 gtaaattctc aatgatggcc gaggcttttt gaatgttagt ctgtattttt cttgaacact
4981 ctttatgaaa ttcagccgta atctcgtctt ccagcaaaaa ttgcgccgct gaagaacaaa
5041 cagcaagagg attacgaatt tcatgagcga taccgcccgc catgacgccc aacgcggtca
5101 acttctgtga ttgacgtaat tgcagttcca atttccgacg ttcagccaaa tcacgaccga
5161 caactacggt gcccaccgtt tgcccatagt catccttcat tggtgaaaat acccaggaaa
5221 tcaatatgct gccaccattg ttaggcgtta ataaatgaaa ttcagtcgta tgtgattcat
5281 tgctgttctt catacgcatg aaaatcagac gcaccgcctc cttttggttg gtatcgcaga
5341 actcaaaaaa gtatcgccca tagacttcat gcgtgggtaa acctgacagc ttttccgccg
5401 ctgaattcca cgtcaatatg catccttcaa tatcggtcga aagaacaatg tcgctggcgt
5461 tttctacaac tgaagcgagg tgccgttcta ctcgatggac ttcttcagac aggcgaacct
5521 gctcggtgac atcctccatg aacagcataa tctggttaat ggggatgggt agaatagaat
5581 agtaataaat ccgaatcggt acgcctggcg cacggtaggt cattttctga ccattaacgg
5641 gtaatccgct acggaataca tcccggatat gtgcttcaag acccgtattt tccaaaatga
5701 cgctcggaaa gacgtccgat aacgggcgac caattgtatc cgggttgccg cgatgaccct
5761 tgattaaaaa attctgattg actgaaacca catgcagttt ctcatccacc attaacacgg
5821 aagaaggaat tgcatccaat agcatgttgt aaagaagttc aaagtggttt tctgactgct
5881 gttcaaaagg attatctgac atggtagcct gtcgttaaca aattagctgg cgaagttata
5941 gggaggccaa ggcccactga taacccctat atccgtcatg ttgttggcca gagtattaaa
6001 ttggtcgcga aacgcctgta acccggttcg tggcaccaga tagctcaaca ggtaagtccg
6061 ctgaccatta aacatactga tttccgcgca gtgtcggcga tacaaccccg taaacgcatt
6121 attcagtagt gcagcctgcc gttcagcgtg ttcaggtacg gcataagcga gttttcgact
6181 ccgcaaatag tccagaccac tcgcttgcgg cgtagtaacg gcattatcct cagccgacgc
6241 cagggataac cgaataccca tttcgtcaca attttccagc tcaaccaatt gcgtttgata
6301 atgtgcctca tggttgagta aatgctgtgt taccgcatct tcatctgcca acacacaacc
6361 ataacgaatc ggaataatcg tggtctgttg atgaacatgc attatctgtt ggccaaaagc
6421 caacgctgcc gcgggctcgc gactgacgtc tgcttcacaa ggcttgacca gcgccgccaa
6481 cgttgccgct ttaacaagct gcatcggctc ttcaccggcg tgcggcgaac cgtctggttc
6541 aagcgccagt tgagtatcct cggcaacaat gccgtacagt agtaagctca tgattcgtca
6601 ggttggcaaa aactgtaggc tgcccacggg cctgtcacgc gaaaactaaa accgtaagct
6661 tcatagcgtt caacaatgtc agcaacctgc tgttgaaacg cggcaacgtc ctccaccggg
6721 agtaaatacg cccaatgcaa aattttatcg tccaacaacc gacgtgaacg gaaatcacgt
6781 accaaaggat gtaattcgtt ctgcatggca gtcaaagcat gtgccaacca gtcactcaac
6841 tcagtggtta acctgcggcg taatttctgc tcttccaaat ggcggcggcc tatggcttcg
6901 ggtaggcaaa agcgaccgga gtccagacct tcggtgaata aaacgtcaac ggcctgttta
6961 cgatccagtg tcgcttccag tgcccattcc tgacagccgg ttatgcgacg tagcgacaca
7021 aatacgtcag tagcgcggct tttcatttcc tgttccaacg cgttctgact ggaaaacagc
7081 gtgccgaagg gcaaaggata gaccggacct tgggccatca ggctgtcaat aatctgcgca
7141 tgccgacaaa tacgcggcgt tagccaagtg acattctgta cattgtcttc acccttttca
7201 cccgtgaaat cggagagtgc cactgggcta atcaccgcat tcaatccggc acattgatgt
7261 gtattaatcg gatgctgatt atcaacgccc aaaattggcg gtgcggctaa atcgggcaac
7321 gtcaggccat acagataaat ggcctgctca gtctgtgctt ctgtgttcat cgtcatgcat
7381 catcgccaaa catttcggca atcacttcag caaaatcatg ctgttcaatg cgtaccgtca
7441 gcgccacagg ggtttctcct accgttgacg ggctcagaac agcctcttca gactgctgga
7501 ttgcacgacg caatgccgct aatgcggcac gattacacac cgcggccaat tctgcaccgc
7561 tggcaccatc acaacgttct gccaaatcgt cagcatgaat attgtcaccc agcggtttat
7621 tacgtaaatg caccgccaga atctgccgac gggcgtcctc gtcgggcagg cccaattcga
7681 ttatttcatc aaaacgtcct ggtcgcagca tcgcgggatc gatcaagtcc gcacgattgg
7741 tcgcccccat aacgaaaaca cctttgagtt cttccagccc atccatttca gataagaatt
7801 ggctcaacac ccgttctgcg acatgtgagt cgatagcgcc ctcgttacgg gcaggaatca
7861 cagaatcgac ttcatcaagg aaaataatgc agggcgcagc ctgacgagct ttgagaaaca
7921 gctctcggac acctttttcc gaatcaccga cataacgcga catcagcgcc ggacctttaa
7981 ctgaaatgac attaacgcca ctttcattag caacggcttt ggcgatcagc gttttcccaa
8041 cgccgggagg cccagctaac agcagacctt taggcggctt aacacccgct tgttcataga
8101 gctcagggta cttgataggc cactcgacgg attcgatcaa tcgacggcgt acgtcatcca
8161 aaccgccgac atcttcccaa cgcacatcgg gaatatcgac gaacagctca cgaatcgctg
8221 acggtgatac ttcacacaag gcggcacgaa aatcatccat catcaccgtt aattcggcca
8281 gacgatcata aggtagctcc gccgaactga aatcgatctc aggcagtaaa cggcgtaacg
8341 cactcatcgc ggcttcacgg cataacgctt ccagatcggc accgacaaaa ccgtgagtga
8401 tatcggcaag cacgttcaga tcgacatcag cattcagcgg cattcccgtg ctatgaattt
8461 cgataatctc gcggcgacct tcacggtctg gaataggaat gctgatttca cggtcaaagc
8521 gtcccggacg gcgcaacgcg ggatcaatcg cattaggtag attggtcgcg gcaatgacaa
8581 tgactttgcc tcgattcttg agaccatcca ttaatgcgag cagttgcgca acgatacgtt
8641 tttctacatc accgacgacc ttgtcgcggt gtggtgcaat agagtcaatt tcatccagaa
8701 aaatgatact tggccctttg cgtccggctt cctcaaaaat tttgcgtaag tgggcttcac
8761 tttcaccata aaacttatgc acaatttccg ggccgctgat ggtaaaaaat tgcgcatcgg
8821 tttcttgcgc aataatgcgg gcgatgaggg ttttaccaca tcccggggga ccagagagca
8881 gtacaccttt gggggcatct atacctaaac gttcaaagac ctcaggatag cgtaatggta
8941 attcgatcat ttctcgaata cgccgcacct ggtttttaag gccgccaaca tcctcatacg
9001 acaaacgttg tgtgctatga ctcgaattcc cgacagcatt tgatttgcca atcaccagtg
9061 tcgtggtcgg atcaatcagt accgcaccat caggtatggt gctttcgact ttaaaatcag
9121 cagaacgact gccaaacagg tgcgcgcgca gcagatcgcc tttctgtacg ggcaacccat
9181 cgagcaggct accgatataa tacaggtcac gttggttggg cgtaatcgtg gtgggaatga
9241 ggacgatctg tgtggcgtgt ttgcaggaag cgggttcaat ctgcactttt tcatctaaac
9301 tcaggctggt attacgccgc gtcaaaccat ccagttgtac gatgccttta ttccgcatat
9361 cgggataagt cggcatcaat ttagcaatcc cggtgccttt tttactggtt aactgcacta
9421 tgtcaccgac agccagattt aacctggcca tatcggccgg atctaagcga gcataaccgc
9481 gcccagcatc ctttggcaga gcctcggcga ccttgagctt gagttcttca gacatgactg
9541 cctcattttc ctaatttgac ttccaaaatg ccattacggc aagaccgttc catcgccttg
9601 tcatcgaatg agcacggcaa cacgatttct ttatgatatt ttttactgcc tctttccgag
9661 tgtaacgtca gaatgtcgcc attcaattcg acttgcacat cttcatcagc gacccccggc
9721 atttctgcca gtaccagcac atgatcactt tcttcgataa catcaaccaa cggttcagac
9781 acttcctgca ctgtagcttc cccggtttgc tcatcccggc gtatgttgcc aaaaggttct
9841 acccgatttt cctggtttcc atccaaaccc atcttgatgg agaaaccata cacggctttg
9901 gcattcttac cgtcgttcgt gtcaatatcg aacgcaccgc tacgctttat ctgttcgccg
9961 gtttcaacca atttttccac cagatcgccc aaaccgcgca aaattccttc aacattccct
10021 gtcacatcat ttttttttta ctactcatga ttgtttacct cggctgcatt gatctttttg
10081 agtttagagt gcagcgtttt atagtcaata ttcaaacgcc gcgctgcttc tgccttgtta
10141 ttgcctgttt tctctaatgt gtcttgaatg ataattcgct ctatttgatc aatattgagt
10201 tgagtgatct ctttaagaga cgaaccttcc agacagaaat tctgtatcgt tctcagatac
10261 ggatttttcg ttgattgcgg aaagtgatct tctttgatga ccaggcctaa atcatcaacc
10321 cctactcttt tgtcggatac caaaacagca cggcggatga cagcgcgtaa ctctcgcacg
10381 ttacccggcc atggataacg taacagtgtt tctttagccg caacggacaa atcaggcggt
10441 gttttgttca attcgacgct ggtttcggcg ataaaccgat tggacaaaaa cagaatatcc
10501 tcgtgccgat tgcgtaacgg cggaatccga atgacatact cattcaatcg ataatagaga
10561 tcctcgcgaa atttcccttt aactaccgcg tccagtagat catcattgct ggcagccagg
10621 actcgcacat tgacaggaat gggtttagtc gcgccaatac gatagacaac gtgctcttgt
10681 agcacacgca gtagcttggc ctgtgcggat aaagacatgt tggcgatttc atccaggaat
10741 aacgtaccgc cgtcagccgc ttcaaactta cccgcatgaa cctgatcagc cccggtgtac
10801 gagccttttt catgaccaaa tagttcattt tcgatcaacg agtcgggtat ggagccacaa
10861 tcaatgggaa caaaaacccc tgtggcatgc tgactggcta aatggatgtt tttcgcaacc
10921 aattctttgc cagttcccgt ttcaccctga ataacgacag aaaaatccgt atgagccacg
10981 ttaacgacat ctctgatcac cgactgaatc tcgcggctat tccccatcat ggcagcgata
11041 cgcgttttag tatccttgct ggctttgcag tttgaatcgt agttacccca acgcatttgc
11101 attgcacgat tcaccagatc taatactcgt ttgttatcaa acggcttggg aatgtaatcc
11161 catgcaccag ctttgatagc accaaccgcg ctaagaatgc cgacattgcc agtaataatg
11221 ataacgggga gttgcggata cagagcatgg gcattggcta acacctccat gccactgtgt
11281 ttagggagaa tactatcaag taacagtaca tccggttcac gctgagacaa caactttatt
11341 gccatgtcgc catcgtaacc ttgctgagtt tcacaaccag ccttttccag caagtgtgat
11401 agcaggtcac aaatatcttt gtcgtcatca attattaaaa catagccagg atcaatactt
11461 ttcatcgact acatccaatt ggttatttca tttttaataa tagttaaaag atacggggaa
11521 gaagaggata aatctccctt ctatcacgta tatatttgac gcaaaaaaag aattattaac
11581 actctagtta tttttttaac acaaccgcct aatcatcgcc tagacggtgc ctaattttaa
11641 tgaatagcat ccttaatact tatagtacaa atttaagata acacaatttt cttttcgatt
11701 ttgtgacaat aatttaggca agtatttcta tttattaacg aatattttaa ctacccggtc
11761 aaataagcga gagactttat tctggacatg ataaacatca tcccatcatc tgtctactga
11821 tggctatcag ccatatcaag ggcgacactc cctttttagc cgattaggtc acatttttag
11881 tatcaaaaga tcatagggct atcgataaaa ctggtaataa ctaagagaca tacatcacat
11941 aaatagctaa taaaccaccg ccaaaaaacg tttgcctatg tcacatctgg catcaattcc
12001 actagaagca ccatatatta tctaacaaaa taatgaatat tgatttcgat ttctttacac
12061 aaaaatgaag gctgaaacag acaaacagga agggtagtat taacaaaatc atcccgtgat
12121 ggaaattccg accacgggtt ccataatatt tatttatcac cgtatacgca ctatattgct
12181 gctatgccga tacttttaca aacaaacata gttggttatt attacgtttt tcatcagtca
12241 aaaaaacgca taaaaatata aaaaaagcaa caaagtccca ttattatgta gtctattaaa
12301 atgacatata aattatattt atactacctc taatattctc ggtgaataaa tccacttcta
12361 atgccaacga taatgcagaa caaggaatgt agtcactaaa tactttcccc tcctgggggt
12421 ttacctaacg gttgaataaa aacaatatct gcgggcagct tcgcttcaat attgtgcata
12481 agccccagga aaagtatcgc catctcgcca tcttgcagcg tttcatcaat gcgctgggca
12541 atataatcat cccgccggtt caataatgtt tttgctgttt ccaaggacgc ttcggtttgc
12601 tccgtagatt gtaacaactg cgtcatcaga tcgcgctctt gcaacaatag ttcaggggac
12661 tctgttccca tcagaattgc gcctttatgt tgcaacgtaa gcaagagctt gtaattctgg
12721 ctaccagcgt tagcaacatc ggtcactatt tcattttctt ttccacaaac cggcagacca
12781 tcctgataaa gtttcagatg tgtatagtcc agatctaaac cctcaatcgt agtggtaatc
12841 attgtccaga acccctcgac ggcagcaaga tacgcacgcg tttgagtttc ggttcgcaaa
12901 tgacttgccg cttcgctcaa tgtccctaaa tctttagggc tatgaataat gggaaaatga
12961 atcagcgctc gcataaaaaa gataatcctt ataaatgagt ttcttctatg acggacggtg
13021 ccgcttccaa acgagcctct agccgctgca gacgttcttc caactctgga gcaggcgctg
13081 atgatgacac tctggctctg gaatcaaacg ccggattgcc actccaccaa tccagcccca
13141 tttctaccgc tttatctacg gaacagataa gcaggcgaat ttccagtgtc agcaattcga
13201 cttcaaccag tctgatgcga atgtcaccgc tgatgacaac ccctttgtcc agtaatcgct
13261 ctaacaaatc cgctacggtg gtcgaatcgg ttgaatgggt tagttttttg ttgccgctca
13321 tggtgcaaat ccttataggg taaacaacct aaagacctca aatcctcatc cccttctctc
13381 taaaatcaaa agcatatata cccaataaat ttcaacacac aggaaggcga caatcgagta
13441 aatccccaag gcttactcca gtaagtgact ggggttagcg tgggcagaca acgcacctgc
13501 aacttgaaag atgatctata ccgtactgat gctcttaatt cttggatcca gggtatcctg
13561 aacatggatc tcactgcgta gaaaacactg ctggcgatat cgcttgcgac ttaatatagg
13621 attcaaacct gataaatagg ccacgttaat atcctgaaac ttatatggat gtcggtgtta
13681 tttccccaac gcgttcaagg atatcaagtg acagccggaa caaaccatct cgttcataac
13741 gttgtgaaat ggtcaacagc gccgtttgcg cggttttact ctccttgctg tcaggatatt
13801 gctcgattat tttcagatag atatccactg cttgtcgcag ttcatcttgt tcaaaccatg
13861 cattacccat acgcagtaga agaccacgcg tagccgcatt tatgtcattg gtagtattaa
13921 ccatggtaca caccctctgc cgaaaacaga ctgataacat ttatatggga actaaaaaat
13981 aatcgtatgt tgttgttcat catacaattt ccttacagca atcggcctag cgggcctaaa
14041 tccaagttca aatcatcatc cttgaagcca aaaacgtcgc acaaatgtgt caactcctcg
14101 gcttggcgca ttagcgccaa acccaaacgc tcgacttctt catcactaag cgaacccgaa
14161 tccatccgcc ttatggcttg acgctccagt agttcgtgca gcagtttgac cagggtcagg
14221 acgagttgcg ccaaaccatc acggacgcgt tctccatcga tgtctatttt atgtcgatta
14281 gcttcagtaa ttggccgttg tattgctggc gatgtcgggc cgaacacagg actatggtgc
14341 gataactgat tggttgtcat agtgttactc cctaccgggt aaacggtttt tcgcttctac
14401 agaagaaacc aaagcctgta ggcttaaata aatcaaatca atattagcga ctgaaatggt
14461 gatatcagcg acaatcacca cacctttgtt cagaacgcgg tccaaggcct cacacagaga
14521 gacctgttgc tgttcatcct gatactgttt gttcactggc ataatgtgta cgctcacagt
14581 ttataaaaac aatgccgaac ccaatgtctt tagcttgctc aatcccgtag gttctgtttg
14641 ccggaaaatc tgcgcgtgtg gttggctgtg gaacatctca tctgcgcact tcagctccaa
14701 cgactctcga ctcgtgattg cctggcacag tgtgcaatcg ctgggaggtg taatctggtt
14761 gataaacaac gcgtttgccg ttaagcctaa tcgctgcaat gcgcatgtca tttcataggt
14821 cttctcaagt gccaaatgcg tcggaacagt aactgcatac aaccccgttt gtttggtatc
14881 ctgcagtaga gcgcgtagat tttttaattc gcgtgaaagc tgtaccaaac gttccgataa
14941 atgcgggaaa cgcatgactt tgcgatattt cagcaataac gagaaaaact gttttagcca
15001 atcccgaatc aattcaggca actctaataa acgtagcaaa tgtccgctag gtgcgccatc
15061 cacgatgacc atgtcataac gaccgctgtc tagatgatcc atgatcgcag tcaatgccat
15121 gatttcatct aatccggggg gcgcgagatc gagcaaatgc tccaacactt cacgatcaaa
15181 agtaatatcc agattgggta acgtatccag caagaaagct tctaattccg cacgatatcc
15241 ctgccggatt ttgtcaaaat cagcttgtgc attaatttcc tgggcatcaa tgttaaccaa
15301 caccgatata ggctgctgct gtagggttac acccagacaa tcactcagcg agtgagcagg
15361 atcggctgag aacaacagaa tacgcaattc tggatattca ctgttcagac gcaatgccgt
15421 agcacaggcc agcgtcgttt tacctacccc acctttacct gcaaaaatca gcagtcggaa
15481 tgtgctggca ggcagcgaaa tggaactctc tacacgtaaa ggcagttgat gatgacacgt
15541 cgtagccatg acttcgtttt catcaagtaa acggaccccg gaccaaaatt cgtgaagcaa
15601 cgcaccacgc ggttcgatgg caagcaaagg caacgtccag aatacgtgcc caggcaaacg
15661 ttggtatccg ttctggagcg caagcatttg tctgttacgc tcaacacaac aggttggaca
15721 gtcattttca ggaaatagtc gattgaccac taaatcggac agaaacaccc gttgctgatt
15781 caacgcgcca gcaagatcaa tgctttcttc cacactcatc gcttccgcca gcatcactaa
15841 cacaaaacaa caacgcgtgg aatccgtcac cagctcgtgc atagctttaa gtgaatcatt
15901 catgtccagc agaaaatgat ccagatgatc caatcgatta tcaccggcaa aacgtttccg
15961 gatataacgc tgtttggcaa gcagggtatc cagagcggtc aaccaccgat aaatcaaatc
16021 cggcatttcc aaaagccgca atgtatgacc tgtaggagca gtatcaataa taatacgata
16081 gtaagtatct ttttgaatcc attcagctat ttcaagataa gcggccaatt catccatgcc
16141 gggaagcgcg gtatccatca atccttgtac gtcgttttga tcgagaacag tgccgcgata
16201 cgctatctct ttcagaacac cttcatgctg cgatttaaat tcatgcaagg aggctgcggc
16261 atttaattca cgtacgtcca gattttttgg caataccaga tcggagagaa tattttgcag
16321 ggaatgtgct ggatcggtag aaaccagcaa aaaatggtgc tgcggctgtt cctgcgccag
16381 ccttaacgcc gtcgacgtcg cacaggtact tttacctact cctcccttac cgccgaagaa
16441 aaccagtttc agcggcgact ccagtaagaa cttgggataa atagcaggtt tcatttcgtc
16501 gtaccctctg gctcatcctc gtcgtcgtca tcctcgtcgt cgtcgtcatc ctcttcgtcg
16561 tcgtcatcct cgtcgtcgtc atcctcgtcg tcgtcatcct cttcgtcgtc gtcctcttcg
16621 tcgtcgtcct cttcgtattc gtcgtcttcg tcatccccag gctcgtcgcc tagcatgtca
16681 tcttcttcat ccagtgcgtc taatctatcc agcagtacag cttcctgttg ttcaaattca
16741 tcttcggtaa tctggccagt ctccagtaac atataaagat cagtcaaact ttcacgaata
16801 cgatccgcct caccagccaa ttcatcttcc gccagttcgt gaatttgccg aaaaatccac
16861 atgacgcctt tgacaggaga aaacaggatg tcatcgatca gcagcatggc ttagcctcag
16921 tcagagagag taccagcgcg ggtcaacgtt ttcttcttcg ctgtaggagc atgtaaatcc
16981 agagtaacga aattatgcgg tgcccatggg ccggtataat cgaacagata aacgttatcg
17041 aataactttg atgcctcaaa aacgcctttg gcaaattcat ccatccgttc tcttgctacc
17101 aaacaggcta gattcattac gtctttttct tttttaaccg ggttttcaat aatctcttcg
17161 caataatcca acagaacttc ttttactttt tccgtcgatt cttttctgtc tccactacgc
17221 aatgactcgt acaaattacc cagccgaatc ttctcttcac ggtgattatt ggcattggaa
17281 tttccattcc agatttcatc gcgcgtttca cttaagactg gatatgtagc aacaaaatat
17341 tcataaatat tagtaacatc ccaggaaaca cgtagcccca tttccacaca gccatccaag
17401 cgttcaaaat gttctctgat cgtatcctga ttagcgacca gcaaattctt tactgcttca
17461 gcattacggg caataacccc aaagcgcatc ggcaatacgg taccttcttc agtcaactta
17521 tgtaatacag cctgatgcgc cgtgatatta cgtctttctg gacgtaaccg ggtactgagc
17581 gtatcgctga ccacagctct taagccatct tgatcaatag aataaaccgg tgctgcttcc
17641 aatccgcgta ttttatagtt aggcgttgag tcatcggaaa cacagagcgc ataaagcacc
17701 ttggcacgat gatttctgga tttatcaata ctcatcatgc actcctcaat gaaatacgtt
17761 tacttttggc gcattgcaca cggttcaatc gtcgatattc ttctaaataa atacgtccaa
17821 aatcagcgag aatttgtgac ttccccgtca tgttgacgca tgtgcgaaag ctcttcacca
17881 tacgcggtgt acggatatga gcgagggtgc gaattttgtt cataatgaca acctcaatac
17941 ttgattcgca aaccagaaga ctttttcttc ttctcggccg cttctgattt atcattattt
18001 tcagccacag cactcgccgc tgtcgcgtct gcaagcagcg ctgctttttc tttatcaata
18061 tccgcgatac gctcatcaat actggcaata cgcgatgagg cagcctcacg ttctcgtgtt
18121 cgccgccaac gctctaattc cagcgcacct atctgaaaat aatcacgata ggtttgatgc
18181 tcaccactca ctcgacctga atgagttttt atgtctgaca gtgttctgag aggacgagta
18241 cttatagcca tgataattac tccactgctt ttattttttg tgtggatgct ttaaccgtgt
18301 ctgaacctat tggtggacag attttacgaa taacgccatc gaccatttca cggtatatag
18361 aacgtccagc gtgttgtatt ttggcagcgt ccatactcaa gacgtcgtag cagatagcat
18421 ggaataattt gtcaccataa cgtggtttta ttccttgctg gtgtaggatg tgggcgatgg
18481 caattcctgc acgtaaccca tgtttggttt cgtgttcgtt accacgcaac tctctgatta
18541 tatcaacgat atatgccgct tcttttaatt ccaattcaga gcgcgcatgc agtatctgta
18601 gttcagtatc ccgatcgaca agaccaatgt tgatagtaat catacgatcc atcagagcgt
18661 cctgggtctt atgcgtaccg gcatactctt ctggattgga ggtaaaaata gcgcgaaatt
18721 ctgggtgtac atcgagatac ccttccccta caccacgtaa accaggaagg tttaaaatcc
18781 cttcggataa tacggataac agcacattat tggtttctgc tttggaacga ttgaactcat
18841 catagatcag catgtcgccg ttacgacacg cggtggtcaa acggttatcc acccacatgc
18901 ggttcatctg ttcttcagtt ttaagcactg aatgaatgta gttatccacc aggctgcttt
18961 tacgatagcc gatgtctttc cccgtcaaat cggagctcac aaattcttcg ttaccttgca
19021 acatggtaac gggacgaccc catagtgctg cgagatgaaa tgccagggtt gttttcccta
19081 tcccagatgg gccggcaaaa tgcacgggat agccagcgcg taaataaacc aatgcgcgtt
19141 ctataatgtc gttcacataa gaagtcgcta caaaatgttc actcgcttcc ggaacgacaa
19201 gatcatcgtc tacggtatac tgcgaaacgg tattttgctt aatcatggaa atactcctgt
19261 gccaaaccgt gtacgcccta gccaacacca gttctacgaa atggccaaat cagtgatcgt
19321 cttagccgct agaatcggtc gattttagcg gattcttata tttcgattta acttagcgat
19381 taatcgaaac tgtatgccca ataatgtcac tccgtgtttg tgagcagact gttcatccac
19441 ctcaccctcc atggacgaca gaggctaata ccgctactta ccttttttca tcttcactac
19501 ttaccgccag agtattatca agaggacgag tggctagact gcgaggctcg tttttttttt
19561 agccagggtc atgccgaaaa atacagcatg ggcagccatt ctgtccttct cagaagcact
19621 aagaaaagca gcgacatcgt ttgccacgtt ggcaatgaaa tcagctcgtt ctttagcaca
19681 gcgttcagcc atacttttac gctcagcatt aaaaccgtct aataaagcta gcgttttctt
19741 cgacatatcc gtcacgaagg tggcacgttc ttcactctgt tgacgactca ttacctcgcg
19801 ggtatgacag aagtcactca acatccgatt aacgcctctg acattatcgg caacgaatgt
19861 cgcgcgtgcg cgtgcgtctt gagccgcgtt tctagcccga gttgtagaaa aaccggcaat
19921 ctgcgcactc acactggaaa cccgcgcatt ttgctgtgct atgcgagatt catgactgtc
19981 atcaatattc ttacgcagct gcgccatacc gtctgttaag catcccatag gattctcctt
20041 atgtgataga gcactcttga agcgttgaaa agtactcaag cagctgaatg actcagccac
20101 ttgggcactt ttattctgct tacttaatta cgctggagtc gctgcactgg ctgtaaggcc
20161 aatcgcttct gcgtatttca aataagtttc aacagaagca ataacaacac gtgcttcgat
20221 ggataacagt tcaataccaa ccaatgagac tttgacccaa gcgtcaatca caatgccttt
20281 gtccagaata cggtctacta cttcagccaa gctggaagag tctgttgatt tttgtacttt
20341 agccatgatg gcgtctccta gaaaatgtta ataaattaca tatgtaatta ggttctacgt
20401 acagacttgt tactgcacgc ccatgtagaa tgcaactcat gcgccaacct ataggcatca
20461 gcgctctatt aatgttaata aatccaccac aatgaaggtt aattcctcct gttagcccaa
20521 aaaaacgggt atggagaaac agtagagagt tgcgataaaa agcgtcaggt aggatccgct
20581 aatcttatgg ataaaaatgc tatggcatag caaagtgtga cgccgtgcaa ataatcaatg
20641 tggacttttc tgccgtgatt atagacactt ttgttacgcg tttttgtcat ggctttggtc
20701 ccgctttgtt acagaatgct tttaataagc ggggttaccg gtttggttag cgagaagagc
20761 cagtaaaaga cgcagtgacg gcaatgtctg atgcaatatg gacaattggt ttcttctctg
20821 aatggcggga gtatgaaaag tatggctgaa gcgcaaaatg atcccctgct gccgggatac
20881 tcgtttaatg cccatctggt ggcgggttta acgccgattg aggccaacgg ttatctcgat
20941 ttttttatcg accgaccgct gggaatgaaa ggttatattc tcaatctcac cattcgcggt
21001 cagggggtgg tgaaaaatca gggacgagaa tttgtttgcc gaccgggtga tattttgctg
21061 ttcccgcaag gagagattca tcactacggt cgtcatccgg aggctcgcga atggtatcac
21121 cagtgggttt actttcgtcc gcgcgcctac tggcatgaat ggcttaactg gccgtcaata
21181 tttgccaata cggggttctt tcgcccggat gaagcgcacc agccgcattt cagcgacctg
21241 tttgggcaaa tcattaacgc cgggcaaggg gaagggcgct attcggagct gctggcgata
21301 aatctgcttg agcaattgtt actgcggcgc atggaagcga ttaacgagtc gctccatcca
21361 ccgatggata atcgggtacg cgaggcttgt cagtacatca gcgatcacct ggcagacagc
21421 aattttgata tcgccagcgt cgcacagcat gtttgcttgt cgccgtcgcg tctgtcacat
21481 cttttccgcc agcagttagg gattagcgtc ttaagctggc gcgaggacca acgtatcagc
21541 caggcgaagc tgcttttgag caccacccgg atgcctatcg ccaccgtcgg tcgcaatgtt
21601 ggttttgacg atcaactcta tttctcgcgg gtatttaaaa aatgcaccgg ggccagcccg
21661 agcgagttcc gtgccggttg tgaagaaaaa gtgaatgatg tagccgtcaa gttgtcataa
21721 ttggtaacga atcagacaat tgacggcttg acggagtagc atagggtttg cagaatccct
21781 gcttcgtcca tttgacaggc acattatgca tcgatgataa gctgtcaaac atgagcagat
21841 catctacgcc ggacgcatcg tggccggcat caccggcgcc acaggtgcgg ttgctggcgc
21901 ctatatcgcc gacatcaccg atggggaaga tcgggctcgc cacttcgggc tcatgagcaa
21961 atccgctaat taactaaaac tagtacaaaa aatttaaaat agattattaa acctattgct
22021 tattctattc agtcatatat ataagcgtca tcctttagta actaacatcc cacactacaa
22081 tagtattata acatatctat tcaacttcaa attgttgaaa tttaagccat ttatatttac
22141 ttttcctcaa ctttagaaat gattttacta accttatatc cgtaatacgc gtaacaaaca
22201 cattcattaa taacaatcaa cactaagcta tattagcttt ttaaatgact agggtaaata
22261 aaataaggat caaaaacttt taagtttctg accctttcct tacttccgat tggttaatag
22321 tgatcttttg cagaataaat aaatatcgtt tcattttcaa ctctatatat cagtctatgt
22381 tcatctgtaa ttcttctgga ccattttcca gataaatcat gctttaatgg ctcaggtttt
22441 cctaatccag caaaggggga acgatcgata tcttttatta acttgttaat cttttttata
22501 ttgcttttgt ttccttgctc atgccaataa agataatcat cccaagcatc atcagaccaa
22561 gccttaatca tcagattcaa cctcgattaa gtcatgtgtt ttaaatgcac ctttggagaa
22621 ttgttcatct cctcgacgaa ttttttccat gacgtaatta ttagaaagtg ttctcaacgt
22681 ttcttgcata gaatcataat ctctttttga taatacaaca actgtatctt ctacatcttt
22741 acttgttaca ataagtgttt cagcatcctc attaacttgt ttcatataac tacgtaaatt
22801 ttggcggaaa tttgaataag ctactgcttc cattcctttc accccttatt tatttctata
22861 aaacaattgt acattatatt gtacaattaa gcaaatgatt aatattctaa cttcttataa
22921 ttaaaagtgc tatcactttt agtttttagg ataaaaagtg acgtcacttt ttgagtgctg
22981 gtacctcaaa tcctgctttt tcttgggcaa tcaataaatc taacgcaata cacataatcc
23041 tggattcact tttgccgtac ttctgcttta gttgctgcaa ttctttttgt ctcttttcat
23101 ccaacgtaac gggtaaccgt ttatttccta ttgcaattgg cattagtact ctctcctttc
23161 atcgaaataa ccctatttta tctgattaat aagatgatct tc
//
| {
"Vector backbone": "pBAD33",
"Backbone size w/o insert (bp)": "5352",
"Vector type": "Bacterial Expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Chloramphenicol, 25 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "NEB Stable",
"Copy number": "Low Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Genomically mined acoustic reporter genes for real-time in vivo monitoring of tumors and tumor-homing bacteria",
"Publication": "Nat Biotechnol. 2023 Jan 2. doi: 10.1038/s41587-022-01581-y.",
"DOI": "10.1038/s41587-022-01581-y.",
"Authors": "Hurt RC, Buss MT, Duan M, Wong K, You MY, Sawyer DP, Swift MB, Dutka P, Barturen-Larrea P, Mittelstein DR, Jin Z, Abedi MH, Farhadi A, Deshpande R, Shapiro MG /s---y",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/36593411",
"PubMed ID": "PubMed 36593411"
} | {
"Gene/Insert name": "bARGSer",
"Species": "Serratia sp. ATCC 39006",
"Insert Size (bp)": "16692",
"Mutation": "the gene Ser39006_001280 was deleted from the cluster, and 3 point mutations were introduced during cloning (GvpG P133P, GvpW P609H, GvpZ D112Y)",
"Entrez Gene": "",
"Promoter": "pBAD",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Gibson Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "CCAATTGTCCATATTGCATC",
"3′ sequencing primer": "GATTTAATCTGTATCAGG"
} | {
"Gene/Insert name": "AxeTxe",
"Species": "Enterococcus faecium",
"Insert Size (bp)": "1212",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "Native promoter from Enterococcus faecium",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Gibson Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "GTTAATGAGGATGCTGAAACAC",
"3′ sequencing primer": "AAGCATCATCAGACCAAGCC"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pLV-Enh eGFP Reporter-muKC2-X41 | 59265 | Lentiviral GFP reporter for testing enhancer activity in different cell types. Insert is an enhancer identified in the exonic region of the keratin type II cluster. | Benjamin Yu | Yu Lab-keratin enhancer GFP reporters (unpublished) | https://media.addgene.org/snapgene-media/v2.0.0/sequences/95728/d347646e-3a44-43b3-b27f-3571e8cf81f4/addgene-plasmid-59265-sequence-95728.gbk | partial | LOCUS sequence-95728-e 713 bp DNA linear UNA 26-AUG-2024
DEFINITION Lentiviral GFP reporter for testing enhancer activity in different
cell types. Insert is an enhancer identified in the exonic region of
the keratin type II cluster. .
ACCESSION .
VERSION .
KEYWORDS .
SOURCE natural DNA sequence
ORGANISM unspecified
REFERENCE 1 (bases 1 to 713)
TITLE Yu Lab-keratin enhancer GFP reporters
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 713)
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..713
/mol_type="genomic DNA"
/organism="unspecified"
protein_bind 143..167
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
ORIGIN
1 ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat acaaactaaa
61 gaattacaaa aacaaattac aaaaattcaa aattttatcg atcacgagac tagcctcgag
121 aagcttgata tcgaattcca tcacaagttt gtacaaaaaa gcaggcttta cctgagcctt
181 ggtatgctcg atctctgctt tcagcctctg gatcaaccgg gtcagttcat tgatctcgtc
241 ccgtgtgcta cgtagattgt cacagtgctg gccagcggtc acccgcatct cctcatactg
301 ggggtggagg tggggtgggg tggggcaaag ttttgaccat acatagtctg ggctgaagac
361 ctcagaccaa aatcttcacc tttggacagg gcacctgtac ccctcccaca gtctcccatt
421 gcccaccttg gtctggtacc acgactccac atctgcacga ctccgcctcg ccacctcctc
481 atactgtgcc ttgacttcag ctatgatccc atctaggttc agatcccggc tgttgtccat
541 cttcacgatg acagatgtct ctgatatgtg tgattgcagc atctcgattt cctgtattgg
601 aaaagcagta gggcagaggg gctcagtggt ccatcagcag ctgggactct gccacagggt
661 cccctgtgaa tcagggacag ggagcattgc ttctcgcttt atgactgtga gag
//
| {
"Vector backbone": "Brachyury-eGFP",
"Backbone size w/o insert (bp)": "7967",
"Vector type": "Lentiviral",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "Stbl3",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "",
"Publication": "",
"DOI": "",
"Authors": "",
"PubMed Link": "",
"PubMed ID": ""
} | {
"Gene/Insert name": "Krt84-ex5-6",
"Species": "M. musculus (mouse)",
"Insert Size (bp)": "599",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "Mu Hsp68 minimal promoter",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Gateway Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "ACCTGAGCCTTGGTATGCTC",
"3′ sequencing primer": "TTAATTGCTTCTCAAGCATGCC"
} | {
"Gene/Insert name": "",
"Species": "",
"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": ""
} |
|
pKCiR5 | 149462 | Broad-Host range expression vector, RK2 and pUC origin, CinR(AM) activator, PCin promoter, mRFP, Gentamicin Resistance Marker. | Christopher Reisch | Schuster et al Nucleic Acids Res. 2021 Jun 14. pii: 6298615. doi: 10.1093/nar/gkab496. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/311916/390691cb-901b-4bb7-a01d-9c7128b57938/addgene-plasmid-149462-sequence-311916.gbk | full | LOCUS sequence-311916- 7398 bp DNA circular SYN 25-AUG-2024
DEFINITION Broad-Host range expression vector, RK2 and pUC origin, CinR(AM)
activator, PCin promoter, mRFP, Gentamicin Resistance Marker. .
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7398)
AUTHORS Schuster LA, Reisch CR
TITLE A plasmid toolbox for controlled gene expression across the
Proteobacteria.
JOURNAL Nucleic Acids Res. 2021 Jun 14. pii: 6298615. doi:
10.1093/nar/gkab496.
PUBMED 34125913
REFERENCE 2 (bases 1 to 7398)
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: "Nucleic
Acids Res. 2021 Jun 14. pii: 6298615. doi: 10.1093/nar/gkab496."
