LOCUS Exported 7443 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION AAV production. ACCESSION . VERSION . KEYWORDS pAAV-EF1a-iDreV SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7443) AUTHORS Yao S, Yuan P, Ouellette B, Zhou T, Mortrud M, Balaram P, Chatterjee S, Wang Y, Daigle TL, Tasic B, Kuang X, Gong H, Luo Q, Zeng S, Curtright A, Dhaka A, Kahan A, Gradinaru V, Chrapkiewicz R, Schnitzer M, Zeng H, Cetin A TITLE RecV recombinase system for in vivo targeted optogenomic modifications of single cells or cell populations. JOURNAL Nat Methods. 2020 Mar 23. pii: 10.1038/s41592-020-0774-3. doi: 10.1038/s41592-020-0774-3. PUBMED 32203389 REFERENCE 2 (bases 1 to 7443) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7443 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..130 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" promoter 231..1409 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 462..1400 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 1357..1377 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" regulatory 1428..1437 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1440..1460 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 2112..2132 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 2148..2204 /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 2214..2234 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" misc_feature 3570..4158 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3623..3643) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(4041..4052) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 4190..4666 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(4389..4408) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 4706..4846 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 4706..4835 /label=AAV2 ITR rep_origin 4921..5376 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5008..5027) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5218..5239 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(5393..5412) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 5512..5534 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(5572..5590) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5658..5762 /gene="bla" /label=AmpR promoter CDS 5763..6623 /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(5981..6000) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6794..7382 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7283..7302 /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 gcgtaagctt tgcaaagatg gataaagttt taaacagaga 181 ggaatctttg cagctaatgg accttctagg tcttgaaagg agtgggaatt ggctccggtg 241 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg 301 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt 361 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg 421 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt 481 cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct 541 ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga 601 ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct 661 ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt 721 ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc 781 ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg 841 acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac 901 cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc 961 cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag 1021 cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct 1081 cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg 1141 tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga 1201 gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc 1261 acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg 1321 aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa 1381 gtttttttct tccatttcag gtgtcgtgag gtaccggatc ttaagccgcc accatggccc 1441 ctaagaagaa gaggaaggtt tctgagctga ttattagtgg ttcatctggt ggattcctgc 1501 gaaacatcgg caaagagtat caggaggccg ctgaaaactt catgaggttt atgaatgacc 1561 agggggcgta cgctcctaac actttgaggg atttgaggtt ggtctttcat agctgggcca 1621 gatggtgcca tgctcggggt acccatacac tgtatgcccc tggaggatat gatattatgg 1681 gatatctgat tcagattatg aaaagaccta atcctcaggt ggaactggga cctgtggata 1741 caagcgtggc cctgattctg tgtgatctga aacagaaaga tacacctatt gtgtatgcca 1801 gcgaagcctt tctgtatatg acaggatata gcaatgccga agtgctggga agaaattgta 1861 gatttctgca gagccctgat ggaatggtga aacctaaaag cacaagaaaa tatgtggata 1921 gcaatacaat taatacaatg agaaaagcca ttgatagaaa tgccgaagtg caggtggaag 1981 tggtgaattt taaaaaaaat ggacagagat ttgtgaattt tctgacaatg attcctgtga 2041 gagatgaaac aggagaatat agatatagca tgggatttca gtgtgaaaca gaaggcggaa 2101 gcggtggcgt gcccaagaag aagaggaaag tcggatccgg cagcggcgcc accaacttca 2161 gcctgctgaa gcaggccggc gacgtggagg agaaccccgg ccccctcgag gtgcccaaga 2221 agaagaggaa agtcggcgga catactctgt acgctcccgg agggtatgac ataatgggtt 2281 acctaatcca gattatgaag cgtcctaatc cccaggttga gttgggacca gtggacacca 2341 gcgtagcctt aattctttgc gaccttaaac agaaagatac accaatagta tacgcgagtg 2401 aggctttttt gtatatgacg ggttattcca acgcggaagt gttaggacga aattgtcggt 2461 ttctacaatc accggatggg atggtcaaac cgaaaagtac tagaaagtat gtggattcga 2521 atacaattaa caccatgaga aaggcaatcg ataggaacgc agaagtacaa gtcgaggttg 2581 