LOCUS       Exported                6315 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  AAV vector to drive the expression of dTomato in PV cortical 
            interneurons under the control of the E2 regulatory element.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-S5E2-dTom-nlsdTom
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6315)
  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 6315)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6315
                     /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"
     regulatory      1089..1098
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1095..1796
                     /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             1818..1874
                     /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             1887..1907
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1908..2606
                     /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    2661..3249
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2714..2734)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3132..3143)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3418..3499
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3455..3474)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3509..3528
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   3578..3718
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3578..3707
                     /label=AAV2 ITR
     rep_origin      3793..4248
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3880..3899)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4090..4111
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4265..4284)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4384..4406
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4444..4462)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4530..4634
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4635..5495
                     /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(4853..4872)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5666..6254
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6155..6174
                     /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 cgatcgctgc caccatggtg agcaagggcg aggaggtcat caaagagttc atgcgcttca
     1141 aggtgcgcat ggagggctcc atgaacggcc acgagttcga gatcgagggc gagggcgagg
     1201 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggc ggccccctgc
     1261 ccttcgcctg ggacatcctg tccccccagt tcatgtacgg ctccaaggcg tacgtgaagc
     1321 accccgccga catccccgat tacaagaagc tgtccttccc cgagggcttc aagtgggagc
     1381 gcgtgatgaa cttcgaggac ggcggtctgg tgaccgtgac ccaggactcc tccctgcagg
     1441 acggcacgct gatctacaag gtgaagatgc gcggcaccaa cttccccccc gacggccccg
     1501 taatgcagaa gaagaccatg ggctgggagg cctccaccga gcgcctgtac ccccgcgacg
     1561 gcgtgctgaa gggcgagatc caccaggccc tgaagctgaa ggacggcggc cactacctgg
     1621 tggagttcaa gaccatctac atggccaaga agcccgtgca actgcccggc tactactacg
     1681 tggacaccaa gctggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg
     1741 agcgctccga gggccgccac cacctgttcc tgtacggcat ggacgagctg tacaagtccg
     1801 gagataacgg aagcggagcc actaacttct ccctgttgaa acaagcaggg gatgtcgaag
     1861 agaatcccgg gccatctaga atggccccca agaagaagag gaaggtggtg agcaagggcg
     1921 aggaggtcat caaagagttc atgcgcttca aggtgcgcat ggagggctcc atgaacggcc
     1981 acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca
     2041 agctgaaggt gaccaagggc ggccccctgc ccttcgcctg ggacatcctg tccccccagt
     2101 tcatgtacgg ctccaaggcg tacgtgaagc accccgccga catccccgat tacaagaagc
     2161 tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggtctgg
     2221 tgaccgtgac ccaggactcc tccctgcagg acggcacgct gatctacaag gtgaagatgc
     2281 gcggcaccaa cttccccccc gacggccccg taatgcagaa gaagaccatg ggctgggagg
     2341 cctccaccga gcgcctgtac ccccgcgacg gcgtgctgaa gggcgagatc caccaggccc
     2401 tgaagctgaa ggacggcggc cactacctgg tggagttcaa gaccatctac atggccaaga
     2461 agcccgtgca actgcccggc tactactacg tggacaccaa gctggacatc acctcccaca
     2521 acgaggacta caccatcgtg gaacagtacg agcgctccga gggccgccac cacctgttcc
     2581 tgtacggcat ggacgagctg tacaagtaag tcgacggcgc gccccacccc tgcagggaat
     2641 tcgatatcaa gcttatcgat aatcaacctc tggattacaa aatttgtgaa agattgactg
     2701 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     2761 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     2821 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     2881 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     2941 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     3001 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat
     3061 cgtcctttcc ttggctgctc gcctatgttg ccacctggat tctgcgcggg acgtccttct
     3121 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     3181 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     3241 cctccccgca tcgataccga gcgctgctcg agagatctgt gatagcggcc atcaagctgg
     3301 ccgcgactct agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     3361 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     3421 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     3481 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcagct
     3541 tatcgatacc gcatgcacgt gcggaccgag cggccgcagg aacccctagt gatggagttg
     3601 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga
     3661 cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg
     3721 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa
     3781 agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     3841 gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt
     3901 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     3961 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt
     4021 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     4081 tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt
     4141 cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt
     4201 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc
     4261 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     4321 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     4381 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     4441 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     4501 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     4561 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     4621 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     4681 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     4741 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     4801 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     4861 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     4921 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     4981 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     5041 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     5101 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     5161 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     5221 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     5281 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     5341 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     5401 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     5461 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     5521 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     5581 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     5641 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     5701 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     5761 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     5821 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     5881 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     5941 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     6001 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     6061 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     6121 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     6181 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     6241 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     6301 cttttgctca catgt
//
