LOCUS Exported 6976 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION AAV vector to drive the expression of dTomato in PV cortical interneuronsunder the control of the E1 regulatory element. ACCESSION . VERSION . KEYWORDS pAAV-S5E1-dTom-nlsdTom SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6976) 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 6976) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6976 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..130 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" intron 1595..1727 /label=chimeric intron /note="chimera between introns from human beta-globin and immunoglobulin heavy chain genes" regulatory 1750..1759 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1756..2457 /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 2479..2535 /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 2548..2568 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 2569..3267 /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 3322..3910 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3375..3395) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3793..3804) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 4079..4160 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4116..4135) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4170..4189 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" repeat_region 4239..4379 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 4239..4368 /label=AAV2 ITR rep_origin 4454..4909 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4541..4560) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4751..4772 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4926..4945) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 5045..5067 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(5105..5123) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5191..5295 /gene="bla" /label=AmpR promoter CDS 5296..6156 /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(5514..5533) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6327..6915 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6816..6835 /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 tttcaaagtg gacagagggg 181 aggggagggg atgcgagggg aggggagggg aatgatcgcg aaaggcttcc aaaccttgtc 241 cttgtttttc accatttctg aaatatatgc tgagtgcaac tatgggaaga ccattttcat 301 aatctataat actcctcctt ttaaaggact ttcgttaacc gcttgcaaag gtgagtgccg 361 ggtagaggac attagctcgc taagtcccta gaaatcacac ttggagacta agcaggcttt 421 cccaggagaa gtccaaagcc aacataagca ggaggctggg ggctggccgt taaccgcaag 481 gcagtggttg agccctcggg atcatcccgg cggggggcgc agcatctccg ccaaggccgc 541 aggctctcac catcagctgc ccgagccacc ctgtacctcg cagtccactc gccctgccca 601 cgccccgcgc cgcccgctca ccttcagccc ctgggagtcc atggccgccg gctacccgga 661 gggtgcccac cgctgcccgc cgcagggtta ggggttcaga cccacttccc gggccctcaa 721 accctaaggg acgcggcgtg cagcacgagg ggcgtggccc gatctccatt ggttgtcggc 781 gtgagggggc ggagcttagt tgtaggacta ggaaggaggg ggccaccgga gcaggcgagg 841 agggaacccc gagggaggac cgcgagggcg actggggctg gaatccgctg agcattgagt 901 gctgccgagt tgtggggcta gaggagggag gtccagcctg gaaacggcgc gaggaggagg 961 gattgggtgg agcaagagat atgagattaa agaataaaga tgatgaagca gcaaatagga 1021 gggagagcca tgccgctttt catccctgca aacaaaggcc gactccattt tctcagcatt 1081 ttttgtggaa gccgatttgc gcaatgcggc ttagtacttg accagggaaa atgatttacc 1141 tgacacgtgt agtaatcgtg tctgggccac aaggtggcgc agaaaaatca cagttcggca 1201 aaaaccttga agcctggctt gggcttgttc taaatctttt caggcgctgc tgtaattttg 1261 ctattcgagt gcttattaaa ctgctccgcc agatttccac ccccaaagtc ttatttaaaa 1321 atatgtggtt acctctttta gatttctatt tcttaagtgt ttgctgtagt ttggatctaa 1381 actgtccctc aaagacacac gtgctgaatg ttccccagcc cgtgtgctgt tggagtggtg 1441 gaaaacttcc tgcagcccgg gctgggcata aaagtcaggg cagagccatc tattgcttac 1501 atttgcttct agcctgcaag tcgaggagcg cagccttcca gaagcagagc gcggcgcctt 1561 aagctgcaga agttggtcgt gaggcactgg gcacgtaagt atcaaggtta caagacaggt 1621 ttaaggagac caatagaaac tgggcttgtc gagacagaga agactcttgc gtttctgata 1681 ggcacctatt ggtcttactg acatccactt tgcctttctc tccacagcga gctcgctagc 1741 gcgatcgctg ccaccatggt gagcaagggc gaggaggtca tcaaagagtt catgcgcttc 1801 aaggtgcgca tggagggctc catgaacggc cacgagttcg agatcgaggg cgagggcgag 1861 ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg cggccccctg 1921 cccttcgcct gggacatcct gtccccccag ttcatgtacg gctccaaggc gtacgtgaag 1981 caccccgccg acatccccga ttacaagaag ctgtccttcc ccgagggctt caagtgggag 2041 cgcgtgatga acttcgagga cggcggtctg gtgaccgtga cccaggactc ctccctgcag 2101 gacggcacgc tgatctacaa ggtgaagatg cgcggcacca acttcccccc cgacggcccc 2161 gtaatgcaga agaagaccat gggctgggag gcctccaccg agcgcctgta cccccgcgac 2221 ggcgtgctga agggcgagat ccaccaggcc ctgaagctga aggacggcgg ccactacctg 2281 gtggagttca agaccatcta catggccaag aagcccgtgc aactgcccgg ctactactac 2341 gtggacacca agctggacat cacctcccac aacgaggact acaccatcgt