LOCUS Exported 6205 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Expresses aa173 to C-terminus of VCre recombinase fused to GAI CID. ACCESSION . VERSION . KEYWORDS pBW2562_pCAG-PV1-GAI-L1-VCre-173C-NLS-BGHpA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6205) AUTHORS Weinberg BH, Cho JH, Agarwal Y, Pham NTH, Caraballo LD, Walkosz M, Ortega C, Trexler M, Tague N, Law B, Benman WKJ, Letendre J, Beal J , Wong WW TITLE High-performance chemical- and light-inducible recombinases in mammalian cells and mice JOURNAL Nature Communications (2019) volume 10, Article number: 4845 REFERENCE 2 (bases 1 to 6205) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6205 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(22..38) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(22..38) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(35..57) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 46..62 /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(70..100) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 115..136 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." rep_origin 221..355 /label=SV40 ori /note="SV40 origin of replication" primer_bind 282..301 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" polyA_signal 375..509 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(412..431) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 466..485 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(577..594) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(748..1336) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(828..847) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1507..2367) /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 2130..2149 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(2368..2472) /gene="bla" /label=AmpR promoter enhancer 2503..2882 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 2884..3159 /label=chicken beta-actin promoter intron 3160..4175 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 4099..4121 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 4183..4202 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" CDS 4322..4339 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" regulatory 5142..5151 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 5252..5272 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" polyA_signal 5282..5506 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(5746..5765) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 5811..5866 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(5865..5884) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" ORIGIN 1 cccaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac 61 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 121 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 181 gtgccagcgg atccgcatct caattagtca gcaaccatag tcccgcccct aactccgccc 241 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag 301 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc 361 ttttgcaaaa agctaacttg tttattgcag cttataatgg ttacaaataa agcaatagca 421 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac 481 tcatcaatgt atcttatcat gtctggatcc gctgcattaa tgaatcggcc aacgcgcggg 541 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc 601 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac 661 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa 721 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca 781 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc 841 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata 901 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta 961 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca 1021 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga 1081 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg 1141 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg 1201 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg 1261 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag 1321 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa 1381 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat 1441 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc 1501 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc 1561 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc 1621 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc 1681 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc 1741 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt 1801 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc 1861 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa 1921 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt 1981 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg 2041 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc 