LOCUS Exported 6303 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION AAV vector to drive the expression of dTomato in VIP cortical interneurons under the control of the E6 regulatory element. ACCESSION . VERSION . KEYWORDS pAAV-S5E6-dTom-nlsdTom SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6303) AUTHORS Vormstein-Schneider DC, Lin JD, Pelkey KA, Chittajallu R, Guo B, Arias Garcia M, Sakopoulos S, Stevenson O, Schneider G, Zhang Q, Sharma J, Franken TP, Smith J, Vogel I, Sanchez V, Ibrahim LA, Burbridge T, Favuzzi E, Saldi GA, Xu Q, Guo L, Yuan X, Zaghloul KA, Sabri E, Goldberg EM, Devinsky O, Batista-Brito R, Reynolds J, Feng G, Fu Z, McBain CJ, Fishell GJ, Dimidschstein J TITLE Viral manipulation of functionally distinct interneurons in mice, non-human primates and humans JOURNAL Nature Neuroscience (2020) REFERENCE 2 (bases 1 to 6303) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6303 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..130 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" CDS complement(255..266) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" RBS 677..688 /note="strong bacterial ribosome binding site (Elowitz and Leibler, 2000)" intron 922..1054 /label=chimeric intron /note="chimera between introns from human beta-globin and immunoglobulin heavy chain genes" regulatory 1077..1086 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1083..1784 /codon_start=1 /product="dimeric variant of DsRed fluorescent protein (Shaner et al., 2004)" /label=dTomato /note="mammalian codon-optimized" /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH HLFLYGMDELYK" CDS 1806..1862 /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 1875..1895 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 1896..2594 /codon_start=1 /product="dimeric variant of DsRed fluorescent protein (Shaner et al., 2004)" /label=dTomato /note="mammalian codon-optimized" /translation="VSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTAK LKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGL VTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIHQ ALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRHH LFLYGMDELYK" misc_feature 2649..3237 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2702..2722) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3120..3131) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 3406..3487 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3443..3462) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3497..3516 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" repeat_region 3566..3706 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 3566..3695 /label=AAV2 ITR rep_origin 3781..4236 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3868..3887) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4078..4099 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4253..4272) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4372..4394 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4432..4450) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4518..4622 /gene="bla" /label=AmpR promoter CDS 4623..5483 /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(4841..4860) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5654..6242 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6143..6162 /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 gcgtttaatt aaactagtgg tttttgtcac tttgttactc 181 tacagtgttg cctggagttc gatacttcat tactctatag tggggtgaag aagttcacct 241 tcttattttc atttccttcc ctcaatgatt tcttcagagc tagctcttac cagctagaaa 301 ttcttcaaac gacactcgtg ccttccttca cacaggttga actatttgtc tctaatgccc 361 taaagtactg gtgttcaatc ttccaggcac ttccaatgat ctgaaatctg acctgcttag 421 gtcagctggc tctgagatta tggtattcta gtcctcaaac caacctgttg gctcgttggt 481 tttgtaccaa acacactgac ttacatagct caaaatacca ctggcctttt aaaaatggca 541 tatcacattc caggggagga tcaaaactgc tggctggtga tatttgtcaa gtctctcaaa 601 gttgcacttt ccaggatttt caattcactg aattcttaga cagacatgtt tatgtgaaag 661 aattctttat atattttttc tcctctttga gtgggcaaat gaaaatcttg acctctgggt 721 tccttatttt atttgactct ctgtagtatt taaatcttaa aattttccta aacttcctgc 781 agcccgggct gggcataaaa gtcagggcag agccatctat tgcttacatt tgcttctagc 841 ctgcaagtcg aggagcgcag ccttccagaa gcagagcgcg gcgccttaag ctgcagaagt 901 tggtcgtgag gcactgggca cgtaagtatc aaggttacaa gacaggttta aggagaccaa 961 tagaaactgg gcttgtcgag acagagaaga ctcttgcgtt tctgataggc acctattggt 1021 cttactgaca tccactttgc ctttctctcc acagcgagct cgctagcgcg atcgctgcca 1081 ccatggtgag caagggcgag gaggtcatca aagagttcat gcgcttcaag gtgcgcatgg 1141 agggctccat gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg 1201 agggcaccca gaccgccaag ctgaaggtga ccaagggcgg ccccctgccc ttcgcctggg 1261 acatcctgtc cccccagttc atgtacggct ccaaggcgta cgtgaagcac cccgccgaca 1321 tccccgatta caagaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact 1381 tcgaggacgg cggtctggtg accgtgaccc aggactcctc cctgcaggac ggcacgctga 1441 tctacaaggt gaagatgcgc ggcaccaact tcccccccga cggccccgta atgcagaaga 1501 agaccatggg ctgggaggcc tccaccgagc gcctgtaccc ccgcgacggc