ggaacagtac 2401 gagcgctccg agggccgcca ccacctgttc ctgtacggca tggacgagct gtacaagtcc 2461 ggagataacg gaagcggagc cactaacttc tccctgttga aacaagcagg ggatgtcgaa 2521 gagaatcccg ggccatctag aatggccccc aagaagaaga ggaaggtggt gagcaagggc 2581 gaggaggtca tcaaagagtt catgcgcttc aaggtgcgca tggagggctc catgaacggc 2641 cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc 2701 aagctgaagg tgaccaaggg cggccccctg cccttcgcct gggacatcct gtccccccag 2761 ttcatgtacg gctccaaggc gtacgtgaag caccccgccg acatccccga ttacaagaag 2821 ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggtctg 2881 gtgaccgtga cccaggactc ctccctgcag gacggcacgc tgatctacaa ggtgaagatg 2941 cgcggcacca acttcccccc cgacggcccc gtaatgcaga agaagaccat gggctgggag 3001 gcctccaccg agcgcctgta cccccgcgac ggcgtgctga agggcgagat ccaccaggcc 3061 ctgaagctga aggacggcgg ccactacctg gtggagttca agaccatcta catggccaag 3121 aagcccgtgc aactgcccgg ctactactac gtggacacca agctggacat cacctcccac 3181 aacgaggact acaccatcgt ggaacagtac gagcgctccg agggccgcca ccacctgttc 3241 ctgtacggca tggacgagct gtacaagtaa gtcgacggcg cgccccaccc ctgcagggaa 3301 ttcgatatca agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact 3361 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg 3421 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg 3481 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg 3541 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg 3601 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc 3661 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca 3721 tcgtcctttc cttggctgct cgcctatgtt gccacctgga ttctgcgcgg gacgtccttc 3781 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct 3841 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc 3901 gcctccccgc atcgataccg agcgctgctc gagagatctg tgatagcggc catcaagctg 3961 gccgcgactc tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa 4021 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa 4081 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa 4141 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatcagc 4201 ttatcgatac cgcatgcacg tgcggaccga gcggccgcag gaacccctag tgatggagtt 4261 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg 4321 acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg 4381 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca 4441 aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg 4501 cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct 4561 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta 4621 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt 4681 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg 4741 ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat 4801 tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt 4861 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact 4921 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc 4981 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc 5041 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga 5101 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag 5161 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa 5221 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat 5281 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg 5341 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa 5401 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt 5461 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt 5521 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat 5581 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg 5641 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta 5701 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat 5761 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag 5821 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa 5881 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca 5941 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc 6001 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt 6061 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc 6121 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat 6181 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt 6241 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac 6301 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc 6361 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca 6421 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta 6481 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct 6541 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg 6601 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc 6661 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta 6721 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg 6781 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt 6841 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg 6901 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg 6961 ccttttgctc acatgt //