FEATURES Location/Qualifiers
source 1..7398
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS complement(249..1397)
/codon_start=1
/product="trans-acting replication protein that binds to
and activates oriV"
/label=trfA
/translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
CDS complement(1669..2040)
/codon_start=1
/gene="traJ"
/product="oriT-recognizing protein"
/label=traJ
/translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSL*AYLLAV
GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
KQDELGKVMMGVVRPRAEP"
oriT 2073..2182
/note="incP origin of transfer"
rep_origin 2335..2966
/label=oriV
/note="incP origin of replication"
rep_origin 3429..4017
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3918..3937
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 4335..4357
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 4349..4366
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 4350..4366
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
misc_RNA 5515..5564
/label=sTRSV HHRz
/note="hammerhead ribozyme from the tobacco ringspot virus
satellite RNA (Khvorova et al., 2003)"
RBS 5609..5620
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 5627..6304
/codon_start=1
/product="monomeric derivative of DsRed (Campbell et al.,
2002)"
/label=mRFP1
/translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
TGA"
promoter 6581..6609
/gene="intI1 (promoter lies within the coding sequence)"
/label=Pc promoter
/note="class 1 integron promoter"
CDS 6798..7331
/codon_start=1
/gene="aacC1"
/product="gentamycin acetyltransferase"
/label=GmR
/note="confers resistance to gentamycin"
/translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
EEVMHFDIDPSTAT"
ORIGIN
1 ggtggcactt ttcggggatc gtggaaacga taggcctatg ccatgcgggt caaggcgact
61 tccggcaagc tatacgcgcc ctagaattgt caattttaat cctctgttta tcggcagttc
121 gtagagcgcg ccgtgcgtcc cgagcgatac tgagcgaagc aagtgcgtcg agcagtgccc
181 gcttgttcct gaaatgccag taaagcgctg gctgctgaac ccccagccgg aactgacccc
241 acaaggccct agcgtttgca atgcaccagg tcatcattga cccaggcgtg ttccaccagg
301 ccgctgcctc gcaactcttc gcaggcttcg ccgacctgct cgcgccactt cttcacgcgg
361 gtggaatccg atccgcacat gaggcggaag gtttccagct tgagcgggta cggctcccgg
421 tgcgagctga aatagtcgaa catccgtcgg gccgtcggcg acagcttgcg gtacttctcc
481 catatgaatt tcgtgtagtg gtcgccagca aacagcacga cgatttcctc gtcgatcagg
541 acctggcaac gggacgtttt cttgccacgg tccaggacgc ggaagcggtg cagcagcgac
601 accgattcca ggtgcccaac gcggtcggac gtgaagccca tcgccgtcgc ctgtaggcgc
661 gacaggcatt cctcggcctt cgtgtaatac cggccattga tcgaccagcc caggtcctgg
721 caaagctcgt agaacgtgaa ggtgatcggc tcgccgatag gggtgcgctt cgcgtactcc
781 aacacctgct gccacaccag ttcgtcatcg tcggcccgca gctcgacgcc ggtgtaggtg
841 atcttcacgt ccttgttgac gtggaaaatg accttgtttt gcagcgcctc gcgcgggatt
901 ttcttgttgc gcgtggtgaa cagggcagag cgggccgtgt cgtttggcat cgctcgcatc
961 gtgtccggcc acggcgcaat atcgaacaag gaaagctgca tttccttgat ctgctgcttc
1021 gtgtgtttca gcaacgcggc ctgcttggcc tcgctgacct gttttgccag gtcctcgccg
1081 gcggtttttc gcttcttggt cgtcatagtt cctcgcgtgt cgatggtcat cgacttcgcc
1141 aaacctgccg cctcctgttc gagacgacgc gaacgctcca cggcggccga tggcgcgggc
1201 agggcagggg gagccagttg cacgctgtcg cgctcgatct tggccgtagc ttgctggacc
1261 atcgagccga cggactggaa ggtttcgcgg ggcgcacgca tgacggtgcg gcttgcgatg
1321 gtttcggcat cctcggcgga aaaccccgcg tcgatcagtt cttgcctgta tgccttccgg
1381 tcaaacgtcc gattcattca ccctccttgc gggattgccc cgactcacgc cggggcaatg
1441 tgcccttatt cctgatttga cccgcctggt gccttggtgt ccagataatc caccttatcg
1501 gcaatgaagt cggtcccgta gaccgtctgg ccgtccttct cgtacttggt attccgaatc
1561 ttgccctgca cgaataccag ctccgcgaag tcgctcttct tgatggagcg catggggacg
1621 tgcttggcaa tcacgcgcac cccccggccg ttttagcggc taaaaaagtc atggctctgc
1681 cctcgggcgg accacgccca tcatgacctt gccaagctcg tcctgcttct cttcgatctt
1741 cgccagcagg gcgaggatcg tggcatcacc gaaccgcgcc gtgcgcgggt cgtcggtgag
1801 ccagagtttc agcaggccgc ccaggcggcc caggtcgcca ttgatgcggg ccagctcgcg
1861 gacgtgctca tagtccacga cgcccgtgat tttgtagccc tggccgacgg ccagcaggta
1921 ggcctacagg ctcatgccgg ccgccgccgc cttttcctca atcgctcttc gttcgtctgg
1981 aaggcagtac accttgatag gtgggctgcc cttcctggtt ggcttggttt catcagccat
2041 ccgcttgccc tcatctgtta cgccggcggt agccggccag cctcgcagag caggattccc
2101 gttgagcacc gccaggtgcg aataagggac agtgaagaag gaacacccgc tcgcgggtgg
2161 gcctacttca cctatcctgc ccggctgacg ccgttggata caccaaggaa agtctacacg
2221 aaccctttgg caaaatcctg tatatcgtgc gaaaaaggat ggatataccg aaaaaatcgc
2281 tataatgacc ccgaagcagg gttatgcagc ggaaaagatc cgtcgaccct ttccgacgct
2341 caccgggctg gttgccctcg ccgctgggct ggcggccgtc tatggccctg caaacgcgcc
2401 agaaacgccg tcgaagccgt gtgcgagaca ccgcggccgc cggcgttgtg gatacctcgc
2461 ggaaaacttg gccctcactg acagatgagg ggcggacgtt gacacttgag gggccgactc
2521 acccggcgcg gcgttgacag atgaggggca ggctcgattt cggccggcga cgtggagctg
2581 gccagcctcg caaatcggcg aaaacgcctg attttacgcg agtttcccac agatgatgtg
2641 gacaagcctg gggataagtg ccctgcggta ttgacacttg aggggcgcga ctactgacag
2701 atgaggggcg cgatccttga cacttgaggg gcagagtgct gacagatgag gggcgcacct
2761 attgacattt gaggggctgt ccacaggcag aaaatccagc atttgcaagg gtttccgccc
2821 gtttttcggc caccgctaac ctgtctttta acctgctttt aaaccaatat ttataaacct
2881 tgtttttaac cagggctgcg ccctgtgcgc gtgaccgcgc acgccgaagg ggggtgcccc
2941 cccttctcga accctcccgg cccgctaacg cgggcctccc atccccccag gggctgcgcc
3001 cctcggccgc gaacggcctc accccaaaaa tggcagccaa gctgaccact tctgcgctcg
3061 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
3121 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
3181 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
3241 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
3301 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
3361 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
3421 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
3481 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
3541 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
3601 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
3661 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
3721 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
3781 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
3841 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
3901 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
3961 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
4021 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttcg tttaagggca
4081 ccaataactg ccttaaaaaa attacgcccc gccctgccac tcatcgcagt acggcctatt
4141 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
4201 ttacaatttc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
4261 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
4321 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgact attaccaatt
4381 acgtcgcgtc atgcggatcc tatagggatt gatgatgcgc aattgaacag cccgcgacgc
4441 ggcggccgag atcgtggtgc agccgagcct gaagcgggcg gttttcaggt aatcgcgtgt
4501 ggtatgctct gatatgccca ggatgaccga aatatccttg taatccttgc cgagggcggt
4561 ccagtgcaga cactcgatct cgcgcggcga caatgccggc accggatcgt tttcgccatg
4621 cagctcatat acggccttgc ggtggatcag atgggcgatc tcgatccact cattgcggca
4681 gcggcgcacg agctcggtcc attcgtcggc cggtatatgg gcattcagcg acagcagggc
4741 gcggcgctgc gccttgtcgg cgacggggat ggagtagcca ttgtcatcga tgccgtgttt
4801 ctgggcgtcg accagcatgg cataggcctc cggcgtcggt tcgacctcgc tccagtcgaa
4861 gggcagctgg cgttcgaagc cctgcttgat gatcggatcg accttcacat agcagttgag
4921 gaggtaacgg gaaacccagg catccggata ggtggtgcgc acgaagggcg aatcgatctt
4981 gctcgcgatt gtctgggcga gatggtaggt gacgaaatcg aggttatatt ccgcctggag
5041 aatacggatg gcggcatcca cgttggccgc cgctttgatc tgttcgaacg cggactcgaa
5101 cttttcgcta taggtattct caatcattcg ccgttctccg ttgacgggcg tctcggacct
5161 gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggcgcg ccgcccaatt
5221 attgaaggcc tccctaacgg ggggcctttt tttgtttctg gtctcccgct taacgatcgt
5281 tggctgccct ttgtgcgtcc aaacggacgc acggcgctct aaagcgggtc gcgatctttc
5341 agattcgctc ctcgcgcttt cagtctttgt tttggcgcat gtcgttatcg caaaaccgct
5401 gcacactttt gcgcgacatg ctctgatccc cctcatctgg gggggcctat ctgagggaat
5461 ttccgatccg gctcgcctga accattctgc tttccacgaa cttgaaaacg ctagctgtca
5521 ccggatgtgc tttccggtct gatgagtccg tgaggacgaa acagcctcta caaataattt
5581 tgtttaagaa ctgggaagag cgcagattaa agaggagaaa tactagatgg cttcctccga
5641 agacgttatc aaagagttca tgcgtttcaa agttcgtatg gaaggttccg ttaacggtca
5701 cgagttcgaa atcgaaggtg aaggtgaagg tcgtccgtac gaaggtaccc agaccgctaa
5761 actgaaagtt accaaaggtg gtccgctgcc gttcgcttgg gacatcctgt ccccgcagtt
5821 ccagtacggt tccaaagctt acgttaaaca cccggctgac atcccggact acctgaaact
5881 gtccttcccg gaaggtttca aatgggaacg tgttatgaac ttcgaagacg gtggtgttgt
5941 taccgttacc caggactcct ccctgcaaga cggtgagttc atctacaaag ttaaactgcg
6001 tggtaccaac ttcccgtccg acggtccggt tatgcagaaa aaaaccatgg gttgggaagc
6061 ttccaccgaa cgtatgtacc cggaagacgg tgctctgaaa ggtgaaatca aaatgcgtct
6121 gaaactgaaa gacggtggtc actacgacgc tgaagttaaa accacctaca tggctaaaaa
6181 accggttcag ctgccgggtg cttacaaaac cgacatcaaa ctggacatca cctcccacaa
6241 cgaagactac accatcgttg aacagtacga acgtgctgaa ggtcgtcact ccaccggtgc
6301 ttaataacgc tgatagtgct agtgtagatc gcgctttcca gtcgggaaac ctgtcgtgcc
6361 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgcatgc
6421 ataaaaactg ttgtaattca ttaagcattc tgccgacatg gaagccatca caaacggcat
6481 gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca
6541 tggacgcaca ccgtggaaac ggatgaaggc acgaacccag ttgacataag cctgttcggt
6601 tcgtaaactg taatgcaagt agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga
6661 acgcagcggt ggtaacggcg cagtggcggt tttcatggct tgttatgact gtttttttgt
6721 acagtctatg cctcgggcat ccaagcagca agcgcgttac gccgtgggtc gatgtttgat
6781 gttatggagc agcaacgatg ttacgcagca gcaacgatgt tacgcagcag ggcagtcgcc
6841 ctaaaacaaa gttaggtggc tcaagtatgg gcatcattcg cacatgtagg ctcggccctg
6901 accaagtcaa atccatgcgg gctgctcttg atcttttcgg tcgtgagttc ggagacgtag
6961 ccacctactc ccaacatcag ccggactccg attacctcgg gaacttgctc cgtagtaaga
7021 cattcatcgc gcttgctgcc ttcgaccaag aagcggttgt tggcgctctc gcggcttacg
7081 ttctgcccaa gtttgagcag ccgcgtagtg agatctatat ctatgatctc gcagtctccg
7141 gagagcaccg gaggcagggc attgccaccg cgctcatcaa tctcctcaag catgaggcca
7201 acgcgcttgg tgcttatgtg atctacgtgc aagcagatta cggtgacgat cccgcagtgg
7261 ctctctatac aaagttgggc atacgggaag aagtgatgca ctttgatatc gacccaagta
7321 ccgccaccta acaattcgtt caagccgaga tcggcttccc ggccgcggag ttgttcggta
7381 aattgtcaca acgccgca
//
| {
"Vector backbone": "Unspecified",
"Backbone size w/o insert (bp)": "",
"Vector type": "Bacterial Expression, Synthetic Biology",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Gentamicin, 10 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": "A plasmid toolbox for controlled gene expression across the Proteobacteria",
"Publication": "Nucleic Acids Res. 2021 Jun 14. pii: 6298615. doi: 10.1093/nar/gkab496.",
"DOI": "10.1093/nar/gkab496.",
"Authors": "Schuster LA, Reisch CR /nar/gkab",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/34125913",
"PubMed ID": "PubMed 34125913"
} | {
"Gene/Insert name": "CinR(AM)-mRFP",
"Species": "Synthetic",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "PCin",
"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": ""
} |
|
R777-E221 Hs.RALGDS | 70505 | Gateway ORF clone of human RALGDS [NM_006266.3] with stop codon (for native or N-terminal fusions) | Dominic Esposito | Esposito lab (unpublished) | https://media.addgene.org/snapgene-media/v2.0.0/sequences/125954/23fe19eb-c273-4823-8aa5-84fc6c46ab0b/addgene-plasmid-70505-sequence-125954.gbk | full | LOCUS sequence-125954- 5438 bp DNA circular SYN 26-AUG-2024
DEFINITION Gateway ORF clone of human RALGDS [NM_006266.3] 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 5438)
TITLE Esposito lab
REFERENCE 2 (bases 1 to 5438)
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..5438
/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(2929..3028)
/gene="mutant version of attL"
/label=attL2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
CDS 3405..4196
/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 4289..4877
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4778..4797
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 5031..5048
/label=L4440
/note="L4440 vector, forward primer"
terminator complement(5207..5234)
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator complement(5326..5412)
/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 gcaccatggt gcagcgcatg tgggccgagg cggccgggcc tgctggcggc gccgagccgc
241 tgtttccggg ctcccggcgg agccgcagcg tgtgggacgc cgtgcgcctg gaggtgggcg
301 tccccgacag ctgcccggtg gtgctgcaca gcttcacgca gctagacccc gacctgccgc
361 gcccggagag ctccacgcag gagatcggtg aggagctgat caacggagtc atctactcca
421 tctccctgcg caaggtgcag ctgcaccacg gaggcaacaa ggggcagcgc tggctcgggt
481 atgagaatga gtcggccctg aacctttatg agacttgcaa ggtgcggacc gtgaaggctg
541 gcacgctgga gaagctggtg gagcacctgg tgccagcctt ccagggcagc gacctctcct
601 acgtcaccat cttcctgtgt acctatagag ccttcaccac cacccaacag gtcctggacc
661 tgctgttcaa aaggtacggt agatgtgacg ccctcacggc ctcctctaga tacggctgca
721 tcctccccta ttccgacgag gatggtggac cccaggacca acttaaaaat gccatctcct
781 ccatcctggg cacctggctg gaccagtact cggaggattt ctgtcaacct ccggactttc
841 cctgcctcaa gcagctggtg gcctacgtgc agctcaacat gccaggctca gacctggagc
901 gccgtgccca ccttctcctg gcccagctgg agcactcgga acccattgag gcagagcctg
961 aggctctgtc accagtgcca gctctaaaac caactccaga gctcgagcta gctctaacac
1021 cagctcgagc acccagccca gtgccggctc cagccccgga gccagagcca gctccaacac
1081 cagctccagg ttcagagcta gaagtagctc cagcaccagc tccggagctc cagcaggctc
1141 cagagccagc tgtgggacta gaatcggctc cagcgccagc tctggaacta gagccagctc
1201 cagaacagga tccagctccc tcacaaactc tagagctgga gccagctcca gcgccagttc
1261 catcattaca gccttcctgg ccttcacctg tggttgcaga gaacgggctg agtgaggaga
1321 agcctcacct cttggtgttc cctccagatc tggtggcaga gcagtttaca ctgatggatg
1381 cggaactgtt caagaaggtg gtgccctacc actgcctggg ctccatctgg tcccagcggg
1441 acaagaaggg caaggagcac ctggcgccca ccatccgcgc cactgtcacc cagttcaaca
1501 gtgtggccaa ctgtgtcatc accacctgcc tcgggaaccg aagcacgaaa gccccagaca
1561 gggccagggt ggtggagcac tggatcgagg tggccaggga gtgccggatc ctcaagaact
1621 tctcgtcact gtatgccatc ctctctgccc tgcagagcaa ctccatccac cgtctgaaga
1681 agacgtggga agacgtttcc agggacagtt tccggatctt tcagaagctg tcagagatct
1741 tctcagatga gaacaactac tcattgagcc gggagctgct catcaaggag ggcacctcca
1801 agtttgccac cctggagatg aaccccaaga gagcccagaa acggccgaag gagacgggca
1861 tcatccaggg caccgttccc tacctgggca cgttcctcac cgacctggtg atgctggaca
1921 ctgccatgaa ggactatctg tatggcagac tcatcaactt tgagaagagg aggaaggagt
1981 tcgaggtgat cgcccagatc aagctgctgc agtcggcctg caacaactac agcatcgcgc
2041 cagatgagca atttggggcc tggttccggg ccgtggagcg gctcagcgag actgagagct
2101 acaacctgtc gtgcgagctg gagcccccat ccgagtcagc cagcaacacc ctcaggacca
2161 agaagaacac agccattgtc aagcgctgga gcgaccgcca ggcccccagc actgagctca
2221 gtaccagtgg cagctcccac tccaagtcct gtgaccagct caggtgtggc ccctacctca
2281 gcagcgggga catcgctgac gcgctcagcg tgcactcggc cggctcctct agctccgacg
2341 tggaggagat caacatcagc ttcgtcccgg agtctcctga tggccaggaa aagaagttct
2401 gggaatcagc ctcacagtca tccccggaga cctccggcat cagctcagcc tccagcagca
2461 cctcgtcctc ctcagcctcc accacgcccg tggctgccac acgcacccac aagcgctctg
2521 tctcagggct ctgcaactcc agctccgcgc tgccgctcta caaccagcag gtgggcgact
2581 gctgtatcat ccgcgtcagc ctggacgtgg acaatggcaa catgtacaag agcatcctgg
2641 tgaccagcca agataaggct ccggctgtaa tccgcaaggc catggacaaa cacaacctgg
2701 aggaggagga gccggaggac tatgagctgc tgcagattct ctcagatgac cggaagctga
2761 agatccctga aaacgccaac gtcttctatg ccatgaactc taccgccaac tatgactttg
2821 tcctcaagaa gcggaccttc accaagggag tgaaggtcaa gcacggagcc agctccaccc
2881 tccctcgcat gaagcagaaa ggactcaaga ttgccaaggg catcttctaa tcaactttct
2941 tgtacaaagt tggcattata ggaaagcatt gcttatcaat ttgttgcaac gaacaggtca
3001 ctatcagtca aaataaaatc attatttgca tgcaagctta gatcctctag accagccagg
3061 acagaaatgc ctcgacttcg ctgctaccca aggttgccgg gtgacgcaca ccgtggaaac
3121 ggatgaaggc acgaacccag tggacataag cctgttcggt tcgtaagctg taatgcaagt
3181 agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga acgcagcggt ggtaacggcg
3241 cagtggcggt tttcatggct tgttatgact gtttttttgg ggtacagtct atgcctcggg
3301 catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg agcagcaacg
3361 atgttacgca gcagggcagt cgccctaaaa caaagttaaa cattatgagg gaagcggtga
3421 tcgccgaagt atcgactcaa ctatcagagg tagttggcgt catcgagcgc catctcgaac
3481 cgacgttgct ggccgtacat ttgtacggct ccgcagtgga tggcggcctg aagccacaca
3541 gtgatattga tttgctggtt acggtgaccg taaggcttga tgaaacaacg cggcgagctt
3601 tgatcaacga ccttttggaa acttcggctt cccctggaga gagcgagatt ctccgcgctg
3661 tagaagtcac cattgttgtg cacgacgaca tcattccgtg gcgttatcca gctaagcgcg
3721 aactgcaatt tggagaatgg cagcgcaatg acattcttgc aggtatcttc gagccagcca
3781 cgatcgacat tgatctggct atcttgctga caaaagcaag agaacatagc gttgccttgg
3841 taggtccagc ggcggaggaa ctctttgatc cggttcctga acaggatcta tttgaggcgc
3901 taaatgaaac cttaacgcta tggaactcgc cgcccgactg ggctggcgat gagcgaaatg
3961 tagtgcttac gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc gcgccgaagg
4021 atgtcgctgc cgactgggca atggagcgcc tgccggccca gtatcagccc gtcatacttg
4081 aagctagaca ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt
4141 tggaagaatt tgtccactac gtgaaaggcg agatcaccaa ggtagtcggc aaataaccct
4201 cgagccaccc atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag
4261 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
4321 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
4381 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
4441 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
4501 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
4561 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
4621 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
4681 attgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
4741 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
4801 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
4861 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
4921 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
4981 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
5041 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
5101 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
5161 caattaatac gcgtaccgct agccaggaag agtttgtaga aacgcaaaaa ggccatccgt
5221 caggatggcc ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca
5281 ccctccgggc cgttgcttca caacgttcaa atccgctccc ggcggatttg tcctactcag
5341 gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtcttccg actgagcctt
5401 tcgttttatt tgatgcctgg cagttcccta ctctcgcg
//
| {
"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": "RALGDS",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "2745",
"Mutation": "",
"Entrez Gene": "RALGDS",
"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": ""
} |
|
Talin1 head-mCherry | 212005 | Expression of mouse talin1 head (1-490)-mCherry in mammalian cells | Vesa Hytönen | Rahikainen et al J Cell Sci. 2019 Apr 3;132(7):jcs226514. doi: 10.1242/jcs.226514. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/423003/88e2b52a-9e7c-43e9-938d-ced50154b779/addgene-plasmid-212005-sequence-423003.gbk | full | LOCUS sequence-423003- 6143 bp DNA circular SYN 27-AUG-2024
DEFINITION Expression of mouse talin1 head (1-490)-mCherry in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6143)
AUTHORS Rahikainen R, Ohman T, Turkki P, Varjosalo M, Hytonen VP
TITLE Talin-mediated force transmission and talin rod domain unfolding
independently regulate adhesion signaling.
JOURNAL J Cell Sci. 2019 Apr 3;132(7):jcs226514. doi: 10.1242/jcs.226514.
PUBMED 30837291
REFERENCE 2 (bases 1 to 6143)
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
Sci."; date: "2019-04-3"; volume: "132(7):jcs226514. doi"; pages: "
10.1242/jcs.226514"
FEATURES Location/Qualifiers
source 1..6143
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 61..364
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 365..568
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 519..539
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
CDS 2080..2790
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
primer_bind complement(2225..2243)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 2493..2512
/label=mCherry-F
/note="mCherry, forward primer"
polyA_signal 2931..3052
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2968..2987)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3022..3041
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(3059..3514)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3196..3217)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 3408..3427
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 3541..3645
/gene="bla"
/label=AmpR promoter
promoter 3647..4004
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
enhancer 3647..3838
/label=SV40 enhancer
/note="enhancer for the SV40 early promoter (Herr, 1993)"
primer_bind complement(3670..3690)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 3855..3990
/label=SV40 ori
/note="SV40 origin of replication"
CDS 4039..4833
/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 4039..4833
/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(4093..4112)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4703..4722
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(5021..5040)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 5065..5112
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 5441..6029
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5930..5949
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
601 ccggtcacca tggttgcgct ttcgctgaag attagcattg ggaatgtggt gaagacgatg
661 caatttgagc catctaccat ggtgtacgac gcctgccgca tgattcgtga gcggatccca
721 gaggccctgg ctggccctcc caacgacttt gggctctttc tgtcagatga tgaccccaaa
781 aaaggcatct ggctagaggc tgggaaagct ttggactact acatgctccg aaatggggac
841 accatggagt acagaaaaaa gcagagaccc ctgaagatcc ggatgttaga cggaacggtg
901 aagactatca tggtggatga ctccaagacc gtcacagaca tgctcatgac catttgtgcc
961 cgaattggta tcaccaacca tgatgagtat tcactggttc gagagctgat ggaagaaaag
1021 aaagatgagg ggacagggac cctgagaaaa gacaagaccc tactgcgaga tgaaaagaag
1081 atggagaagc taaagcagaa gttgcacaca gacgatgagt tgaactggct ggaccatggg
1141 cggacactga gggaacaggg agtggaagag catgagacgt tgctgctgcg gagaaagttc
1201 ttctactcag accagaatgt ggattcccga gaccctgtac agctgaacct tctctatgtg
1261 caggcacgag atgacatcct gaatggctcc catcctgtct cctttgacaa ggcctgtgaa
1321 tttgcaggct tccagtgcca gatccagttt ggacctcaca atgaacagaa gcacaaggct
1381 ggcttccttg acctgaagga cttcctgccc aaggagtatg tgaagcagaa gggagagcgt
1441 aagatctttc aggcacacaa gaattgtggg cagatgagtg aaattgaggc caaggtacgc
1501 tacgtgaagc tggcccgttc cctcaagact tacggtgtct ccttcttcct agtcaaggaa
1561 aagatgaagg ggaagaataa actggtgccc aggctgttgg gcatcactaa ggagtgtgtg
1621 atgcgtgtgg atgagaagac caaggaggtg attcaggaat ggagcctcac caacatcaaa
1681 cgctgggctg cctctcccaa gagctttact ctggactttg gagactatca ggatggctac
1741 tactcagtac agacgacaga aggcgagcag atcgcacagc tcattgctgg ctacatagat
1801 atcatcctta agaagaaaaa aagcaaggac cattttgggc tggagggaga tgaagagtct
1861 actatgttgg aggactcagt gtctcccaaa aagtcaacag tccttcagca gcagtacaac
1921 cgagtgggga aagtggagca cggctctgtg gctctgccag ccatcatgcg ctctggagcc
1981 tctggtcctg agaatttcca ggtggggagc atgccacctg cccagcagca gatcaccagt
2041 ggccagatgc accgaggaca catgccacct ctgactagta tggtgagcaa gggcgaggag
2101 gataacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga gggctccgtg
2161 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag
2221 accgccaagc tgaaggtgac caagggtggc cccctgccct tcgcctggga catcctgtcc
2281 cctcagttca tgtacggctc caaggcctac gtgaagcacc ccgccgacat ccccgactac
2341 ttgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc
2401 ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg
2461 aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaagaa gaccatgggc
2521 tgggaggcct cctccgagcg gatgtacccc gaggacggcg ccctgaaggg cgagatcaag
2581 cagaggctga agctgaagga cggcggccac tacgacgctg aggtcaagac cacctacaag
2641 gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt ggacatcacc
2701 tcccacaacg aggactacac catcgtggaa cagtacgaac gcgccgaggg ccgccactcc
2761 accggcggca tggacgagct gtacaagtaa ccgcgggccc gggatccacc ggatctagat
2821 aactgatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
2881 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
2941 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
3001 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct taacgcgtaa
3061 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
3121 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
3181 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
3241 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
3301 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
3361 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
3421 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
3481 cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg
3541 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
3601 caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga
3661 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
3721 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
3781 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
3841 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
3901 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
3961 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca
4021 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
4081 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
4141 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
4201 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc
4261 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
4321 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
4381 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
4441 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
4501 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
4561 aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
4621 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
4681 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
4741 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
4801 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
4861 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
4921 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
4981 tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa
5041 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg
5101 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
5161 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
5221 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct
5281 caggttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
5341 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
5401 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
5461 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
5521 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
5581 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
5641 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
5701 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
5761 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
5821 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
5881 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
5941 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
6001 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
6061 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
6121 ataaccgtat taccgccatg cat
//
| {
"Vector backbone": "pEGFP-C1",
"Backbone size w/o insert (bp)": "3962",
"Vector type": "Mammalian Expression",
"Selectable markers": "Neomycin (select with G418)"
} | {
"Bacterial Resistance(s)": "Kanamycin, 50 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA ; Takara Bio Limited Use Label License (formerly Clontech)",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Talin-mediated force transmission and talin rod domain unfolding independently regulate adhesion signaling",
"Publication": "J Cell Sci. 2019 Apr 3;132(7):jcs226514. doi: 10.1242/jcs.226514.",
"DOI": "10.1242/jcs.226514.",
"Authors": "Rahikainen R, Ohman T, Turkki P, Varjosalo M, Hytonen VP /jcs",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/30837291",
"PubMed ID": "PubMed 30837291"
} | {
"Gene/Insert name": "Mouse talin1 head domain (1-490)",
"Species": "M. musculus (mouse)",
"Insert Size (bp)": "2181",
"Mutation": "",
"Entrez Gene": "Tln1",
"Promoter": "CMV",
"Tag / Fusion Protein": "mCherry fluorescent protein (C terminal on insert)"
} | {
"Cloning method": "Unknown",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "acgcaaatgggcggtagg",
"3′ sequencing primer": "GATGAGTTTGGACAAACCAC"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pD649-HAsp-TREM1-Fc(DAPA)-AviTag-6xHis | 156593 | Mammalian expression of cell-surface protein extracellular domain fused to Fc(DAPA)-Avi-6xHis. Protein is secreted from cells. | Chris Garcia | Wojtowicz et al Cell. 2020 Aug 20;182(4):1027-1043.e17. doi: 10.1016/j.cell.2020.07.025. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/420989/1ad8018b-9ba4-4eac-a960-07f3f327c9e9/addgene-plasmid-156593-sequence-420989.gbk | full | LOCUS sequence-420989- 6661 bp DNA circular SYN 26-AUG-2024
DEFINITION Mammalian expression of cell-surface protein extracellular domain
fused to Fc(DAPA)-Avi-6xHis. Protein is secreted from cells..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6661)
AUTHORS Wojtowicz WM, Vielmetter J, Fernandes RA, Siepe DH, Eastman CL,
Chisholm GB, Cox S, Klock H, Anderson PW, Rue SM, Miller JJ, Glaser
SM, Bragstad ML, Vance J, Lam AW, Lesley SA, Zinn K, Garcia KC
TITLE A Human IgSF Cell-Surface Interactome Reveals a Complex Network of
Protein-Protein Interactions.
JOURNAL Cell. 2020 Aug 20;182(4):1027-1043.e17. doi:
10.1016/j.cell.2020.07.025.
PUBMED 32822567
REFERENCE 2 (bases 1 to 6661)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1016/j.cell.2020.07"; journalName: "Cell"; date: "2020-08-20-
20"; volume: "182"; issue: "4"; pages: "1027-1043.e17"
FEATURES Location/Qualifiers
source 1..6661
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 96..116
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
polyA_signal 234..345
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind 334..351
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
CDS complement(491..508)
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS complement(509..553)
/codon_start=1
/product="peptide tag that allows for enzymatic
biotinylation"
/label=AviTag(TM)
/translation="GLNDIFEAQKIEWHE"
primer_bind complement(1925..1944)
/label=pMT2-F
/note="Synthetic intron, forward primer"
promoter complement(1957..1975)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(1958..1975)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(2185..2209)
/label=LNCX
/note="Human CMV promoter, forward primer"
promoter complement(2206..2409)
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind complement(2235..2255)
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
enhancer 2410..2789
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
protein_bind 2956..2972
/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(3428..3445)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(3428..3444)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3543..3560)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3714..4302)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3794..3813)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind complement(4388..4407)
/label=pENTR-R
/note="pENTR vectors, reverse primer"
primer_bind complement(4413..4432)
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
promoter 4474..4565
/gene="bla"
/label=AmpR promoter
CDS 4566..5426
/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(4784..4803)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
polyA_signal 5479..5600
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5490..5509)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 5544..5563
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
CDS complement(5730..6329)
/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(5813..5833)
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
primer_bind 6310..6329
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
promoter complement(6348..6395)
/label=EM7 promoter
/note="synthetic bacterial promoter "
promoter complement(join(6443..6661,1..111))
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 6457..6592
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(6511..6530)
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
ORIGIN
1 tggggagcct ggggactttc cacacctggt tgctgactaa ttgagatgca tgctttgcat
61 acttctgcct gctggggagc ctggggactt tccacaccct aactgacaca cattccacag
121 aattaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
181 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgtgatgcaa
241 tttcctcatt ttattaggaa aggacagtgg gagtggcacc ttccagggtc aaggaaggca
301 cgggggaggg gcaaacaaca gatggctggc aactagaagg cacagtcgag gctgattttc
361 tctatctatt tattttacta ctcaagacta tcttgcaaag tagaagaacc ctgtctgcag
421 tatgacgacc gtgtataccc gattggctta ccgggtaacg cgttgtgcca gtgactacag
481 gccggcctta gtggtgatgg tggtggtgtt catgccactc aatcttctga gcctcgaaaa
541 tatcattaag accgctgccg ccggggctca ggctcaggga cttctgggtg tagtggttgt
601 gcagggcctc gtgcatcacg ctgcagctga acacgttgcc ctgctgccac ctggacttgt
661 ccacggtcag cttgctgtac aggaagaagc tgccgtcgct gtccagcact gggggggtgg
721 tcttgtagtt gttctcgggc tggccgttgc tctcccactc cacggcgata tcgctggggt
781 agaagccctt caccagacag gtcagggaca cctggttctt ggtcatctcc tcccggctgg
841 ggggcagggt gtacacctgg ggctcccgtg gctggccctt ggccttgctg attgtctttt
901 cgattggggc agccagggcc ttgttggaga ctttgcactt gtattctttg ccgttcagcc
961 agtcctggtg cagcacggtc agcacggaca ccaccctgta ggtgctgttg tactgctcct
1021 ctctgggctt ggtcttggcg ttgtgcacct ccacgccgtc cacgtaccag ttgaacttca
1081 cctctgggtc ctcgtgggac acggccacca ccacgcaggt cacctcgggg gtcctgctga
1141 tcatcagggt gtccttgggc ttggggggga acaggaacac ggaaggccct cccagcagtt
1201 ctggagctgg gcaggggggg caggtgtggg tcttgtcgcc gcccggcgcg ccattgaaca
1261 cgggcaccct gatgatgtcg gtcacgttgg tcaggttgat ctcgctgtcg ggtgtagaca
1321 catcggcggt gctcttaggt ggggcctgtg tcactgttct ggggcttgtg tacagagggc
1381 acagagcctt ggtggtggta ggagggatct tgtacacgtt ctgggtgctg ttctcgttgc
1441 tgccaggggt gccagaaaag cccttggtga ccaccagtct gatccggtcg aacagcatgt
1501 ggggttcttt aggaggctgg tagatcacac actggtacag gccgctatcc tcgacctgca
1561 gattgaccat ccgcactctc agcaggccgt gatcgtggta atcttccagg atgatgcggc
1621 ccacttgcac agggtgagaa ttcttgctgg gcctctcggt gcaggccagt gtcttaggca
1681 tctcgccatc tctgatgatc tgccaggcct tctgagagct ggcgaacttt tccagggtgt
1741 agtcgcactt cacgtccagt gtctggccct ctttcagctc gtacttctct tctgtcagct
1801 ttgttgcggc ggccgcagcg aacaccaggc agaagatgta gctcagggca atgatggtct
1861 tcatggtggc gctagcaggc gcgttttcga tagaaccgag gtgcagttgg acctgggagt
1921 ggacacctgt ggagagaaag gcaaagtgga tgttattcta tagtgtcacc taaatcgtat
1981 gtgtatgata cataaggtta tgtattaatt gtagccgcgt tctaacgaag ccaagggggt
2041 gggcctatag actctatagg cggtacttac gtcactcttg gcacggggaa tccgcgttcc
2101 aatgcaccgt tcccggccgc ggaggctgga tcggtcccgg tgtcttctat ggaggtcaaa
2161 acagcgtgga tggcgtctcc aggcgatctg acggttcact aaacgagctc tgcttatata
2221 gacctcccac cgtacacgcc taccgcccat ttgcgtcaat ggggcggagt tgttacgaca
2281 ttttggaaag tcccgttgat tttggtgcca aaacaaactc ccattgacgt caatggggtg
2341 gagacttgga aatccccgtg agtcaaaccg ctatccacgc ccattgatgt actgccaaaa
2401 ccgcatcacc atggtaatag cgatgactaa tacgtagatg tactgccaag taggaaagtc
2461 ccataaggtc atgtactggg cataatgcca ggcgggccat ttaccgtcat tgacgtcaat
2521 agggggcgta cttggcatat gatacacttg atgtactgcc aagtgggcag tttaccgtaa
2581 atactccacc cattgacgtc aatggaaagt ccctattggc gttactatgg gaacatacgt
2641 cattattgac gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc catttaccgt
2701 aagttatgta acgcggaact ccatatatgg gctatgaact aatgaccccg taattgatta
2761 ctattaataa ctagtcaata atcaatgcca acatggcggt catattggac atgagccaat
2821 ataaatgtac atattatgat atagatacaa cgtatgcaat ggccaatagc caatattgat
2881 ttatgctata taaccaatga ataatatggc taatggccaa tattgagtca acgccactgt
2941 gcatgaagct cgtaattgtt atccgctcac aattaagggc gacacaaaat ttattctaaa
3001 tgataataaa tactgataac atcttatagt ttgtattata ttttgtatta tcgttgacat
3061 gtataatttt gatatcaaaa actgattttc cctttattat tttcgagatt tattttctta
3121 attctcttta acaaactaga aatattgtat atacaaaaaa tcataaataa tagatgaata
3181 gtttaattat aggtgttcat caatcgaaaa agcaacgtat cttatttaaa gtgcgttgct
3241 tttttctcat ttataaggtt aaataattct catatatcaa gcaaagtgac aggcgccctt
3301 aaatattctg acaaatgctc tttccctaaa ctccccccat aaaaaaaccc gccgaagcgg
3361 gtttttacgt tatttgcgga ttaacgatta ctcgttatca gaaccgccca gggggcccga
3421 gcttaagact ggccgtcgtt ttacaacaca gaaagagttt gtagaaacgc aaaaaggcca
3481 tccgtcaggg gccttctgct tagtttgatg cctggcagtt ccctactctc gccttccgct
3541 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
3601 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
3661 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
3721 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
3781 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
3841 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
3901 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
3961 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
4021 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
4081 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg ggctaactac
4141 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
4201 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
4261 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
4321 tctacggggt ctgacgctca gtggaacgac gcgcgcgtaa ctcacgttaa gggattttgg
4381 tcatgagcaa ggggtgttat gagccatatt caggtataaa tgggctcgcg ataatgttca
4441 gaattggtta attggttgta acactgaccc ctatttgttt atttttctaa atacattcaa
4501 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
4561 agaatatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
4621 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
4681 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
4741 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
4801 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
4861 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
4921 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
4981 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
5041 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
5101 cgatgcctgt agcgatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
5161 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
5221 tgcgctcggc ccttccggct ggctggttta ttgctgataa atccggagcc ggtgagcgtg
5281 gttctcgcgg tatcatcgca gcgctggggc cagatggtaa gccctcccgt atcgtagtta
5341 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
5401 gtgcctcact gattaagcat tggtaatgac catgattacg ccaagctcta gctagaggtc
5461 gacggtatac agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
5521 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
5581 taagctgcaa taaacaagtt ggggtgggcg aagaactcca gcatgagatc cccgcgctgg
5641 aggatcatcc agccggcgtc ccggaaaacg attccgaagc ccaacctttc atagaaggcg
5701 gcggtggaat cgaaatctcg tagcacgtgt caggcaccgg gcttgcgggt catgcaccag
5761 gtgcgcggtc cttcgggcac ctcgacgtcg gcggtgacgg tgaagccgag ccgctcgtag
5821 aaggggaggt tgcggggcgc ggatgtctcc aggaaggcgg gcaccccggc gcgctcggcc
5881 gcctccactc cggggagcac gacggcgctg cccagaccct tgccctggtg gtcgggcgag
5941 acgccgacgg tggccaggaa ccacgcgggc tccttgggcc ggtgcggcgc caggaggcct
6001 tccatctgtt gctgcgcggc cagccgggaa ccgctcaact cggccatgcg cgggccgatc
6061 tcggcgaaca ccgcccccgc ttcgacgctc tccggcgtgg tccagaccgc caccgcggcg
6121 ccgtcgtccg cgacccacac cttgccgatg tcgagcccga cgcgcgtgag gaagagttct
6181 tgcagctcgg tgacccgctc gatgtggcgg tccggatcga cggtgtggcg cgtggcgggg
6241 tagtcggcga acgcggcggc gagggtgcgt acggccctgg ggacgtcgtc gcgggtggcg
6301 aggcgcaccg tgggcttgta ctcggtcatg gtttagttcc tcaccttgtc gtattatact
6361 atgccgatat actatgccga tgattaattg tcaacacgtg ctgatcagat ccgaaaatgg
6421 atatacaagc tcccgggagc tttttgcaaa agcctaggcc tccaaaaaag cctcctcact
6481 acttctggaa tagctcagag gcagaggcgg cctcggcctc tgcataaata aaaaaaatta
6541 gtcagccatg gggcggagaa tgggcggaac tgggcggagt taggggcggg atgggcggag
6601 ttaggggcgg gactatggtt gctgactaat tgagatgcat gctttgcata cttctgcctg
6661 c
//
| {
"Vector backbone": "pD649",
"Backbone size w/o insert (bp)": "6106",
"Vector type": "Mammalian Expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "A Human IgSF Cell-Surface Interactome Reveals a Complex Network of Protein-Protein Interactions",
"Publication": "Cell. 2020 Aug 20;182(4):1027-1043.e17. doi: 10.1016/j.cell.2020.07.025.",
"DOI": "10.1016/j.cell.2020.07.025.",
"Authors": "Wojtowicz WM, Vielmetter J, Fernandes RA, Siepe DH, Eastman CL, Chisholm GB, Cox S, Klock H, Anderson PW, Rue SM, Miller JJ, Glaser SM, Bragstad ML, Vance J, Lam AW, Lesley SA, Zinn K, Garcia KC /jcell",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/32822567",
"PubMed ID": "PubMed 32822567"
} | {
"Gene/Insert name": "TREM1",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "555",
"Mutation": "Fc region of human IgG contains D265A and P329A mutations (DAPA)",
"Entrez Gene": "TREM1",
"Promoter": "CMV/SP6",
"Tag / Fusion Protein": "Fc(DAPA)-AviTag-6xHis (C terminal on backbone)"
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "GCGGCCGC (not destroyed)",
"3′ cloning site": "GGCGCGCC (not destroyed)",
"5′ sequencing primer": "ATTTAGGTGACACTATAG",
"3′ sequencing primer": "CACGTACCAGTTGAACTTCACC"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
WRB_pCSdest | 53856 | Roger Reeves | Edie et al G3 (Bethesda). 2018 Jul 2;8(7):2215-2223. doi: 10.1534/g3.118.200144. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/165457/1d443af5-70ca-4fda-8782-4586b35b6b23/addgene-plasmid-53856-sequence-165457.gbk | partial | LOCUS sequence-165457- 931 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 931)
AUTHORS Edie S, Zaghloul NA, Leitch CC, Klinedinst DK, Lebron J, Thole JF,
McCallion AS, Katsanis N, Reeves RH
TITLE Survey of Human Chromosome 21 Gene Expression Effects on Early
Development in Danio rerio.