gtgctgaagg 1561 gcgagatcca ccaggccctg aagctgaagg acggcggcca ctacctggtg gagttcaaga 1621 ccatctacat ggccaagaag cccgtgcaac tgcccggcta ctactacgtg gacaccaagc 1681 tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgag cgctccgagg 1741 gccgccacca cctgttcctg tacggcatgg acgagctgta caagtccgga gataacggaa 1801 gcggagccac taacttctcc ctgttgaaac aagcagggga tgtcgaagag aatcccgggc 1861 catctagaat ggcccccaag aagaagagga aggtggtgag caagggcgag gaggtcatca 1921 aagagttcat gcgcttcaag gtgcgcatgg agggctccat gaacggccac gagttcgaga 1981 tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga 2041 ccaagggcgg ccccctgccc ttcgcctggg acatcctgtc cccccagttc atgtacggct 2101 ccaaggcgta cgtgaagcac cccgccgaca tccccgatta caagaagctg tccttccccg 2161 agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggtctggtg accgtgaccc 2221 aggactcctc cctgcaggac ggcacgctga tctacaaggt gaagatgcgc ggcaccaact 2281 tcccccccga cggccccgta atgcagaaga agaccatggg ctgggaggcc tccaccgagc 2341 gcctgtaccc ccgcgacggc gtgctgaagg gcgagatcca ccaggccctg aagctgaagg 2401 acggcggcca ctacctggtg gagttcaaga ccatctacat ggccaagaag cccgtgcaac 2461 tgcccggcta ctactacgtg gacaccaagc tggacatcac ctcccacaac gaggactaca 2521 ccatcgtgga acagtacgag cgctccgagg gccgccacca cctgttcctg tacggcatgg 2581 acgagctgta caagtaagtc gacggcgcgc cccacccctg cagggaattc gatatcaagc 2641 ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact 2701 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg 2761 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg 2821 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa 2881 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc 2941 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg 3001 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt 3061 ggctgctcgc ctatgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt 3121 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc 3181 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc 3241 gataccgagc gctgctcgag agatctgtga tagcggccat caagctggcc gcgactctag 3301 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac 3361 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca 3421 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt 3481 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcagctta tcgataccgc 3541 atgcacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct 3601 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt 3661 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg 3721 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt 3781 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg 3841 ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca 3901 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta 3961 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc 4021 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg 4081 gactcttgtt ccaaactgga acaacactca actctatctc gggctattct tttgatttat 4141 aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta 4201 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct 4261 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct 4321 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct 4381 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga 4441 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca 4501 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata 4561 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga 4621 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc 4681 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg 4741 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc 4801 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat 4861 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact 4921 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat 4981 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga 5041 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc 5101 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga 5161 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag 5221 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc 5281 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt 5341 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct 5401 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg 5461 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg 5521 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca 5581 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga 5641 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa 5701 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga 5761 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt 5821 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt 5881 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat 5941 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct 6001 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca 6061 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag 6121 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc 6181 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga 6241 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca 6301 tgt //