LOCUS Exported 7261 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION AAV vector to drive the expression of Gq-DREADD-P2A-dTomato in PV cortical interneuronsunder the control of the E2 regulatory element. ACCESSION . VERSION . KEYWORDS pAAV-S5E2-Gq-P2A-dTomato SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7261) 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 7261) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7261 /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 934..1066 /label=chimeric intron /note="chimera between introns from human beta-globin and immunoglobulin heavy chain genes" CDS 1089..1115 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 2904..2960 /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 2973..2993 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 2994..3692 /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" primer_bind complement(3124..3142) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 3392..3411 /label=mCherry-F /note="mCherry, forward primer" primer_bind 3586..3609 /label=DsRed1-C /note="DsRed1, forward primer" misc_feature 3747..4335 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3800..3820) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(4218..4229) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 4364..4445 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4401..4420) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4455..4474 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" repeat_region 4524..4664 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 4524..4653 /label=AAV2 ITR rep_origin 4739..5194 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4826..4845) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5036..5057 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(5211..5230) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 5330..5352 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(5390..5408) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5476..5580 /gene="bla" /label=AmpR promoter CDS 5581..6441 /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(5799..5818) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6612..7200 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7101..7120 /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 tttaatctaa catggctgct 181 atagcttact gactagaagt taagtgcaca cttcctaaaa gaaggctttg acacaagcca 241 cttcagttcc ctcctcattt tcttgtcccc attcctctct ctgtagaatt ctgagatttc 301 aattcagttt tatacagaaa ccacattact gtaagcccta caaagttatg gcaatatagc 361 tatatggagt caagtaatgt aggttatttt tttcccaatg gtgctggtga aggtggcaat 421 tatgtagcta tacttagcag actgaggaaa ttctgctaga gtcagcattt gtctcttcat 481 tgctatgaaa cagtaatgga aaaataaaca aaaacaaaag gcaaacacta tgcataattc 541 cctcagatca tattaacatg tgatgttgga gtaaattgtt ataaccccat tttggaaata 601 cttaccttaa ttaactatga tttccttaaa ataatgcagt atttacaatc tatatgaaag 661 cactatatgg gacacatggt atgatggaac agtgcaccca agagacacca agaacattcc 721 tgtctgtggc agtcttttct ctatacagag gcatttagtc tcaattgctc agagttattt 781 taaacttcct gcagcccggg ctgggcataa aagtcagggc agagccatct attgcttaca 841 tttgcttcta gcctgcaagt cgaggagcgc agccttccag aagcagagcg cggcgcctta 901 agctgcagaa gttggtcgtg aggcactggg cacgtaagta tcaaggttac aagacaggtt 961 taaggagacc aatagaaact gggcttgtcg agacagagaa gactcttgcg tttctgatag 1021 gcacctattg gtcttactga catccacttt gcctttctct ccacagcgag ctcgctagcg 1081 ccaccatgta cccatacgat gttccagatt acgctacctt gcacaataac agtacaacct 1141 cgcctttgtt tccaaacatc agctcctcct ggatacacag cccctccgat gcagggctgc 1201 ccccgggaac cgtcactcat ttcggcagct acaatgtttc tcgagcagct ggcaatttct 1261 cctctccaga cggtaccacc gatgaccctc tgggaggtca taccgtctgg caagtggtct 1321 tcatcgcttt cttaacgggc atcctggcct tggtgaccat catcggcaac atcctggtaa 1381 ttgtgtcatt taaggtcaac aagcagctga agacggtcaa caactacttc ctcttaagcc 1441 tggcctgtgc cgatctgatt atcggggtca tttcaatgaa tctgtttacg acctacatca 1501 tcatgaatcg atgggcctta gggaacttgg cctgtgacct ctggcttgcc attgactgcg 1561 tagccagcaa tgcctctgtt atgaatcttc tggtcatcag ctttgacaga tacttttcca 1621 tcacgaggcc gctcacgtac cgagccaaac gaacaacaaa gagagccggt gtgatgatcg 1681 gtctggcttg ggtcatctcc tttgtccttt gggctcctgc catcttgttc tggcaatact 1741 ttgttggaaa gagaactgtg cctccgggag agtgcttcat tcagttcctc agtgagccca 1801 ccattacttt tggcacagcc atcgctggtt tttatatgcc tgtcaccatt atgactattt 1861 tatactggag gatctataag gaaactgaaa agcgtaccaa agagcttgct ggcctgcaag 1921 cctctgggac agaggcagag acagaaaact ttgtccaccc cacgggcagt tctcgaagct 1981 gcagcagtta cgaacttcaa cagcaaagca tgaaacgctc caacaggagg aagtatggcc 2041 gctgccactt ctggttcaca accaagagct ggaaacccag ctccgagcag atggaccaag 2101 accacagcag cagtgacagt tggaacaaca atgatgctgc tgcctccctg gagaactccg 2161 cctcctccga cgaggaggac attggctccg agacgagagc catctactcc atcgtgctca 2221 agcttccggg tcacagcacc atcctcaact ccaccaagtt accctcatcg gacaacctgc 2281 aggtgcctga ggaggagctg gggatggtgg acttggagag gaaagccgac aagctgcagg 2341 cccagaagag cgtggacgat ggaggcagtt ttccaaaaag cttctccaag cttcccatcc 2401 agctagagtc agccgtggac acagctaaga cttctgacgt caactcctca gtgggtaaga 2461 gcacggccac tctacctctg tccttcaagg aagccactct