JOURNAL G3 (Bethesda). 2018 Jul 2;8(7):2215-2223. doi:
10.1534/g3.118.200144.
PUBMED 29760202
REFERENCE 2 (bases 1 to 931)
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: "10.1534/g3.118";
journalName: "G3 (Bethesda)"; date: "2018-07-2- 2"; volume: "8";
issue: "7"; pages: "2215-2223"
FEATURES Location/Qualifiers
source 1..931
/mol_type="genomic DNA"
/organism="unspecified"
protein_bind 38..62
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
protein_bind complement(591..615)
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
promoter complement(669..687)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(670..687)
/label=SP6
/note="SP6 promoter, forward primer"
ORIGIN
1 atacgactca ctatagttct agaggctcga gaggatcacc actttgtaca agaaagctgg
61 gtttcagctg aacggatgaa gcacaatagc gacaactttg ttacagacta aaatccaaca
121 ggtaattcca acaccacctg ctactctagt aggaaaggct accaggcggt ctaggggggt
181 tatccattta ctcggcacga cagccacagg gacagaataa tacttccaaa tgagtgagat
241 catcagggca gcctgcaata cgtagaaagc gacacttatc acccatttta tcttggctaa
301 ttgagctgtc cgagctttca catgggtttt gagcttatcc gtcatcttgt tgatctttct
361 ttccagcctg gcatatctgg caaactcgtc catcatgttg actgtggaga gctcctgctt
421 catgtcctgg atctccgctc tcatctgtga ctcctgctcc gcgtccttct gcagcaccct
481 ggacatgaag gatgagaagg acgggaggag gatcctaaga acattgcatc caaacacgaa
541 gctgagcacc agcaaccacg cccagtggtc ggccgcggct gagctcatgg tgcctgcttt
601 tttgtacaaa cttgtgatcc ttgaattcga atcgatggga tcctgcaaaa agaacaagta
661 gcttgtattc tatagtgtca cctaaatcaa gcttgctccg acgtccccag gcagaatggc
721 ggttccctaa acgagcattg cttatataga cctcccatta ggcacgccta ccgcccattt
781 acgtcaatgg aacgcccatt tgcgtcattg cccctcccca ttgacgtcaa tggggatgta
841 cttggcagcc atcgcgggcc atttaccgcc attgacgtca atgggagtac tgccaatgta
901 ccctggcgta cttccaatag taatgtactt g
//
| {
"Vector backbone": "pCSDest",
"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)": "Top10",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Industry MTA"
} | {
"Title": "Survey of Human Chromosome 21 Gene Expression Effects on Early Development in Danio rerio",
"Publication": "G3 (Bethesda). 2018 Jul 2;8(7):2215-2223. doi: 10.1534/g3.118.200144.",
"DOI": "10.1534/g3.118.200144.",
"Authors": "Edie S, Zaghloul NA, Leitch CC, Klinedinst DK, Lebron J, Thole JF, McCallion AS, Katsanis N, Reeves RH /g",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/29760202",
"PubMed ID": "PubMed 29760202"
} | {
"Gene/Insert name": "WRB",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "GET1",
"Promoter": "SP6 and CMV",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Gateway Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "CMV-F",
"3′ sequencing primer": "EBV-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": ""
} |
||
pCASP-SptP120-TD-HilA | 153330 | Triggers secretion of SptP120 fused to the HA4-7c12 Tandem Monobody from Salmonella upon induction with arabinose | Josef Penninger | Chabloz et al Commun Biol. 2020 Jul 3;3(1):342. doi: 10.1038/s42003-020-1072-4. | partial | {
"Vector backbone": "pCASP-HilA",
"Backbone size w/o insert (bp)": "",
"Vector type": "Salmonella expression",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Chloramphenicol, 25 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Salmonella-based platform for efficient delivery of functional binding proteins to the cytosol",
"Publication": "Commun Biol. 2020 Jul 3;3(1):342. doi: 10.1038/s42003-020-1072-4.",
"DOI": "10.1038/s42003-020-1072-4.",
"Authors": "Chabloz A, Schaefer JV, Kozieradzki I, Cronin SJF, Strebinger D, Macaluso F, Wald J, Rabbitts TH, Pluckthun A, Marlovits TC, Penninger JM /s---",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/32620833",
"PubMed ID": "PubMed 32620833"
} | {
"Gene/Insert name": "SptP120-HA4-7c12 Tandem Monobody",
"Species": "Synthetic",
"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": ""
} |
|||
pFETCh_KDM1A | 86260 | Donor vector for 3' FLAG tag of human KDM1A | Savic et al Genome Res. 2015 Sep 9. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/168141/3e70b91d-1a99-4b3f-bf8b-3eca51a481d9/addgene-plasmid-86260-sequence-168141.gbk | full | LOCUS sequence-168141- 7154 bp DNA circular SYN 25-AUG-2024
DEFINITION Donor vector for 3' FLAG tag of human KDM1A.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7154)
AUTHORS Savic D, Partridge EC, Newberry KM, Smith SB, Meadows SK, Roberts
BS, Mackiewicz M, Mendenhall EM, Myers RM
TITLE CETCh-seq: CRISPR epitope tagging ChIP-seq of DNA-binding proteins.
JOURNAL Genome Res. 2015 Sep 9.
PUBMED 26355004
REFERENCE 2 (bases 1 to 7154)
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: "Genome Res.
2015 Sep 9."
FEATURES Location/Qualifiers
source 1..7154
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 837..860
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS 864..887
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS 891..914
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS 930..986
/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 999..1793
/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(1053..1072)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 1663..1682
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(2656..2672)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2665..2687)
/label=M13/pUC Forward
/note="In lacZ gene"
CDS complement(2891..3706)
/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="MSHIQRETSCSKPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind 3614..3633
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
primer_bind 3698..3717
/label=pENTR-R
/note="pENTR vectors, reverse primer"
rep_origin 4086..4674
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4575..4594
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 4828..4845
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 5145..5166
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 5181..5211
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5219..5235
/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 5224..5246
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 5243..5259
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 5243..5259
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
promoter 5436..5638
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 5589..5609
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 5635..5659
/label=LNCX
/note="Human CMV promoter, forward primer"
CDS 5735..6865
/codon_start=1
/product="herpes simplex virus thymidine kinase"
/label=HSV TK
/note="confers sensitivity to nucleoside analogs such as
acyclovir or ganciclovir"
/translation="MASYPCHQHASAFDQAARSRGHSNRRTALRPRRQQEATEVRLEQK
MPTLLRVYIDGPHGMGKTTTTQLLVALGSRDDIVYVPEPMTYWQVLGASETIANIYTTQ
HRLDQGEISAGDAAVVMTSAQITMGMPYAVTDAVLAPHIGGEAGSSHAPPPALTLIFDR
HPIAALLCYPAARYLMGSMTPQAVLAFVALIPPTLPGTNIVLGALPEDRHIDRLAKRQR
PGERLDLAMLAAIRRVYGLLANTVRYLQGGGSWREDWGQLSGTAVPPQGAEPQSNAGPR
PHIGDTLFTLFRAPELLAPNGDLYNVFAWALDVLAKRLRPMHVFILDYDQSPAGCRDAL
LQLTSGMVQTHVTTPGSIPTICDLARTFAREMGEAN"
ORIGIN
1 ttcagaatgg aggagtccag agtataagtc aaataacatt tttccttatt caggttttct
61 ccctaaaaaa caaaaccatt tttaattgca cttccatttt gtaatggctc ataggattgc
121 tgtaaggtct caattactag gggcttccag ggcatttctg agaaataacc ctgggtcctt
181 gtctagaccc ttatgccaga ccccactcca aagagcggta agaattcctt agtgtcatag
241 cccagacctg ctgagctgcg aggcttaagt gtccctgatc accaaatgtc ctgtgcttca
301 gggtagcgag gctccctccc agaaggtagt tatacatgag ggaaggacgc tttacaactg
361 ggtatctaaa ctgatgagaa cacatgttaa gcatcacttt aggactgagc ctaggtagag
421 ttttattgtc tcatttctac ttgtcaattc tgggaaagtg cctactgata agggagactc
481 ttcgatagaa tgatgaatag taattggggg ggtcagcctt taaaaaggtc aacagcaatt
541 taagtactta gcaatttaag tacaagaata aaggtatatg tgcagcctgc caattttctc
601 tttttcccct aaaatagccg attccacgac tcttctttgc gggagaacat acgatccgta
661 actacccagc cacagtgcat ggtgctctgc tgagtgggct gcgagaagcg ggaagaattg
721 cagaccagtt tttgggggcc atgtatacgc tgcctcgcca ggccacacca ggtgttcctg
781 cacagcagtc cccaagcatg gggaacggag gaggttccgg tggaggtggt tctggagatt
841 acaaggatga cgacgataag ggcgattaca aggatgacga cgataaggga gattacaagg
901 atgacgacga taaggtttca ggaagcggag ctactaactt cagcctgctg aagcaggctg
961 gagacgtgga ggagaaccct ggacctggat cgtttcgcat gattgaacaa gatggattgc
1021 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
1081 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
1141 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
1201 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
1261 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
1321 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
1381 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
1441 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
1501 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
1561 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
1621 actgtggccg gctgggtgtg gccgaccgct atcaggacat agcgttggct acccgtgata
1681 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
1741 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgattcgaac
1801 atctaaggga agaggcccat gtgcctgttt ctgccatgta aggaaggctc ttctagcaat
1861 actagatccc actgagaaaa tccaccctgg catctgggct cctgatcagc tgatggagct
1921 cctgatttga caaaggagct tgcctccttt gaatgaccta gagcacaggg aggaacttgt
1981 ccattagttt ggaattgtgt tcttcgtaaa gactgaggca agcaagtgct gtgaaataac
2041 atcatcttag tcccttggtg tgtggggttt ttgttttttt tttatatttt gagaataaaa
2101 cttcatataa aattggccct ctcttttgtt cctttgagtt ggagttacat acaagatggg
2161 ggcattactc ctttggaaat gccaagttta tcctatggca atagtaatac atgttttctc
2221 catgtttaaa taaggaagaa gatgttcata taaaagagaa aaatggtttg atcaggtcag
2281 aagataaggt aggggggccg gtcctgtaga agcagtggac ttcctggtca gtcggactag
2341 gcttcctcac ttgggtgggg gtggagtgtg gtggtgtgcc aagaacatgg ttattaggaa
2401 actttggatg cactttcaag atagaacaca aaagagctga aacaaagtct gagcctttgg
2461 gagaccagtc tgcctttatt cctgattccc tccatgggta tgtgctaaaa cttcccaatt
2521 ctactaaagg tggtggtgta cattttggga agcgcactac ataggatcaa gggcttaggt
2581 aacagctggt ttaacagatt gcccttggaa agtcctctcc actgactgta gcctccaatt
2641 cactggccgt cgttcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
2701 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
2761 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gcgatttatt
2821 caacaaagcc gccgtcccgt caagtcagcg taatgctctg ccagtgttac aaccaattaa
2881 ccaattctga ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag
2941 gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga
3001 ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat
3061 caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat
3121 gagtgacgac tgaatccggt gagaatggca aaagcttatg catttctttc cagacttgtt
3181 caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca
3241 ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct gttaaaagga caattacaaa
3301 caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg
3361 aatcaggata ttcttctaat acctggaatg ctgttttccc ggggatcgca gtggtgagta
3421 accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg
3481 tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat
3541 gtttcagaaa caactctggc gcatcgggct tcccatacaa tcgatagatt gtcgcacctg
3601 attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat
3661 ttaatcgcgg cttcgagcaa gacgtttccc gttgaatatg gctcataaca ccccttgtat
3721 tactgtttat gtaagcagac agttttattg ttcatgatga tatattttta tcttgtgcaa
3781 tgtaacatca gagattttga gacacaacgt ggctttgttg aataaatcga acttttgctg
3841 agttgaagga tcagatcacg catcttcccg acaacgcaga ccgttccgtg gcaaagcaaa
3901 agttcaaaat caccaactgg tccacctaca acaaagctct catcaaccgt ggctccctca
3961 ctttctggct ggatgatggg gcgattcagg cctggtatga gtcagcaaca ccttcttcac
4021 gaggcagacc tctcgacgga gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
4081 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
4141 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
4201 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
4261 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
4321 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
4381 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
4441 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
4501 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
4561 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
4621 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
4681 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
4741 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
4801 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagaagct
4861 cgcacattca gcagcgtttt tcagcgcgtt ttcgatcaac gtttcaatgt tggtatcaac
4921 accaggttta actttgaact tatcggcact gacggttact gattttgaac ttttgctttg
4981 ccacggaacg gtctgcgttg tcgggaagat gcgtgatctg atccttcaac tcagcaaaag
5041 ttcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
5101 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
5161 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
5221 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcttgca
5281 tgcctgcagg tcgactctag aggatccccg ggtaccgagc tcgaattagc ttgcatgcct
5341 gcaggtcaat tccctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
5401 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca
5461 atgggcgtgg atagcggttt gactcacggg gatttcaagt ctccacccca ttgacgtcaa
5521 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
5581 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg
5641 tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccataggag
5701 acaccgggac cgatccagcc tggggatccc gaccatggct tcgtacccct gccatcaaca
5761 cgcgtctgcg ttcgaccagg ctgcgcgttc tcgcggccat agcaaccgac gtacggcgtt
5821 gcgccctcgc cggcagcaag aagccacgga agtccgcctg gagcagaaaa tgcccacgct
5881 actgcgggtt tatatagacg gtcctcacgg gatggggaaa accaccacca cgcaactgct
5941 ggtggccctg ggttcgcgcg acgatatcgt ctacgtaccc gagccgatga cttactggca
6001 ggtgctgggg gcttccgaga caatcgcgaa catctacacc acacaacacc gcctcgacca
6061 gggtgagata tcggccgggg acgcggcggt ggtaatgaca agcgcccaga taacaatggg
6121 catgccttat gccgtgaccg acgccgttct ggctcctcat atcggggggg aggctgggag
6181 ctcacatgcc ccgcccccgg ccctcaccct catcttcgac cgccatccca tcgccgccct
6241 cctgtgctac ccggccgcgc gataccttat gggcagcatg accccccagg ccgtgctggc
6301 gttcgtggcc ctcatcccgc cgaccttgcc cggcacaaac atcgtgttgg gggcccttcc
6361 ggaggacaga cacatcgacc gcctggccaa acgccagcgc cccggcgagc ggcttgacct
6421 ggctatgctg gccgcgattc gccgcgttta cgggctgctt gccaatacgg tgcggtatct
6481 gcagggcggc gggtcgtggc gggaggattg gggacagctt tcggggacgg ccgtgccgcc
6541 ccagggtgcc gagccccaga gcaacgcggg cccacgaccc catatcgggg acacgttatt
6601 taccctgttt cgggcccccg agttgctggc ccccaacggc gacctgtaca acgtgtttgc
6661 ctgggccttg gacgtcttgg ccaaacgcct ccgtcccatg cacgtcttta tcctggatta
6721 cgaccaatcg cccgccggct gccgggacgc cctgctgcaa cttacctccg ggatggtcca
6781 gacccacgtc accacccccg gctccatacc gacgatctgc gacctggcgc gcacgtttgc
6841 ccgggagatg ggggaggcta actgaggatc ccacgtcact attgtatact ctatattata
6901 ctctatgtta tactctgtaa tcctactcaa taaacgtgtc acgcctgtga aaccgtacta
6961 agtctcccgt gtattcttat caccatcagg tgacatcctc gcccaggctg tcaatcatgc
7021 cggtatcgat tccagtagca ccggccccac gctgacaacc cactcttgca gcgttagcag
7081 cgcccctctt aacaagccga cccccaccag cgtcgcggtt actaacactc ctctccccga
7141 cctgcagccc agct
//
| {
"Vector backbone": "pFETCh_Donor",
"Backbone size w/o insert (bp)": "",
"Vector type": "CRISPR",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Kanamycin, 50 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "CETCh-seq: CRISPR epitope tagging ChIP-seq of DNA-binding proteins",
"Publication": "Genome Res. 2015 Sep 9.",
"DOI": "10.1101/gr.193540.115",
"Authors": "Savic D, Partridge EC, Newberry KM, Smith SB, Meadows SK, Roberts BS, Mackiewicz M, Mendenhall EM, Myers RM",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/26355004",
"PubMed ID": "PubMed 26355004"
} | {
"Gene/Insert name": "KDM1A homology arms",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "KDM1A",
"Promoter": "",
"Tag / Fusion Protein": "3XFLAG-P2A-NeoR (C terminal on insert)"
} | {
"Cloning method": "Gibson Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "acgcctgtgaaaccgtacta",
"3′ sequencing primer": "aactgttgggaagggcgatc"
} | {
"Gene/Insert name": "",
"Species": "",
"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": ""
} |
||
pAG425GAL-EYFP-ccdB | 14345 | (Empty Backbone) | Susan Lindquist | Alberti et al Yeast. 2007 Oct . 24(10):913-9. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/8008/b28009f3-ddbb-431b-8a59-2e6891fe10a5/addgene-plasmid-14345-sequence-8008.gbk | full | LOCUS sequence-8008-ex 9950 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 9950)
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 9950)
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..9950
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(10..28)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
rep_origin 69..1411
/label=2u ori
/note="yeast 2u plasmid origin of replication"
protein_bind 401..448
/label=FRT
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
promoter 1438..1542
/gene="bla"
/label=AmpR promoter
CDS 1543..2403
/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(1761..1780)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 2574..3162
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3063..3082
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 3316..3333
/label=L4440
/note="L4440 vector, forward primer"
promoter 3486..3516
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3524..3540
/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 3529..3551
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 3548..3564
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 3548..3564
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 3583..3603
/label=T3
/note="T3 promoter, forward primer"
promoter 3585..3603
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 3627..4068
/gene="S. cerevisiae GAL1"
/label=GAL1 promoter
/note="inducible promoter, regulated by Gal4"
protein_bind 3627..3744
/label=UAS
/bound_moiety="Gal4"
/note="upstream activating sequence mediating
Gal4-dependent induction"
primer_bind 4039..4062
/label=GAL1
/note="S. cerevisiae GAL1 promoter, forward primer"
primer_bind 4083..4099
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 4083..4099
/label=pBluescriptSK
/note="For pBluescript vector"
CDS 4112..4828
/codon_start=1
/product="enhanced YFP"
/label=EYFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(4157..4178)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(4418..4437)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 4765..4786
/label=EGFP-C
/note="EGFP, forward primer"
protein_bind 4853..4977
/gene="mutant version of attR"
/label=attR1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
promoter 5002..5032
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
CDS 5086..5745
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind complement(5152..5171)
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
CDS 6090..6395
/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 6274..6293
/label=ccdB-fwd
/note="ccdB gene, forward primer"
protein_bind complement(6436..6560)
/gene="mutant version of attR"
/label=attR2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
terminator 6567..6814
/gene="S. cerevisiae CYC1"
/label=CYC1 terminator
/note="transcription terminator for CYC1"
primer_bind complement(6583..6601)
/label=CYC1
/note="CYC1 transcription termination signal, reverse
primer"
primer_bind complement(6832..6851)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(6833..6851)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(6861..6878)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(6861..6877)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6870..6892)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 7022..7477
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(7109..7128)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 7319..7340
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 7777..8181
/gene="S. cerevisiae LEU2"
/label=LEU2 promoter
CDS 8194..9288
/codon_start=1
/gene="S. cerevisiae LEU2"
/product="3-isopropylmalate dehydrogenase, required for
leucine biosynthesis"
/label=LEU2
/note="yeast auxotrophic marker"
/translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
IGGAAIDATGVPLPDEALEASKKVDAVLLGAVAGPKWGTGSVRPEQGLLKIRKELQLYA
NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
VPEVQRITRMAAFMALQHEPPLPIWSLDKANLLASSRLWRKTVEETIKNEFPTLKVQHQ
LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
DLGGSNSTTEVGDAVAEEVKKILA"
primer_bind complement(9781..9800)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 9900..9922
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
ORIGIN
1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
61 cttagtatga tccaatatca aaggaaatga tagcattgaa ggatgagact aatccaattg
121 aggagtggca gcatatagaa cagctaaagg gtagtgctga aggaagcata cgataccccg
181 catggaatgg gataatatca caggaggtac tagactacct ttcatcctac ataaatagac
241 gcatataagt acgcatttaa gcataaacac gcactatgcc gttcttctca tgtatatata
301 tatacaggca acacgcagat ataggtgcga cgtgaacagt gagctgtatg tgcgcagctc
361 gcgttgcatt ttcggaagcg ctcgttttcg gaaacgcttt gaagttccta ttccgaagtt
421 cctattctct agaaagtata ggaacttcag agcgcttttg aaaaccaaaa gcgctctgaa
481 gacgcacttt caaaaaacca aaaacgcacc ggactgtaac gagctactaa aatattgcga
541 ataccgcttc cacaaacatt gctcaaaagt atctctttgc tatatatctc tgtgctatat
601 ccctatataa cctacccatc cacctttcgc tccttgaact tgcatctaaa ctcgacctct
661 acatttttta tgtttatctc tagtattact ctttagacaa aaaaattgta gtaagaacta
721 ttcatagagt gaatcgaaaa caatacgaaa atgtaaacat ttcctatacg tagtatatag
781 agacaaaata gaagaaaccg ttcataattt tctgaccaat gaagaatcat caacgctatc
841 actttctgtt cacaaagtat gcgcaatcca catcggtata gaatataatc ggggatgcct
901 ttatcttgaa aaaatgcacc cgcagcttcg ctagtaatca gtaaacgcgg gaagtggagt
961 caggcttttt ttatggaaga gaaaatagac accaaagtag ccttcttcta accttaacgg
1021 acctacagtg caaaaagtta tcaagagact gcattataga gcgcacaaag gagaaaaaaa
1081 gtaatctaag atgctttgtt agaaaaatag cgctctcggg atgcattttt gtagaacaaa
1141 aaagaagtat agattctttg ttggtaaaat agcgctctcg cgttgcattt ctgttctgta
1201 aaaatgcagc tcagattctt tgtttgaaaa attagcgctc tcgcgttgca tttttgtttt
1261 acaaaaatga agcacagatt cttcgttggt aaaatagcgc tttcgcgttg catttctgtt
1321 ctgtaaaaat gcagctcaga ttctttgttt gaaaaattag cgctctcgcg ttgcattttt
1381 gttctacaaa atgaagcaca gatgcttcgt tcaggtggca cttttcgggg aaatgtgcgc
1441 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
1501 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
1561 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
1621 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
1681 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
1741 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
1801 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
1861 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
1921 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
1981 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
2041 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
2101 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
2161 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
2221 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
2281 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
2341 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
2401 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
2461 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
2521 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
2581 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
2641 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
2701 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
2761 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
2821 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
2881 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
2941 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
3001 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
3061 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
3121 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
3181 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
3241 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
3301 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
3361 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
3421 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt acctcactca ttaggcaccc
3481 caggctttac actttatgct tccggctcct atgttgtgtg gaattgtgag cggataacaa
3541 tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa ccctcactaa
3601 agggaacaaa agctggagct ctagtacgga ttagaagccg ccgagcgggc gacagccctc
3661 cgacggaaga ctctcctccg tgcgtcctcg tcttcaccgg tcgcgttcct gaaacgcaga
3721 tgtgcctcgc gccgcactgc tccgaacaat aaagattcta caatactagc ttttatggtt
3781 atgaagagga aaaattggca gtaacctggc cccacaaacc ttcaaattaa cgaatcaaat
3841 taacaaccat aggatgataa tgcgattagt tttttagcct tatttctggg gtaattaatc
3901 agcgaagcga tgatttttga tctattaaca gatatataaa tggaaaagct gcataaccac
3961 tttaactaat actttcaaca ttttcagttt gtattacttc ttattcaaat gtcataaaag
4021 tatcaacaaa aaattgttaa tatacctcta tactttaacg tcaaggagaa aaaaccccgg
4081 attctagaac tagtggatcc cccgggacaa aatggtgagc aagggcgagg agctgttcac
4141 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
4201 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
4261 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accttcggct acggcctgca
4321 gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
4381 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
4441 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
4501 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
4561 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
4621 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
4681 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc taccagtccg ccctgagcaa
4741 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
4801 cactctcggc atggacgagc tgtacaaggg tggaccaggt ggtggacatc aaacaagttt
4861 gtacaaaaaa gctgaacgag aaacgtaaaa tgatataaat atcaatatat taaattagat
4921 tttgcataaa aaacagacta cataatactg taaaacacaa catatccagt cactatggcg
4981 gccgcattag gcaccccagg ctttacactt tatgcttccg gctcgtataa tgtgtggatt
5041 ttgagttagg atccgtcgag attttcagga gctaaggaag ctaaaatgga gaaaaaaatc
5101 actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt tgaggcattt
5161 cagtcagttg ctcaatgtac ctataaccag accgttcagc tggatattac ggccttttta
5221 aagaccgtaa agaaaaataa gcacaagttt tatccggcct ttattcacat tcttgcccgc
5281 ctgatgaatg ctcatccgga attccgtatg gcaatgaaag acggtgagct ggtgatatgg
5341 gatagtgttc acccttgtta caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc
5401 tggagtgaat accacgacga tttccggcag tttctacaca tatattcgca agatgtggcg
5461 tgttacggtg aaaacctggc ctatttccct aaagggttta ttgagaatat gtttttcgtc
5521 tcagccaatc cctgggtgag tttcaccagt tttgatttaa acgtggccaa tatggacaac
5581 ttcttcgccc ccgttttcac catgggcaaa tattatacgc aaggcgacaa ggtgctgatg
5641 ccgctggcga ttcaggttca tcatgccgtc tgtgatggct tccatgtcgg cagaatgctt
5701 aatgaattac aacagtactg cgatgagtgg cagggcgggg cgtaaacgcc gcgtggatcc
5761 ggcttactaa aagccagata acagtatgcg tatttgcgcg ctgatttttg cggtataaga
5821 atatatactg atatgtatac ccgaagtatg tcaaaaagag gtatgctatg aagcagcgta
5881 ttacagtgac agttgacagc gacagctatc agttgctcaa ggcatatatg atgtcaatat
5941 ctccggtctg gtaagcacaa ccatgcagaa tgaagcccgt cgtctgcgtg ccgaacgctg
6001 gaaagcggaa aatcaggaag ggatggctga ggtcgcccgg tttattgaaa tgaacggctc
6061 ttttgctgac gagaacaggg gctggtgaaa tgcagtttaa ggtttacacc tataaaagag
6121 agagccgtta tcgtctgttt gtggatgtac agagtgatat tattgacacg cccgggcgac
6181 ggatggtgat ccccctggcc agtgcacgtc tgctgtcaga taaagtctcc cgtgaacttt
6241 acccggtggt gcatatcggg gatgaaagct ggcgcatgat gaccaccgat atggccagtg
6301 tgccggtctc cgttatcggg gaagaagtgg ctgatctcag ccaccgcgaa aatgacatca
6361 aaaacgccat taacctgatg ttctggggaa tataaatgtc aggctccctt atacacagcc
6421 agtctgcagg tcgaccatag tgactggata tgttgtgttt tacagtatta tgtagtctgt
6481 tttttatgca aaatctaatt taatatattg atatttatat cattttacgt ttctcgttca
6541 gctttcttgt acaaagtggt tcgatgtcat gtaattagtt atgtcacgct tacattcacg
6601 ccctcccccc acatccgctc taaccgaaaa ggaaggagtt agacaacctg aagtctaggt
6661 ccctatttat ttttttatag ttatgttagt attaagaacg ttatttatat ttcaaatttt
6721 tctttttttt ctgtacagac gcgtgtacgc atgtaacatt atactgaaaa ccttgcttga
6781 gaaggttttg ggacgctcga aggctttaat ttgcggccgg tacccaattc gccctatagt
6841 gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
6901 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
6961 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc
7021 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
7081 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
7141 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
7201 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
7261 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
7321 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
7381 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
7441 aacgcgaatt ttaacaaaat attaacgttt acaatttcct gatgcggtat tttctcctta
7501 cgcatctgtg cggtatttca caccgcatat cgacggtcga ggagaacttc tagtatatcc
7561 acatacctaa tattattgcc ttattaaaaa tggaatccca acaattacat caaaatccac
7621 attctcttca aaatcaattg tcctgtactt ccttgttcat gtgtgttcaa aaacgttata
7681 tttataggat aattatactc tatttctcaa caagtaattg gttgtttggc cgagcggtct
7741 aaggcgcctg attcaagaaa tatcttgacc gcagttaact gtgggaatac tcaggtatcg
7801 taagatgcaa gagttcgaat ctcttagcaa ccattatttt tttcctcaac ataacgagaa
7861 cacacagggg cgctatcgca cagaatcaaa ttcgatgact ggaaattttt tgttaatttc
7921 agaggtcgcc tgacgcatat acctttttca actgaaaaat tgggagaaaa aggaaaggtg
7981 agaggccgga accggctttt catatagaat agagaagcgt tcatgactaa atgcttgcat
8041 cacaatactt gaagttgaca atattattta aggacctatt gttttttcca ataggtggtt
8101 agcaatcgtc ttactttcta acttttctta ccttttacat ttcagcaata tatatatata
8161 tttcaaggat ataccattct aatgtctgcc cctatgtctg cccctaagaa gatcgtcgtt
8221 ttgccaggtg accacgttgg tcaagaaatc acagccgaag ccattaaggt tcttaaagct
8281 atttctgatg ttcgttccaa tgtcaagttc gatttcgaaa atcatttaat tggtggtgct
8341 gctatcgatg ctacaggtgt cccacttcca gatgaggcgc tggaagcctc caagaaggtt
8401 gatgccgttt tgttaggtgc tgtggctggt cctaaatggg gtaccggtag tgttagacct
8461 gaacaaggtt tactaaaaat ccgtaaagaa cttcaattgt acgccaactt aagaccatgt
8521 aactttgcat ccgactctct tttagactta tctccaatca agccacaatt tgctaaaggt
8581 actgacttcg ttgttgtcag agaattagtg ggaggtattt actttggtaa gagaaaggaa
8641 gacgatggtg atggtgtcgc ttgggatagt gaacaataca ccgttccaga agtgcaaaga
8701 atcacaagaa tggccgcttt catggcccta caacatgagc caccattgcc tatttggtcc
8761 ttggataaag ctaatctttt ggcctcttca agattatgga gaaaaactgt ggaggaaacc
8821 atcaagaacg aattccctac attgaaggtt caacatcaat tgattgattc tgccgccatg
8881 atcctagtta agaacccaac ccacctaaat ggtattataa tcaccagcaa catgtttggt
8941 gatatcatct ccgatgaagc ctccgttatc ccaggttcct tgggtttgtt gccatctgcg
9001 tccttggcct ctttgccaga caagaacacc gcatttggtt tgtacgaacc atgccacggt
9061 tctgctccag atttgccaaa gaataaggtt gaccctatcg ccactatctt gtctgctgca
9121 atgatgttga aattgtcatt gaacttgcct gaagaaggta aggccattga agatgcagtt
9181 aaaaaggttt tggatgcagg tatcagaact ggtgatttag gtggttccaa cagtaccacc
9241 gaagtcggtg atgctgtcgc cgaagaagtt aagaaaatcc ttgcttaaaa agattctctt
9301 tttttatgat atttgtacat aaactttata aatgaaattc ataatagaaa cgacacgaaa
9361 ttacaaaatg gaatatgttc atagggtaga cgaaactata tacgcaatct acatacattt
9421 atcaagaagg agaaaaagga ggatagtaaa ggaatacagg taagcaaatt gatactaatg
9481 gctcaacgtg ataaggaaaa agaattgcac tttaacatta atattgacaa ggaggagggc
9541 accacacaaa aagttaggtg taacagaaaa tcatgaaact acgattccta atttgatatt
9601 ggaggatttt ctctaaaaaa aaaaaaatac aacaaataaa aaacactcaa tgacctgacc
9661 atttgatgga gtttaagtca ataccttctt gaagcatttc ccataatggt gaaagttccc
9721 tcaagaattt tactctgtca gaaacggcct tacgacgtag tcgatatggt gcactctcag
9781 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
9841 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
9901 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
//
| {
"Vector backbone": "pRS425",
"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": "EYFP (N terminal on backbone)"
} | {
"Cloning method": "Gateway Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "EGFP-C",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pHRW-CRY2-VHH(GFP) | 126022 | express RFP-tagged CRY2 PHR domain fused to anti-GFP nanobody for optogenetics | Eurico Morais-de-Sá | Osswald et al Biomolecules. 2019 Feb 12;9(2). pii: biom9020061. doi: 10.3390/biom9020061. | partial | {
"Vector backbone": "pHRW",
"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 ; Takara Bio Limited Use Label License (formerly Clontech)",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Light-Induced Protein Clustering for Optogenetic Interference and Protein Interaction Analysis in Drosophila S2 Cells",
"Publication": "Biomolecules. 2019 Feb 12;9(2). pii: biom9020061. doi: 10.3390/biom9020061.",
"DOI": "10.3390/biom9020061.",
"Authors": "Osswald M, Santos AF, Morais-de-Sa E /biom",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/30759894",
"PubMed ID": "PubMed 30759894"
} | {
"Gene/Insert name": "CRY2 PHR-VHH(GFP)",
"Species": "A. thaliana (mustard weed)",
"Insert Size (bp)": "",
"Mutation": "WT",
"Entrez Gene": "",
"Promoter": "Hsp70",
"Tag / Fusion Protein": "RFP (N terminal on insert)"
} | {
"Cloning method": "Gateway 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": ""
} |
|||
pmiReport PU.1 3' UTR | 26530 | David Baltimore | O'Connell et al J Exp Med. 2008 Mar 17. 205(3):585-94. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/32642/30be2927-1ac3-40e7-9a7e-1a98f39f0098/addgene-plasmid-26530-sequence-32642.gbk | partial | LOCUS sequence-32642-e 903 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 903)
AUTHORS O'Connell RM, Rao DS, Chaudhuri AA, Boldin MP, Taganov KD, Nicoll J,
Paquette RL, Baltimore D
TITLE Sustained expression of microRNA-155 in hematopoietic stem cells
causes a myeloproliferative disorder.
JOURNAL J Exp Med. 2008 Mar 17. 205(3):585-94.
PUBMED 18299402
REFERENCE 2 (bases 1 to 903)
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 Exp Med.
2008 Mar 17. 205(3):585-94."
FEATURES Location/Qualifiers
source 1..903
/mol_type="genomic DNA"
/organism="unspecified"
ORIGIN
1 gcggccgcat acaaaaacca acacacagat ccaatgaaaa taaaagatcc tttattaagc
61 ttgggagaat agctgtcaat aattttactt gtctttagtg actaatgagg gggtggggag
121 gaagagcttt ttaaaaagca ttcaatatac aaaaaggtgc cagcgccaga ctcccaagga
181 agcaccgtgg ccactaagcc aggctgaccc tcggttgagg tccgaggccc aggcggccct
241 tggagcgggc tagagtcctg gtgggaggca aagcgggtgg ggcgggccag gccgggcatg
301 ggagggcgag gccgggcccc cagtcctgcc tcggccctgg gaatgtcctc cctgtgtccg
361 ggccgggcga cgggttaatg ctatggccgg actagtgcgc agctttcatc actcgagcaa
421 tttggacttt ccgcccttct tggcctttat gaggatctct ctgatttttc ttgcgtcgag
481 ttttccggta agacctttcg gtacttcgtc cacaaacaca actcctccgc gcaacttttt
541 cgcggttgtt acttgactgg cgacgtaatc cacgatctct ttttccgtca tcgtctttcc
601 gtgctccaaa acaacaacgg cggcgggaag ttcaccggcg tcatcgtcgg gaagacctgc
661 cacgcccgcg tcgaagatgt tggggtgttg taacaatatc gattccaatt cagcgggggc
721 cacctgatat cctttgtatt taattaaaga cttcaagcgg tcaactatga agaagtgttc
781 gtcttcgtcc cagtaagcta tgtctccaga atgtagccat ccatccttgt caatcaaggc
841 gttggtcgct tccggattgt ttacataacc ggacataatc ataggtcctc tgacacataa
901 ttc
//
| {
"Vector backbone": "pmiReport",
"Backbone size w/o insert (bp)": "6470",
"Vector type": "Mammalian Expression, Luciferase",
"Selectable markers": "Puromycin"
} | {
"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": "Sustained expression of microRNA-155 in hematopoietic stem cells causes a myeloproliferative disorder",
"Publication": "J Exp Med. 2008 Mar 17. 205(3):585-94.",
"DOI": "10.1084/jem.20072108",
"Authors": "O'Connell RM, Rao DS, Chaudhuri AA, Boldin MP, Taganov KD, Nicoll J, Paquette RL, Baltimore D",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/18299402",
"PubMed ID": "PubMed 18299402"
} | {
"Gene/Insert name": "PU.1 3' UTR",
"Species": "M. musculus (mouse)",
"Insert Size (bp)": "333",
"Mutation": "",
"Entrez Gene": "Spi1",
"Promoter": "",
"Tag / Fusion Protein": "Luciferase (N terminal on backbone)"
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "SpeI (not destroyed)",
"3′ cloning site": "HindIII (not destroyed)",
"5′ sequencing primer": "Luc-F",
"3′ sequencing primer": "M13pUC-fwd"
} | {
"Gene/Insert name": "",
"Species": "",
"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": ""
} |
||
eSpCas9(1.1)_No_FLAG_LMNA_G2 | 178090 | Expresses the LMNA G2 sgRNA in combination with FLAGless eSpCas9(1.1). This vector can be used in combination with LMNA_mScarlet-I_Donor to tag LMNA with mScarlet-I. pX330-like plasmid. | Yannick Doyon | Levesque et al Nat Commun. 2022 Oct 7;13(1):5909. doi: 10.1038/s41467-022-33669-z. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/350140/7730e353-ff31-4fee-a37e-d7a97fb307af/addgene-plasmid-178090-sequence-350140.gbk | full | LOCUS sequence-350140- 8439 bp DNA circular SYN 26-AUG-2024
DEFINITION Expresses the LMNA G2 sgRNA in combination with FLAGless
eSpCas9(1.1). This vector can be used in combination with
LMNA_mScarlet-I_Donor to tag LMNA with mScarlet-I. pX330-like
plasmid..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8439)
AUTHORS Levesque S, Mayorga D, Fiset JP, Goupil C, Duringer A, Loiselle A,
Bouchard E, Agudelo D, Doyon Y
TITLE Marker-free co-selection for successive rounds of prime editing in
human cells.
JOURNAL Nat Commun. 2022 Oct 7;13(1):5909. doi: 10.1038/s41467-022-33669-z.