tcaatttcaa aaagaatggg caacgcttcg taaactttct cactatgata cctgttaggg 2641 acgaaacggg cgagtatcga tactctatgg gcttccaatg tgaaacagaa ggtacccagc 2701 ttgcatggtt tccaattagt cctgaaatgg cacgcgaata ctttcttcag ttgcacgatg 2761 cagacctggc ctccactacc atcgacaagc actatgctat gcttaatatg cttctgtccc 2821 actgcggact gccacccttg tccgacgaca agtcagtgag tcttgccatg agaagaatta 2881 gaagagaagc cgcaaccgaa aagggtgaga ggacaggaca ggcaatcccc ctgcgctggg 2941 acgacctgaa gctgctggat gtgctgctca gcaggagcga gcggctggtc gacctgcgca 3001 acagggcttt cctgttcgta gcctataaca ccctcatgag aatgtctgaa atatcacgca 3061 tcagggttgg ggacttggat cagacaggag acacagtgac cctgcacatc agtcacacta 3121 agacaatcac cacagctgcg ggccttgaca aagtgctctc ccggcgaacc acagcagtgc 3181 tcaatgactg gctggacgtc agtgggctta gagaacatcc agacgctgtg ctcttcccac 3241 ctatacaccg gtcaaacaaa gcccgcatta ctaccacgcc cctgaccgcc cctgccatgg 3301 agaagatttt cagtgatgcc tgggtgctgc tgaacaaacg ggacgccacc cccaataaag 3361 ggaggtatag gacctggacc ggccattccg ccagggtggg tgccgcaata gacatggccg 3421 agaaacaggt gtctatggtc gagattatgc aggaagggac atggaagaag cctgaaacac 3481 tgatgcggta tctcagaagg ggcggagtgt ccgtgggagc caattctcga ctgatggata 3541 gctaagaatt cgatatcaag cttatcgata atcaacctct ggattacaaa atttgtgaaa 3601 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa 3661 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat 3721 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt 3781 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc 3841 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc 3901 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg 3961 ggaaatcatc gtcctttcct tggctgctcg cctatgttgc cacctggatt ctgcgcggga 4021 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc 4081 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc 4141 tttgggccgc ctccccgcat cgataccgag cgctgctcga gagatctacg ggtggcatcc 4201 ctgtgacccc tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag 4261 ccttgtccta ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat 4321 tatggggtgg aggggggtgg tatggagcaa ggggcaagtt gggaagacaa cctgtagggc 4381 ctgcggggtc tattgggaac caagctggag tgcagtggca caatcttggc tcactgcaat 4441 ctccgcctcc tgggttcaag cgattctcct gcctcagcct cccgagttgt tgggattcca 4501 ggcatgcatg accaggctca gctaattttt gtttttttgg tagagacggg gtttcaccat 4561 attggccagg ctggtctcca actcctaatc tcaggtgatc tacccacctt ggcctcccaa 4621 attgctggga ttacaggcgt gaaccactgc tcccttccct gtccttctga ttttgtaggt 4681 aaccacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct 4741 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt 4801 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg 4861 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt 4921 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg 4981 ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca 5041 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta 5101 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc 5161 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg 5221 gactcttgtt ccaaactgga acaacactca actctatctc gggctattct tttgatttat 5281 aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta 5341 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct 5401 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct 5461 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct 5521 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga 5581 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca 5641 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata 5701 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga 5761 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc 5821 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg 5881 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc 5941 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat 6001 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact 6061 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat 6121 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga 6181 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc 6241 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga 6301 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag 6361 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc 6421 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt 6481 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct 6541 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg 6601 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg 6661 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca 6721 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga 6781 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa 6841 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga 6901 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt 6961 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt 7021 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat 7081 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct 7141 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca 7201 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag 7261 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc 7321 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga 7381 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca 7441 tgt //