ggccaagagg tttgctctga 2521 agaccagaag tcagatcact aagcggaaaa ggatgtccct ggtcaaggag aagaaagcgg 2581 cccagaccct cagtgcgatc ttgcttgcct tcatcatcac ttggacccca tacaacatca 2641 tggttctggt gaacaccttt tgtgacagct gcatacccaa aaccttttgg aatctgggct 2701 actggctgtg ctacatcaac agcaccgtga accccgtgtg ctatgctctg tgcaacaaaa 2761 cattcagaac cactttcaag atgctgctgc tgtgccagtg tgacaaaaaa aagaggcgca 2821 agcagcagta ccagcagaga cagtcggtca tttttcacaa gcgcgcaccc gagcaggcct 2881 tgtccggaga taacggaagc ggagccacta acttctccct gttgaaacaa gcaggggatg 2941 tcgaagagaa tcccgggcca tctagaatgg cccccaagaa gaagaggaag gtggtgagca 3001 agggcgagga ggtcatcaaa gagttcatgc gcttcaaggt gcgcatggag ggctccatga 3061 acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga 3121 ccgccaagct gaaggtgacc aagggcggcc ccctgccctt cgcctgggac atcctgtccc 3181 cccagttcat gtacggctcc aaggcgtacg tgaagcaccc cgccgacatc cccgattaca 3241 agaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg 3301 gtctggtgac cgtgacccag gactcctccc tgcaggacgg cacgctgatc tacaaggtga 3361 agatgcgcgg caccaacttc ccccccgacg gccccgtaat gcagaagaag accatgggct 3421 gggaggcctc caccgagcgc ctgtaccccc gcgacggcgt gctgaagggc gagatccacc 3481 aggccctgaa gctgaaggac ggcggccact acctggtgga gttcaagacc atctacatgg 3541 ccaagaagcc cgtgcaactg cccggctact actacgtgga caccaagctg gacatcacct 3601 cccacaacga ggactacacc atcgtggaac agtacgagcg ctccgagggc cgccaccacc 3661 tgttcctgta cggcatggac gagctgtaca agtaagtcga cggcgcgccc cacccctgca 3721 gggaattcga tatcaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat 3781 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc 3841 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct 3901 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca 3961 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt 4021 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg 4081 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga 4141 aatcatcgtc ctttccttgg ctgctcgcct atgttgccac ctggattctg cgcgggacgt 4201 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc 4261 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt 4321 gggccgcctc cccgcatcga taccgagcgc tgctcgagag atcaacttgt ttattgcagc 4381 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc 4441 actgcattct agttgtggtt tgtccaaact catcaatgta tcagcttatc gataccgcat 4501 gcacgtgcgg accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct 4561 gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc 4621 ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta 4681 ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac 4741 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct 4801 acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg 4861 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt 4921 gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca 4981 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga 5041 ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa 5101 gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac 5161 gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc 5221 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga 5281 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc 5341 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata 5401 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact 5461 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg 5521 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt 5581 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct 5641 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca 5701 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc 5761 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc 5821 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg 5881 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta 5941 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc 6001 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt 6061 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg 6121 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct 6181 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc 6241 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct 6301 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac 6361 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc 6421 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat 6481 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg 6541 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc 6601 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa 6661 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag 6721 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta 6781 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta 6841 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag 6901 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg 6961 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg 7021 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag 7081 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc 7141 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa 7201 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg 7261 t //