PUBMED 36207338
REFERENCE 2 (bases 1 to 8439)
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: "2022-10-7"; volume: "13(1):5909. doi"; pages: "
10.1038/s41467-022-33669-z"
FEATURES Location/Qualifiers
source 1..8439
/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"
promoter 730..1007
/label=chicken beta-actin promoter
intron 1008..1235
/label=hybrid intron
/note="hybrid between chicken beta-actin (CBA) and minute
virus of mice (MMV) introns (Gray et al., 2011)"
regulatory 1247..1256
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 1259..1279
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 1304..5404
/codon_start=1
/product="Cas9 endonuclease from the Streptococcus pyogenes
Type II CRISPR/Cas system, mutated to improve targeting
specificity (Slaymaker et al., 2016)"
/label=eSpCas9(1.1)
/note="carries the mutations K848A, K1003A, and R1060A"
/translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLADDSIDNKVLTRSDKNRGKSDNVPS
EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
AVVGTALIKKYPALESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
ITLANGEIRKAPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
ATLIHQSITGLYETRIDLSQLGGD"
CDS 5405..5452
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
primer_bind complement(5480..5497)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 5486..5693
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
repeat_region 5702..5842
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 5702..5831
/label=AAV2 ITR
rep_origin 5917..6372
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(6004..6023)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 6214..6235
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(6389..6408)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 6508..6530
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(6568..6586)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 6654..6758
/gene="bla"
/label=AmpR promoter
CDS 6759..7619
/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(6977..6996)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 7790..8378
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 8279..8298
/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 ccatggagac cccgtcccag 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 gggggggggg gcgcgcgccr ggsggggsgg ggsggggsgr ggggsggggs ggggsgaggc
901 ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct tttatggcga
961 ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga gtcgctgcgc
1021 gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac
1081 tgaccgcgtt actcccacag gtgagcgggc gggacggccc ttctcctccg ggctgtaatt
1141 agctgagcaa gaggtaaggg tttaagggat ggttggttgg tggggtatta atgtttaatt
1201 acctggagca cctgcctgaa atcacttttt ttcaggttgg accggtgcca ccatggcccc
1261 aaagaagaag cggaaggtcg gtatccacgg agtcccagca gccgacaaga agtacagcat
1321 cggcctggac atcggcacca actctgtggg ctgggccgtg atcaccgacg agtacaaggt
1381 gcccagcaag aaattcaagg tgctgggcaa caccgaccgg cacagcatca agaagaacct
1441 gatcggagcc ctgctgttcg acagcggcga aacagccgag gccacccggc tgaagagaac
1501 cgccagaaga agatacacca gacggaagaa ccggatctgc tatctgcaag agatcttcag
1561 caacgagatg gccaaggtgg acgacagctt cttccacaga ctggaagagt ccttcctggt
1621 ggaagaggat aagaagcacg agcggcaccc catcttcggc aacatcgtgg acgaggtggc
1681 ctaccacgag aagtacccca ccatctacca cctgagaaag aaactggtgg acagcaccga
1741 caaggccgac ctgcggctga tctatctggc cctggcccac atgatcaagt tccggggcca
1801 cttcctgatc gagggcgacc tgaaccccga caacagcgac gtggacaagc tgttcatcca
1861 gctggtgcag acctacaacc agctgttcga ggaaaacccc atcaacgcca gcggcgtgga
1921 cgccaaggcc atcctgtctg ccagactgag caagagcaga cggctggaaa atctgatcgc
1981 ccagctgccc ggcgagaaga agaatggcct gttcggaaac ctgattgccc tgagcctggg
2041 cctgaccccc aacttcaaga gcaacttcga cctggccgag gatgccaaac tgcagctgag
2101 caaggacacc tacgacgacg acctggacaa cctgctggcc cagatcggcg accagtacgc
2161 cgacctgttt ctggccgcca agaacctgtc cgacgccatc ctgctgagcg acatcctgag
2221 agtgaacacc gagatcacca aggcccccct gagcgcctct atgatcaaga gatacgacga
2281 gcaccaccag gacctgaccc tgctgaaagc tctcgtgcgg cagcagctgc ctgagaagta
2341 caaagagatt ttcttcgacc agagcaagaa cggctacgcc ggctacattg acggcggagc
2401 cagccaggaa gagttctaca agttcatcaa gcccatcctg gaaaagatgg acggcaccga
2461 ggaactgctc gtgaagctga acagagagga cctgctgcgg aagcagcgga ccttcgacaa
2521 cggcagcatc ccccaccaga tccacctggg agagctgcac gccattctgc ggcggcagga
2581 agatttttac ccattcctga aggacaaccg ggaaaagatc gagaagatcc tgaccttccg
2641 catcccctac tacgtgggcc ctctggccag gggaaacagc agattcgcct ggatgaccag
2701 aaagagcgag gaaaccatca ccccctggaa cttcgaggaa gtggtggaca agggcgcttc
2761 cgcccagagc ttcatcgagc ggatgaccaa cttcgataag aacctgccca acgagaaggt
2821 gctgcccaag cacagcctgc tgtacgagta cttcaccgtg tataacgagc tgaccaaagt
2881 gaaatacgtg accgagggaa tgagaaagcc cgccttcctg agcggcgagc agaaaaaggc
2941 catcgtggac ctgctgttca agaccaaccg gaaagtgacc gtgaagcagc tgaaagagga
3001 ctacttcaag aaaatcgagt gcttcgactc cgtggaaatc tccggcgtgg aagatcggtt
3061 caacgcctcc ctgggcacat accacgatct gctgaaaatt atcaaggaca aggacttcct
3121 ggacaatgag gaaaacgagg acattctgga agatatcgtg ctgaccctga cactgtttga
3181 ggacagagag atgatcgagg aacggctgaa aacctatgcc cacctgttcg acgacaaagt
3241 gatgaagcag ctgaagcggc ggagatacac cggctggggc aggctgagcc ggaagctgat
3301 caacggcatc cgggacaagc agtccggcaa gacaatcctg gatttcctga agtccgacgg
3361 cttcgccaac agaaacttca tgcagctgat ccacgacgac agcctgacct ttaaagagga
3421 catccagaaa gcccaggtgt ccggccaggg cgatagcctg cacgagcaca ttgccaatct
3481 ggccggcagc cccgccatta agaagggcat cctgcagaca gtgaaggtgg tggacgagct
3541 cgtgaaagtg atgggccggc acaagcccga gaacatcgtg atcgaaatgg ccagagagaa
3601 ccagaccacc cagaagggac agaagaacag ccgcgagaga atgaagcgga tcgaagaggg
3661 catcaaagag ctgggcagcc agatcctgaa agaacacccc gtggaaaaca cccagctgca
3721 gaacgagaag ctgtacctgt actacctgca gaatgggcgg gatatgtacg tggaccagga
3781 actggacatc aaccggctgt ccgactacga tgtggaccat atcgtgcctc agagctttct
3841 ggccgacgac tccatcgaca acaaggtgct gaccagaagc gacaagaacc ggggcaagag
3901 cgacaacgtg ccctccgaag aggtcgtgaa gaagatgaag aactactggc ggcagctgct
3961 gaacgccaag ctgattaccc agagaaagtt cgacaatctg accaaggccg agagaggcgg
4021 cctgagcgaa ctggataagg ccggcttcat caagagacag ctggtggaaa cccggcagat
4081 cacaaagcac gtggcacaga tcctggactc ccggatgaac actaagtacg acgagaatga
4141 caagctgatc cgggaagtga aagtgatcac cctgaagtcc aagctggtgt ccgatttccg
4201 gaaggatttc cagttttaca aagtgcgcga gatcaacaac taccaccacg cccacgacgc
4261 ctacctgaac gccgtcgtgg gaaccgccct gatcaaaaag taccctgcgc tggaaagcga
4321 gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag atgatcgcca agagcgagca
4381 ggaaatcggc aaggctaccg ccaagtactt cttctacagc aacatcatga actttttcaa
4441 gaccgagatt accctggcca acggcgagat ccggaaggcg cctctgatcg agacaaacgg
4501 cgaaaccggg gagatcgtgt gggataaggg ccgggatttt gccaccgtgc ggaaagtgct
4561 gagcatgccc caagtgaata tcgtgaaaaa gaccgaggtg cagacaggcg gcttcagcaa
4621 agagtctatc ctgcccaaga ggaacagcga taagctgatc gccagaaaga aggactggga
4681 ccctaagaag tacggcggct tcgacagccc caccgtggcc tattctgtgc tggtggtggc
4741 caaagtggaa aagggcaagt ccaagaaact gaagagtgtg aaagagctgc tggggatcac
4801 catcatggaa agaagcagct tcgagaagaa tcccatcgac tttctggaag ccaagggcta
4861 caaagaagtg aaaaaggacc tgatcatcaa gctgcctaag tactccctgt tcgagctgga
4921 aaacggccgg aagagaatgc tggcctctgc cggcgaactg cagaagggaa acgaactggc
4981 cctgccctcc aaatatgtga acttcctgta cctggccagc cactatgaga agctgaaggg
5041 ctcccccgag gataatgagc agaaacagct gtttgtggaa cagcacaagc actacctgga
5101 cgagatcatc gagcagatca gcgagttctc caagagagtg atcctggccg acgctaatct
5161 ggacaaagtg ctgtccgcct acaacaagca ccgggataag cccatcagag agcaggccga
5221 gaatatcatc cacctgttta ccctgaccaa tctgggagcc cctgccgcct tcaagtactt
5281 tgacaccacc atcgaccgga agaggtacac cagcaccaaa gaggtgctgg acgccaccct
5341 gatccaccag agcatcaccg gcctgtacga gacacggatc gacctgtctc agctgggagg
5401 cgacaaaagg ccggcggcca cgaaaaaggc cggccaggca aaaaagaaaa agtaagaatt
5461 cctagagctc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
5521 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
5581 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
5641 gggcaggaca gcaaggggga ggattgggaa gagaatagca ggcatgctgg ggagcggccg
5701 caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
5761 gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag
5821 cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc
5881 ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa
5941 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccttagcgc
6001 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
6061 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
6121 aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
6181 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
6241 cactcaactc tatctcgggc tattcttttg atttataagg gattttgccg atttcggtct
6301 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
6361 cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
6421 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
6481 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
6541 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
6601 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
6661 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
6721 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
6781 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
6841 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
6901 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
6961 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
7021 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
7081 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
7141 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
7201 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
7261 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
7321 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
7381 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
7441 ttattgctga taaatctgga gccggtgagc gtggaagccg cggtatcatt gcagcactgg
7501 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
7561 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
7621 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
7681 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
7741 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
7801 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
7861 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
7921 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
7981 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
8041 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
8101 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
8161 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
8221 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
8281 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
8341 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
8401 tacggttcct ggccttttgc tggccttttg ctcacatgt
//
| {
"Vector backbone": "pX330-U6-Chimeric_BB-CBh-hSpCas9",
"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": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Marker-free co-selection for successive rounds of prime editing in human cells",
"Publication": "Nat Commun. 2022 Oct 7;13(1):5909. doi: 10.1038/s41467-022-33669-z.",
"DOI": "10.1038/s41467-022-33669-z.",
"Authors": "Levesque S, Mayorga D, Fiset JP, Goupil C, Duringer A, Loiselle A, Bouchard E, Agudelo D, Doyon Y /s---z",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/36207338",
"PubMed ID": "PubMed 36207338"
} | {
"Gene/Insert name": "LMNA G2 sgRNA + FLAGless enhanced specificity Cas9 (1.1)",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "U6 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": "GAGGGCCTATTTCCCATGATT",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"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": ""
} |
|
psiCHECK2-let-7 4x | 20930 | Low | Yukihide Tomari | Iwasaki et al Mol Cell. 2009 Mar 4. ():. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/390788/31662a5a-361b-4e18-b8e7-63c5c408780b/addgene-plasmid-20930-sequence-390788.gbk | full | LOCUS sequence-390788- 6357 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 6357)
AUTHORS Iwasaki S, Kawamata T, Tomari Y
TITLE Drosophila Argonaute1 and Argonaute2 Employ Distinct Mechanisms for
Translational Repression.
JOURNAL Mol Cell. 2009 Mar 4. ():.
PUBMED 19268617
REFERENCE 2 (bases 1 to 6357)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Mol Cell.
2009 Mar 4. ():."
FEATURES Location/Qualifiers
source 1..6357
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 62..419
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
enhancer 62..253
/label=SV40 enhancer
/note="enhancer for the SV40 early promoter (Herr, 1993)"
primer_bind complement(85..105)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 270..405
/label=SV40 ori
/note="SV40 origin of replication"
intron 489..621
/label=chimeric intron
/note="chimera between introns from human beta-globin and
immunoglobulin heavy chain genes"
primer_bind 604..623
/label=pMT2-F
/note="Synthetic intron, forward primer"
promoter 666..684
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory 688..697
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 694..1629
/codon_start=1
/product="Renilla luciferase"
/label=hRluc
/note="human codon-optimized"
/translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
polyA_signal 1772..1820
/note="synthetic polyadenylation signal"
promoter 1829..2580
/label=HSV TK promoter
/note="herpes simplex virus thymidine kinase promoter"
regulatory 2610..2619
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 2616..4268
/codon_start=1
/gene="luc+"
/product="firefly luciferase"
/label=luciferase
/note="enhanced luc+ version of the luciferase gene"
/db_xref="GI:42718124"
/protein_id="AAS38485.1"
/translation="MADAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
RBS 3575..3583
/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 4312..4433
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4323..4342)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 4377..4396
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
promoter 4566..4670
/gene="bla"
/label=AmpR promoter
CDS 4671..5531
/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(4889..4908)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 5702..6290
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6191..6210
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 agatctgcgc agcaccatgg cctgaaataa cctctgaaag aggaacttgg ttaggtacct
61 tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
121 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
181 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
241 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
301 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
361 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
421 agcttgattc ttctgacaca acagtctcga acttaagctg cagaagttgg tcgtgaggca
481 ctgggcaggt aagtatcaag gttacaagac aggtttaagg agaccaatag aaactgggct
541 tgtcgagaca gagaagactc ttgcgtttct gataggcacc tattggtctt actgacatcc
601 actttgcctt tctctccaca ggtgtccact cccagttcaa ttacagctct taaggctaga
661 gtacttaata cgactcacta taggctagcc accatggctt ccaaggtgta cgaccccgag
721 caacgcaaac gcatgatcac tgggcctcag tggtgggctc gctgcaagca aatgaacgtg
781 ctggactcct tcatcaacta ctatgattcc gagaagcacg ccgagaacgc cgtgattttt
841 ctgcatggta acgctgcctc cagctacctg tggaggcacg tcgtgcctca catcgagccc
901 gtggctagat gcatcatccc tgatctgatc ggaatgggta agtccggcaa gagcgggaat
961 ggctcatatc gcctcctgga tcactacaag tacctcaccg cttggttcga gctgctgaac
1021 cttccaaaga aaatcatctt tgtgggccac gactgggggg cttgtctggc ctttcactac
1081 tcctacgagc accaagacaa gatcaaggcc atcgtccatg ctgagagtgt cgtggacgtg
1141 atcgagtcct gggacgagtg gcctgacatc gaggaggata tcgccctgat caagagcgaa
1201 gagggcgaga aaatggtgct tgagaataac ttcttcgtcg agaccatgct cccaagcaag
1261 atcatgcgga aactggagcc tgaggagttc gctgcctacc tggagccatt caaggagaag
1321 ggcgaggtta gacggcctac cctctcctgg cctcgcgaga tccctctcgt taagggaggc
1381 aagcccgacg tcgtccagat tgtccgcaac tacaacgcct accttcgggc cagcgacgat
1441 ctgcctaaga tgttcatcga gtccgaccct gggttctttt ccaacgctat tgtcgaggga
1501 gctaagaagt tccctaacac cgagttcgtg aaggtgaagg gcctccactt cagccaggag
1561 gacgctccag atgaaatggg taagtacatc aagagcttcg tggagcgcgt gctgaagaac
1621 gagcagtaat tctaggcgat cgctcgagac tatacaagga tctacctcag tcgagactat
1681 acaaggatct acctcagtcg agactataca aggatctacc tcagtcgaga ctatacaagg
1741 atctacctca gtcgacgcgg ccgctggccg caataaaata tctttatttt cattacatct
1801 gtgtgttggt tttttgtgtg aggatctaaa tgagtcttcg gacctcgcgg gggccgctta
1861 agcggtggtt agggtttgtc tgacgcgggg ggagggggaa ggaacgaaac actctcattc
1921 ggaggcggct cggggtttgg tcttggtggc cacgggcacg cagaagagcg ccgcgatcct
1981 cttaagcacc cccccgccct ccgtggaggc gggggtttgg tcggcgggtg gtaactggcg
2041 ggccgctgac tcgggcgggt cgcgcgcccc agagtgtgac cttttcggtc tgctcgcaga
2101 cccccgggcg gcgccgccgc ggcggcgacg ggctcgctgg gtcctaggct ccatggggac
2161 cgtatacgtg gacaggctct ggagcatccg cacgactgcg gtgatattac cggagacctt
2221 ctgcgggacg agccgggtca cgcggctgac gcggagcgtc cgttgggcga caaacaccag
2281 gacggggcac aggtacacta tcttgtcacc cggaggcgcg agggactgca ggagcttcag
2341 ggagtggcgc agctgcttca tccccgtggc ccgttgctcg cgtttgctgg cggtgtcccc
2401 ggaagaaata tatttgcatg tctttagttc tatgatgaca caaaccccgc ccagcgtctt
2461 gtcattggcg aattcgaaca cgcagatgca gtcggggcgg cgcggtccca ggtccacttc
2521 gcatattaag gtgacgcgtg tggcctcgaa caccgagcga ccctgcagcg acccgcttaa
2581 aagcttggca ttccggtact gttggtaaag ccaccatggc cgatgctaag aacattaaga
2641 agggccctgc tcccttctac cctctggagg atggcaccgc tggcgagcag ctgcacaagg
2701 ccatgaagag gtatgccctg gtgcctggca ccattgcctt caccgatgcc cacattgagg
2761 tggacatcac ctatgccgag tacttcgaga tgtctgtgcg cctggccgag gccatgaaga
2821 ggtacggcct gaacaccaac caccgcatcg tggtgtgctc tgagaactct ctgcagttct
2881 tcatgccagt gctgggcgcc ctgttcatcg gagtggccgt ggcccctgct aacgacattt
2941 acaacgagcg cgagctgctg aacagcatgg gcatttctca gcctaccgtg gtgttcgtgt
3001 ctaagaaggg cctgcagaag atcctgaacg tgcagaagaa gctgcctatc atccagaaga
3061 tcatcatcat ggactctaag accgactacc agggcttcca gagcatgtac acattcgtga
3121 catctcatct gcctcctggc ttcaacgagt acgacttcgt gccagagtct ttcgacaggg
3181 acaaaaccat tgccctgatc atgaacagct ctgggtctac cggcctgcct aagggcgtgg
3241 ccctgcctca tcgcaccgcc tgtgtgcgct tctctcacgc ccgcgaccct attttcggca
3301 accagatcat ccccgacacc gctattctga gcgtggtgcc attccaccac ggcttcggca
3361 tgttcaccac cctgggctac ctgatttgcg gctttcgggt ggtgctgatg taccgcttcg
3421 aggaggagct gttcctgcgc agcctgcaag actacaaaat tcagtctgcc ctgctggtgc
3481 caaccctgtt cagcttcttc gctaagagca ccctgatcga caagtacgac ctgtctaacc
3541 tgcacgagat tgcctctggc ggcgccccac tgtctaagga ggtgggcgaa gccgtggcca
3601 agcgctttca tctgccaggc atccgccagg gctacggcct gaccgagaca accagcgcca
3661 ttctgattac cccagagggc gacgacaagc ctggcgccgt gggcaaggtg gtgccattct
3721 tcgaggccaa ggtggtggac ctggacaccg gcaagaccct gggagtgaac cagcgcggcg
3781 agctgtgtgt gcgcggccct atgattatgt ccggctacgt gaataaccct gaggccacaa
3841 acgccctgat cgacaaggac ggctggctgc actctggcga cattgcctac tgggacgagg
3901 acgagcactt cttcatcgtg gaccgcctga agtctctgat caagtacaag ggctaccagg
3961 tggccccagc cgagctggag tctatcctgc tgcagcaccc taacattttc gacgccggag
4021 tggccggcct gcccgacgac gatgccggcg agctgcctgc cgccgtcgtc gtgctggaac
4081 acggcaagac catgaccgag aaggagatcg tggactatgt ggccagccag gtgacaaccg
4141 ccaagaagct gcgcggcgga gtggtgttcg tggacgaggt gcccaagggc ctgaccggca
4201 agctggacgc ccgcaagatc cgcgagatcc tgatcaaggc taagaaaggc ggcaagatcg
4261 ccgtgtaata attctagagt cggggcggcc ggccgcttcg agcagacatg ataagataca
4321 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
4381 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
4441 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca
4501 agtaaaacct ctacaaatgt ggtaaaatcg ataaggatcc aggtggcact tttcggggaa
4561 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
4621 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
4681 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
4741 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
4801 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
4861 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
4921 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
4981 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
5041 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
5101 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
5161 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
5221 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
5281 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
5341 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
5401 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
5461 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
5521 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
5581 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
5641 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
5701 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
5761 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
5821 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
5881 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
5941 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
6001 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
6061 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
6121 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
6181 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
6241 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
6301 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg gctcgac
//
| {
"Vector backbone": "psiCHECK-2",
"Backbone size w/o insert (bp)": "6273",
"Vector type": "Mammalian Expression, 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": "Drosophila Argonaute1 and Argonaute2 Employ Distinct Mechanisms for Translational Repression",
"Publication": "Mol Cell. 2009 Mar 4. ():.",
"DOI": "10.1016/j.molcel.2009.02.010",
"Authors": "Iwasaki S, Kawamata T, Tomari Y",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/19268617",
"PubMed ID": "PubMed 19268617"
} | {
"Gene/Insert name": "4x let-7 target sites",
"Species": "",
"Insert Size (bp)": "108",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "XhoI (not destroyed)",
"3′ cloning site": "NotI (not destroyed)",
"5′ sequencing primer": "TAAGTACATCAAGAGCTTCG",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"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": ""
} |
|
pKF294 | 72858 | template for N-terminal 1x Strep tag, inducible Blasticidin resistance | Klaus Foerstemann | Kunzelmann et al G3 (Bethesda). 2016 Jun 1;6(6):1777-85. doi: 10.1534/g3.116.028241. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/136538/4fd056f0-8e55-4e42-9efa-a90c1a383ccc/addgene-plasmid-72858-sequence-136538.gbk | full | LOCUS sequence-136538- 3986 bp DNA circular SYN 26-AUG-2024
DEFINITION template for N-terminal 1x Strep tag, inducible Blasticidin
resistance.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3986)
AUTHORS Kunzelmann S, Bottcher R, Schmidts I, Forstemann K
TITLE A Comprehensive Toolbox for Genome Editing in Cultured Drosophila
melanogaster Cells.
JOURNAL G3 (Bethesda). 2016 Jun 1;6(6):1777-85. doi: 10.1534/g3.116.028241.
PUBMED 27172193
REFERENCE 2 (bases 1 to 3986)
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: "10.1534/g3.116";
journalName: "G3 (Bethesda)"; date: "2016-06-1- 1"; volume: "6";
issue: "6"; pages: "1777-85"
FEATURES Location/Qualifiers
source 1..3986
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 305..324
/label=T7
/note="T7 promoter, forward primer"
promoter 305..323
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind complement(372..404)
/label=FRT (minimal)
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="supports FLP-mediated excision but not integration
(Turan and Bode, 2011)"
CDS complement(453..851)
/codon_start=1
/gene="Aspergillus terreus BSD"
/product="blasticidin S deaminase"
/label=BSD
/note="confers resistance to blasticidin"
/translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
KAIVKDSDGQPTAVGIRELLPSGYVWEG"
protein_bind complement(1254..1287)
/label=FRT (minimal)
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="supports FLP-mediated excision but not integration
(Turan and Bode, 2011)"
CDS 1312..1335
/codon_start=1
/product="peptide that binds Strep-Tactin(R), an engineered
form of streptavidin"
/label=Strep-Tag II
/translation="WSHPQFEK"
promoter complement(1818..1848)
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
protein_bind 1863..1884
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(2004..2021)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2175..2763)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2255..2274)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(2934..3794)
/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 3557..3576
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(3795..3899)
/gene="bla"
/label=AmpR promoter
primer_bind 3967..3985
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
ORIGIN
1 gcccctgcag ccgaattata ttatttttgc caaataattt ttaacaaaag ctctgaagtc
61 ttcttcattt aaattcttag atgatacttc atctggaaaa ttgtcccaat tagtagcatc
121 acgctgtgag taagttctaa accatttttt tattgttgta ttatctctaa tcttactact
181 cgatgagttt tcggtattat ctctattttt aacttggagc aggttccatt cattgttttt
241 ttcatcatag tgaataaaat caactgcttt aacacttgtg cctgaacacc atatccatcc
301 ggcgtaatac gactcactat agggagagcg gccgccagat cttccggatg gctcgagttt
361 ttcagcaaga taagttccta tactttctag agaataggaa cttccatatg ttagaaacaa
421 atttattttt aaagttttat ttttaataat ttctagccct cccacacata accagagggc
481 agcaattcac gaatcccaac tgccgtcggc tgtccatcac tgtccttcac tatggctttg
541 atcccaggat gcagatcgag aagaacctgt cggcaccgtc cgcaggggct caagatgccc
601 ctgttctcat ttccgatcgc gacgatacaa gtcaggttgc cagctgccgc agcagcagca
661 gtgcccagca ccacgagttc tgcgcaaggt cccccagtaa aatgatatac attgacacca
721 gtgaagatgc ggccgtcgct agagagagct gcgctggcga cgctgtagtc ttcagagatg
781 gggatgctgt tgattgtagc cgttgctctt tcaatgaggg tggattcttc ttgagacaaa
841 ggcttggcca tatgcctagg cttgttgatt tgttttttgt ttatttactg cggtaattcc
901 tagtcttaga agcttattga acttgttgat gccttctggt tgggaaatcc tcctctttat
961 acccgatcca gtccaatcga aactgtgggc cgtggtgccc gataagataa gaaagccgtg
1021 tgcaagtcga tcggcgtgcg caaaagcata atgctttttc tgcccggcga tgataagcga
1081 tttgcacacg ccctgataac aggaattgat ttttttggtg cgcaaagcag tgaaagctct
1141 ataagaatgc tggaccgcca gtcgacaatc atcaaacagt gctagctaac ggctaagtga
1201 atcaagttta ttttaatata atcaacaaca aaatatttta acacaaacct agggaagttc
1261 ctatactttc tagagaatag gaacttctaa tccaaaatgg gatccagcgc ttggagccac
1321 ccgcagttcg aaaaaaccgg tgaattcggt acctctccag ctgctccaag cggcggtact
1381 agtatctttc tagaagatct cctacaatat tctcagctgc catggaaaat cgatgttctt
1441 cttttattct ctcaagattt tcaggctgta tattaaaact tatattaaga actatgctaa
1501 ccacctcatc aggaaccgtt gtaggtggcg tgggttttct tggcaatcga ctctcatgaa
1561 aactacgagc taaatattca atatgttcct cttgaccaac tttattctgc attttttttg
1621 aacgaggttt agagcaagct tcaggaaact gagacaggaa ttttattaaa aatttaaatt
1681 ttgaagaaag ttcagggtta atagcatcca ttttttgctt tgcaagttcc tcagcattct
1741 taacaaaaga cgtctctttt gacatgttta aagtttaaac ctcctgtgtg aaattattat
1801 ccgctcataa ttccacacat tatacgagcc ggaagcataa agtgtaaagc ctggggtgcc
1861 taatgagtga gctaactcac attaattgcg ttgcgctcac tgccaattgc tttccagtcg
1921 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
1981 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
2041 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
2101 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
2161 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
2221 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
2281 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
2341 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
2401 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
2461 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
2521 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
2581 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
2641 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
2701 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
2761 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
2821 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
2881 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
2941 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
3001 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
3061 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
3121 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
3181 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
3241 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
3301 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
3361 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
3421 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
3481 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
3541 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
3601 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
3661 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
3721 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
3781 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
3841 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
3901 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
3961 tataaaaata ggcgtatcac gaggcc
//
| {
"Vector backbone": "pJet1.2",
"Backbone size w/o insert (bp)": "2974",
"Vector type": "Insect Expression",
"Selectable markers": "Blasticidin"
} | {
"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 Comprehensive Toolbox for Genome Editing in Cultured Drosophila melanogaster Cells",
"Publication": "G3 (Bethesda). 2016 Jun 1;6(6):1777-85. doi: 10.1534/g3.116.028241.",
"DOI": "10.1534/g3.116.028241.",
"Authors": "Kunzelmann S, Bottcher R, Schmidts I, Forstemann K /g",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/27172193",
"PubMed ID": "PubMed 27172193"
} | {
"Gene/Insert name": "single Strep tag",
"Species": "synthetic",
"Insert Size (bp)": "63",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "XhoI (not destroyed)",
"3′ cloning site": "SpeI (not destroyed)",
"5′ sequencing primer": "pJet fwd",
"3′ sequencing primer": "pJet 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": ""
} |
|
3XFlagNICD2 | 20184 | High | Raphael Kopan | Ong et al J Biol Chem. 2006 Feb 24. 281(8):5106-19. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/219884/548f7380-b5dc-480d-bed8-64d0653405bb/addgene-plasmid-20184-sequence-219884.gbk | full | LOCUS sequence-219884- 7012 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 7012)
AUTHORS Ong CT, Cheng HT, Chang LW, Ohtsuka T, Kageyama R, Stormo GD, Kopan
R
TITLE Target selectivity of vertebrate notch proteins. Collaboration
between discrete domains and CSL-binding site architecture
determines activation probability.
JOURNAL J Biol Chem. 2006 Feb 24. 281(8):5106-19.
PUBMED 16365048
REFERENCE 2 (bases 1 to 7012)
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: "J Biol
Chem. 2006 Feb 24. 281(8):5106-19."
FEATURES Location/Qualifiers
source 1..7012
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 42..421
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 422..625
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 576..596
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
CDS 655..720
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
polyA_signal 3077..3699
/label=hGH poly(A) signal
/note="human growth hormone polyadenylation signal"
primer_bind complement(3276..3295)
/label=hGH-PA-R
/note="Human growth hormone terminator, reverse primer"
primer_bind complement(3723..3743)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 3728..4057
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3908..4043
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(4095..4114)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(4096..4114)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(4128..4144)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4128..4144)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4141..4163)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4152..4168
/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(4176..4206)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4221..4242
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4359..4376)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4530..5118)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4610..4629)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5289..6149)
/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 5912..5931
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6150..6254)
/gene="bla"
/label=AmpR promoter
rep_origin 6281..6736
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(6368..6387)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 6578..6599
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 6862..6884
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 6876..6893
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 6877..6893
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
ORIGIN
1 gtacatttat attggctcat gtccaatatg accgccatgt tgacattgat tattgactag
61 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
121 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
181 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
241 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
301 tccgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
361 gaccttacgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
421 ggtgatgcgg ttttggcagt acaccaatgg gcgtggatag cggtttgact cacggggatt
481 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
541 ctttccaaaa tgtcgtaata accccgcccc gttgacgcaa atgggcggta ggcgtgtacg
601 gtgggaggtc tatataagca gagctcgttt agtgaaccgt cagaattcac catggactac
661 aaagaccatg acggtgatta taaagatcat gatatcgatt acaaggatga cgatgacaag
721 cttgcatgcc tgcaggtcga ctctagaatg gccaagcgga agcgcaagca tggcttcctc
781 tggctgccgg aagggttcac cctccgccga gactctagca atcacaagcg ccgtgaacct
841 gtgggacagg atgccgtggg gctgaaaaat ctctccgtgc aagtgtcaga ggctaacctg
901 attggttctg ggacaagtga acattgggtt gatgatgaag gaccccagcc aaagaaagcc
961 aaggctgagg atgaggcttt gctgtcggaa gatgacccca tcgatcgacg gccctggaca
1021 cagcagcacc ttgaagctgc agacatccgc cacactccat ccctggcact cactcctcct
1081 caggcagaac aggaggtgga cgtgctggac gtgaatgtcc gaggcccaga tgggtgtact
1141 ccactgatgc tggcttctct ccgaggaggc agctcagacc tgagtgatga agacgaagat
1201 gctgaggact cttctgccaa catcatcaca gacttggtct accaaggtgc cagccttcaa
1261 gcacagacag accgcactgg cgagatggcc ctgcaccttg cagcccgcta ttcgagagct
1321 gatgctgcca aacgcctcct ggatgctggt gcggatgcaa atgcccagga caacatgggc
1381 cgatgtcctc ttcacgctgc ggtggcagca gacgcccaag gtgtctttca gattctgatc
1441 cgcaaccgtg taaccgatct ggatgccaga atgaacgatg gtactacccc cctgatcctg
1501 gctgcccgcc tggctgtgga aggaatggtg gcagagttga tcaattgcca agcagatgtc
1561 aatgcagtgg atgatcatgg aaaatctgcc ctccactggg cagctgctgt caataatgtg
1621 gaggcgactc ttctgctgtt gaagaatggg gccaacagag atatgcagga caataaggaa
1681 gagacacctt tgtttcttgc tgcccgagag ggaagttatg aagcggccaa aatcctgtta
1741 gaccattttg ccaaccggga catcactgac cacatggacc gccttccccg ggatgtggct
1801 cgggaccgca tgcaccatga catcgttcgc ctcctggacg agtataacgt gactcccagc
1861 cctccgggaa cggtcttgac ttctgcgctc tcacctgtcc tctgtgggcc caacaggtct
1921 ttcctcagtc tgaagcacac cccaatgggt aagaaggcta ggcggcccaa caccaagagc
1981 accatgccca cgagcctgcc taaccttgcc aaggaggcca aggatgccaa gggcagcagg
2041 aggaagaagt gtctgaacga gaaggtccag ctgtccgaga gctcagtgac tctatccccc
2101 gtcgattcgc tcgagtctcc tcacacgtat gtctccgatg ccacatcctc tcccatgatc
2161 acatcccctg gaatcttaca ggcctcgccc acccccctgc tggctgctgc cgccccggct
2221 gccccagtgc acacacagca tgcgctgtct ttctctaacc ttcatgacat gcagcctttg
2281 gctcctggag ccagcaccgt gctcccctcg gtcagccagc tgctatccca ccaccacatc
2341 gcgcccccag gtagtagcag tgcaggaagc ttgggcaggt tacatccagt tcctgtccca
2401 gcagactgga tgaaccgtgt ggagatgaac gagacccagt acagtgaaat gtttggcatg
2461 gtcctggctc ctgcagaggg agcccaccct ggcatagcag ctccccagag cagacctccg
2521 gaagggaagc acatgtccac ccagcgggag cccttgcctc ccatcgtgac tttccagctt
2581 atcccaaaag gcagcattgc ccaggcagcc ggagctcccc agacgcagtc cagttgccct
2641 ccagctgttg caggcccctt gccctctatg taccagatcc cagagatgcc ccgtttgccc
2701 agtgtggctt tcccacctac catgatgccc cagcaggagg ggcaggtagc tcagaccatt
2761 gtgccaacct atcatccttt cccagcctct gtgggcaagt accccacacc cccttcccaa
2821 cacagttacg cctcctcaaa tgctgctgag cgaaccccca gtcatggtgg tcacctccag
2881 ggcgagcacc catacctgac accatcccca gagtctcctg accaatggtc aagctcttca
2941 ccacactctg catctgactg gtcagatgtg accaccagcc caactcctgg aggtggtgga
3001 ggcggtcagc ggggacccgg aacacacatg tccgagccac cacacagcaa catgcaggtg
3061 tatgcatagg gatcccgggt ggcatccctg tgacccctcc ccagtgcctc tcctggccct
3121 ggaagttgcc actccagtgc ccaccagcct tgtcctaata aaattaagtt gcatcatttt
3181 gtctgactag gtgtccttct ataatattat ggggtggagg ggggtggtat ggagcaaggg
3241 gcaagttggg aagacaacct gtagggcctg cggggtctat tgggaaccaa gctggagtgc
3301 agtggcacaa tcttggctca ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc
3361 tcagcctccc gagttgttgg gattccaggc atgcatgacc aggctcagct aatttttgtt
3421 tttttggtag agacggggtt tcaccatatt ggccaggctg gtctccaact cctaatctca
3481 ggtgatctac ccaccttggc ctcccaaatt gctgggatta caggcgtgaa ccactgctcc
3541 cttccctgtc cttctgattt taaaataact ataccagcag gaggacgtcc agacacagca
3601 taggctacct ggccatgccc aaccggtggg acatttgagt tgcttgcttg gcactgtcct
3661 ctcatgcgtt gggtccactc agtagatgcc tgttgaattg ggtacgcggc cagcttggct
3721 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
3781 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
3841 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
3901 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
3961 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
4021 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcctcgagg aactgaaaaa
4081 ccagaaagtt aattccctat agtgagtcgt attaaattcg taatcatgtc atagctgttt
4141 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
4201 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
4261 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
4321 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
4381 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
4441 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
4501 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
4561 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
4621 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
4681 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
4741 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
4801 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
4861 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
4921 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
4981 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
5041 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
5101 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
5161 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
5221 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
5281 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
5341 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
5401 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
5461 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
5521 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
5581 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
5641 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
5701 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
5761 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
5821 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
5881 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
5941 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
6001 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
6061 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
6121 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
6181 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
6241 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca
6301 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
6361 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
6421 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
6481 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
6541 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
6601 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
6661 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
6721 ttaacgctta caatttccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
6781 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
6841 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgccaagc
6901 tgatctatac attgaatcaa tattggcaat tagccatatt agtcattggt tatatagcat
6961 aaatcaatat tggctattgg ccattgcata cgttgtatct atatcataat at
//
| {
"Vector backbone": "p3XFLAG-CMV-7",
"Backbone size w/o insert (bp)": "4700",
"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": "Target selectivity of vertebrate notch proteins. Collaboration between discrete domains and CSL-binding site architecture determines activation probability",
"Publication": "J Biol Chem. 2006 Feb 24. 281(8):5106-19.",
"DOI": "10.1074/jbc.M506108200",
"Authors": "Ong CT, Cheng HT, Chang LW, Ohtsuka T, Kageyama R, Stormo GD, Kopan R",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/16365048",
"PubMed ID": "PubMed 16365048"
} | {
"Gene/Insert name": "Notch 2 intracellular domain",
"Species": "M. musculus (mouse)",
"Insert Size (bp)": "2403",
"Mutation": "Contains murine Notch 2 Intracellular Domain (Met-1699 to Ala-2470)",
"Entrez Gene": "Notch2",
"Promoter": "CMV",
"Tag / Fusion Protein": "3X FLAG (N terminal on backbone)"
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "Xba1 (not destroyed)",
"3′ cloning site": "BamH1 (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": ""
} |
|
pET-CVm-VAMP8 | 168028 | Expresses eCFP-VAMP8(G7-R67)-mVenus in bacterial cells | David Liu | Blum et al Science. 2021 Feb 19;371(6531):803-810. doi: 10.1126/science.abf5972. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/329626/643cc35e-a9ce-484e-b2dc-52eb9a319d85/addgene-plasmid-168028-sequence-329626.gbk | full | LOCUS sequence-329626- 6932 bp DNA circular SYN 26-AUG-2024
DEFINITION Expresses eCFP-VAMP8(G7-R67)-mVenus in bacterial cells.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6932)
AUTHORS Blum TR, Liu H, Packer MS, Xiong X, Lee PG, Zhang S, Richter M,
Minasov G, Satchell KJF, Dong M, Liu DR
TITLE Phage-assisted evolution of botulinum neurotoxin proteases with
reprogrammed specificity.
JOURNAL Science. 2021 Feb 19;371(6531):803-810. doi:
10.1126/science.abf5972.