2101 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa 2161 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt 2221 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt 2281 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag 2341 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta 2401 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat 2461 aggggttccg cgcacatttc cccgaaaagt gccacctggg tcgacattga ttattgacta 2521 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg 2581 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga 2641 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat 2701 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa 2761 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca 2821 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca 2881 tggtcgaggt gagccccacg ttctgcttca ctctccccat ctcccccccc tccccacccc 2941 caattttgta tttatttatt ttttaattat tttgtgcagc gatgggggcg gggggggggg 3001 gggcgcgcgc caggcggggc ggggcggggc gaggggcggg gcggggcgag gcggaaaggt 3061 gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg 3121 cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc gttgccttcg 3181 ccccgtgccc cgctccgcgc cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta 3241 ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta gcgcttggtt 3301 taatgacggc tcgtttcttt tctgtggctg cgtgaaagcc ttaaagggct ccgggagggc 3361 cctttgtgcg ggggggagcg gctcgggggg tgcgtgcgtg tgtgtgtgcg tggggagcgc 3421 cgcgtgcggc tccgcgctgc ccggcggctg tgagcgctgc gggcgcggcg cggggctttg 3481 tgcgctccgc agtgtgcgcg aggggagcgc ggccgggggc ggtgccccgc ggtgcggggg 3541 gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc gtgggggggt gagcaggggg 3601 tgtgggcgcg gcggtcgggc tgtaaccccc ccctgcaccc ccctccccga gttgctgagc 3661 acggcccggc ttcgggtgcg gggctccgta cggggcgtgg cgcggggctc gccgtgccgg 3721 gcggggggtg gcggcaggtg ggggtgccgg gcggggcggg gccgcctcgg gccggggagg 3781 gctcggggga ggggcgcggc ggcccccgga gcgccggcgg ctgtcgaggc gcggcgagcc 3841 gcagccattg ccttttatgg taatcgtgcg agagggcgca gggacttcct ttgtcccaaa 3901 tctgtgcgga gccgaaatct gggaggcgcc gccgcacccc ctctagcggg cgcggggcga 3961 agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg ccttcgtgcg tcgccgcgcc 4021 gccgtcccct tctccctctc cagcctcggg gctgtccgcg gggggacggc tgccttcggg 4081 ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac cggcggctct agagcctctg 4141 ctaaccatgt tcatgccttc ttctttttcc tacagctcct gggcaacgtg ctggttattg 4201 tgctgtctca tcattttggc aaagaattga ttaattcgag cgaaggcgcg cccattactc 4261 gcatccattc tcaggctgtc tcgtctcgtc tcgcacgcgt gccgccacca tgaagcggga 4321 ccaccaccat caccatcatc aggacaagaa aaccatgatg atgaacgaag aggacgacgg 4381 caacggcatg gacgagctgc tggctgtgct gggctacaaa gtgcggagca gcgagatggc 4441 cgacgtggcc cagaaactgg aacagctgga agtgatgatg agcaacgtgc aggaagatga 4501 cctgtcccag ctggccaccg agacagtgca ctacaacccc gccgagctgt acacctggct 4561 ggactccatg ctgaccgacc tgaactccgg agggtctggc tccggatcaa gtggtggcag 4621 cggtaccgac aagctgctgc tgcggcggaa cctggctctg ctggctgtgg cctacgagag 4681 catgctgaga gccagcgagc tggccaacat ccgggtgtcc gatatggaac tggccggcga 4741 cggaaccgcc atcctgacca tccctatcac caagaccaac cactccggcg agcccgatac 4801 ctgcatcctg tcccaggatg tggtgtccct gctgatggac tacaccgagg ccggcaagct 4861 ggatatgagc agcgacggct tcctgttcgt gggcgtgtcc aagcacaaca cctgtatcaa 4921 gcccaagaag gacaagcaga ccggcgaggt gctgcacaag cccatcacca ccaagacagt 4981 ggaaggcgtg ttctacagcg cctgggagac actggacctg ggcagacagg gcgtgaagcc 5041 tttcacagcc cacagcgcca gagtgggagc cgctcaggac ctgctgaaga agggctacaa 5101 taccctgcag atccagcagt ccggccggtg gtctagcgga gccatggtgg ccagatacgg 5161 cagagccatc ctggctaggg atggcgctat ggcccacagc agagtgaaaa ccagatccgc 5221 ccccatgcag tggggcaagg acgagaagga ccccaagaaa aagcggaagg tgtgagaatt 5281 cctgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc 5341 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt 5401 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat 5461 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa 5521 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcgtgta ccgctgggag 5581 ttcgtagacg gaaacaaacg cagaatccaa gcgcggccgc ccaggataca tagattacca 5641 caactccgag cccttccacc gctagcgcac tcctcaggtg caggctgcct atcagaaggt 5701 ggtggctggt gtggccaatg ccctggctca caaataccac tgagatcttt ttccctctgc 5761 caaaaattat ggggacatca tgaagcccct tgagcatctg acttctggct aataaaggaa 5821 atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata 5881 tgggagggca aatcatttaa aacatcagaa tgagtatttg gtttagagtt tggcaacata 5941 tgcccatatg ctggctgcca tgaacaaagg ttggctataa agaggtcatc agtatatgaa 6001 acagccccct gctgtccatt ccttattcca tagaaaagcc ttgacttgag gttagatttt 6061 ttttatattt tgttttgtgt tatttttttc tttaacatcc ctaaaatttt ccttacatgt 6121 tttactagcc agatttttcc tcctctcctg actactccca gtcatagctg tccctcttct 6181 cttatggaga tccctcgacc tgcag //