PUBMED 33602850
REFERENCE 2 (bases 1 to 6932)
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: "2021-02-19- 19"; volume: "371";
issue: "6531"; pages: "803-810"
FEATURES Location/Qualifiers
source 1..6932
/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(158..874)
/codon_start=1
/product="yellow fluorescent protein (YFP) with fast and
efficient maturation (Nagai et al., 2002)"
/label=Venus
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(200..221)
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind 549..568
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 808..829
/label=EGFP-N
/note="EGFP, reverse primer"
CDS complement(881..904)
/codon_start=1
/product="Xpress(TM) epitope tag, including an enterokinase
recognition and cleavage site"
/label=Xpress(TM) tag
/translation="DLYDDDDK"
CDS complement(1835..1864)
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
RBS 1873..1895
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
protein_bind 1910..1934
/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(1934..1953)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(1935..1953)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(2037..2056)
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
promoter 2262..2339
/gene="lacI"
/label=lacI promoter
CDS 2340..3422
/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(2359..2378)
/label=LacI-R
/note="LacI, reverse primer"
CDS 4231..4422
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
primer_bind complement(4416..4438)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
misc_feature 4524..4663
/label=bom
/note="basis of mobility region from pBR322"
primer_bind 4579..4598
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(4678..4695)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4849..5437)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4929..4948)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind complement(5548..5567)
/label=pENTR-R
/note="pENTR vectors, reverse primer"
CDS 5559..6374
/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 5559..6374
/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(5632..5651)
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
rep_origin complement(6466..6921)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(6603..6624)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 6815..6834
/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 ggtggtgctt gtacagctcg tccatgccga
181 gagtgatccc ggcggcggtc acgaactcca gcaggaccat gtgatcgcgc ttctcgttgg
241 ggtctttgct cagggcggac tggtagctca ggtagtggtt gtcgggcagc agcacggggc
301 cgtcgccgat gggggtgttc tgctggtagt ggtcggcgag ctgcacgccg ccgtcctcga
361 tgttgtggcg gatcttgaag ttggccttga tgccgttctt ctgcttgtcg gcggtgatat
421 agacgttgtg gctgttgtag ttgtactcca gcttgtgccc caggatgttg ccgtcctcct
481 tgaagtcgat gcccttcagc tcgatgcggt tcaccagggt gtcgccctcg aacttcacct
541 cggcgcgggt cttgtagttg ccgtcgtcct tgaagaagat ggtgcgctcc tggacgtagc
601 cttcgggcat ggcggacttg aagaagtcgt gctgcttcat gtggtcgggg tagcgggcga
661 agcactgcag gccgtagccc agggtggtca cgagggtggg ccagggcacg ggcagcttgc
721 cggtggtgca gatcagcttc agggtcagct tgccgtaggt ggcatcgccc tcgccctcgc
781 cggacacgct gaacttgtgg ccgtttacgt cgccgtccag ctcgaccagg atgggcacca
841 ccccggtgaa cagctcctcg cccttgctca ccatcggatc cttatcgtca tcgtcgtaca
901 gatcacttcc accactattt gaacgcgcaa ccttttgact ggtagtctta aaatgctctg
961 aagtggcctc taagtcttcg gttttattgc gcagatgctc aaggttctcg ccacgggcca
1021 aaatacgctc tacattttgg gtcatgatat ttttcacgcc ttcgacctct gattgtaagt
1081 tacgaacgcg gtcgttacca cctccacgac tacgcccacc tttatacagt tcatccatac
1141 catgagtaat accagcggcg gtaacgaact ctaaaaggac catatggtcg cgtttctcat
1201 tggggtcctt agacagggct gactgtgtgc tcaaatagtg attatcaggt aaaaggacgg
1261 ggccatctcc aattggtgtg ttttgctggt aatggtctgc aagctggacg cttccatcct
1321 caatgttgtg acgaatcttg aaatttgcct tgattccgtt tttttgttta tcagccgtga
1381 tgtagacgtt atgagagata tagttgtatt ctaatttatg tcctaagatg ttgccatcct
1441 ctttaaagtc gatacccttc aactcaatgc gattaacaag agtatctccc tcaaacttaa
1501 cctcggcgcg ggtcttatag tttccatcat ctttaaaaaa aatggtgcgt tcctggacgt
1561 aaccttctgg catcgcggac ttgaaaaagt cgtgttgttt catatggtca gggtagcgag
1621 cgaaacactg cacgccccag gcgaacgtag taaccaatgt aggccacggt actggaagct
1681 tgccggtagt gcaaataaat tttaatgtca actttccgta cgtcgcatcg ccctctcctt
1741 ccccagatac ggaaaacttg tgaccattca catcgccatc cagttctaca agaatgggga
1801 ccacgccggt aaacaattcc tcccccttag acatcaagtc ttcctcagag ataagttttt
1861 gttccatgta tatctccttc ttaaagttaa acaaaattat ttctagaggg gaattgttat
1921 ccgctcacaa ttcccctata gtgagtcgta ttaatttcgc gggatcgaga tctcgatcct
1981 ctacgccgga cgcatcgtgg ccggcatcac cggcgccaca ggtgcggttg ctggcgccta
2041 tatcgccgac atcaccgatg gggaagatcg ggctcgccac ttcgggctca tgagcgcttg
2101 tttcggcgtg ggtatggtgg caggccccgt ggccggggga ctgttgggcg ccatctcctt
2161 gcatgcacca ttccttgcgg cggcggtgct caacggcctc aacctactac tgggctgctt
2221 cctaatgcag gagtcgcata agggagagcg tcgagatccc ggacaccatc gaatggcgca
2281 aaacctttcg cggtatggca tgatagcgcc cggaagagag tcaattcagg gtggtgaatg
2341 tgaaaccagt aacgttatac gatgtcgcag agtatgccgg tgtctcttat cagaccgttt
2401 cccgcgtggt gaaccaggcc agccacgttt ctgcgaaaac gcgggaaaaa gtggaagcgg
2461 cgatggcgga gctgaattac attcccaacc gcgtggcaca acaactggcg ggcaaacagt
2521 cgttgctgat tggcgttgcc acctccagtc tggccctgca cgcgccgtcg caaattgtcg
2581 cggcgattaa atctcgcgcc gatcaactgg gtgccagcgt ggtggtgtcg atggtagaac
2641 gaagcggcgt cgaagcctgt aaagcggcgg tgcacaatct tctcgcgcaa cgcgtcagtg
2701 ggctgatcat taactatccg ctggatgacc aggatgccat tgctgtggaa gctgcctgca
2761 ctaatgttcc ggcgttattt cttgatgtct ctgaccagac acccatcaac agtattattt
2821 tctcccatga agacggtacg cgactgggcg tggagcatct ggtcgcattg ggtcaccagc
2881 aaatcgcgct gttagcgggc ccattaagtt ctgtctcggc gcgtctgcgt ctggctggct
2941 ggcataaata tctcactcgc aatcaaattc agccgatagc ggaacgggaa ggcgactgga
3001 gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa tgagggcatc gttcccactg
3061 cgatgctggt tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt accgagtccg
3121 ggctgcgcgt tggtgcggat atctcggtag tgggatacga cgataccgaa gacagctcat
3181 gttatatccc gccgttaacc accatcaaac aggattttcg cctgctgggg caaaccagcg
3241 tggaccgctt gctgcaactc tctcagggcc aggcggtgaa gggcaatcag ctgttgcccg
3301 tctcactggt gaaaagaaaa accaccctgg cgcccaatac gcaaaccgcc tctccccgcg
3361 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
3421 gagcgcaacg caattaatgt aagttagctc actcattagg caccgggatc tcgaccgatg
3481 cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat gactatcgtc
3541 gccgcactta tgactgtctt ctttatcatg caactcgtag gacaggtgcc ggcagcgctc
3601 tgggtcattt tcggcgagga ccgctttcgc tggagcgcga cgatgatcgg cctgtcgctt
3661 gcggtattcg gaatcttgca cgccctcgct caagccttcg tcactggtcc cgccaccaaa
3721 cgtttcggcg agaagcaggc cattatcgcc ggcatggcgg ccccacgggt gcgcatgatc
3781 gtgctcctgt cgttgaggac tcggctaggc tggcggggtt gccttactgg ttagcagaat
3841 gaatcaccga tacgcgagcg aacgtgaagc gactgctgct gcaaaacttc tgcgacctga
3901 gcaacaacat gaatggtctt cggtttccgt gtttcgtaaa gtctggaaac gcggaagtca
3961 gcgccctgca ccattatgtt ccggatctgc atcgcaggat gctgctggct accctgtgga
4021 acacctacat ctgtattaac gaagcgctgg cattgaccct gagtgatttt tctctggtcc
4081 cgccgcatcc ataccgccag ttgtttaccc tcacaacgtt ccagtaaccg ggcatgttca
4141 tcatcagtaa cccgtatcgt gagcatcctc tctcgtttca tcggtatcat tacccccatg
4201 aacagaaatc ccccttacac gtaggcatca gtgaccaaac aggaaaaaac cgcccttaac
4261 atggcccgct ttatcagaag ccagacatta acgcttctgg agaaactcaa cgagctggac
4321 gcggatgaac aggcagacat ctgtgaatcg cttcacgacc acgctgatga gctttaccgc
4381 agctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
4441 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
4501 gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga tagcggagtg
4561 tatactggct taactatgcg gcatcagagc agattgtact gagagtgcac cattgcggtg
4621 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgctctt ccgcttcctc
4681 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
4741 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
4801 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
4861 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
4921 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
4981 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
5041 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
5101 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
5161 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
5221 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
5281 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
5341 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
5401 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
5461 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgaacaataa
5521 aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa
5581 cgtcttgctc taggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat
5641 gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgattgtat gggaagcccg
5701 atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg
5761 agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta
5821 tccgtactcc tgatgatgca tggttactca ccactgcgat ccccgggaaa acagcattcc
5881 aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc
5941 tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc
6001 gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg
6061 acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat
6121 tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg
6181 aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg
6241 atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt
6301 ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg
6361 atgagttttt ctaagaatta attcatgagc ggatacatat ttgaatgtat ttagaaaaat
6421 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta
6481 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
6541 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
6601 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
6661 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
6721 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
6781 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
6841 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
6901 atgcgccgct acagggcgcg tcccattcgc ca
//
| {
"Vector backbone": "pET (pBR322)",
"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": "Phage-assisted evolution of botulinum neurotoxin proteases with reprogrammed specificity",
"Publication": "Science. 2021 Feb 19;371(6531):803-810. doi: 10.1126/science.abf5972.",
"DOI": "10.1126/science.abf5972.",
"Authors": "Blum TR, Liu H, Packer MS, Xiong X, Lee PG, Zhang S, Richter M, Minasov G, Satchell KJF, Dong M, Liu DR /scienceabf",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/33602850",
"PubMed ID": "PubMed 33602850"
} | {
"Gene/Insert name": "eCFP-VAMP8(G7-R67)-mVenus",
"Species": "Synthetic",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Ligation Independent Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pLV-TRE3G-hYAP1-CMV-mCherry-T2A-Puro | 176850 | lentiviral vector for doxycycline-inducible YAP1 overexpression | Roland Friedel | Junqueira Alves et al Nat Commun. 2021 Oct 14;12(1):6019. doi: 10.1038/s41467-021-26296-7. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/347164/898d29f7-02b4-45e7-80a6-d4ce95f470e1/addgene-plasmid-176850-sequence-347164.gbk | full | LOCUS sequence-347164- 10089 bp DNA circular SYN 25-AUG-2024
DEFINITION lentiviral vector for doxycycline-inducible YAP1 overexpression.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10089)
AUTHORS Junqueira Alves C, Dariolli R, Haydak J, Kang S, Hannah T, Wiener
RJ, DeFronzo S, Tejero R, Gusella GL, Ramakrishnan A, Alves Dias R,
Wojcinski A, Kesari S, Shen L, Sobie EA, Rodrigues Furtado de
Mendonca JP, Azeloglu EU, Zou H, Friedel RH
TITLE Plexin-B2 orchestrates collective stem cell dynamics via actomyosin
contractility, cytoskeletal tension and adhesion.
JOURNAL Nat Commun. 2021 Oct 14;12(1):6019. doi: 10.1038/s41467-021-26296-7.
PUBMED 34650052
REFERENCE 2 (bases 1 to 10089)
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: "2021-10-14"; volume: "12(1):6019. doi"; pages: "
10.1038/s41467-021-26296-7"
FEATURES Location/Qualifiers
source 1..10089
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 3..229
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 230..410
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 457..582
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1075..1308
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1493..1537
/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 1803..1920
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
protein_bind 1938..1958
/gene="mutant version of attB"
/label=attB4
/bound_moiety="BP Clonase(TM)"
/note="core recombination site for the Gateway(R) BP
reaction"
protein_bind 1964..1982
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 2000..2018
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 2036..2054
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 2072..2090
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 2108..2126
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 2144..2162
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 2180..2198
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
primer_bind 2243..2267
/label=LNCX
/note="Human CMV promoter, forward primer"
protein_bind 2334..2358
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
regulatory 2359..2368
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
protein_bind complement(3892..3916)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
misc_feature 3934..4522
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(3987..4007)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(4405..4416)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
enhancer 4552..4931
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 4932..5135
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 5086..5106
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 5162..5171
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 5168..5875
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
primer_bind complement(5313..5331)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 5581..5600
/label=mCherry-F
/note="mCherry, forward primer"
CDS 5885..5938
/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 5939..6538
/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(5939..5958)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 6435..6455
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
LTR 6610..6843
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 6915..7036
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(6952..6971)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 7006..7025
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 7076..7211
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(7231..7250)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(7232..7250)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(7260..7277)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(7260..7276)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(7269..7291)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 7418..7873
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(7505..7524)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 7715..7736
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 7899..8003
/gene="bla"
/label=AmpR promoter
CDS 8004..8864
/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(8222..8241)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 9035..9623
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 9524..9543
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 9777..9794
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 9911..9932
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 9947..9977
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 9985..10001
/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 9990..10012
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 10009..10025
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 10009..10025
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 10044..10064
/label=T3
/note="T3 promoter, forward primer"
promoter 10046..10064
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
1 aatgtagtct tatgcaatac tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca
61 tgccttacaa ggagagaaaa agcaccgtgc atgccgattg gtggaagtaa ggtggtacga
121 tcgtgcctta ttaggaaggc aacagacggg tctgacatgg attggacgaa ccactgaatt
181 gccgcattgc agagatattg tatttaagtg cctagctcga tacataaacg ggtctctctg
241 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
301 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
361 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg
421 aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt
481 gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg
541 actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga
601 attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta
661 aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta
721 gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga
781 tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg
841 atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt
901 aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga
961 caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc
1021 acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc
1081 tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct
1141 gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag
1201 ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca
1261 ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg
1321 ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa
1381 atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa
1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcgcta
1801 gcttttaaaa gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata
1861 atagcaacag acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt
1921 actagtgatt atcggatcaa ctttgtatag aaaagttgtt tactccctat cagtgataga
1981 gaacgtatga agagtttact ccctatcagt gatagagaac gtatgcagac tttactccct
2041 atcagtgata gagaacgtat aaggagttta ctccctatca gtgatagaga acgtatgacc
2101 agtttactcc ctatcagtga tagagaacgt atctacagtt tactccctat cagtgataga
2161 gaacgtatat ccagtttact ccctatcagt gatagagaac gtataagctt taggcgtgta
2221 cggtgggcgc ctataaaagc agagctcgtt tagtgaaccg tcagatcgcc tggagcaatt
2281 ccacaacact tttgtcttat accaactttc cgtaccactt cctaccctcg taaacaagtt
2341 tgtacaaaaa agcaggctgc caccatggat cccgggcagc agccgccgcc tcaaccggcc
2401 ccccagggcc aagggcagcc gccttcgcag cccccgcagg ggcagggccc gccgtccgga
2461 cccgggcaac cggcacccgc ggcgacccag gcggcgccgc aggcaccccc cgccgggcat
2521 cagatcgtgc acgtccgcgg ggactcggag accgacctgg aggcgctctt caacgccgtc
2581 atgaacccca agacggccaa cgtgccccag accgtgccca tgaggctccg gaagctgccc
2641 gactccttct tcaagccgcc ggagcccaaa tcccactccc gacaggccag tactgatgca
2701 ggcactgcag gagccctgac tccacagcat gttcgagctc attcctctcc agcttctctg
2761 cagttgggag ctgtttctcc tgggacactg acccccactg gagtagtctc tggcccagca
2821 gctacaccca cagctcagca tcttcgacag tcttcttttg agatacctga tgatgtacct
2881 ctgccagcag gttgggagat ggcaaagaca tcttctggtc agagatactt cttaaatcac
2941 atcgatcaga caacaacatg gcaggacccc aggaaggcca tgctgtccca gatgaacgtc
3001 acagccccca ccagtccacc agtgcagcag aatatgatga actcggcttc aggtcctctt
3061 cctgatggat gggaacaagc catgactcag gatggagaaa tttactatat aaaccataag
3121 aacaagacca cctcttggct agacccaagg cttgaccctc gttttgccat gaaccagaga
3181 atcagtcaga gtgctccagt gaaacagcca ccacccctgg ctccccagag cccacaggga
3241 ggcgtcatgg gtggcagcaa ctccaaccag cagcaacaga tgcgactgca gcaactgcag
3301 atggagaagg agaggctgcg gctgaaacag caagaactgc ttcggcaggt gaggccacag
3361 gcaatgcgga atatcaatcc cagcacagca aattctccaa aatgtcagga gttagccctg
3421 cgtagccagt taccaacact ggagcaggat ggtgggactc aaaatccagt gtcttctccc
3481 gggatgtctc aggaattgag aacaatgacg accaatagct cagatccttt ccttaacagt
3541 ggcacctatc actctcgaga tgagagtaca gacagtggac taagcatgag cagctacagt
3601 gtccctcgaa ccccagatga cttcctgaac agtgtggatg agatggatac aggtgatact
3661 atcaaccaaa gcaccctgcc ctcacagcag aaccgtttcc cagactacct tgaagccatt
3721 cctgggacaa atgtggacct tggaacactg gaaggagatg gaatgaacat agaaggagag
3781 gagctgatgc caagtctgca ggaagctttg agttctgaca tccttaatga catggagtct
3841 gttttggctg ccaccaagct agataaagaa agctttctta catggttata gacccagctt
3901 tcttgtacaa agtggtgata atcgaattcc gataatcaac ctctggatta caaaatttgt
3961 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
4021 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
4081 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
4141 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
4201 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
4261 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
4321 tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc
4381 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
4441 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
4501 tccctttggg ccgcctcccc gcatcgggaa ttcccgcggt tcgaacgcgt tgacattgat
4561 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
4621 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
4681 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
4741 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
4801 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
4861 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
4921 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
4981 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
5041 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
5101 ggcgtgtacg gtgggaggtc tatataagca gagctctctg gctaactaga gaacccactg
5161 cgccaccatg gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg
5221 cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg
5281 cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc
5341 cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacgt
5401 gaagcacccc gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg
5461 ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct
5521 gcaggacggc gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg
5581 ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc tccgagcgga tgtaccccga
5641 ggacggcgcc ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcggccacta
5701 cgacgctgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta
5761 caacgtcaac atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca
5821 gtacgaacgc gccgagggcc gccactccac cggcggcatg gacgagctgt acaagggctc
5881 cggagagggc aggggaagtc ttctaacatg cggggacgtg gaggaaaatc ccggccccat
5941 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg
6001 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg
6061 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat
6121 cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag
6181 cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc
6241 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc
6301 cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag
6361 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga
6421 gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga
6481 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgagg
6541 tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa
6601 aggggggact ggaagggcta attcactccc aacgaagaca agatctgctt tttgcttgta
6661 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
6721 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
6781 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
6841 gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat
6901 cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
6961 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
7021 tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc cgcccatccc
7081 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
7141 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
7201 ttttggaggc ctagggacgt acccaattcg ccctatagtg agtcgtatta cgcgcgctca
7261 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
7321 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
7381 ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta
7441 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
7501 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
7561 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
7621 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
7681 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
7741 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
7801 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
7861 acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
7921 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
7981 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
8041 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
8101 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
8161 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
8221 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
8281 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
8341 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
8401 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
8461 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
8521 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
8581 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
8641 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
8701 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
8761 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
8821 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
8881 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
8941 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
9001 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
9061 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
9121 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
9181 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
9241 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
9301 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
9361 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
9421 gtgagctatg agaaagcgcc acgcttcccg aagagagaaa ggcggacagg tatccggtaa
9481 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
9541 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
9601 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
9661 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
9721 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
9781 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
9841 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
9901 gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc
9961 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
10021 atgaccatga ttacgccaag cgcgcaatta accctcacta aagggaacaa aagctggagc
10081 tgcaagctt
//
| {
"Vector backbone": "pLV[Tet]-mCherry:T2A:Puro-TRE3G",
"Backbone size w/o insert (bp)": "8562",
"Vector type": "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 ; Takara Bio Limited Use Label License (formerly Clontech) ; Tet Systems Limited Use Label License",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Plexin-B2 orchestrates collective stem cell dynamics via actomyosin contractility, cytoskeletal tension and adhesion",
"Publication": "Nat Commun. 2021 Oct 14;12(1):6019. doi: 10.1038/s41467-021-26296-7.",
"DOI": "10.1038/s41467-021-26296-7.",
"Authors": "Junqueira Alves C, Dariolli R, Haydak J, Kang S, Hannah T, Wiener RJ, DeFronzo S, Tejero R, Gusella GL, Ramakrishnan A, Alves Dias R, Wojcinski A, Kesari S, Shen L, Sobie EA, Rodrigues Furtado de Mendonca JP, Azeloglu EU, Zou H, Friedel RH /s---",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/34650052",
"PubMed ID": "PubMed 34650052"
} | {
"Gene/Insert name": "hYAP1",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "1527",
"Mutation": "full length YAP1",
"Entrez Gene": "YAP1",
"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": ""
} |
|
Lenti-(BB)-hPGK-KRAB-dCas9-P2A-BlastR rs7732130-guide4 | 125447 | CRISPR-mediated repression of human islet enhancer containing T2D-associated variant rs7732130 (ZBED3 GWAS locus) | Jorge Ferrer | Miguel-Escalada et al Nat Genet. 2019 Jun 28. pii: 10.1038/s41588-019-0457-0. doi: 10.1038/s41588-019-0457-0. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/251363/94756888-c8b7-480b-b4f1-c6fef6482829/addgene-plasmid-125447-sequence-251363.gbk | full | LOCUS sequence-251363- 13383 bp DNA circular SYN 26-AUG-2024
DEFINITION CRISPR-mediated repression of human islet enhancer containing
T2D-associated variant rs7732130 (ZBED3 GWAS locus).
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 13383)
AUTHORS Miguel-Escalada I, Bonas-Guarch S, Cebola I, Ponsa-Cobas J,
Mendieta-Esteban J, Atla G, Javierre BM, Rolando DMY, Farabella I,
Morgan CC, Garcia-Hurtado J, Beucher A, Moran I, Pasquali L,
Ramos-Rodriguez M, Appel EVR, Linneberg A, Gjesing AP, Witte DR,
Pedersen O, Grarup N, Ravassard P, Torrents D, Mercader JM, Piemonti
L, Berney T, de Koning EJP, Kerr-Conte J, Pattou F, Fedko IO, Groop
L, Prokopenko I, Hansen T, Marti-Renom MA, Fraser P, Ferrer J
TITLE Human pancreatic islet three-dimensional chromatin architecture
provides insights into the genetics of type 2 diabetes.
JOURNAL Nat Genet. 2019 Jun 28. pii: 10.1038/s41588-019-0457-0. doi:
10.1038/s41588-019-0457-0.
PUBMED 31253982
REFERENCE 2 (bases 1 to 13383)
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 Genet.
2019 Jun 28. pii: 10.1038/s41588-019-0457-0. doi:
10.1038/s41588-019-0457-0."
FEATURES Location/Qualifiers
source 1..13383
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_RNA 1..76
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
promoter 107..617
/label=hPGK promoter
/note="human phosphoglycerate kinase 1 promoter"
primer_bind 536..554
/label=hPGK-F
/note="Human PGK promoter, forward primer"
regulatory 630..639
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 666..851
/codon_start=1
/product="Kruppel-associated box (KRAB) transcriptional
repression domain from the human zinc finger protein ZNF10
(Margolin et al., 1994)"
/label=KRAB
/translation="RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGY
QLTKPDVILRLEKGEEP"
CDS 873..4976
/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="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTSLGAPAAFKYFDTTIDRKRYTSTKEVL
DATLIHQSITGLYETRIDLSQLGGD"
CDS 4980..5006
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 4986..5012
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/note="This is a variant of the standard HA tag, which has
sequence YPYDVPDYA. Some anti-HA antibodies may not
recognize this variant."
/translation="YDVPDYASL"
CDS 5025..5045
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 5052..5072
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 5118..5174
/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 5175..5573
/codon_start=1
/gene="Aspergillus terreus BSD"
/product="blasticidin S deaminase"
/label=BSD
/note="confers resistance to blasticidin"
/translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
KAIVKDSDGQPTAVGIRELLPSGYVWEG"
misc_feature 5598..6186
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(5651..5671)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(6069..6080)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 6258..6491
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
primer_bind complement(6517..6534)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 6523..6747
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 6793..7221
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(6880..6899)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 7090..7111
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(7230..7250)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 7235..7564
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 7415..7550
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 7477..7496
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter 7612..7659
/label=EM7 promoter
/note="synthetic bacterial promoter "
CDS 7678..8052
/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 8182..8303
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(8219..8238)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 8273..8292
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(8352..8368)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(8352..8368)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(8365..8387)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 8376..8392
/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(8400..8430)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 8445..8466
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(8583..8600)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(8754..9342)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(8834..8853)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(9513..10373)
/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 10136..10155
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(10374..10478)
/gene="bla"
/label=AmpR promoter
primer_bind complement(10553..10572)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 10744..11123
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 11125..11323
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 11278..11298
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
LTR 11341..11521
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 11568..11693
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 12186..12419
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 12604..12648
/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 12946..13063
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 13114..13362
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA
(Domitrovich & Kunkel, 2003)"
primer_bind 13114..13134
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 13285..13304
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
ORIGIN
1 gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt
61 ggcaccgagt cggtgctttt ttgaattcgc tagctaggtc ttgaaagggg ttggggttgc
121 gccttttcca aggcagccct gggtttgcgc agggacgcgg ctgctctggg cgtggttccg
181 ggaaacgcag cggcgccgac cctgggtctc gcacattctt cacgtccgtt cgcagcgtca
241 cccggatctt cgccgctacc cttgtgggcc ccccggcgac gcttcctgct ccgcccctaa
301 gtcgggaagg ttccttgcgg ttcgcggcgt gccggacgtg acaaacggaa gccgcacgtc
361 tcactagtac cctcgcagac ggacagcgcc agggagcaat ggcagcgcgc cgaccgcgat
421 gggctgtggc caatagcggc tgctcagcag ggcgcgccga gagcagcggc cgggaagggg
481 cggtgcggga ggcggggtgt ggggcggtag tgtgggccct gttcctgccc gcgcggtgtt
541 ccgcattctg caagcctccg gagcgcacgt cggcagtcgg ctccctcgtt gaccgaatca
601 ccgacctctc tccccaggac cggtgatccg ccaccatgga tgctaagtca ctaactgcct
661 ggtcccggac actggtgacc ttcaaggatg tatttgtgga cttcaccagg gaggagtgga
721 agctgctgga cactgctcag cagatcgtgt acagaaatgt gatgctggag aactataaga
781 acctggtttc cttgggttat cagcttacta agccagatgt gatcctccgg ttggagaagg
841 gagaagagcc cggcggttcc ggcggagggt cgatggacaa gaagtacagc atcggcctgg
901 ccatcggcac caactctgtc ggctgggccg tgatcaccga cgagtacaag gtgcccagca
961 agaaattcaa ggtgctgggc aacaccgacc ggcacagcat caagaagaac ctgatcggcg
1021 ccctgctgtt cgacagcgga gaaacagccg aggccacccg gctgaagaga accgccagaa
1081 gaagatacac cagacggaag aaccggatct gctatctgca agagatcttc agcaacgaga
1141 tggccaaggt ggacgacagc ttcttccaca gactggaaga gtccttcctg gtggaagagg
1201 ataagaagca cgagcggcac cccatcttcg gcaacatcgt ggacgaggtg gcctaccacg
1261 agaagtaccc caccatctac cacctgagaa agaaactggt ggacagcacc gacaaggccg
1321 acctgcggct gatctatctg gccctggccc acatgatcaa gttccggggc cacttcctga
1381 tcgagggcga cctgaacccc gacaacagcg acgtggacaa gctgttcatc cagctggtgc
1441 agacctacaa ccagctgttc gaggaaaacc ccatcaacgc cagcggcgtg gacgccaagg
1501 ccatcctgtc tgccagactg agcaagagca gacggctgga aaatctgatc gcccagctgc
1561 ccggcgagaa gaagaatggc ctgttcggca acctgattgc cctgagcctg ggcctgaccc
1621 ccaacttcaa gagcaacttc gacctggccg aggatgccaa actgcagctg agcaaggaca
1681 cctacgacga cgacctggac aacctgctgg cccagatcgg cgaccagtac gccgacctgt
1741 ttctggccgc caagaacctg tccgacgcca tcctgctgag cgacatcctg agagtgaaca
1801 ccgagatcac caaggccccc ctgagcgcct ctatgatcaa gagatacgac gagcaccacc
1861 aggacctgac cctgctgaaa gctctcgtgc ggcagcagct gcctgagaag tacaaagaga
1921 ttttcttcga ccagagcaag aacggctacg ccggctacat cgatggcgga gccagccagg
1981 aagagttcta caagttcatc aagcccatcc tggaaaagat ggacggcacc gaggaactgc
2041 tcgtgaagct gaacagagag gacctgctgc ggaagcagcg gaccttcgac aacggcagca
2101 tcccccacca gatccacctg ggagagctgc acgccattct gcggcggcag gaagattttt
2161 acccattcct gaaggacaac cgggaaaaga tcgagaagat cctgaccttc cgcatcccct
2221 actacgtggg ccctctggcc aggggaaaca gcagattcgc ctggatgacc agaaagagcg
2281 aggaaaccat caccccctgg aacttcgagg aagtggtgga caagggcgcc agcgcccaga
2341 gcttcatcga gcggatgacc aacttcgata agaacctgcc caacgagaag gtgctgccca
2401 agcacagcct gctgtacgag tacttcaccg tgtacaacga gctgaccaaa gtgaaatacg
2461 tgaccgaggg aatgagaaag cccgccttcc tgagcggcga gcagaaaaaa gccatcgtgg
2521 acctgctgtt caagaccaac cggaaagtga ccgtgaagca gctgaaagag gactacttca
2581 agaaaatcga gtgcttcgac tccgtggaaa tctccggcgt ggaagatcgg ttcaacgcct
2641 ccctgggcac ataccacgat ctgctgaaaa ttatcaagga caaggacttc ctggacaatg
2701 aggaaaacga ggacattctg gaagatatcg tgctgaccct gacactgttt gaggacagag
2761 agatgatcga ggaacggctg aaaacctatg cccacctgtt cgacgacaaa gtgatgaagc
2821 agctgaagcg gcggagatac accggctggg gcaggctgag ccggaagctg atcaacggca
2881 tccgggacaa gcagtccggc aagacaatcc tggatttcct gaagtccgac ggcttcgcca
2941 acagaaactt catgcagctg atccacgacg acagcctgac ctttaaagag gacatccaga
3001 aagcccaggt gtccggccag ggcgatagcc tgcacgagca cattgccaat ctggccggca
3061 gccccgccat taagaagggc atcctgcaga cagtgaaggt ggtggacgag ctcgtgaaag
3121 tgatgggccg gcacaagccc gagaacatcg tgatcgaaat ggccagagag aaccagacca
3181 cccagaaggg acagaagaac agccgcgaga gaatgaagcg gatcgaagag ggcatcaaag
3241 agctgggcag ccagatcctg aaagaacacc ccgtggaaaa cacccagctg cagaacgaga
3301 agctgtacct gtactacctg cagaatgggc gggatatgta cgtggaccag gaactggaca
3361 tcaaccggct gtccgactac gatgtggacg ctatcgtgcc tcagagcttt ctgaaggacg
3421 actccatcga taacaaagtg ctgactcgga gcgacaagaa ccggggcaag agcgacaacg
3481 tgccctccga agaggtcgtg aagaagatga agaactactg gcgccagctg ctgaatgcca
3541 agctgattac ccagaggaag ttcgacaatc tgaccaaggc cgagagaggc ggcctgagcg
3601 aactggataa ggccggcttc atcaagagac agctggtgga aacccggcag atcacaaagc
3661 acgtggcaca gatcctggac tcccggatga acactaagta cgacgagaac gacaaactga
3721 tccgggaagt gaaagtgatc accctgaagt ccaagctggt gtccgatttc cggaaggatt
3781 tccagtttta caaagtgcgc gagatcaaca actaccacca cgcccacgac gcctacctga
3841 acgccgtcgt gggaaccgcc ctgatcaaaa agtaccctaa gctggaaagc gagttcgtgt
3901 acggcgacta caaggtgtac gacgtgcgga agatgatcgc caagagcgag caggaaatcg
3961 gcaaggctac cgccaagtac ttcttctaca gcaacatcat gaactttttc aagaccgaga
4021 ttaccctggc caacggcgag atccggaagc ggcctctgat cgagacaaac ggcgaaacag
4081 gcgagatcgt gtgggataag ggccgggact ttgccaccgt gcggaaagtg ctgtctatgc
4141 cccaagtgaa tatcgtgaaa aagaccgagg tgcagacagg cggcttcagc aaagagtcta
4201 tcctgcccaa gaggaacagc gacaagctga tcgccagaaa gaaggactgg gaccctaaga
4261 agtacggcgg cttcgacagc cccaccgtgg cctattctgt gctggtggtg gccaaagtgg
4321 aaaagggcaa gtccaagaaa ctgaagagtg tgaaagagct gctggggatc accatcatgg
4381 aaagaagcag cttcgagaag aatcccatcg actttctgga agccaagggc tacaaagaag
4441 tgaaaaagga cctgatcatc aagctgccta agtactccct gttcgagctg gaaaacggcc
4501 ggaagagaat gctggcctct gccggcgaac tgcagaaggg aaacgaactg gccctgccct
4561 ccaaatatgt gaacttcctg tacctggcca gccactatga gaagctgaag ggctcccccg
4621 aggataatga gcagaaacag ctgtttgtgg aacagcacaa acactacctg gacgagatca
4681 tcgagcagat cagcgagttc tccaagagag tgatcctggc cgacgctaat ctggacaagg
4741 tgctgagcgc ctacaacaag cacagagaca agcctatcag agagcaggcc gagaatatca
4801 tccacctgtt taccctgacc agtctgggag cccctgccgc cttcaagtac tttgacacca
4861 ccatcgaccg gaagaggtac accagcacca aagaggtgct ggacgccacc ctgatccacc
4921 agagcatcac cggcctgtac gagacacgga tcgacctgtc tcagctggga ggcgacgcct
4981 atccctatga cgtgcccgat tatgccagcc tgggcagcgg ctcccccaag aaaaaacgca
5041 aggtggaaga tcctaagaaa aagcggaaag tggacggcat tggtagtggg agcaacggca
5101 gcagcgggga tccggccgca acaaacttct ctctgctgaa acaagccgga gatgtcgaag
5161 agaatcctgg accgatggcc aagcctttgt ctcaagaaga atccaccctc attgaaagag
5221 caacggctac aatcaacagc atccccatct ctgaagacta cagcgtcgcc agcgcagctc
5281 tctctagcga cggccgcatc ttcactggtg tcaatgtata tcattttact gggggacctt
5341 gtgcagaact cgtggtgctg ggcactgctg ctgctgcggc agctggcaac ctgacttgta
5401 tcgtcgcgat cggaaatgag aacaggggca tcttgagccc ctgcggacgg tgccgacagg
5461 tgcttctcga tctgcatcct gggatcaaag ccatagtgaa ggacagtgat ggacagccga
5521 cggcagttgg gattcgtgaa ttgctgccct ctggttatgt gtgggagggc taagaattcg
5581 atatcaagct tatcggtaat caacctctgg attacaaaat ttgtgaaaga ttgactggta
5641 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
5701 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
5761 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
5821 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
5881 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
5941 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt
6001 cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct
6061 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
6121 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
6181 ccccgcgtcg actttaagac caatgactta caaggcagct gtagatctta gccacttttt
6241 aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag atctgctttt
6301 tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
6361 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
6421 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
6481 aatctctagc agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc
6541 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
6601 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
6661 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
6721 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta
6781 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
6841 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
6901 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
6961 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
7021 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
7081 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
7141 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
7201 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
7261 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
7321 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
7381 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
7441 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
7501 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
7561 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcagcacg tgttgacaat
7621 taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa ctaaaccatg
7681 gccaagttga ccagtgccgt tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag
7741 ttctggaccg accggctcgg gttctcccgg gacttcgtgg aggacgactt cgccggtgtg
7801 gtccgggacg acgtgaccct gttcatcagc gcggtccagg accaggtggt gccggacaac
7861 accctggcct gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc
7921 gtgtccacga acttccggga cgcctccggg ccggccatga ccgagatcgg cgagcagccg
7981 tgggggcggg agttcgccct gcgcgacccg gccggcaact gcgtgcactt cgtggccgag
8041 gagcaggact gacacgtgct acgagatttc gattccaccg ccgccttcta tgaaaggttg
8101 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
8161 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
8221 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
8281 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
8341 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
8401 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
8461 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
8521 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
8581 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
8641 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
8701 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
8761 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
8821 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
8881 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
8941 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
9001 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
9061 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
9121 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
9181 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
9241 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
9301 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
9361 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
9421 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
9481 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
9541 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
9601 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
9661 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
9721 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
9781 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
9841 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
9901 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
9961 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
10021 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
10081 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
10141 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
10201 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
10261 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
10321 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
10381 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
10441 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
10501 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct
10561 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
10621 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
10681 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc
10741 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
10801 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
10861 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
10921 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
10981 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
11041 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
11101 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
11161 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
11221 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
11281 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagcgcgttt tgcctgtact
11341 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
11401 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
11461 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
11521 agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca
11581 ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc
11641 caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta
11701 agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa
11761 aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc
11821 ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc
11881 ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg
11941 tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc
12001 aaaacaaaag taagaccacc gcacagcaag cggccgctga tcttcagacc tggaggagga
12061 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
12121 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
12181 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
12241 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
12301 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
12361 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
12421 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
12481 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga
12541 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
12601 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
12661 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
12721 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca
12781 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata
12841 gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa cggatcggca
12901 ctgcgtgcgc caattctgca gacaaatggc agtattcatc cacaatttta aaagaaaagg
12961 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca
13021 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga
13081 cagcagagat ccagtttggt taattaaggt accgagggcc tatttcccat gattccttca
13141 tatttgcata tacgatacaa ggctgttaga gagataatta gaattaattt gactgtaaac
13201 acaaagatat tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg gtagtttgca
13261 gttttaaaat tatgttttaa aatggactat catatgctta ccgtaacttg aaagtatttc
13321 gatttcttgg ctttatatat cttgtggaaa ggacgaaaca ccgcgaaaca gagcattctc
13381 tgc
//
| {
"Vector backbone": "Lenti-(BB)-hPGK-KRAB-dCas9-P2A-BlastR",
"Backbone size w/o insert (bp)": "",
"Vector type": "Mammalian Expression, Lentiviral, CRISPR",
"Selectable markers": "Blasticidin"
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "Stbl3",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Human pancreatic islet three-dimensional chromatin architecture provides insights into the genetics of type 2 diabetes",
"Publication": "Nat Genet. 2019 Jun 28. pii: 10.1038/s41588-019-0457-0. doi: 10.1038/s41588-019-0457-0.",
"DOI": "10.1038/s41588-019-0457-0.",
"Authors": "Miguel-Escalada I, Bonas-Guarch S, Cebola I, Ponsa-Cobas J, Mendieta-Esteban J, Atla G, Javierre BM, Rolando DMY, Farabella I, Morgan CC, Garcia-Hurtado J, Beucher A, Moran I, Pasquali L, Ramos-Rodriguez M, Appel EVR, Linneberg A, Gjesing AP, Witte DR, Pedersen O, Grarup N, Ravassard P, Torrents D, Mercader JM, Piemonti L, Berney T, de Koning EJP, Kerr-Conte J, Pattou F, Fedko IO, Groop L, Prokopenko I, Hansen T, Marti-Renom MA, Fraser P, Ferrer J /s---",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/31253982",
"PubMed ID": "PubMed 31253982"
} | {
"Gene/Insert name": "KRAB-dCas9-2A-BlastR",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "hPGK and U6",
"Tag / Fusion Protein": "HA (C terminal on backbone)"
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "Unknown (destroyed during cloning)",
"3′ cloning site": "Unknown (destroyed during cloning)",
"5′ sequencing primer": "GATCCGCCACCATGGATGC",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"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": ""
} |
|
Pmyo3::tag:sl1(G16C) | 79003 | Expression of exogenous SL1 in C. elegans muscle | Ma et al Nucleic Acids Res. 2016 Aug 23. pii: gkw734. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/147765/3a59cf2d-f5af-41de-b8eb-e449eb74c6e5/addgene-plasmid-79003-sequence-147765.gbk | full | LOCUS sequence-147765- 6209 bp DNA circular SYN 26-AUG-2024
DEFINITION Expression of exogenous SL1 in C. elegans muscle.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6209)
AUTHORS Ma X, Zhan G, Sleumer MC, Chen S, Liu W, Zhang MQ, Liu X
TITLE Analysis of C. elegans muscle transcriptome using
trans-splicing-based RNA tagging (SRT).
JOURNAL Nucleic Acids Res. 2016 Aug 23. pii: gkw734.
PUBMED 27557708
REFERENCE 2 (bases 1 to 6209)
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. 2016 Aug 23. pii: gkw734."
FEATURES Location/Qualifiers
source 1..6209
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(11..28)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(182..770)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(262..281)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(941..1801)
/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 1564..1583
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
CDS complement(1819..2613)
/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(1930..1949)
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind 2540..2559
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
rep_origin complement(2957..3385)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3067..3088)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 3279..3298
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3511..3533
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 3525..3542
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 3526..3542
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 3549..3568
/label=T7
/note="T7 promoter, forward primer"
promoter 3549..3567
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(5868..5884)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5869..5885)
/label=pBluescriptKS
/note="For pBluescript vector"
primer_bind complement(5910..5930)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(5910..5928)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(5949..5965)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5989..6005)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6002..6024)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 6013..6029
/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(6037..6067)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6082..6103
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
ORIGIN
1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
1801 tactcttcct ttttcaattc agaagaactc gtcaagaagg cgatagaagg cgatgcgctg
1861 cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag
1921 ctcttcagca atatcacggg tagccaacgc tatgtcctga tagcggtccg ccacacccag
1981 ccggccacag tcgatgaatc cagaaaagcg gccattttcc accatgatat tcggcaagca
2041 ggcatcgccg tgggtcacga cgagatcctc gccgtcgggc atgcgcgcct tgagcctggc
2101 gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc agatcatcct gatcgacaag
2161 accggcttcc atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg
2221 gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca tcagccatga tggatacttt
2281 ctcggcagga gcaaggtggg atgacaggag atcctgcccc ggcacttcgc ccaatagcag
2341 ccagtccctt cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt
2401 ggccagccac gatagccgcg ctgcctcgtc ctgcagttca ttcagggcac cggacaggtc
2461 ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg cggcatcaga
2521 gcagccgatt gtctgttgtg cccagtcata gccgaatagc ctctccaccc aagcggccgg
2581 agaacctgcg tgcaatccat cttgttcaat catgcgaaac gatcctcatc ctgtctcttg
2641 atcagatctt gatcccctgc gccatcagat ccttggcggc aagaaagcca tccagtttac
2701 tttgcagggc ttcccaacct taccagaggg cgccccagct ggcaattccg gttcgcttgc
2761 tgtccataaa accgcccagt ctagctatcg ccatgtaagc ccactgcaag ctacctgctt
2821 tctctttgcg cttgcgtttt cccttgtcca gatagcccag tagctgacat tcatccgggg
2881 tcagcaccgt ttctgcggac tggctttcta cgtgttccgc ttcctttagc agcccttgcg
2941 ccctgaattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
3001 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
3061 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
3121 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
3181 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
3241 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
3301 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
3361 cttaatgcgc cgctacaggg cgcgtccatt cgccattcag gctgcgcaac tgttgggaag
3421 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
3481 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
3541 gtgaattgta atacgactca ctatagggcg aattgaagct gcccttgctc gagcggctat
3601 aataagttct tgaataaaat aattttcccg acaaaacatg agtatttctt tcgaaaataa
3661 aagtgcaggc taattagaga ttattctgta attaactgca taatttgtca cgtgccatag
3721 ttttacattc cactacgtca tagttcttaa aatactaatc tcctgaaaat agaagtaggt
3781 gaagaaagtt taattatcag ttctaaaatg acaattgatc tttggaatat gttctgaaac
3841 taccgatcat tgaacagatg ctatttgaat gatatagaat tgtatatttg caatttctga
3901 aacgcgttct taaaggcaca cagattaatt caaaagggtc tggccgcaaa aaggtttatg
3961 gtggccgatt ttgagttttg tgtgtgattg ctttttcaca atcagtgttt tcaggattat
4021 gtgatgaact agatcttcaa gtttcgttac atttcatatg ttttcggaac tcacgaagta
4081 catattgggt attgtgctca aaaaattcag caatcagctt cgctccgctg actttagaac
4141 ccaaaaaaat agtatggcca aactgactgt gttacgatca tttcaatttt tcaatacata
4201 tttaagattt ctaagagtaa gaaggtcaaa aactgttctg gaatacatat atatttttca
4261 ggttacaatt agtcaaaaag tgcactgaaa tatacgtttt aatttcacga ataacccaat
4321 tagttcaatg tatttttggt caaccaacgt taaagtttgg cttccaacca attatcattt
4381 ctgatcaacc acaatgtttt ttctttatct gcaagttaat tttttatttt tatccagatg
4441 tttggcatat ttttcaattc ttcactagcg cccacttctt gcacttccgg cgccctgaat
4501 ctaatgcatc tgttgcaaga attgaaagac caatcaacac attgttttct tcacgagata
4561 ctgaagaaaa tgaataaaaa cagagaaaaa gagccatgtg attagtgaca actgttgcta
4621 acagatacca tagcttggac ttggtacgtg atggcaacgt atgggtcaac aaaaatgatt
4681 gcagaggggg tgcaaaacag tcaagtcgag aaaatatgaa aaacagaaaa caaagaacag
4741 aaaaatgggt ttgagagtca gtataattta taaaagaaaa attgtacata gaaattaacc
4801 atttttgtag aagaagttat ttttcaagca tcgttaaaaa ttattcaaag caccttattt
4861 catatttaat tttaaacatg gttaaatgaa caacacggtg cgcaatcagg aaaacttgaa
4921 atctgaaact gttgttgtga tcttcttcgc aactgttcag atagcactag tgtaatgtta
4981 agagtgcgcg aatataatgg aatataatgg atcacacctc ctgccatcag gtaaacgtct
5041 ctgttatcac atatttccaa ctattaaatt tttacctttt acagttttac atttttttga
5101 aaaaagtaac tttttgtctt caaaatccct gacgaaaata tcaaatattt taatcgagac
5161 tgcagaggaa ccgattgatg atttggaaaa tccagcttta cctgtgtaag aactgaaaag
5221 tttcataacc ctagggtatt cccagttaca ttccccactg gctaacaata gcacccagtt
5281 tttcatcacc tttcttcaaa tttctcggcg atttgttaaa aacaaaattt gtgtcccttc
5341 tctgatatct ctatgtctct aaacacaagt tcatcggaaa acgaaggagg gtaggtgttg
5401 gttgggctcc cgaagtgaaa atagaagagc aagaatagaa tattagagag agagtgcaga
5461 gagggcggga tagctcccgg gattccgttt tcttcttctt tatcttcaac gatgatgtgt
5521 gtgcgtgttg tatagattct gttgctcccc cacaactcgc tccgaaggct caatacaatt
5581 caattacacg acgctcttcc gatctggttt aattacccaa ctttgaggta aacattgaaa
5641 ctgacccaaa gaaatttggc gttagctata aattttggaa cgtctcctct cggggagaca
5701 aaaatactaa tagggataac agggtaatga aattcgaaaa tcggttcatc catgttggag
5761 aacggtccat tcgaaaatat gtggagatga gggagcatgg ctgtccgtat gtggagatgt
5821 gggaaggtga tgatattatt ggggaacgaa ttcgatatca agcttatcga taccgtcgac
5881 ctcgagggcc cggtacccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg
5941 taatcatggt catagctgtt tcctgaaggg cagcttggcg taatcatggt catagctgtt
6001 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
6061 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
6121 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
6181 ggggagaggc ggtttgcgta ttgggcgct
//
| {
"Vector backbone": "pEASY Blunt Simple Cloning Kit",
"Backbone size w/o insert (bp)": "3830",
"Vector type": "Worm 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": "Analysis of C. elegans muscle transcriptome using trans-splicing-based RNA tagging (SRT)",
"Publication": "Nucleic Acids Res. 2016 Aug 23. pii: gkw734.",
"DOI": "10.1093/nar/gkw734",
"Authors": "Ma X, Zhan G, Sleumer MC, Chen S, Liu W, Zhang MQ, Liu X",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/27557708",
"PubMed ID": "PubMed 27557708"
} | {
"Gene/Insert name": "sls-1.2",
"Species": "C. elegans (nematode)",
"Insert Size (bp)": "2379",
"Mutation": "G16C",
"Entrez Gene": "sls-1.2",
"Promoter": "myo-3",
"Tag / Fusion Protein": "illumina-adapter (N terminal on insert)"
} | {
"Cloning method": "TOPO Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "M13-F",
"3′ sequencing primer": "T3"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
||
pFudio-Cpg15-IRES2-TdTomatoW | 134309 | Expresses cpg15 and TdTomato upon coexpression with Cre recombinase | Elly Nedivi | Subramanian et al Cell Rep. 2019 Aug 6;28(6):1584-1595.e5. doi: 10.1016/j.celrep.2019.07.012. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/264807/5fd96e64-4ff2-482c-badd-1e3ab234d415/addgene-plasmid-134309-sequence-264807.gbk | full | LOCUS sequence-264807- 11982 bp DNA circular SYN 27-AUG-2024
DEFINITION Expresses cpg15 and TdTomato upon coexpression with Cre recombinase.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11982)
AUTHORS Subramanian J, Michel K, Benoit M, Nedivi E
TITLE CPG15/Neuritin Mimics Experience in Selecting Excitatory Synapses
for Stabilization by Facilitating PSD95 Recruitment.
JOURNAL Cell Rep. 2019 Aug 6;28(6):1584-1595.e5. doi:
10.1016/j.celrep.2019.07.012.
PUBMED 31390571
REFERENCE 2 (bases 1 to 11982)
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:
"10.1016/j.celrep.2019.07"; journalName: "Cell Rep"; date:
"2019-08-6- 6"; volume: "28"; issue: "6"; pages: "1584-1595.e5"
FEATURES Location/Qualifiers
source 1..11982
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 41..420
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 422..620
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 575..595
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
LTR 638..818
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 865..990
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1483..1716
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1901..1945
/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 2243..2360
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2425..3636
/label=UbC promoter
/note="human ubiquitin C promoter"
primer_bind 3473..3492
/label=hUBCpro-F
/note="Human Ubiquitin C (UbC) promoter, forward primer"
protein_bind complement(3689..3722)
/label=lox2272
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are
compatible with each other, but incompatible with loxP or
loxN sites (Lee and Saito, 1988)."
protein_bind complement(3773..3806)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
CDS complement(3815..5245)
/codon_start=1
/product="tandem dimeric (pseudo-monomeric) derivative of
DsRed (Shaner et al., 2004)"
/label=tdTomato
/note="mammalian codon-optimized"
/translation="MVSKGEEVIKEFMRFKVCMEGSMNGHEFEIEGEGEGRPYEGTQTA
KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
LKGEIHQALKLKDGGRYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
RSEGRHHLFLYGMDELYK"
misc_feature 5246..5835
/label=IRES2
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(5422..5441)
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
primer_bind 5651..5668
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
CDS complement(6203..6226)
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
regulatory 6316..6325
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
protein_bind 6335..6368
/label=lox2272
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are
compatible with each other, but incompatible with loxP or
loxN sites (Lee and Saito, 1988)."
protein_bind 6419..6452
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
misc_feature 6477..7065
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(6530..6550)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(6948..6959)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(7068..7084)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(7069..7085)
/label=pBluescriptKS
/note="For pBluescript vector"
LTR 7590..7770
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
primer_bind complement(7796..7813)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 7802..8026
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 8072..8500
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(8159..8178)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 8369..8390
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(8509..8529)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 8514..8843
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 8694..8829
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 8756..8775
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter 8891..8938
/label=EM7 promoter
/note="synthetic bacterial promoter "
CDS 8957..9331
/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 9461..9582
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(9498..9517)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 9552..9571
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(9631..9647)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(9631..9647)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(9644..9666)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 9655..9671
/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(9679..9709)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 9724..9745
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(9862..9879)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(10033..10621)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(10113..10132)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(10792..11652)
/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 11415..11434
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(11653..11757)
/gene="bla"
/label=AmpR promoter
primer_bind complement(11832..11851)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt
61 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
121 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
181 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
241 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
301 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
361 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
421 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
481 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
541 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
601 tgggaggtct atataagcag cgcgttttgc ctgtactggg tctctctggt tagaccagat
661 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
721 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
781 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg
841 aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc
901 acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc
961 tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga
1021 tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta
1081 tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa
1141 ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt
1201 agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa
1261 gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca
1321 cagcaagcgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
1381 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
1441 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
1501 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
1561 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
1621 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
1681 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
1741 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
1801 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
1861 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
1921 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
1981 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
2041 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
2101 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
2161 cagagacaga tccattcgat tagtgaacgg atcggcactg cgtgcgccaa ttctgcagac
2221 aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag
2281 gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa
2341 ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca gtttggttaa
2401 ttaacccgtg tcggctccag atctggcctc cgcgccgggt tttggcgcct cccgcgggcg
2461 cccccctcct cacggcgagc gctgccacgt cagacgaagg gcgcagcgag cgtcctgatc
2521 cttccgcccg gacgctcagg acagcggccc gctgctcata agactcggcc ttagaacccc
2581 agtatcagca gaaggacatt ttaggacggg acttgggtga ctctagggca ctggttttct
2641 ttccagagag cggaacaggc gaggaaaagt agtcccttct cggcgattct gcggagggat
2701 ctccgtgggg cggtgaacgc cgatgattat ataaggacgc gccgggtgtg gcacagctag
2761 ttccgtcgca gccgggattt gggtcgcggt tcttgtttgt ggatcgctgt gatcgtcact
2821 tggtgagtag cgggctgctg ggctggccgg ggctttcgtg gccgccgggc cgctcggtgg
2881 gacggaagcg tgtggagaga ccgccaaggg ctgtagtctg ggtccgcgag caaggttgcc
2941 ctgaactggg ggttgggggg agcgcagcaa aatggcggct gttcccgagt cttgaatgga
3001 agacgcttgt gaggcgggct gtgaggtcgt tgaaacaagg tggggggcat ggtgggcggc
3061 aagaacccaa ggtcttgagg ccttcgctaa tgcgggaaag ctcttattcg ggtgagatgg
3121 gctggggcac catctgggga ccctgacgtg aagtttgtca ctgactggag aactcggttt
3181 gtcgtctgtt gcgggggcgg cagttatggc ggtgccgttg ggcagtgcac ccgtaccttt
3241 gggagcgcgc gccctcgtcg tgtcgtgacg tcacccgttc tgttggctta taatgcaggg
3301 tggggccacc tgccggtagg tgtgcggtag gcttttctcc gtcgcaggac gcagggttcg
3361 ggcctagggt aggctctcct gaatcgacag gcgccggacc tctggtgagg ggagggataa
3421 gtgaggcgtc agtttctttg gtcggtttta tgtacctatc ttcttaagta gctgaagctc
3481 cggttttgaa ctatgcgctc ggggttggcg agtgtgtttt gtgaagtttt ttaggcacct
3541 tttgaaatgt aatcatttgg gtcaatatgt aattttcagt gttagactag taaattgtcc
3601 gctaaattct ggccgttttt ggcttttttg ttagacgaag cttgggctgc aggtcgactc
3661 tagaggatcc tctagagtcg actccggaat aacttcgtat aggatacttt atacgaagtt
3721 atgcagaatg gtagctggat tgtagctgct attagcaata tgaaacctct taataacttc
3781 gtatagcata cattatacga agttatggcg cgccttactt gtacagctcg tccatgccgt
3841 acaggaacag gtggtggcgg ccctcggagc gctcgtactg ttccacgatg gtgtagtcct
3901 cgttgtggga ggtgatgtcc agcttggtgt ccacgtagta gtagccgggc agttgcacgg
3961 gcttcttggc catgtagatg gtcttgaact ccaccaggta gcggccgccg tccttcagct
4021 tcagggcctg gtggatctcg cccttcagca cgccgtcgcg ggggtacagg cgctcggtgg
4081 aggcctccca gcccatggtc ttcttctgca ttacggggcc gtcggggggg aagttggtgc
4141 cgcgcatctt caccttgtag atcagcgtgc cgtcctgcag ggaggagtcc tgggtcacgg
4201 tcaccagacc gccgtcctcg aagttcatca cgcgctccca cttgaagccc tcggggaagg
4261 acagcttctt gtaatcgggg atgtcggcgg ggtgcttcac gtacgccttg gagccgtaca
4321 tgaactgggg ggacaggatg tcccaggcga agggcagggg gccgcccttg gtcaccttca
4381 gcttggcggt ctgggtgccc tcgtaggggc ggccctcgcc ctcgccctcg atctcgaact
4441 cgtggccgtt catggagccc tccatgcgca ccttgaagcg catgaactct ttgatgacgg
4501 ccatgttgtt gtcctcggag gaggcggtgc cggagctgcc gctgccggtg ctgccggtgc
4561 catgccccag gaacaggtgg tggcggccct cggagcgctc gtactgttcc acgatggtgt
4621 agtcctcgtt gtgggaggtg atgtccagct tggtgtccac gtagtagtag ccgggcagtt
4681 gcacgggctt cttggccatg tagatggtct tgaactccac caggtagtgg ccgccgtcct
4741 tcagcttcag ggcctggtgg atctcgccct tcagcacgcc gtcgcggggg tacaggcgct
4801 cggtggaggc ctcccagccc atggtcttct tctgcattac ggggccgtcg ggggggaagt
4861 tggtgccgcg catcttcacc ttgtagatca gcgtgccgtc ctgcagggag gagtcctggg
4921 tcacggtcac cagaccgccg tcctcgaagt tcatcacgcg ctcccacttg aagccctcgg
4981 ggaaggacag cttcttgtaa tcggggatgt cggcggggtg cttcacgtac gccttggagc
5041 cgtacatgaa ctggggggac aggatgtccc aggcgaaggg cagggggccg cccttggtca
5101 ccttcagctt ggcggtctgg gtgccctcgt aggggcggcc ctcgccctcg ccctcgatct
5161 cgaactcgtg gccgttcatg gagccctcca tgcacacctt gaagcgcatg aactctttga
5221 tgacctcctc gcccttgctc accatggttg tggcaccggt attatcatcg tgtttttcaa
5281 aggaaaacca cgtccccgtg gttcgggggg cctagacgtt tttttaacct cgactaaaca
5341 catgtaaagc atgtgtaccg aggccccaga tcagatccca tacaatgggg taccttctgg
5401 gcatccttca gccccttgtt gaatacgctt gaggagagcc atttgactct ttccacaact
5461 atccaactca caacgtggca ctggggttgt gccgcctttg caggtgtatc ttatacacgt
5521 ggcttttggc cgcagaggca cctgtcgcca ggtggggggt tccgctgcct gcaaagggtc
5581 gctacagacg ttgtttgtct tcaagaagct tccagaggaa ctgcttcctt cacgacattc
5641 aacagacctt gcattccttt ggcgagaggg gaaagacccc taggaatgct cgtcaagaag
5701 acagggccag gtttccgggc cctcacattg ccaaaagacg gcaatatggt ggaaaataac
5761 atatagacaa acgcacaccg gccttattcc aagcggcttc ggccagtaac gttagggggg
5821 ggggagggag aggggcggat cccgggcccg cggtaccgtc gactcagaag gaaagccagg
5881 tcgctaaagc tgccgagaga gacaccagga gcacggaaag cgccgggagc agggaccccg
5941 ccgccccgtt gccgctgccg cagagttcga ataagctgcc ttggatattg aggtttttcg
6001 attcttttct cagtttatcc cacatatctt tcgccccttc ctggcaatcc gtaagagctg
6061 tgaccgtgca gctgtggaaa tcctcccagt atgtgcacac ggtcttgatg ttcgtcttgt
6121 cgtccaggcc ctgcgggtag ttggccatgc tgtcacccag cttgagcaaa cagtctgaaa
6181 agcccttaaa gactgcatcg cacttgtcat catcgtcctt atagtcggta ccgcctgctg
6241 ctctcacggc ctgcaccagg taagctattt gcaccgcgag gatcagtgaa atatatctgc
6301 cgttcaactt aagtcccatg gtggcggcgc tagcataact tcgtataaag tatcctatac
6361 gaagttattt gccttaaccc agaaattatc actgttattc tttagaatgg tgcaaagaat
6421 aacttcgtat aatgtatgct atacgaagtt atgaattcga tatcaagctt atcgataatc
6481 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
6541 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
6601 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
6661 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
6721 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
6781 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
6841 gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct
6901 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
6961 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
7021 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcga taccgtcgac
7081 ctcgagacct agaaaaacat ggagcaatca caagtagcaa tacagcagct accaatgctg
7141 attgtgcctg gctagaagca caagaggagg aggaggtggg ttttccagtc acacctcagg
7201 tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa
7261 aggggggact ggaagggcta attcactccc aacgaagaca agatatcctt gatctgtgga
7321 tctaccacac acaaggctac ttccctgatt ggcagaacta cacaccaggg ccagggatca
7381 gatatccact gacctttgga tggtgctaca agctagtacc agttgagcaa gagaaggtag
7441 aagaagccaa tgaaggagag aacacccgct tgttacaccc tgtgagcctg catgggatgg
7501 atgacccgga gagagaagta ttagagtgga ggtttgacag ccgcctagca tttcatcaca
7561 tggcccgaga gctgcatccg gactgtactg ggtctctctg gttagaccag atctgagcct
7621 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
7681 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
7741 ccttttagtc agtgtggaaa atctctagca gggcccgttt aaacccgctg atcagcctcg
7801 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
7861 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
7921 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
7981 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
8041 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
8101 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
8161 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
8221 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
8281 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
8341 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
8401 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
8461 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
8521 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
8581 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
8641 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
8701 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
8761 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
8821 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
8881 atcagcacgt gttgacaatt aatcatcggc atagtatatc ggcatagtat aatacgacaa
8941 ggtgaggaac taaaccatgg ccaagttgac cagtgccgtt ccggtgctca ccgcgcgcga
9001 cgtcgccgga gcggtcgagt tctggaccga ccggctcggg ttctcccggg acttcgtgga
9061 ggacgacttc gccggtgtgg tccgggacga cgtgaccctg ttcatcagcg cggtccagga
9121 ccaggtggtg ccggacaaca ccctggcctg ggtgtgggtg cgcggcctgg acgagctgta
9181 cgccgagtgg tcggaggtcg tgtccacgaa cttccgggac gcctccgggc cggccatgac
9241 cgagatcggc gagcagccgt gggggcggga gttcgccctg cgcgacccgg ccggcaactg
9301 cgtgcacttc gtggccgagg agcaggactg acacgtgcta cgagatttcg attccaccgc
9361 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
9421 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
9481 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
9541 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
9601 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
9661 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
9721 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
9781 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
9841 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
9901 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
9961 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
10021 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
10081 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
10141 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
10201 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
10261 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
10321 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
10381 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
10441 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
10501 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
10561 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
10621 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
10681 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
10741 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
10801 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
10861 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
10921 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
10981 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
11041 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
11101 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
11161 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
11221 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
11281 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
11341 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
11401 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
11461 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
11521 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
11581 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
11641 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
11701 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
11761 atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg
11821 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct
11881 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
11941 ttgaccgaca attgcatgaa gaatctgctt agggttaggc gt
//
| {
"Vector backbone": "pFUW",
"Backbone size w/o insert (bp)": "9534",
"Vector type": "Mammalian Expression, Lentiviral, Cre/Lox",
"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": "CPG15/Neuritin Mimics Experience in Selecting Excitatory Synapses for Stabilization by Facilitating PSD95 Recruitment",
"Publication": "Cell Rep. 2019 Aug 6;28(6):1584-1595.e5. doi: 10.1016/j.celrep.2019.07.012.",
"DOI": "10.1016/j.celrep.2019.07.012.",
"Authors": "Subramanian J, Michel K, Benoit M, Nedivi E /jcelrep",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/31390571",
"PubMed ID": "PubMed 31390571"
} | {
"Gene/Insert name": "cpg15",
"Species": "M. musculus (mouse), R. norvegicus (rat)",
"Insert Size (bp)": "465",
"Mutation": "",
"Entrez Gene": "Nrn1",
"Promoter": "UbC",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "BamH1 (not destroyed)",
"3′ cloning site": "Nhe1 (not destroyed)",
"5′ sequencing primer": "GGACCCCGCCGCCCC",
"3′ sequencing primer": "CCGCCACCATGGGACTTAAGTTGAACGG"
} | {
"Gene/Insert name": "TdTomato",
"Species": "Synthetic",
"Insert Size (bp)": "1431",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "UbC",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "Asc1 (not destroyed)",
"3′ cloning site": "Age1 (not destroyed)",
"5′ sequencing primer": "TTACTTGTACAGCTCGTCCATGCCG",
"3′ sequencing primer": "ACCGGTGCCACAACCATGGTGA"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pEF5-FRT-V5-DEST-GFP-Kif26b | 102862 | Low | Expresses GFP-Kif26b in mammalian cells | Henry Ho | Susman et al Elife. 2017 Sep 8;6. pii: e26509. doi: 10.7554/eLife.26509. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/200036/e38cffa5-d180-48b8-83f7-dbd5388a3363/addgene-plasmid-102862-sequence-200036.gbk | full | LOCUS sequence-200036- 13228 bp DNA circular SYN 25-AUG-2024
DEFINITION Expresses GFP-Kif26b in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 13228)
AUTHORS Susman MW, Karuna EP, Kunz RC, Gujral TS, Cantu AV, Choi SS, Jong
BY, Okada K, Scales MK, Hum J, Hu LS, Kirschner MW, Nishinakamura R,
Yamada S, Laird DJ, Jao LE, Gygi SP, Greenberg ME, Ho HH
TITLE Kinesin superfamily protein Kif26b links Wnt5a-Ror signaling to the
control of cell and tissue behaviors in vertebrates.
JOURNAL Elife. 2017 Sep 8;6. pii: e26509. doi: 10.7554/eLife.26509.
PUBMED 28885975
REFERENCE 2 (bases 1 to 13228)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Elife.";
date: "2017-09-8"; volume: "6. pii: e26509. doi"; pages: "
10.7554/eLife.26509"
FEATURES Location/Qualifiers
source 1..13228
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(42..61)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
promoter 349..1527
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 580..1518
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 1475..1495
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
promoter 1544..1562
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 1641..1665
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
CDS 1693..2409
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 1693..2409
/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(1738..1759)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1999..2018)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 2346..2367
/label=EGFP-C
/note="EGFP, forward primer"
polyA_signal 8798..8932
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
protein_bind complement(9022..9046)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
CDS 9099..9140
/codon_start=1
/product="epitope tag from simian virus 5"
/label=V5 tag
/translation="GKPIPNPLLGLDST"
primer_bind complement(9178..9195)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 9184..9408
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(9541..9560)
/label=F1ori-R
/note="F1 origin, reverse primer"
protein_bind 9692..9739
/label=FRT
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
CDS 9747..10769
/codon_start=1
/product="hygromycin B phosphotransferase"
/label=HygR
/note="confers resistance to hygromycin"
/translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
"
CDS 9747..10769
/codon_start=1
/gene="aph(4)-Ia"
/product="aminoglycoside phosphotransferase from E. coli"
/label=HygR
/note="confers resistance to hygromycin"
/translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
"
polyA_signal 10899..11020
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(11069..11085)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(11069..11085)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(11082..11104)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 11093..11109
/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(11117..11147)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 11162..11183
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(11300..11317)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(11471..12059)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(11551..11570)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(12230..13090)
/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 12853..12872
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(13091..13195)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc
61 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
121 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
181 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga
241 ttattgacta ggcttttgca aaaagctttg caaagatgga taaagtttta aacagagagg
301 aatctttgca gctaatggac cttctaggtc ttgaaaggag tgggaattgg ctccggtgcc
361 cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc
421 aattgaaccg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac
481 tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg
541 aacgttcttt ttcgcaacgg gtttgccgcc agaacacagg taagtgccgt gtgtggttcc
601 cgcgggcctg gcctctttac gggttatggc ccttgcgtgc cttgaattac ttccacctgg
661 ctgcagtacg tgattcttga tcccgagctt cgggttggaa gtgggtggga gagttcgagg
721 ccttgcgctt aaggagcccc ttcgcctcgt gcttgagttg aggcctggcc tgggcgctgg
781 ggccgccgcg tgcgaatctg gtggcacctt cgcgcctgtc tcgctgcttt cgataagtct
841 ctagccattt aaaatttttg atgacctgct gcgacgcttt ttttctggca agatagtctt
901 gtaaatgcgg gccaagatct gcacactggt atttcggttt ttggggccgc gggcggcgac
961 ggggcccgtg cgtcccagcg cacatgttcg gcgaggcggg gcctgcgagc gcggccaccg
1021 agaatcggac gggggtagtc tcaagctggc cggcctgctc tggtgcctgg cctcgcgccg
1081 ccgtgtatcg ccccgccctg ggcggcaagg ctggcccggt cggcaccagt tgcgtgagcg
1141 gaaagatggc cgcttcccgg ccctgctgca gggagctcaa aatggaggac gcggcgctcg
1201 ggagagcggg cgggtgagtc acccacacaa aggaaaaggg cctttccgtc ctcagccgtc
1261 gcttcatgtg actccacgga gtaccgggcg ccgtccaggc acctcgatta gttctcgagc
1321 ttttggagta cgtcgtcttt aggttggggg gaggggtttt atgcgatgga gtttccccac
1381 actgagtggg tggagactga agttaggcca gcttggcact tgatgtaatt ctccttggaa
1441 tttgcccttt ttgagtttgg atcttggttc attctcaagc ctcagacagt ggttcaaagt
1501 ttttttcttc catttcaggt gtcgtgagga attagcttgg tactaatacg actcactata
1561 gggagaccca agctggctag gtaagcttgg taccgagctc ggatccacta gtccagtgtg
1621 gtggaattct gcagatatca acaagtttgt acaaaaaagc aggctggcgc cggaaccaat
1681 tcagtcgaca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
1741 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
1801 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
1861 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
1921 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
1981 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
2041 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
2101 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
2161 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
2221 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
2281 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
2341 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
2401 ctgtacaagg gccggcctac catgaattcg gtagccggaa ataaagagag gcttgcggtc
2461 tccaccaggg gcaagaaata cggggtgaac gaaatgtgct cgcccaccaa gccctcagcg
2521 ccctgctctc ctgagagctg gtaccgcaag gcttacgagg agtcgcgcgc cggcagccgg
2581 cccaccccgg agggagcagg ctctgcgctg ggctcatcgg gcaccccgtc cccgggctcg
2641 ggcacctcgt ccccgagctc cttcaccggc tctccgggac ctgcatctcc gggcattggt
2701 accagctcgc ccggctctct gggcggctca cccggcttcg gcaccggctc ccctggctcg
2761 ggcagcggcg gcggctcctc ccccggctcc gaccgcggcg tctggtgcga gaactgtaac
2821 gcccgcttgg tggagctgaa gaggcaggct ctcaagctgc tgctgccggg acccctcccg
2881 ggaaaggacc ctgctttctc tgctgtgatt cacgacaaac tccaggttcc caatactatc
2941 aggaagtcgt ggaatgaccg agacaaccgc tgtgacattt gtgccacaca cctgaaccag
3001 ctaaagcagg aagccatcca aatggtgctg accttggagc aggcagctgg cagcgagcac
3061 tatgacacct ctctgggctc accccctccg atctccagca ttcccacact ggtggggtct
3121 cgccacatgg gtgggctcca gcagcccagg gagtgggcct ttgtgcctgc cccctatgcc
3181 acttctacct acacaggcct tgtcaacaaa cacagcggca aacccaacag ccttggggtc
3241 agcaacggag cggaaaagaa gagtggctcc ccaacccacc aggccaaggt cagcctccag
3301 atggccacca gtcccagcaa tggaaacatc ctcaactcgg tggccattca ggctcatcaa
3361 tatctagacg gcacctggtc cctttcaaga accaatgggg ttaccctgta cccctaccag
3421 atctcccagc tgatgacaga gaccggccgc gagggactga cagagtcagc gctcaaccgc
3481 tacaatgctg acaagcctgc agccagcagt gtcccagccc cacagggttc ctgtgtggcc
3541 agtgagactt ccacagggac ctcagtggcc gcttccttct tcgcacgagc tgcccagaag
3601 ttaaatctgt catctaagaa gaagaaacac cggccttcga ctccatctgt tgcggaggct
3661 cccctctttg cgaccagctt cagtgggatc ctgcagacct ctcctccccc agccccaccc
3721 tgtctcctga gggctgtcaa caaggtgaag gacactcctg gcatgggcaa ggtgaaagtc
3781 atgctgcgta tctgttccac atcagctcga gatacctcag aatccagctc tttcctgaag
3841 gtggacccac ggaagaagca gatcaccttg tatgatcccc tgacttgcgg aggtcaaaat
3901 gccttccaga aaagaagcag ccaggttcct ccgaagatgt ttgccttcga tgctgttttc
3961 ccacaagatg cttctcaggc tgaagtttgt gctggaactg tggccgaagt catccagtct
4021 gtggtgaacg gggcagatgg ctgtgtgttc tgttttggcc atgccaagct agggaaatcc
4081 tacaccatga tcggaaggga cgactctatg cagaacctgg gcattattcc atgtgctatt
4141 tcttggctgt tcaagcttat caatgagcgc aaagagaaga cgggggcccg attctcagtc
4201 cgcatctcag ccgttgaggt gtggggaaaa gaggagaacc ttcgagacct gttgtctgag
4261 gtggccacgg gcagcctaca ggacgggcag tccccaggcg tgtacctctg tgaagacccc
4321 atctgtggca cacagctgca gaaccagagt gagctccggg cccccaccgc agagaaagct
4381 gccttcttcc tggacgccgc cattgcctcg aggcgaagca accaacagga ttgtgacgag
4441 gatgaccacc gcaactccca tatgctcttc accctgcaca tctaccagta ccggatggag
4501 aagagcggga agggtggaat gtctggaggt cgaagccgcc tgcatctcat cgacctcggg
4561 agctgtgtga aagctctcag caaaaaccga gaaggaggct cggggctgtg cctctcactg
4621 tctgctctgg gcaacgtcat cctggctttg gtcaacggga gcaagcacat cccatacaag
4681 gagagcaagc tcaccatgct tctgagggag tctctgggga atgtgaactg tcgcaccacc
4741 atgatcgcac acatctcggc cgcagccagc agctacgcgg agaccctgtc caccatccag
4801 atcgcgtcga gagtcttaag gatgaagaag aagaagacca agtacacatc aagctcctct
4861 gggggagaaa gctcctgtga ggaaggcaga atgcgcaggc ccacccagct gagacccttc
4921 catgcccggg ctccggtgga tccagaattc ccccttgccc cactgtccag tgatcctgac
4981 tactcctcca gcagtgagca gtcctgcgac accgtcatct acatcgggcc caatggcaca
5041 gccctgtctg acaaggaact cactgacaat gaggggcccc cagactttgt gcccattgtg
5101 ccagccctac agaagaccag aggtgacagc cggccggggg aggcagcaga ggctgcagcc
5161 agcaagtcag aaagggactg tctgaagtgc aatacgtttg ctgagctgca ggaaagacta
5221 gattgcatag atggcagcga agaacccagt aagtttccct tcgaagagct gcccatccag
5281 ttcgggccgg agcaagctgg cagatgtgct cctttaagcc aagcagtggg gccagacaca
5341 ccctctgagt ctgaaaagga agataacggc tcagacaatg gccaattgac cgagagggaa
5401 ggcacagagc ttccagcctc caaggcacag cggaatcggt cccctgcccc tactgtgaca
5461 cgcagcagca gccccagtcc agcctcacca cggagcatcc cgggcagcag tagccagcac
5521 agtacctccc agctcacaca gagccccagt ctccagagca gccgggagag tctgaactca
5581 tgtggcttcg tggagggcaa gcccagaccc atgggctccc cgaggctagg cattgccagt
5641 ctgtccaaga cctcagagta caaacctccc agttctccct ctcagaggtg caaagtctac
5701 acccagaagg gggtcctgcc ctcctctgcc ccaccacctt ccctgagcaa ggactctggc
5761 atggtctcca gtgaatcctt gcttcagccc gacgtgcgca ctcccccagt tggaatgagt
5821 ccccaggttt tgaagaaatc catgtctgcc gggagcgaag ggtttcccgg aactctagtt
5881 gacggtgagg accaggaagg ccctccagcg gactctaaga aggagattct gagcaccaca
5941 atggtgactg tgcaacagcc cctggagctg aatggtgagg atgagttggt cttcacactg
6001 gtggaggagc tgaccatcag cggggtgctc gacagtggac gccctaccag catcatcagc
6061 ttcaacagtg actgctccgt gcaggccctg gcctcaggct cccggcccgt cagcatcatc
6121 agcagcatca gcgaggatct ggagtgctac tccagcatgg cccccgtctc cgaggtgagc
6181 atcacgcagt tcctgcctct cccaaagctg ggcctggatg agaaggctcg ggacgcgggc
6241 agccggcgtt cctccatcag ctcctggctg agcgagatga gcgcgggcag tgatggcgag
6301 cagtcatgcc acagcttcat agcccagacg tgctttgggc atggggaagc catggcagaa
6361 cccccagcct cggagtttgt cagcagcatc cagaacacgg cagtggtgtg cagagagaag
6421 cccgaggtgg gccctgacaa cctgctcatc ttatctgaga tgggggaaga gtctggcaac
6481 aaagccgctc ccatcaaagg ctgcaaaatc tcaacggtgg gcaaggccat ggtcaccatc
6541 tccaacacag ccagcctgag cagctgcgaa gggtacatcc ccatgaagac caacatcacc
6601 gtttacccct gcatcgccat gagcccccga aacgtgcaag agcctgagtc ctccactgcc
6661 accccaaagg tgagccccaa agcatcccag gcccaggaaa gcaaggaacc cagtacaagg
6721 agagagatga aatttgagga tccttggctt aaacgagaag aagaggtgaa aagggagaac
6781 gcttactcca gcgaggaagg ggttaagtgt gagccgctcc caagctccct gaagactgaa
6841 ggtaagtcag agcaggaact ggatggcagg cccagctcag gcaacaggct gagcagcagc
6901 agctcagagg cagccgcctt tcaagggact gataacgtta ggagagtcgt ggatggctgt
6961 gagatggccc tgcctggctt ggtggcccaa agtcccctcc atgtcaaccg aaacctaaag
7021 tcaagcagcc tcccaagggc atttcagaaa gctgacaggc acgaggagct ggacagcttc
7081 taccattgcc tggcagacag caatggcttc agtgctgctt cgggcatccc atcctcgaag
7141 acaaccctag agaggaaagt tgcttcaccc aagcattgcg tcctggcaag gcccaaaggg
7201 accccgcctc tgcctcctgt ccgcaagtct agcttggacc agaagaacag ggccagccct
7261 caacacggtg gtggcagcag caacaccagc agccccctga accagccagc taccttcttg
7321 gcttgcttcc ctgatgagtc caacggcaag actaaggatg tcagtagcag cagtaagctc
7381 ttcagcgcca agctggaaca gctggccagc agaagcaact ctcttggcag gacaacagtc
7441 agccactacg aatgcttgtc gttggagagg gccgaaagcc tgtcgtcggt gagctcccgc
7501 atgcatgccg gcaaagatag taccatgccc cgcactggga ggagcctggg tcgcagcact
7561 ggcgcctcac cacccagctg tggcatcact cagtccacag gggcctctcc caaggccagc
7621 cagtctaaga tctctgcagt gagcaagctc ctgctggcca gtcctaaatc ccgcagcagc
7681 ctgtccacct ccaccaccaa aaccctcagc ttctccacca agtccctgcc acagtctgtg
7741 ggccagagct ccaacctacc gcccagcggg aaacacatgt cctggtccac ccagtcgctc
7801 agcaggaacc ggggctctgg gctggcttcc aaactgccgc taagagcggt caacgggcgc
7861 atttcggagc tgctccaggg cagtgcgggc ccgaggagtg cacagctgcg ggcagaggcg
7921 gaagagcgca gtggggctcc ctcggaggac aagcccgcag ccgcccacct gctgccctcg
7981 ccctacagca agatcacccc tccgaggaag ccgcaccgct gcagcagcgg gcacggtagt
8041 gataacagca gtgtcctgag tggggagcta ccacctgcca tggggaagac tgccctcttc
8101 taccacagcg gaggcagcag tggctacgag agcatgatga gagacagcga ggccaccggc
8161 agtgcatcct cagcccagga ctccatgagc gagaacagca gctccgtggg agggagatgc
8221 aggagcctca aaaaccaaaa gaaaaggtcc aattcaggtt cccagagacg gaggctcatt
8281 ccagcactgt ctctcgatac cccctcacct gtgaggaaaa ctgccagcag cacaggggtc
8341 cgctgggtgg acggcccctt gaggagcacc cagaggagcc ttggggaacc attcgaaatt
8401 aaagtctatg agattgatga cgtggagcgc ctgcagcgac gtcgaggagc tacaagcaag
8461 gaggtcatgt gcttcaatgc caagctgaaa atcctagaac atcggcagca gagaatcgct
8521 gaagtccgag ccaagtacga gtggctgatg aaggagctag aagcgaccaa gcagtacctg
8581 atgctggacc ccaacaagtg gcttcgtgaa tttgacttgg agcaggtcct gcagctggat
8641 tccctggagt acctggaggc actggagggt gtgacagagc gcttggagag ccgcgtcaac
8701 ttctgcaagg cccatctcat gatgatcacc tgctttgaca tcacctccag gcgccgataa
8761 ggcgcgcctc tagaactata gtgagtcgta ttacgtagat ccagacatga taagatacat
8821 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
8881 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
8941 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaattcgc
9001 ggccgcactc gagatatcta gacccagctt tcttgtacaa agtggttgat atccagcaca
9061 gtggcggccg ctcgagtcta gagggcccgc ggttcgaagg taagcctatc cctaaccctc
9121 tcctcggtct cgattctacg cgtaccggtt agtaatgagt ttaaacccgc tgatcagcct
9181 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
9241 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
9301 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
9361 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
9421 aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
9481 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
9541 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
9601 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
9661 aacttgatta gggtgatggt tcacgtacct agaagttcct attccgaagt tcctattctc
9721 tagaaagtat aggaacttcc ttggccaaaa agcctgaact caccgcgacg tctgtcgaga
9781 agtttctgat cgaaaagttc gacagcgtct ccgacctgat gcagctctcg gagggcgaag
9841 aatctcgtgc tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct
9901 gcgccgatgg tttctacaaa gatcgttatg tttatcggca ctttgcatcg gccgcgctcc
9961 cgattccgga agtgcttgac attggggaat tcagcgagag cctgacctat tgcatctccc
10021 gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc
10081 agccggtcgc ggaggccatg gatgcgatcg ctgcggccga tcttagccag acgagcgggt
10141 tcggcccatt cggaccgcaa ggaatcggtc aatacactac atggcgtgat ttcatatgcg
10201 cgattgctga tccccatgtg tatcactggc aaactgtgat ggacgacacc gtcagtgcgt
10261 ccgtcgcgca ggctctcgat gagctgatgc tttgggccga ggactgcccc gaagtccggc
10321 acctcgtgca cgcggatttc ggctccaaca atgtcctgac ggacaatggc cgcataacag
10381 cggtcattga ctggagcgag gcgatgttcg gggattccca atacgaggtc gccaacatct
10441 tcttctggag gccgtggttg gcttgtatgg agcagcagac gcgctacttc gagcggaggc
10501 atccggagct tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc
10561 aactctatca gagcttggtt gacggcaatt tcgatgatgc agcttgggcg cagggtcgat
10621 gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa
10681 gcgcggccgt ctggaccgat ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc
10741 ccagcactcg tccgagggca aaggaatagc acgtactacg agatttcgat tccaccgccg
10801 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
10861 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
10921 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
10981 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
11041 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
11101 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
11161 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
11221 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
11281 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
11341 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
11401 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
11461 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
11521 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
11581 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
11641 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
11701 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
11761 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
11821 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
11881 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
11941 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
12001 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
12061 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
12121 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
12181 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
12241 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
12301 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
12361 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
12421 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
12481 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
12541 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
12601 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
12661 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
12721 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
12781 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
12841 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
12901 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
12961 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
13021 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
13081 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
13141 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
13201 ttccccgaaa agtgccacct gacgtcga
//
| {
"Vector backbone": "pEF5-FRT-V5-DEST",
"Backbone size w/o insert (bp)": "7528",
"Vector type": "Mammalian Expression",
"Selectable markers": "Hygromycin"
} | {
"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": "Kinesin superfamily protein Kif26b links Wnt5a-Ror signaling to the control of cell and tissue behaviors in vertebrates",
"Publication": "Elife. 2017 Sep 8;6. pii: e26509. doi: 10.7554/eLife.26509.",
"DOI": "10.7554/eLife.26509.",
"Authors": "Susman MW, Karuna EP, Kunz RC, Gujral TS, Cantu AV, Choi SS, Jong BY, Okada K, Scales MK, Hum J, Hu LS, Kirschner MW, Nishinakamura R, Yamada S, Laird DJ, Jao LE, Gygi SP, Greenberg ME, Ho HH /eLife",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/28885975",
"PubMed ID": "PubMed 28885975"
} | {
"Gene/Insert name": "Mouse Kif26b",
"Species": "M. musculus (mouse)",
"Insert Size (bp)": "6341",
"Mutation": "",
"Entrez Gene": "Kif26b",
"Promoter": "EF1A",
"Tag / Fusion Protein": "GFP (N terminal on insert)"
} | {
"Cloning method": "Gateway Cloning",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "EF1A promoter forward (TCAAGCCTCAGACAGTGGTTC)",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"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": ""
} |
pETM-11-6His-DmBelle_299-694_B | 145952 | Bacterial Expression of DmBelle_299-694 | Elisa Izaurralde | Izaurralde lab plasmids (unpublished) | https://media.addgene.org/snapgene-media/v2.0.0/sequences/288397/edbd5f68-9765-4399-bcfb-3b7eca090a57/addgene-plasmid-145952-sequence-288397.gbk | full | LOCUS sequence-288397- 6552 bp DNA circular SYN 26-AUG-2024
DEFINITION Bacterial Expression of DmBelle_299-694.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6552)
TITLE Izaurralde lab plasmids
REFERENCE 2 (bases 1 to 6552)
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..6552
/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)."
CDS 5078..5095
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 5126..5146
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQG"
CDS 6396..6413
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(6466..6484)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 6480..6527
/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 ttttgattta actttaagaa ggagatatac catgaaacat caccatcacc atcaccccat
5101 gagcgattac gacatcccca ctactgagaa tctttatttt cagggcgcca tggatgtgca
5161 gctgacggag attatccgca acaacgtggc cttagcacgt tatgacaaac cgacaccggt
5221 gcagaagcac gccattccga tcattatcaa tggccgggat ctaatggcct gcgcccagac
5281 tggatccggc aagacggccg ccttcctggt gccgattctc aaccagatgt acgagctggg
5341 acatgtgccc ccgccgcaaa gcacccggca gtacagccgg cgcaagcagt atccgttggg
5401 tctggtgctg gcgccgaccc gcgaactggc cacccagatc tttgaggagg ccaagaagtt
5461 cgcctatcgc tcccggatgc gcccggcagt gctgtacggc ggaaacaata ctagcgagca
5521 gatgcgcgag ctggaccgcg gttgccatct gattgtggcc acacccggtc gcctagagga
5581 catgatcaca cgcggaaagg tgggactgga gaacattcga ttccttgtac tggatgaggc
5641 tgatcgtatg ttggacatgg gtttcgagcc gcagatccgt cgcatcgtcg aacagctaaa
5701 catgccgcca accggacagc gccagacgct tatgttctcg gccacattcc ccaagcagat
5761 ccaggagtta gccagcgatt tcctcagcaa ctacattttc ttggccgttg gtcgtgtggg
5821 ctccacatct gagaacatta cgcagacaat cctgtgggtc tatgaacccg acaagcgctc
5881 gtatctgctg gatctgctct cgtctatccg cgatggcccc gaatacacca aagacagcct
5941 aacactgatc tttgtggaga ctaagaaggg tgcagactcg ctggaggagt tcctgtacca
6001 gtgcaatcat cccgtcacta gcatccacgg tgatcgcacg cagaaggagc gcgaggaggc
6061 actgcgctgc ttccgatcgg gcgactgccc catcctggtg gccacagccg tggccgctcg
6121 tggcctggac attccgcacg ttaagcacgt tattaacttt gacctgccct cggatgtgga
6181 ggagtatgtc caccgtatcg ggcgtaccgg acgtatgggt aatcttggtg tggccacctc
6241 ctttttcaac gaaaagaacc gcaacatttg ctccgatctc ttggagctgc tgatcgaaac
6301 gaagcaggag atccccagtt tcatggagga tatgagttcg tagggtaccg gatccgaatt
6361 cgagctccgt cgacaagctt gcggccgcac tcgagcacca ccaccaccac cactgagatc
6421 cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac
6481 tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa
6541 ctatatccgg at
//
| {
"Vector backbone": "Unknown",
"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": "DmBelle_299-694",
"Species": "D. melanogaster (fly)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "bel",
"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": ""
} |
|
pMpGWB417 | 68682 | (Empty Backbone)Gateway binary vector designed for transgenic research with Marchantia polymorpha as well as other plants | Takayuki Kohchi | Ishizaki et al PLoS One. 2015 Sep 25;10(9):e0138876. doi: 10.1371/journal.pone.0138876. eCollection 2015. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/121809/e874ad09-c8ed-4469-94bb-52a8b54e6d89/addgene-plasmid-68682-sequence-121809.gbk | full | LOCUS sequence-121809- 11035 bp DNA circular SYN 26-AUG-2024
DEFINITION Gateway binary vector designed for transgenic research with
Marchantia polymorpha as well as other plants.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11035)
AUTHORS Ishizaki K, Nishihama R, Ueda M, Inoue K, Ishida S, Nishimura Y,
Shikanai T, Kohchi T
TITLE Development of Gateway Binary Vector Series with Four Different
Selection Markers for the Liverwort Marchantia polymorpha.
JOURNAL PLoS One. 2015 Sep 25;10(9):e0138876. doi:
10.1371/journal.pone.0138876. eCollection 2015.
PUBMED 26406247
REFERENCE 2 (bases 1 to 11035)
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: "2015-09-25"; volume: "10(9):e0138876. doi"; pages: "
10.1371/journal.pone.0138876. eCollection 2015"
FEATURES Location/Qualifiers
source 1..11035
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 54..78
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
primer_bind 266..288
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 280..297
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 281..297
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 323..447
/gene="mutant version of attR"
/label=attR1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
promoter 484..514
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
CDS 568..1227
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind complement(634..653)
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
CDS 1569..1874
/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 1753..1772
/label=ccdB-fwd
/note="ccdB gene, forward primer"
protein_bind complement(1915..2039)
/gene="mutant version of attR"
/label=attR2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
terminator 2110..2362
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
primer_bind complement(2396..2412)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2396..2412)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(2409..2431)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 2420..2436
/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(2444..2474)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2489..2510
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
polyA_signal 2595..2769
/label=CaMV poly(A) signal
/note="cauliflower mosaic virus polyadenylation signal"
CDS complement(2826..3623)
/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="MGIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLL
SSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEH
QGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDI
ALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(2937..2956)
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind 3547..3566
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
promoter complement(3686..4362)
/label=CaMV 35S promoter (enhanced)
/note="cauliflower mosaic virus 35S promoter with a
duplicated enhancer region"
primer_bind complement(3729..3749)
/label=35S promoter
/note="CaMV 35S promoter, forward primer"
misc_feature 4557..4581
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
CDS 5102..5893
/codon_start=1
/gene="aadA"
/product="aminoglycoside adenylyltransferase (Murphy,
1985)"
/label=SmR
/note="confers resistance to spectinomycin and
streptomycin"
/translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
rep_origin 6139..6727
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6628..6647
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 6881..6898
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 6913..7053
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(6979..6998)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 7139..7161
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
rep_origin 7397..7591
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
CDS complement(7657..8730)
/codon_start=1
/product="replication protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/label=pVS1 RepA
/translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
LVMRYRNLIEGEASAGS"
CDS complement(9159..9788)
/codon_start=1
/product="stability protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/label=pVS1 StaA
/translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
ORIGIN
1 tttcacgccc ttttaaatat ccgattattc taataaacgc tcttttctct taggtttacc
61 cgccaatata tcctgtcaaa cactgatagt ttaaactgaa ggcgggaaac gacaatctga
121 tccaagctca agctgctcta gcattcgcca ttcaggctgc gcaactgttg ggaagggcga
181 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
241 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc
301 aagcttgggt ctagagttat caacaagttt gtacaaaaaa gctgaacgag aaacgtaaaa
361 tgatataaat atcaatatat taaattagat tttgcataaa aaacagacta cataatactg
421 taaaacacaa catatccagt cactatggcg gcccctcgag gggccgcatt aggcacccca
481 ggctttacac tttatgcttc cggctcgtat aatgtgtgga ttttgagtta ggatccggcg
541 agattttcag gagctaagga agctaaaatg gagaaaaaaa tcactggata taccaccgtt
601 gatatatccc aatggcatcg taaagaacat tttgaggcat ttcagtcagt tgctcaatgt
661 acctataacc agaccgttca gctggatatt acggcctttt taaagaccgt aaagaaaaat
721 aagcacaagt tttatccggc ctttattcac attcttgccc gcctgatgaa tgctcatccg
781 gaattccgta tggcaatgaa agacggtgag ctggtgatat gggatagtgt tcacccttgt
841 tacaccgttt tccatgagca aactgaaacg ttttcatcgc tctggagtga ataccacgac
901 gatttccggc agtttctaca catatattcg caagatgtgg cgtgttacgg tgaaaacctg
961 gcctatttcc ctaaagggtt tattgagaat atgtttttcg tctcagccaa tccctgggtg
1021 agtttcacca gttttgattt aaacgtggcc aatatggaca acttcttcgc ccccgttttc
1081 accatgggca aatattatac gcaaggcgac aaggtgctga tgccgctggc gattcaggtt
1141 catcatgccg tctgtgatgg cttccatgtc ggcagaatgc ttaatgaatt acaacagtac
1201 tgcgatgagt ggcagggcgg ggcgtaaacg cgtggatccg gcttactaaa agccagataa
1261 cagtatgcgt atttgcgcgc tgatttttgc ggtataagaa tatatactga tatgtatacc
1321 cgaagtatgt caaaaagagg tgtgctatga agcagcgtat tacagtgaca gttgacagcg
1381 acagctatca gttgctcaag gcatatatga tgtcaatatc tccggtctgg taagcacaac
1441 catgcagaat gaagcccgtc gtctgcgtgc cgaacgctgg aaagcggaaa atcaggaagg
1501 gatggctgag gtcgcccggt ttattgaaat gaacggctct tttgctgacg agaacaggga
1561 ctggtgaaat gcagtttaag gtttacacct ataaaagaga gagccgttat cgtctgtttg
1621 tggatgtaca gagtgatatt attgacacgc ccgggcgacg gatggtgatc cccctggcca
1681 gtgcacgtct gctgtcagat aaagtctccc gtgaacttta cccggtggtg catatcgggg
1741 atgaaagctg gcgcatgatg accaccgata tggccagtgt gccggtctcc gttatcgggg
1801 aagaagtggc tgatctcagc caccgcgaaa atgacatcaa aaacgccatt aacctgatgt
1861 tctggggaat ataaatgtca ggctccctta tacacagcca gtctgcaggt cgaccatagt
1921 gactggatat gttgtgtttt acagtattat gtagtctgtt ttttatgcaa aatctaattt
1981 aatatattga tatttatatc attttacgtt tctcgttcag ctttcttgta caaagtggtt
2041 gataacagct tggatttgga tttggagttg cgcttgggat tcgcctaggc ttagagctcg
2101 aatttccccg atcgttcaaa catttggcaa taaagtttct taagattgaa tcctgttgcc
2161 ggtcttgcga tgattatcat ataatttctg ttgaattacg ttaagcatgt aataattaac
2221 atgtaatgca tgacgttatt tatgagatgg gtttttatga ttagagtccc gcaattatac
2281 atttaatacg cgatagaaaa caaaatatag cgcgcaaact aggataaatt atcgcgcgcg
2341 gtgtcatcta tgttactaga tcgggaattg gttccggaac caattcgtaa tcatggtcat
2401 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
2461 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta ggctgaatta
2521 ggcgcgccta tttctgaatt caacacattg cggacgtttt taatgtactg aattaacgcc
2581 gaattaattc gggggatctg gattttagta ctggattttg gttttaggaa ttagaaattt
2641 tattgataga agtattttac aaatacaaat acatactaag ggtttcttat atgctcaaca
2701 catgagcgaa accctatagg aaccctaatt cccttatctg ggaactactc acacattatt
2761 atggagaaac tcgagcttgt cgatcgactc tagctagagg atcgatccga accccagagt
2821 cccgctcaga agaactcgtc aagaaggcga tagaaggcga tgcgctgcga atcgggagcg
2881 gcgataccgt aaagcacgag gaagcggtca gcccattcgc cgccaagctc ttcagcaata
2941 tcacgggtag ccaacgctat gtcctgatag cggtccgcca cacccagccg gccacagtcg
3001 atgaatccag aaaagcggcc attttccacc atgatattcg gcaagcaggc atcgccatgt
3061 gtcacgacga gatcctcgcc gtcgggcatg cgcgccttga gcctggcgaa cagttcggct
3121 ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat cgacaagacc ggcttccatc
3181 cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca ggtagccgga
3241 tcaagcgtat gcagccgccg cattgcatca gccatgatgg atactttctc ggcaggagca
3301 aggtgagatg acaggagatc ctgccccggc acttcgccca atagcagcca gtcccttccc
3361 gcttcagtga caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc cagccacgat
3421 agccgcgctg cctcgtcctg gagttcattc agggcaccgg acaggtcggt cttgacaaaa
3481 agaaccgggc gcccctgcgc tgacagccgg aacacggcgg catcagagca gccgattgtc
3541 tgttgtgccc agtcatagcc gaatagcctc tccacccaag cggccggaga acctgcgtgc
3601 aatccatctt gttcaatccc catggtcgat cgacagatct gcgaaagctc gagagagata
3661 gatttgtaga gagagactgg tgatttcagc gtgtcctctc caaatgaaat gaacttcctt
3721 atatagagga agggtcttgc gaaggatagt gggattgtgc gtcatccctt acgtcagtgg
3781 agatatcaca tcaatccact tgctttgaag acgtggttgg aacgtcttct ttttccacga
3841 tgctcctcgt gggtgggggt ccatctttgg gaccactgtc ggcagaggca tcttgaacga
3901 tagcctttcc tttatcgcaa tgatggcatt tgtaggtgcc accttccttt tctactgtcc
3961 ttttgatgaa gtgacagata gctgggcaat ggaatccgag gaggtttccc gatattaccc
4021 tttgttgaaa agtctcaata gccctttggt cttctgagac tgtatctttg atattcttgg
4081 agtagacgag agtgtcgtgc tccaccatgt tcacatcaat ccacttgctt tgaagacgtg
4141 gttggaacgt cttctttttc cacgatgctc ctcgtgggtg ggggtccatc tttgggacca
4201 ctgtcggcag aggcatcttg aacgatagcc tttcctttat cgcaatgatg gcatttgtag
4261 gtgccacctt ccttttctac tgtccttttg atgaagtgac agatagctgg gcaatggaat
4321 ccgaggaggt ttcccgatat taccctttgt tgaaaagtct caatagccct ttggtcttct
4381 gagactgtat ctttgatatt cttggagtag acgagagtgt cgtgctccac catgttggca
4441 agctgctcta gccaatacgc aaaccgaatt cagaaataaa ttcagcctaa ttcggcgtta
4501 attcagtaca ttaaaaacgt ccgcaatgtg ttattaagtt gtctaagcgt caatttgttt
4561 acaccacaat atatcctgcc accagccagc caacagctcc ccgaccggca gctcggcaca
4621 aaatcaccac tcgatacagg cagcccatca gtccgggacg gcgtcagcgg gagagccgtt
4681 gtaaggcggc agactttgct catgttaccg atgctattcg gaagaacggc aactaagctg
4741 ccgggtttga aacacggatg atctcgcgga gggtagcatg ttgattgtaa cgatgacaga
4801 gcgttgctgc ctgtgatcaa ttcgggcacg aacccagtgg acataagcct gttcggttcg
4861 taagctgtaa tgcaagtagc gtatgcgctc acgcaactgg tccagaacct tgaccgaacg
4921 cagcggtggt aacggcgcag tggcggtttt catggcttgt tatgactgtt tttttggggt
4981 acagtctatg cctcgggcat ccaagcagca agcgcgttac gccgtgggtc gatgtttgat
5041 gttatggagc agcaacgatg ttacgcagca gggcagtcgc cctaaaacaa agttaaacat
5101 catgggggaa gcggtgatcg ccgaagtatc gactcaacta tcagaggtag ttggcgtcat
5161 cgagcgccat ctcgaaccga cgttgctggc cgtacatttg tacggctccg cagtggatgg
5221 cggcctgaag ccacacagtg atattgattt gctggttacg gtgaccgtaa ggcttgatga
5281 aacaacgcgg cgagctttga tcaacgacct tttggaaact tcggcttccc ctggagagag
5341 cgagattctc cgcgctgtag aagtcaccat tgttgtgcac gacgacatca ttccgtggcg
5401 ttatccagct aagcgcgaac tgcaatttgg agaatggcag cgcaatgaca ttcttgcagg
5461 tatcttcgag ccagccacga tcgacattga tctggctatc ttgctgacaa aagcaagaga
5521 acatagcgtt gccttggtag gtccagcggc ggaggaactc tttgatccgg ttcctgaaca
5581 ggatctattt gaggcgctaa atgaaacctt aacgctatgg aactcgccgc ccgactgggc
5641 tggcgatgag cgaaatgtag tgcttacgtt gtcccgcatt tggtacagcg cagtaaccgg
5701 caaaatcgcg ccgaaggatg tcgctgccga ctgggcaatg gagcgcctgc cggcccagta
5761 tcagcccgtc atacttgaag ctagacaggc ttatcttgga caagaagaag atcgcttggc
5821 ctcgcgcgca gatcagttgg aagaatttgt ccactacgtg aaaggcgaga tcaccaaggt
5881 agtcggcaaa taatgtctag ctagaaattc gttcaagccg acgccgcttc gcggcgcggc
5941 ttaactcaag cgttagatgc actaagcaca taattgctca cagccaaact atcaggtcaa
6001 gtctgctttt attattttta agcgtgcata ataagcccta cacaaattgg gagatatatc
6061 atgcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
6121 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
6181 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
6241 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
6301 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
6361 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
6421 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
6481 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
6541 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
6601 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
6661 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
6721 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
6781 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
6841 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
6901 agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
6961 catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact
7021 ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac
7081 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
7141 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagggtgcc
7201 ttgatgtggg cgccggcggt cgagtggcga cggcgcggct tgtccgcgcc ctggtagatt
7261 gcctggccgt aggccagcca tttttgagcg gccagcggcc gcgataggcc gacgcgaagc
7321 ggcggggcgt agggagcgca gcgaccgaag ggtaggcgct ttttgcagct cttcggctgt
7381 gcgctggcca gacagttatg cacaggccag gcgggtttta agagttttaa taagttttaa
7441 agagttttag gcggaaaaat cgcctttttt ctcttttata tcagtcactt acatgtgtga
7501 ccggttccca atgtacggct ttgggttccc aatgtacggg ttccggttcc caatgtacgg
7561 ctttgggttc ccaatgtacg tgctatccac aggaaagaga ccttttcgac ctttttcccc
7621 tgctagggca atttgcccta gcatctgctc cgtacattag gaaccggcgg atgcttcgcc
7681 ctcgatcagg ttgcggtagc gcatgactag gatcgggcca gcctgccccg cctcctcctt
7741 caaatcgtac tccggcaggt catttgaccc gatcagcttg cgcacggtga aacagaactt
7801 cttgaactct ccggcgctgc cactgcgttc gtagatcgtc ttgaacaacc atctggcttc
7861 tgccttgcct gcggcgcggc gtgccaggcg gtagagaaaa cggccgatgc cgggatcgat
7921 caaaaagtaa tcggggtgaa ccgtcagcac gtccgggttc ttgccttctg tgatctcgcg
7981 gtacatccaa tcagctagct cgatctcgat gtactccggc cgcccggttt cgctctttac
8041 gatcttgtag cggctaatca aggcttcacc ctcggatacc gtcaccaggc ggccgttctt
8101 ggccttcttc gtacgctgca tggcaacgtg cgtggtgttt aaccgaatgc aggtttctac
8161 caggtcgtct ttctgctttc cgccatcggc tcgccggcag aacttgagta cgtccgcaac
8221 gtgtggacgg aacacgcggc cgggcttgtc tcccttccct tcccggtatc ggttcatgga
8281 ttcggttaga tgggaaaccg ccatcagtac caggtcgtaa tcccacacac tggccatgcc
8341 ggccggccct gcggaaacct ctacgtgccc gtctggaagc tcgtagcgga tcacctcgcc
8401 agctcgtcgg tcacgcttcg acagacggaa aacggccacg tccatgatgc tgcgactatc
8461 gcgggtgccc acgtcataga gcatcggaac gaaaaaatct ggttgctcgt cgcccttggg
8521 cggcttccta atcgacggcg caccggctgc cggcggttgc cgggattctt tgcggattcg
8581 atcagcggcc gcttgccacg attcaccggg gcgtgcttct gcctcgatgc gttgccgctg
8641 ggcggcctgc gcggccttca acttctccac caggtcatca cccagcgccg cgccgatttg
8701 taccgggccg gatggtttgc gaccgctcac gccgattcct cgggcttggg ggttccagtg
8761 ccattgcagg gccggcagac aacccagccg cttacgcctg gccaaccgcc cgttcctcca
8821 cacatggggc attccacggc gtcggtgcct ggttgttctt gattttccat gccgcctcct
8881 ttagccgcta aaattcatct actcatttat tcatttgctc atttactctg gtagctgcgc
8941 gatgtattca gatagcagct cggtaatggt cttgccttgg cgtaccgcgt acatcttcag
9001 cttggtgtga tcctccgccg gcaactgaaa gttgacccgc ttcatggctg gcgtgtctgc
9061 caggctggcc aacgttgcag ccttgctgct gcgtgcgctc ggacggccgg cacttagcgt
9121 gtttgtgctt ttgctcattt tctctttacc tcattaactc aaatgagttt tgatttaatt
9181 tcagcggcca gcgcctggac ctcgcgggca gcgtcgccct cgggttctga ttcaagaacg
9241 gttgtgccgg cggcggcagt gcctgggtag ctcacgcgct gcgtgatacg ggactcaaga
9301 atgggcagct cgtacccggc cagcgcctcg gcaacctcac cgccgatgcg cgtgcctttg
9361 atcgcccgcg acacgacaaa ggccgcttgt agccttccat ccgtgacctc aatgcgctgc
9421 ttaaccagct ccaccaggtc ggcggtggcc catatgtcgt aagggcttgg ctgcaccgga
9481 atcagcacga agtcggctgc cttgatcgcg gacacagcca agtccgccgc ctggggcgct
9541 ccgtcgatca ctacgaagtc gcgccggccg atggccttca cgtcgcggtc aatcgtcggg
9601 cggtcgatgc cgacaacggt tagcggttga tcttcccgca cggccgccca atcgcgggca
9661 ctgccctggg gatcggaatc gactaacaga acatcggccc cggcgagttg cagggcgcgg
9721 gctagatggg ttgcgatggt cgtcttgcct gacccgcctt tctggttaag tacagcgata
9781 accttcatgc gttccccttg cgtatttgtt tatttactca tcgcatcata tacgcagcga
9841 ccgcatgacg caagctgttt tactcaaata cacatcacct ttttagacgg cggcgctcgg
9901 tttcttcagc ggccaagctg gccggccagg ccgccagctt ggcatcagac aaaccggcca
9961 ggatttcatg cagccgcacg gttgagacgt gcgcgggcgg ctcgaacacg tacccggccg
10021 cgatcatctc cgcctcgatc tcttcggtaa tgaaaaacgg ttcgtcctgg ccgtcctggt
10081 gcggtttcat gcttgttcct cttggcgttc attctcggcg gccgccaggg cgtcggcctc
10141 ggtcaatgcg tcctcacgga aggcaccgcg ccgcctggcc tcggtgggcg tcacttcctc
10201 gctgcgctca agtgcgcggt acagggtcga gcgatgcacg ccaagcagtg cagccgcctc
10261 tttcacggtg cggccttcct ggtcgatcag ctcgcgggcg tgcgcgatct gtgccggggt
10321 gagggtaggg cgggggccaa acttcacgcc tcgggccttg gcggcctcgc gcccgctccg
10381 ggtgcggtcg atgattaggg aacgctcgaa ctcggcaatg ccggcgaaca cggtcaacac
10441 catgcggccg gccggcgtgg tggtgtcggc ccacggctct gccaggctac gcaggcccgc
10501 gccggcctcc tggatgcgct cggcaatgtc cagtaggtcg cgggtgctgc gggccaggcg
10561 gtctagcctg gtcactgtca caacgtcgcc agggcgtagg tggtcaagca tcctggccag
10621 ctccgggcgg tcgcgcctgg tgccggtgat cttctcggaa aacagcttgg tgcagccggc
10681 cgcgtgcagt tcggcccgtt ggttggtcaa gtcctggtcg tcggtgctga cgcgggcata
10741 gcccagcagg ccagcggcgg cgctcttgtt catggcgtaa tgtctccggt tctagtcgca
10801 agtattctac tttatgcgac taaaacacgc gacaagaaaa cgccaggaaa agggcagggc
10861 ggcagcctgt cgcgtaactt aggacttgtg cgacatgtcg ttttcagaag acggctgcac
10921 tgaacgtcag aagccgactg cactatagca gcggaggggt tggatcaaag tactttgatc
10981 ccgaggggaa ccctgtggtt ggcatgcaca tacaaatgga cgaacggata aacct
//
| {
"Vector backbone": "pMpGWB400",
"Backbone size w/o insert (bp)": "",
"Vector type": "Plant Expression",
"Selectable markers": "Neomycin (select with G418)"
} | {
"Bacterial Resistance(s)": "Chloramphenicol and Spectinomycin, 25 & 50 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": "Development of Gateway Binary Vector Series with Four Different Selection Markers for the Liverwort Marchantia polymorpha",
"Publication": "PLoS One. 2015 Sep 25;10(9):e0138876. doi: 10.1371/journal.pone.0138876. eCollection 2015.",
"DOI": "10.1371/journal.pone.0138876.",
"Authors": "Ishizaki K, Nishihama R, Ueda M, Inoue K, Ishida S, Nishimura Y, Shikanai T, Kohchi T PONE-D-- [pii]",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/26406247",
"PubMed ID": "PubMed 26406247"
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"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": ""
} |
|
pMuLE ENTR SFFV L5-L2 | 62098 | (Empty Backbone)MuLE (Multiple Lentiviral Expression) Entry vector containing a SFFV promoter and multiple cloning site module. Compatible with MultiSite Gateway cloning | Ian Frew | Albers et al J Clin Invest. 2015 Mar 9. pii: 79743. doi: 10.1172/JCI79743. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/101387/85362848-d71d-42e8-b17f-cc43c8cc76ee/addgene-plasmid-62098-sequence-101387.gbk | full | LOCUS sequence-101387- 3098 bp DNA circular SYN 26-AUG-2024
DEFINITION MuLE (Multiple Lentiviral Expression) Entry vector containing a SFFV
promoter and multiple cloning site module. Compatible with MultiSite
Gateway cloning.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3098)
AUTHORS Albers J, Danzer C, Rechsteiner M, Lehmann H, Brandt LP, Hejhal T,
Catalano A, Busenhart P, Goncalves AF, Brandt S, Bode PK,
Bode-Lesniewska B, Wild PJ, Frew IJ
TITLE A versatile modular vector system for rapid combinatorial mammalian
genetics.
JOURNAL J Clin Invest. 2015 Mar 9. pii: 79743. doi: 10.1172/JCI79743.
PUBMED 25751063
REFERENCE 2 (bases 1 to 3098)
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 Clin
Invest. 2015 Mar 9. pii: 79743. doi: 10.1172/JCI79743."
FEATURES Location/Qualifiers
source 1..3098
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind complement(85..184)
/gene="mutant version of attL"
/label=attL2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
primer_bind complement(214..233)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(215..233)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(238..254)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(238..254)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(356..375)
/label=pENTR-R
/note="pENTR vectors, reverse primer"
CDS 367..1176
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind complement(434..453)
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
rep_origin 1323..1911
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 1812..1831
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 2065..2082
/label=L4440
/note="L4440 vector, forward primer"
terminator complement(2241..2268)
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator complement(2360..2446)
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
primer_bind 2495..2517
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 2509..2526
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 2510..2526
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 2543..2638
/gene="mutant version of attL"
/label=attL5
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
ORIGIN
1 gatccactag taacggccgc cagtgtgctg gaattctgca gatatccatc acactggcgg
61 ccgctcgagc atgcatctag aggtacccag ctttcttgta caaagttggc attataagaa
121 agcattgctt atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta
181 tttgccatcc agctgcaggg cggccgcgat atcccctata gtgagtcgta ttacatggtc
241 atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa
301 gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag taatacaagg
361 ggtgttatga gccatattca acgggaaacg tcgaggccgc gattaaattc caacatggat
421 gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc
481 tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc
541 gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct
601 cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg
661 atccccggaa aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt
721 gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct
781 tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg
841 gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa
901 gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca
961 cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc
1021 ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct
1081 ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa
1141 ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa ttggttgtaa
1201 cactggcaga gcattacgct gacttgacgg gacggcgcaa gctcatgacc aaaatccctt
1261 aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
1321 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
1381 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
1441 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
1501 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
1561 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
1621 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
1681 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
1741 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
1801 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
1861 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
1921 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
1981 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
2041 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
2101 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
2161 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atacgcgtac cgctagccag
2221 gaagagtttg tagaaacgca aaaaggccat ccgtcaggat ggccttctgc ttagtttgat
2281 gcctggcagt ttatggcggg cgtcctgccc gccaccctcc gggccgttgc ttcacaacgt
2341 tcaaatccgc tcccggcgga tttgtcctac tcaggagagc gttcaccgac aaacaacaga
2401 taaaacgaaa ggcccagtct tccgactgag cctttcgttt tatttgatgc ctggcagttc
2461 cctactctcg cgttaacgct agcatggatg ttttcccagt cacgacgttg taaaacgacg
2521 gccagtctta agctcgggcc ccaaataatg attttatttt gactgatagt gacctgttcg
2581 ttgcaacaaa ttgatgagca atgctttttt ataatgccaa ctttgtatac aaaagttggg
2641 agacccaagc ttcgctgaaa gaccccacct gtaggtttgg caagctagct gcagtaacgc
2701 cattttgcaa ggcatggaaa aataccaaac caagaataga gaagttcaga tcaagggcgg
2761 gtacatgaaa atagctaacg ttgggccaaa caggatatct gcggtgagca gtttcggccc
2821 cggcccgggg ccaagaacag atggtcaccg cagtttcggc cccggcccga ggccaagaac
2881 agatggtccc cagatatggc ccaaccctca gcagtttctt aagacccatc agatgtttcc
2941 aggctccccc aaggacctga aatgaccctg cgccttattt gaattaacca atcagcctgc
3001 ttctcgcttc tgttcgcgcg cttctgcttc ccgagctcta taaaagagct cacaacccct
3061 cactcggcgc gccagtcctc cgattgactg agtcgccg
//
| {
"Vector backbone": "N/A",
"Backbone size w/o insert (bp)": "",
"Vector type": "Mammalian Expression ; MuLE Gateway Entry Vector",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Kanamycin, 50 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "DH5alpha",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "A versatile modular vector system for rapid combinatorial mammalian genetics",
"Publication": "J Clin Invest. 2015 Mar 9. pii: 79743. doi: 10.1172/JCI79743.",
"DOI": "10.1172/JCI79743.",
"Authors": "Albers J, Danzer C, Rechsteiner M, Lehmann H, Brandt LP, Hejhal T, Catalano A, Busenhart P, Goncalves AF, Brandt S, Bode PK, Bode-Lesniewska B, Wild PJ, Frew IJ /JCI",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/25751063",
"PubMed ID": "PubMed 25751063"
} | {
"Gene/Insert name": "None",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
LEP-shTAF4.2818 | 105563 | retrovirally express TAF4 shRNA with puro resistance and GFP marker | Christopher Vakoc | Xu et al Cancer Cell. 2018 Jan 8;33(1):13-28.e8. doi: 10.1016/j.ccell.2017.12.002. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/203676/d1fffeb9-fddc-468e-ac34-75bc524d7a67/addgene-plasmid-105563-sequence-203676.gbk | full | LOCUS sequence-203676- 7979 bp DNA circular SYN 26-AUG-2024
DEFINITION retrovirally express TAF4 shRNA with puro resistance and GFP marker.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7979)
AUTHORS Xu Y, Milazzo JP, Somerville TDD, Tarumoto Y, Huang YH, Ostrander
EL, Wilkinson JE, Challen GA, Vakoc CR
TITLE A TFIID-SAGA Perturbation that Targets MYB and Suppresses Acute
Myeloid Leukemia.
JOURNAL Cancer Cell. 2018 Jan 8;33(1):13-28.e8. doi:
10.1016/j.ccell.2017.12.002.
PUBMED 29316427
REFERENCE 2 (bases 1 to 7979)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1016/j.ccell.2017.12"; journalName: "Cancer Cell"; date:
"2018-01-8- 8"; volume: "33"; issue: "1"; pages: "13-28.e8"
FEATURES Location/Qualifiers
source 1..7979
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 579..920
/label=MESV Psi
/note="packaging signal of murine embryonic stem cell
virus"
CDS 987..1403
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
primer_bind 1333..1355
/label=pLXSN 5'
/note="Murine stem cell virus, forward primer. Also called
MSCV"
primer_bind 1371..1387
/label=pBABE 5'
/note="Psi packaging signal, forward primer for pBABE
vectors"
ncRNA 1592..1606
/label=miR-30a loop
/note="loop from the the 71-nt precursor of the human
miR-30a microRNA (Zeng et al., 2002)"
/ncRNA_class="miRNA"
promoter 1790..2289
/label=PGK promoter
/note="mouse phosphoglycerate kinase 1 promoter"
primer_bind complement(1825..1848)
/label=MSCV-rev
/note="Murine stem cell virus, reverse primer"
primer_bind 2222..2241
/label=mPGK-F
/note="Mouse PGK promoter, forward primer"
CDS 2310..2909
/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(2310..2329)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 2806..2826
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
misc_feature 2970..3537
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
misc_feature 2987..3539
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(3131..3148)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 3358..3377
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 3551..4270
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(3596..3617)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(3857..3876)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 4204..4225
/label=EGFP-C
/note="EGFP, forward primer"
LTR 4372..4886
/label=3' LTR
/note="3' long terminal repeat from murine embryonic stem
cell virus"
protein_bind 5048..5064
/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(5072..5102)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5117..5138
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(5255..5272)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5426..6014)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5506..5525)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6185..7045)
/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 6808..6827
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7046..7150)
/gene="bla"
/label=AmpR promoter
primer_bind 7218..7236
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(7274..7296)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 7396..7415
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 7609..7631
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 7759..7778
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
LTR join(7978..7979,1..515)
/label=5' LTR
/note="5' long terminal repeat from murine embryonic stem
cell virus"
ORIGIN
1 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
61 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca
121 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
181 acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt
241 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc
301 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc
361 ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt attcccaata
421 aagcctcttg ctgtttgcat ccgaatcgtg gactcgctga tccttgggag ggtctcctca
481 gattgattga ctgcccacct cgggggtctt tcatttggag gttccaccga gatttggaga
541 cccctgccca gggaccaccg acccccccgc cgggaggtaa gctggccagc ggtcgtttcg
601 tgtctgtctc tgtctttgtg cgtgtttgtg ccggcatcta atgtttgcgc ctgcgtctgt
661 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa
721 cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga
781 cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag
841 acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa
901 gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg
961 tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct
1021 taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga
1081 gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag
1141 acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc
1201 cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc
1261 cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg
1321 ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag
1381 ccctcactcc ttctctaggc gccggaatta gatctctcga ctagggataa cagggtaatt
1441 gtttgaatga ggcttcagta ctttacagaa tcgttgcctg cacatcttgg aaacacttgc
1501 tgggattact tcgacttctt aacccaacag aaggctcgag aaggtatatt gctgttgaca
1561 gtgagcgcaa ggttgaagca gaaagcaaat agtgaagcca cagatgtatt tgctttctgc
1621 ttcaacctta tgcctactgc ctcggacttc aaggggctag aattcgagca attatcttgt
1681 ttactaaaac tgaatacctt gctatctctt tgatacattt ttacaaagct gaattaaaat
1741 ggtataaatt aaatcacttt tttcaattga cgcgtgatct aattctaccg ggtaggggag
1801 gcgcttttcc caaggcagtc tggagcatgc gctttagcag ccccgctggg cacttggcgc
1861 tacacaagtg gcctctggcc tcgcacacat tccacatcca ccggtaggcg ccaaccggct
1921 ccgttctttg gtggcccctt cgcgccacct tctactcctc ccctagtcag gaagttcccc
1981 cccgccccgc agctcgcgtc gtgcaggacg tgacaaatgg aagtagcacg tctcactagt
2041 ctcgtgcaga tggacagcac cgctgagcaa tggaagcggg taggcctttg gggcagcggc
2101 caatagcagc tttgctcctt cgctttctgg gctcagaggc tgggaagggg tgggtccggg
2161 ggcgggctca ggggcgggct caggggcggg gcgggcgccc gaaggtcctc cggaggcccg
2221 gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg
2281 ggcctttcga cctgcagccc aagcttacca tgaccgagta caagcccacg gtgcgcctcg
2341 ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc
2401 ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag
2461 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg
2521 ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga
2581 tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag
2641 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct
2701 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg
2761 ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct
2821 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct
2881 ggtgcatgac ccgcaagccc ggtgcctgac gcccgcccca cgacccgcag cgcccgaccg
2941 aaaggagcgc acgaccccat gcatccaatt ccgccccccc cccctaacgt tactggccga
3001 agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg
3061 tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg
3121 ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt
3181 cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac
3241 cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca
3301 aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg
3361 ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg
3421 ggatctgatc tggggcctcg gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa
3481 cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg atgataatat
3541 ggccacaacc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
3601 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tctggcgagg gcgagggcga
3661 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
3721 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
3781 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
3841 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
3901 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
3961 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
4021 gcagaagaac ggcatcaagg cgaacttcaa gatccgccac aacatcgagg acggcagcgt
4081 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
4141 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga
4201 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
4261 gtacaagtaa tgaattaatt aagaattatc aagcttatcg ataccgtcga cctgcagcca
4321 agcttatcga taaaataaaa gattttattt agtctccaga aaaagggggg aatgaaagac
4381 cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc atggaaaata
4441 cataactgag aatagagaag ttcagatcaa ggttaggaac agagagacag cagaatatgg
4501 gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg
4561 tccccagatg cggtcccgcc ctcagcagtt tctagagaac catcagatgt ttccagggtg
4621 ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt tcgcttctcg
4681 cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac ccctcactcg
4741 gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc gtgtatccaa taaaccctct
4801 tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc tgagtgattg
4861 actacccgtc agcgggggtc tttcatgggt aacagtttct tgaagttgga gaacaacatt
4921 ctgagggtag gagtcgaata ttaagtaatc ctgactcaat tagccactgt tttgaatcca
4981 catactccaa tactcctgaa atagttcatt atggacagcg cagaaagagc tggggagaat
5041 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
5101 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
5161 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
5221 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
5281 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
5341 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
5401 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
5461 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
5521 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
5581 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
5641 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
5701 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
5761 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
5821 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
5881 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
5941 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
6001 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
6061 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
6121 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
6181 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
6241 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
6301 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
6361 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
6421 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
6481 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
6541 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
6601 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
6661 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
6721 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
6781 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
6841 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
6901 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
6961 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
7021 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
7081 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
7141 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
7201 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
7261 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
7321 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
7381 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
7441 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc
7501 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
7561 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
7621 gttgtaaaac gacggcgcaa ggaatggtgc atgcaaggag atggcgccca acagtccccc
7681 ggccacgggg cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg
7741 agcccgatct tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc
7801 gccggtgatg ccggccacga tgcgtccggc gtagaggcga ttagtccaat ttgttaaaga
7861 caggatatca gtggtccagg ctctagtttt gactcaacaa tatcaccagc tgaagcctat
7921 agagtacgag ccatagataa aataaaagat tttatttagt ctccagaaaa aggggggaa
//
| {
"Vector backbone": "LEP (MSCV-LTR-mirE-shRNA-PGK-puro-IRES-GFP)",
"Backbone size w/o insert (bp)": "",
"Vector type": "Mammalian Expression, Retroviral, RNAi",
"Selectable markers": "Puromycin"
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "NEB Stable",
"Copy number": "Unknown"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "A TFIID-SAGA Perturbation that Targets MYB and Suppresses Acute Myeloid Leukemia",
"Publication": "Cancer Cell. 2018 Jan 8;33(1):13-28.e8. doi: 10.1016/j.ccell.2017.12.002.",
"DOI": "10.1016/j.ccell.2017.12.002.",
"Authors": "Xu Y, Milazzo JP, Somerville TDD, Tarumoto Y, Huang YH, Ostrander EL, Wilkinson JE, Challen GA, Vakoc CR /jccell",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/29316427",
"PubMed ID": "PubMed 29316427"
} | {
"Gene/Insert name": "TAF4 shRNA",
"Species": "M. musculus (mouse)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "MSCV-LTR",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "XhoI (unknown if destroyed)",
"3′ cloning site": "EcoRI (unknown if destroyed)",
"5′ sequencing primer": "TGT TTG AAT GAG GCT TCA GTA C",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
FLAG-MBP1-E322TAG-E309C-W340A.pcDNA3-k | 127408 | Expresses MBP in mammalian cells. Amber codon at AA# 322 as a noncanonical amino acid incorporation site | Sharona Gordon | Gordon et al Elife. 2018 Jun 20;7. pii: 37248. doi: 10.7554/eLife.37248. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/247971/c1541869-3b0c-4b8c-84c9-505a7470cac3/addgene-plasmid-127408-sequence-247971.gbk | full | LOCUS sequence-247971- 6650 bp DNA circular SYN 26-AUG-2024
DEFINITION Expresses MBP in mammalian cells. Amber codon at AA# 322 as a
noncanonical amino acid incorporation site.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6650)
AUTHORS Gordon SE, Munari M, Zagotta WN
TITLE Visualizing conformational dynamics of proteins in solution and at
the cell membrane.
JOURNAL Elife. 2018 Jun 20;7. pii: 37248. doi: 10.7554/eLife.37248.
PUBMED 29923827
REFERENCE 2 (bases 1 to 6650)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Elife.";
date: "2018-06-20"; volume: "7. pii: 37248. doi"; pages: "
10.7554/eLife.37248"
FEATURES Location/Qualifiers
source 1..6650
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 74..453
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 454..657
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 608..628
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 702..721
/label=T7
/note="T7 promoter, forward primer"
promoter 702..720
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory 862..871
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 871..894
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
promoter complement(2042..2060)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(2043..2060)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(2080..2097)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2086..2310
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2356..2784
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2443..2462)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2653..2674
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2793..2813)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2798..3127
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2978..3113
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3040..3059
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3194..3988
/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(3248..3267)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3858..3877
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 4162..4283
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4199..4218)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4253..4272
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4332..4348)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4332..4348)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4345..4367)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4356..4372
/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(4380..4410)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4425..4446
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4563..4580)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4734..5322)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4814..4833)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5493..6353)
/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 6116..6135
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6354..6458)
/gene="bla"
/label=AmpR promoter
primer_bind complement(6533..6552)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc
61 agatatacgc gttgacattg attattgact agttattaat agtaatcaat tacggggtca
121 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
181 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
241 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
301 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
361 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
421 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
481 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
541 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
601 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc
661 tggctaacta gagaacccac tgcttactgg cttatcgaaa ttaatacgac tcactatagg
721 gagacccaag cttgatagcg cactcaggct cactagtaac ggccgccagt gtgctggaaa
781 gcagctccaa ggcacttgct ccatttgggg tgtgcctgca cctagctggt tgcaaattgg
841 gccacagagg atctggccac cgccaccatg gactacaagg acgatgatga caagaagatc
901 gaggagggga agctggtcat ctggatcaac ggcgacaagg gctacaacgg gcttgctgag
961 gtcggaaaga agttcgagaa ggacaccgga atcaaggtca ccgtggagca ccctgataag
1021 ctggaggaga agttcccaca ggtggcagct accggagacg gacctgatat catcttctgg
1081 gcacacgaca gattcggagg ctacgctcag tccggccttc ttgctgaaat cactcctgac
1141 aaggcattcc aggacaagct gtaccctttc acctgggacg cagtcaggta caacggcaag
1201 ttgatcgctt accctatcgc cgtggaggct ttgtccttga tctacaacaa agacctcctg
1261 cctaaccccc caaaaacctg ggaggagatc cccgctctgg acaaggagct gaaggccaag
1321 ggaaagtctg ccctgatgtt caacctgcag gagccatact ttacctggcc actgatcgct
1381 gccgatgggg ggtacgcttt taagtacgag aacggcaaat acgacattaa ggacgtgggc
1441 gtggacaacg ccggcgccaa agccgggttg actttcctgg tggatctgat taagaacaag
1501 catatgaacg ccgataccga ttactccatt gccgaggccg cctttaacaa gggcgagaca
1561 gccatgacaa ttaacgggcc ctgggcctgg tcaaacatcg atacaagcaa ggtgaactac
1621 ggggtcacag tcctgcccac atttaagggg cagcccagca aaccctttgt gggggtgctg
1681 agcgcaggga ttaacgccgc cagtccaaat aaggagctcg ccaaggaatt tctcgaaaac
1741 tacctcctca cagatgaggg gctcgaggcc gtgaacaagg ataagcccct cggcgccgtc
1801 gccctcaaaa gctatgaatg cgagctggtg aaggatccaa ggattgccgc cactatgtag
1861 aacgcccaga agggggaaat tatgcccaac attccccaga tgtccgcctt tgcgtatgcc
1921 gtgcggactg ccgtgattaa tgccgccagc ggccggcaaa ctgtcgatga ggccctgaaa
1981 gatgcccaga cacgcattac aaaatgagcg gccgctcgag catgcatcta gagggcccta
2041 ttctatagtg tcacctaaat gctagagctc gctgatcagc ctcgactgtg ccttctagtt
2101 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
2161 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
2221 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
2281 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
2341 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
2401 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
2461 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
2521 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
2581 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
2641 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
2701 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
2761 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
2821 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
2881 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
2941 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
3001 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
3061 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
3121 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg
3181 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
3241 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
3301 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
3361 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
3421 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
3481 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
3541 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
3601 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
3661 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
3721 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
3781 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
3841 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
3901 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
3961 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
4021 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
4081 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
4141 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
4201 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
4261 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
4321 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
4381 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
4441 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
4501 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
4561 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
4621 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
4681 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
4741 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
4801 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
4861 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
4921 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
4981 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
5041 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
5101 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
5161 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
5221 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
5281 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
5341 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
5401 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
5461 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
5521 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
5581 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
5641 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
5701 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
5761 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
5821 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
5881 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
5941 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
6001 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
6061 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
6121 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
6181 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
6241 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
6301 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
6361 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
6421 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
6481 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct
6541 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
6601 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac
//
| {
"Vector backbone": "pcDNA3",
"Backbone size w/o insert (bp)": "5370",
"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": "Visualizing conformational dynamics of proteins in solution and at the cell membrane",
"Publication": "Elife. 2018 Jun 20;7. pii: 37248. doi: 10.7554/eLife.37248.",
"DOI": "10.7554/eLife.37248.",
"Authors": "Gordon SE, Munari M, Zagotta WN [pii]",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/29923827",
"PubMed ID": "PubMed 29923827"
} | {
"Gene/Insert name": "Maltose Binding Protein",
"Species": "",
"Insert Size (bp)": "1281",
"Mutation": "Amino acid mutations: E322 to Amber codon, E309C and W340A",
"Entrez Gene": "",
"Promoter": "CMV",
"Tag / Fusion Protein": "FLAG (N terminal on insert)"
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "Hind III (not destroyed)",
"3′ cloning site": "Not I (not destroyed)",
"5′ sequencing primer": "TAATACGACTCACTATAGGG",
"3′ sequencing primer": "GACAGTGGGAGTGGCACCTT"
} | {
"Gene/Insert name": "",
"Species": "",
"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": ""
} |
|
qTAG-N-Blast-V5 | 207686 | (Empty Backbone)Empty N-terminus donor cassette. Integrate homology arms to target the N-terminus insertion of a Blast-2A-V5 cassette | Laurence Pelletier | Philip et al bioRxiv | https://media.addgene.org/snapgene-media/v2.0.0/sequences/413750/a3941b55-5580-4cee-bb70-8f53e80ff4aa/addgene-plasmid-207686-sequence-413750.gbk | full | LOCUS sequence-413750- 3319 bp DNA circular SYN 27-AUG-2024
DEFINITION Empty N-terminus donor cassette. Integrate homology arms to target
the N-terminus insertion of a Blast-2A-V5 cassette.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3319)
AUTHORS Philip R, Sharma A, Matellan L, Erpf AC, Hsu W, Tkach JM, Wyatt HDM,
Pelletier L
TITLE qTAG: An adaptable CRISPR-based endogenous tagging protocol using
optimized repair cassettes
JOURNAL bioRxiv
REFERENCE 2 (bases 1 to 3319)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 27, 2024 from SnapGene Server 7.0.3
https://www.snapgene.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv"
FEATURES Location/Qualifiers
source 1..3319
/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"
protein_bind complement(423..456)
/label=lox71
/bound_moiety="Cre recombinase"
/note="Left element (LE) mutant of loxP (Araki et al.,
2010). Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
CDS 465..860
/codon_start=1
/gene="Aspergillus terreus BSD"
/product="blasticidin S deaminase"
/label=BSD
/note="confers resistance to blasticidin"
/translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
KAIVKDSDGQPTAVGIRELLPSGYVWEG"
CDS 870..926
/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"
protein_bind complement(933..966)
/label=lox66
/bound_moiety="Cre recombinase"
/note="Right element (RE) mutant of loxP (Araki et al.,
2010). Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
CDS 990..1031
/codon_start=1
/product="epitope tag from simian virus 5"
/label=V5 tag
/translation="GKPIPNPLLGLDST"
primer_bind complement(1098..1114)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1098..1114)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(1111..1133)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 1122..1138
/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(1146..1176)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 1191..1212
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(1329..1346)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(1500..2088)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1580..1599)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(2259..3119)
/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 2882..2901
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(3120..3224)
/gene="bla"
/label=AmpR promoter
primer_bind 3292..3310
/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 cggcaagagc aagaccggta
421 gctaccgttc gtatagcata cattatacga agttatctgc tagcatggcc aagcctttgt
481 ctcaagaaga atccaccctc attgaaagag caacggctac aatcaacagc atccccatct
541 ctgaagacta cagcgtcgcc agcgcagctc tctctagcga cggccgcatc ttcactggtg
601 tcaatgtata tcattttact gggggacctt gtgcagaact cgtggtgctg ggcactgctg
661 ctgctgcggc agctggcaac ctgacttgta tcgtcgcgat cggaaatgag aacaggggca
721 tcttgagccc ctgcggacgg tgccgacagg tgcttctcga tctgcatcct gggatcaaag
781 ccatagtgaa ggacagtgat ggacagccga cggcagttgg gattcgtgaa ttgctgccct
841 ctggttatgt gtgggagggc ggatccggag ctactaactt cagcctgctg aagcaggccg
901 gagacgtgga ggagaaccct ggaccttcta gaataacttc gtatagcata cattatacga
961 acggtagcac gcgtggaggt ggcggttccg gcaagcccat ccccaacccc ctgctgggcc
1021 tggacagcac cggtggaggc ggatctctcg agggaggaag cggaggatct ggcggtacca
1081 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
1141 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
1201 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
1261 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
1321 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
1381 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
1441 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
1501 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
1561 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
1621 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
1681 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
1741 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
1801 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
1861 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
1921 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
1981 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
2041 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
2101 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
2161 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
2221 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
2281 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
2341 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
2401 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
2461 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
2521 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
2581 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
2641 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
2701 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
2761 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
2821 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
2881 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
2941 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
3001 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
3061 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
3121 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
3181 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
3241 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
3301 atcacgaggc cctttcgtc
//
| {
"Vector backbone": "pUC19",
"Backbone size w/o insert (bp)": "",
"Vector type": "Mammalian Expression, CRISPR ; Donor Cassette",
"Selectable markers": "Blasticidin"
} | {
"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": "qTAG: An adaptable CRISPR-based endogenous tagging protocol using optimized repair cassettes",
"Publication": "bioRxiv",
"DOI": "10.1101/2023.11.01.565029",
"Authors": "Philip R, Sharma A, Matellan L, Erpf AC, Hsu W, Tkach JM, Wyatt HDM, Pelletier L",
"PubMed Link": "",
"PubMed ID": ""
} | {
"Gene/Insert name": "None",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pX330-sgRNA_Dgcr8_2 | 73526 | Expresses a human codon-optimized SpCas9 and sgRNA targeting Dgcr8 | Constance Ciaudo | Cirera-Salinas et al J Cell Biol. 2017 Jan 18. pii: jcb.201606073. doi: 10.1083/jcb.201606073. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/139836/82e8ee70-36e2-4d5c-86b6-f7ca14d3bb67/addgene-plasmid-73526-sequence-139836.gbk | partial | LOCUS sequence-139836- 769 bp DNA linear UNA 26-AUG-2024
DEFINITION Expresses a human codon-optimized SpCas9 and sgRNA targeting Dgcr8.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE natural DNA sequence
ORGANISM unspecified
REFERENCE 1 (bases 1 to 769)
AUTHORS Cirera-Salinas D, Yu J, Bodak M, Ngondo RP, Herbert KM, Ciaudo C
TITLE Noncanonical function of DGCR8 controls mESC exit from pluripotency.
JOURNAL J Cell Biol. 2017 Jan 18. pii: jcb.201606073. doi:
10.1083/jcb.201606073.
PUBMED 28100686
REFERENCE 2 (bases 1 to 769)
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
Biol. 2017 Jan 18. pii: jcb.201606073. doi: 10.1083/jcb.201606073."
FEATURES Location/Qualifiers
source 1..769
/mol_type="genomic DNA"
/organism="unspecified"
CDS complement(1..48)
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
ORIGIN
1 ctttttcttt tttgcctggc cggccttttt cgtggccgcc ggccttttgt cgcctcccag
61 ctgagacagg tcgatccgtg tctcgtacag gccggtgatg ctctggtgga tcagggtggc
121 gtccagcacc tctttggtgc tggtgtacct cttccggtcg atggtggtgt caaagtactt
181 gaaggcggca ggggctccca gattggtcag ggtaaacagg tggatgatat tctcggcctg
241 ctctctgatg ggcttatccc ggtgcttgtt gtaggcggac agcactttgt ccagattagc
301 gtcggccagg atcactctct tggagaactc gctgatctgc tcgatgatct cgtccaggta
361 gtgcttgtgc tgttccacaa acagctgttt ctgctcatta tcctcggggg agcccttcag
421 cttctcatag tggctggcca ggtacaggaa gttcacatat ttggagggca gggccagttc
481 gtttcccttc tgcagttcgc cggcagaggc cagcattctc ttccggccgt tttccagctc
541 gaacagggag tacttaggca gcttgatgat caggtccttt ttcacttctt tgtagccctt
601 ggcttccaga aagtcgatgg gattcttctc gaagctgctt ctttccatga tggtgatccc
661 cagcagctct ttcacactct tcagtttctt ggacttgccc ttttccactt tggccaccac
721 cagcacagaa taggccacgg tggggctgtc gaagccgccg tacttctta
//
| {
"Vector backbone": "X330-U6-Chimeric_BB-CBh-hSpCas9",
"Backbone size w/o insert (bp)": "8506",
"Vector type": "Mammalian Expression, 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": "Noncanonical function of DGCR8 controls mESC exit from pluripotency",
"Publication": "J Cell Biol. 2017 Jan 18. pii: jcb.201606073. doi: 10.1083/jcb.201606073.",
"DOI": "10.1083/jcb.201606073.",
"Authors": "Cirera-Salinas D, Yu J, Bodak M, Ngondo RP, Herbert KM, Ciaudo C /jcb",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/28100686",
"PubMed ID": "PubMed 28100686"
} | {
"Gene/Insert name": "DGCR8",
"Species": "M. musculus (mouse)",
"Insert Size (bp)": "19",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "hU6",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "BbsI (destroyed during cloning)",
"3′ cloning site": "BbsI (destroyed during cloning)",
"5′ sequencing primer": "agggatggttggttggtggg",
"3′ sequencing primer": "CCAATCCTCCCCCTTGCTGT"
} | {
"Gene/Insert name": "",
"Species": "",
"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": ""
} |
|
pCMV5 HA-TGFBRII | 19158 | Low | Joan Massague | Joan Massague lab plasmids (unpublished) | https://media.addgene.org/snapgene-media/v2.0.0/sequences/364716/5f2148c6-2c7f-4440-9d3f-a19c834a34f4/addgene-plasmid-19158-sequence-364716.gbk | full | LOCUS sequence-364716- 6458 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 6458)
TITLE Joan Massague lab plasmids
REFERENCE 2 (bases 1 to 6458)
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..6458
/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"
CDS 763..789
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/note="This is a variant of the standard HA tag, which has
sequence YPYDVPDYA. Some anti-HA antibodies may not
recognize this variant."
/translation="YDVPDYASL"
CDS 1967..1978
/codon_start=1
/product="WELQut protease recognition and cleavage site"
/label=WELQut site
/translation="WELQ"
polyA_signal 2483..3105
/label=hGH poly(A) signal
/note="human growth hormone polyadenylation signal"
primer_bind complement(2682..2701)
/label=hGH-PA-R
/note="Human growth hormone terminator, reverse primer"
primer_bind complement(3129..3149)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 3134..3463
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3314..3449
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(3501..3520)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(3502..3520)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(3534..3550)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3534..3550)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3547..3569)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3558..3574
/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(3582..3612)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3627..3648
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(3765..3782)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3936..4524)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4016..4035)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4695..5555)
/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 5318..5337
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5556..5660)
/gene="bla"
/label=AmpR promoter
rep_origin 5687..6141
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5774..5793)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 5984..6005
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 6267..6289
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 6281..6298
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 6282..6298
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
ORIGIN
1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
601 agaattcaga tctggtacca cgcgtatcga taagcttgca tgcctgcagg tcgagtcggt
661 ctatgacgag cagcggggtc tgccatgggt cgggggctgc tcaggggcct gtggccgctg
721 cacatcgtcc tgtggacgcg tatcgccagc acgatcccac cgtatgatgt tcctgattat
781 gctagcctcc acgttcagaa gtcggttaat aacgacatga tagtcactga caacaacggt
841 gcagtcaagt ttccacaact gtgtaaattt tgtgatgtga gattttccac ctgtgacaac
901 cagaaatcct gcatgagcaa ctgcagcatc acctccatct gtgagaagcc acaggaagtc
961 tgtgtggctg tatggagaaa gaatgacgag aacataacac tagagacagt ttgccatgac
1021 cccaagctcc cctaccatga ctttattctg gaagatgctg cttctccaaa gtgcattatg
1081 aaggaaaaaa aaaagcctgg tgagactttc ttcatgtgtt cctgtagctc tgatgagtgc
1141 aatgacaaca tcatcttctc agaagaatat aacaccagca atcctgactt gttgctagtc
1201 atatttcaag tgacaggcat cagcctcctg ccaccactgg gagttgccat atctgtcatc
1261 atcatcttct actgctaccg cgttaaccgg cagcagaagc tgagttcaac ctgggaaacc
1321 ggcaagacgc ggaagctcat ggagttcagc gagcactgtg ccatcatcct ggaagatgac
1381 cgctctgaca tcagctccac gtgtgccaac aacatcaacc acaacacaga gctgctgccc
1441 attgagctgg acaccctggt ggggaaaggt cgctttgctg aggtctataa ggccaagctg
1501 aagcagaaca cttcagagca gtttgagaca gtggcagtca agatctttcc ctatgaggag
1561 tatgcctctt ggaagacaga gaaggacatc ttctcagaca tcaatctgaa gcatgagaac
1621 atactccagt tcctgacggc tgaggagcgg aagacggagt tggggaaaca atactggctg
1681 atcaccgcct tccacgccaa gggcaaccta caggagtacc tgacgcggca tgtcatcagc
1741 tgggaggacc tgcgcaagct gggcagctcc ctcgcccggg ggattgctca cctccacagt
1801 gatcacactc catgtgggag gcccaagatg cccatcgtgc acagggacct caagagctcc
1861 aatatcctcg tgaagaacga cctaacctgc tgcctgtgtg actttgggct ttccctgcgt
1921 ctggacccta ctctgtctgt ggatgacctg gctaacagtg ggcaggtggg aactgcaaga
1981 tacatggctc cagaagtcct agaatccagg atgaatttgg agaatgttga gtccttcaag
2041 cagaccgatg tctactccat ggctctggtg ctctgggaaa tgacatctcg ctgtaatgca
2101 gtgggagaag taaaagatta tgagcctcca tttggttcca aggtgcggga gcacccctgt
2161 gtcgaaagca tgaaggacaa cgtgttgaga gatcgagggc gaccagaaat tcccagcttc
2221 tggctcaacc accagggcat ccagatggtg tgtgagacgt tgactgagtg ctgggaccac
2281 gacccagagg cccgtctcac agcccagtgt gtggcagaac gcttcagtga gctggagcat
2341 ctggacaggc tctcggggag gagctgctcg gaggagaaga ttcctgaaga cggctcccta
2401 aacactacca aatagctctt ctggggcagg ctgggccatg tccaaagagg ctgccccgaa
2461 ttcctgcagc ccgggggatc ccgggtggca tccctgtgac ccctccccag tgcctctcct
2521 ggccctggaa gttgccactc cagtgcccac cagccttgtc ctaataaaat taagttgcat
2581 cattttgtct gactaggtgt ccttctataa tattatgggg tggagggggg tggtatggag
2641 caaggggcaa gttgggaaga caacctgtag ggcctgcggg gtctattggg aaccaagctg
2701 gagtgcagtg gcacaatctt ggctcactgc aatctccgcc tcctgggttc aagcgattct
2761 cctgcctcag cctcccgagt tgttgggatt ccaggcatgc atgaccaggc tcagctaatt
2821 tttgtttttt tggtagagac ggggtttcac catattggcc aggctggtct ccaactccta
2881 atctcaggtg atctacccac cttggcctcc caaattgctg ggattacagg cgtgaaccac
2941 tgctcccttc cctgtccttc tgattttaaa ataactatac cagcaggagg acgtccagac
3001 acagcatagg ctacctggcc atgcccaacc ggtgggacat ttgagttgct tgcttggcac
3061 tgtcctctca tgcgttgggt ccactcagta gatgcctgtt gaattgggta cgcggccagc
3121 ttggctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
3181 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
3241 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
3301 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
3361 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
3421 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc tcgaggaact
3481 gaaaaaccag aaagttaatt ccctatagtg agtcgtatta aattcgtaat catgtcatag
3541 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
3601 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
3661 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
3721 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
3781 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
3841 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
3901 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
3961 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
4021 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
4081 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
4141 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
4201 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
4261 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
4321 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
4381 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
4441 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
4501 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
4561 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
4621 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
4681 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
4741 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
4801 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
4861 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
4921 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
4981 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
5041 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
5101 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
5161 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
5221 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
5281 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
5341 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
5401 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
5461 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
5521 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
5581 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
5641 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgc gccctgtagc
5701 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
5761 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
5821 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
5881 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
5941 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
6001 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
6061 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaatttaaca
6121 aaatattaac gcttacaatt tccattcgcc attcaggctg cgcaactgtt gggaagggcg
6181 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
6241 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc
6301 caagctgatc tatacattga atcaatattg gcaattagcc atattagtca ttggttatat
6361 agcataaatc aatattggct attggccatt gcatacgttg tatctatatc ataatatgta
6421 catttatatt ggctcatgtc caatatgacc gccatgtt
//
| {
"Vector backbone": "pCMV5",
"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": "",
"Publication": "",
"DOI": "",
"Authors": "",
"PubMed Link": "",
"PubMed ID": ""
} | {
"Gene/Insert name": "Transforming growth factor, beta receptor II",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "PCR based mutagenesis",
"Entrez Gene": "TGFBR2",
"Promoter": "",
"Tag / Fusion Protein": "HA (YDVPDYASL) (N terminal on insert)"
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "SalI (destroyed during cloning)",
"3′ cloning site": "BamHI (not destroyed)",
"5′ sequencing primer": "CMV Forward",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} | {
"Gene/Insert name": "",
"Species": "",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "",
"Promoter": "",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "",
"5′ cloning site": "",
"3′ cloning site": "",
"5′ sequencing primer": "",
"3′ sequencing primer": ""
} |
|
pcDNA5/FRT/TO_HaloTag9_SKL | 175542 | Peroxysomal expression of HaloTag9 in mammalian cells | Kai Johnsson | Frei et al Nat Methods. 2021 Dec 16. pii: 10.1038/s41592-021-01341-x. doi: 10.1038/s41592-021-01341-x. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/343742/2bef2c96-28ab-4e84-bf01-23a46e043162/addgene-plasmid-175542-sequence-343742.gbk | full | LOCUS sequence-343742- 6070 bp DNA circular SYN 26-AUG-2024
DEFINITION Peroxysomal expression of HaloTag9 in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6070)
AUTHORS Frei MS, Tarnawski M, Roberti MJ, Koch B, Hiblot J, Johnsson K
TITLE Engineered HaloTag variants for fluorescence lifetime multiplexing.
JOURNAL Nat Methods. 2021 Dec 16. pii: 10.1038/s41592-021-01341-x. doi:
10.1038/s41592-021-01341-x.
PUBMED 34916672
REFERENCE 2 (bases 1 to 6070)
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. 2021 Dec 16. pii: 10.1038/s41592-021-01341-x. doi:
10.1038/s41592-021-01341-x."
FEATURES Location/Qualifiers
source 1..6070
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(62..81)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 253..632
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 633..836
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 787..807
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
protein_bind 838..856
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 859..877
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
primer_bind 887..911
/label=LNCX
/note="Human CMV promoter, forward primer"
CDS 1070..1954
/codon_start=1
/product="modified bacterial dehalogenase that forms
covalent bonds with chloroalkane derivatives"
/label=HaloTag(R)
/translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDH
NVFIEGTLRMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
SEIARWLSTLEISG"
primer_bind complement(2040..2057)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2046..2270
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(2403..2422)
/label=F1ori-R
/note="F1 origin, reverse primer"
protein_bind 2554..2601
/label=FRT
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
CDS 2609..3631
/codon_start=1
/product="hygromycin B phosphotransferase"
/label=HygR
/note="confers resistance to hygromycin"
/translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
"
CDS 2609..3631
/codon_start=1
/gene="aph(4)-Ia"
/product="aminoglycoside phosphotransferase from E. coli"
/label=HygR
/note="confers resistance to hygromycin"
/translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
"
polyA_signal 3761..3882
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3798..3817)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3852..3871
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(3931..3947)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3931..3947)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3944..3966)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3955..3971
/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(3979..4009)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4024..4045
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4162..4179)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4333..4921)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4413..4432)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5092..5952)
/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 5715..5734
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5953..6057)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca
61 gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg
121 aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca
181 attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca
241 gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat
301 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
361 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
421 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
481 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
541 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
601 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg
661 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
721 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc
781 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctctcc
841 ctatcagtga tagagatctc cctatcagtg atagagatcg tcgacgagct cgtttagtga
901 accgtcagat cgcctggaga cgccatccac gctgttttga cctccataga agacaccggg
961 accgatccag cctccggact ctagcgttta aacttaagct tggtaccgag ctcggatcca
1021 ctagtccagt gtggtggaat tctgcagata tccagcaacc atgggatccg aaatcggtac
1081 tggctttcca ttcgaccccc attatgtgga agtcctgggc gagcgcatgc actacgtcga
1141 tgttggtccg cgcgatggca cccctgtgct gttcctgcac ggtaacccga cctcctccta
1201 cgtgtggcgc aacatcatcc cgcatgttgc accgacccat cgctgcattg ctccagacct
1261 gatcggtatg ggcaaatccg acaaaccaga cctgggttat ttcttcgacg accacgtccg
1321 cttcatggat gccttcatcg aagccctggg tctggaagag gtcgtcctgg tcattcacga
1381 ctggggctcc gctctgggtt tccactgggc caagcgcaat ccagagcgcg tcaaaggtat
1441 tgcatttatg gagttcatcc gccctatccc gacctgggac gaatggccag aatttgcccg
1501 cgagaccttc caggccttcc gcaccaccga cgtcggccgc aagctgatca tcgatcataa
1561 cgtttttatc gagggtacgc tgcgtatggg tgtcgtccgc ccgctgactg aagtcgagat
1621 ggaccattac cgcgagccgt tcctgaatcc tgttgaccgc gagccactgt ggcgcttccc
1681 aaacgagctg ccaatcgccg gtgagccagc gaacatcgtc gcgctggtcg aagaatacat
1741 ggactggctg caccagtccc ctgtcccgaa gctgctgttc tggggcaccc caggcgttct
1801 gatcccaccg gccgaagccg ctcgcctggc caaaagcctg cctaactgca aggctgtgga
1861 catcggcccg ggtctgaatc tgctgcaaga agacaacccg gacctgatcg gcagcgagat
1921 cgcgcgctgg ctgtctactc tggagatttc cggtggcgct ccaggattct cctcaatttc
1981 cgcttccaaa ctgtagtaac cgctcgagtc tagagggccc gtttaaaccc gctgatcagc
2041 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
2101 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
2161 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
2221 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
2281 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
2341 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
2401 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
2461 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
2521 aaaacttgat tagggtgatg gttcacgtac ctagaagttc ctattccgaa gttcctattc
2581 tctagaaagt ataggaactt ccttggccaa aaagcctgaa ctcaccgcga cgtctgtcga
2641 gaagtttctg atcgaaaagt tcgacagcgt ctccgacctg atgcagctct cggagggcga
2701 agaatctcgt gctttcagct tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag
2761 ctgcgccgat ggtttctaca aagatcgtta tgtttatcgg cactttgcat cggccgcgct
2821 cccgattccg gaagtgcttg acattgggga attcagcgag agcctgacct attgcatctc
2881 ccgccgtgca cagggtgtca cgttgcaaga cctgcctgaa accgaactgc ccgctgttct
2941 gcagccggtc gcggaggcca tggatgcgat cgctgcggcc gatcttagcc agacgagcgg
3001 gttcggccca ttcggaccgc aaggaatcgg tcaatacact acatggcgtg atttcatatg
3061 cgcgattgct gatccccatg tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc
3121 gtccgtcgcg caggctctcg atgagctgat gctttgggcc gaggactgcc ccgaagtccg
3181 gcacctcgtg cacgcggatt tcggctccaa caatgtcctg acggacaatg gccgcataac
3241 agcggtcatt gactggagcg aggcgatgtt cggggattcc caatacgagg tcgccaacat
3301 cttcttctgg aggccgtggt tggcttgtat ggagcagcag acgcgctact tcgagcggag
3361 gcatccggag cttgcaggat cgccgcggct ccgggcgtat atgctccgca ttggtcttga
3421 ccaactctat cagagcttgg ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg
3481 atgcgacgca atcgtccgat ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag
3541 aagcgcggcc gtctggaccg atggctgtgt agaagtactc gccgatagtg gaaaccgacg
3601 ccccagcact cgtccgaggg caaaggaata gcacgtacta cgagatttcg attccaccgc
3661 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
3721 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
3781 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
3841 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
3901 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
3961 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
4021 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
4081 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
4141 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
4201 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
4261 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
4321 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
4381 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
4441 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
4501 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
4561 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
4621 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
4681 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
4741 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
4801 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
4861 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
4921 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
4981 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
5041 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
5101 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
5161 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
5221 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
5281 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
5341 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
5401 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
5461 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
5521 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
5581 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
5641 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
5701 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
5761 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
5821 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
5881 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
5941 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
6001 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
6061 atttccccga
//
| {
"Vector backbone": "pCDNA5/FR/TO",
"Backbone size w/o insert (bp)": "",
"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 ; HaloTag Vector Limited Use Label License",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Engineered HaloTag variants for fluorescence lifetime multiplexing",
"Publication": "Nat Methods. 2021 Dec 16. pii: 10.1038/s41592-021-01341-x. doi: 10.1038/s41592-021-01341-x.",
"DOI": "10.1038/s41592-021-01341-x.",
"Authors": "Frei MS, Tarnawski M, Roberti MJ, Koch B, Hiblot J, Johnsson K /s---x",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/34916672",
"PubMed ID": "PubMed 34916672"
} | {
"Gene/Insert name": "HaloTag9-SKL",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "HaloTag9 = HaloTag7-Q165H-P174R",
"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": ""
} |
|
FYN gRNA (BRDN0001147012) | 77242 | 3rd generation lentiviral gRNA plasmid targeting human FYN | Doench et al Nat Biotechnol. 2016 Jan 18. doi: 10.1038/nbt.3437. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/187386/d4050f27-875d-4bb7-88cf-175f6cbc4c42/addgene-plasmid-77242-sequence-187386.gbk | full | LOCUS sequence-187386- 8323 bp DNA circular SYN 26-AUG-2024
DEFINITION 3rd generation lentiviral gRNA plasmid targeting human FYN.
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 cgttgtcctt tggaaaccca 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": "FYN",
"Species": "H. sapiens (human)",
"Insert Size (bp)": "",
"Mutation": "",
"Entrez Gene": "FYN",
"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": ""
} |
||
TAL3495 | 47214 | Keith Joung | Sander et al Nat Biotechnol. 2011 Aug 5;29(8):697-8. doi: 10.1038/nbt.1934. | https://media.addgene.org/snapgene-media/v2.0.0/sequences/69174/64f16458-27ea-4b7d-b05d-90a3b94a0a69/addgene-plasmid-47214-sequence-69174.gbk | partial | LOCUS sequence-69174-e 938 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 938)
AUTHORS Sander JD, Cade L, Khayter C, Reyon D, Peterson RT, Joung JK, Yeh JR
TITLE Targeted gene disruption in somatic zebrafish cells using engineered
TALENs.
JOURNAL Nat Biotechnol. 2011 Aug 5;29(8):697-8. doi: 10.1038/nbt.1934.
PUBMED 21822241
REFERENCE 2 (bases 1 to 938)
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";
journalName: "Nat Biotechnol"; date: "2011-08-5- 5"; volume: "29";
issue: "8"; pages: "697-8"
FEATURES Location/Qualifiers
source 1..938
/mol_type="genomic DNA"
/organism="unspecified"
ORIGIN
1 ggcgggtgag cttagggggc ctccgctcca gctcgacacc gggcagctgc tgaagatcgc
61 gaagagaggg ggagtaacag cggtagaggc agtgcacgcc tggcgcaatg cgctcaccgg
121 ggcccccttg aacctgaccc cagaccaggt agtcgcaatc gcgaacaata atgggggaaa
181 gcaagccctg gaaaccgtgc aaaggttgtt gccggtcctt tgtcaagacc acggccttac
241 accggagcaa gtcgtggcca ttgcatccca cgacggtggc aaacaggctc ttgagacggt
301 tcagagactt ctcccagttc tctgtcaagc ccacgggctg actcccgatc aagttgtagc
361 gattgcgaat aacaatggag ggaaacaagc attggagact gtccaacggc tccttcccgt
421 gttgtgtcaa gcccacggtt tgacgcctgc acaagtggtc gccatcgcct ccaatattgg
481 cggtaagcag gcgctggaaa cagtacagcg cctgctgcct gtactgtgcc aggatcatgg
541 actgacccca gaccaggtag tcgcaatcgc gtcacatgac gggggaaagc aagccctgga
601 aaccgtgcaa aggttgttgc cggtcctttg tcaagaccac ggccttacac cggagcaagt
661 cgtggccatt gcaaataata acggtggcaa acaggctctt gagacggttc agagacttct
721 cccagttctc tgtcaagccc acgggctgac tcccgatcaa gttgtagcga ttgcgtcgca
781 tgacggaggg aaacaagcat tggagactgt ccaacggctc cttcccgtgt tgtgtcaagc
841 ccacggtttg acgcctgcac aagtggtcgc catcgccaac aacaacggcg gtaagcaggc
901 gctggaaaca gtacagcgcc tgctgcctgt actgtgcc
//
| {
"Vector backbone": "JDS71",
"Backbone size w/o insert (bp)": "6447",
"Vector type": "Mammalian Expression ; T7",
"Selectable markers": ""
} | {
"Bacterial Resistance(s)": "Ampicillin, 100 μg/mL",
"Growth Temperature": "37°C",
"Growth Strain(s)": "XL1 Blue",
"Copy number": "High Copy"
} | {
"Academic/Nonprofit Terms": "UBMTA",
"Industry Terms": "Not Available to Industry"
} | {
"Title": "Targeted gene disruption in somatic zebrafish cells using engineered TALENs",
"Publication": "Nat Biotechnol. 2011 Aug 5;29(8):697-8. doi: 10.1038/nbt.1934.",
"DOI": "10.1038/nbt.1934.",
"Authors": "Sander JD, Cade L, Khayter C, Reyon D, Peterson RT, Joung JK, Yeh JR /nbt",
"PubMed Link": "https://www.ncbi.nlm.nih.gov/pubmed/21822241",
"PubMed ID": "PubMed 21822241"
} | {
"Gene/Insert name": "ZebrafishCommunity-mtdhb-right",
"Species": "D. rerio (zebrafish)",
"Insert Size (bp)": "1430",
"Mutation": "",
"Entrez Gene": "mtdhb",
"Promoter": "CMV",
"Tag / Fusion Protein": ""
} | {
"Cloning method": "Restriction Enzyme",
"5′ cloning site": "BsmBI (destroyed during cloning)",
"3′ cloning site": "BsmBI (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": ""
} |