LOCUS       Exported                7341 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  AAV vector to drive the expression of dTomato in PV cortical 
            interneuronsunder the control of the E8 regulatory element.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-S5E8-dTom-nlsdTom
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7341)
  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 7341)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7341
                     /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          1960..2092
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     regulatory      2115..2124
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2121..2822
                     /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             2844..2900
                     /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             2913..2933
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2934..3632
                     /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    3687..4275
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3740..3760)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4158..4169)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4444..4525
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4481..4500)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4535..4554
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   4604..4744
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4604..4733
                     /label=AAV2 ITR
     rep_origin      4819..5274
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4906..4925)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5116..5137
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5291..5310)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5410..5432
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5470..5488)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5556..5660
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5661..6521
                     /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(5879..5898)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6692..7280
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7181..7200
                     /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 tttattgatc tccaactttt
      181 taaatccctc tgtcatttta aatgaggtgc actcggttgt gtcatcatct cggttttaat
      241 tgtgtgaaaa tttcctgcca atctcacacc gctgcggcaa cctcaccttg ctacttgccc
      301 tgcaagttct gcagtgtgcc gttctgagta tgccgtttta actagttctt gcagcaggac
      361 acaaagcgag atagtctgat agaaccagtt ctcctctggt tttaccttta ctcttagatg
      421 agttaagggt cacatcaaac cagggctcag cccgccagat ctcctaagca cagcccctcc
      481 tgacccaatg cagttaaccc aacctcattc agcgctagta tcaaatgaca ctggagctgc
      541 tgcagtatgc atcccgagac taagtaggca ggatttatta tcagcagaag tcccctaact
      601 accaggttat tcaagctccg ttcttgtcac aaacaggcgc ggcggaagac acagtgcagc
      661 agactcagag ctcatttaca agacaagcga attctcagtt agagacaagg gcagcgcggc
      721 agcgaactgc agtaaatctt ttcacgctca cagcaacatc taacaatgct ctcctgcaac
      781 gcctcagatc aaacgaatcc tacttggttt aaacatcaaa tcaacaccat aaaaaaggct
      841 tcattagcaa agttcaattt aggatgtttt taatcgtgtc ttaattctag aaccagtgcg
      901 agactttcca tgcttattca agcatgctga cagaattgga acctcttaga attgcctacc
      961 tgcacctatc agcctggctg acaggagccc gccaaaggat taaaaaaaaa caaaacccaa
     1021 aacataaaat catgcaaaaa aatatttacc cccgaaagat gtatgtagtt aaagctcagc
     1081 ttcctgcagc ctcgatagcc cctgaagtgt taatctgaag aaacagttcc atgagtttcc
     1141 acaggccggt agtgagtctc ctacacttga cctagacaga cttacataat gaagcatcag
     1201 tgctggggag cttgcacgat gtcatcacca gcaagagtaa gaagtattgg cagcagcaag
     1261 caggcgggca ggctgagatc ttgcatggaa atcatgaacc aggtcttgct tttcgttttt
     1321 gaaacgtttt ggaaggagag ttatgaatag cccagaaata ggtctcattt tgtgggtagg
     1381 aagaatgacc agaagcatga aagctaaatc tcctggcaag tgcaggggac ctctcttgga
     1441 gtgtgcagta aacccgaggg gacgacttct cctgctgtca actcctgaac catcacatct
     1501 ggagtgaagg aaggggctgg tgaagccttg taataaatgc aaaggatgct gctgagagct
     1561 ttggtctgcc tttaactcat tgtggtgagt agaggggatg tggcagtatg caatgagagt
     1621 tggttgtgta ggttgctttg cagagtaata accaaaaaaa aaaaaatctg tgaagtgctc
     1681 aatactttag acacatttta ataaacaaga tgatagtaaa attactcttc tccatcaaat
     1741 tgagactgtg ctgggttaaa ctgttttaat gcattttaac tcctgatgtt catccaagta
     1801 ataagagaaa cttcctgcag cccgggctgg gcataaaagt cagggcagag ccatctattg
     1861 cttacatttg cttctagcct gcaagtcgag gagcgcagcc ttccagaagc agagcgcggc
     1921 gccttaagct gcagaagttg gtcgtgaggc actgggcacg taagtatcaa ggttacaaga
     1981 caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact cttgcgtttc
     2041 tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac agcgagctcg
     2101 ctagcgcgat cgctgccacc atggtgagca agggcgagga ggtcatcaaa gagttcatgc
     2161 gcttcaaggt gcgcatggag ggctccatga acggccacga gttcgagatc gagggcgagg
     2221 gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggcggcc
     2281 ccctgccctt cgcctgggac atcctgtccc cccagttcat gtacggctcc aaggcgtacg
     2341 tgaagcaccc cgccgacatc cccgattaca agaagctgtc cttccccgag ggcttcaagt
     2401 gggagcgcgt gatgaacttc gaggacggcg gtctggtgac cgtgacccag gactcctccc
     2461 tgcaggacgg cacgctgatc tacaaggtga agatgcgcgg caccaacttc ccccccgacg
     2521 gccccgtaat gcagaagaag accatgggct gggaggcctc caccgagcgc ctgtaccccc
     2581 gcgacggcgt gctgaagggc gagatccacc aggccctgaa gctgaaggac ggcggccact
     2641 acctggtgga gttcaagacc atctacatgg ccaagaagcc cgtgcaactg cccggctact
     2701 actacgtgga caccaagctg gacatcacct cccacaacga ggactacacc atcgtggaac
     2761 agtacgagcg ctccgagggc cgccaccacc tgttcctgta cggcatggac gagctgtaca
     2821 agtccggaga taacggaagc ggagccacta acttctccct gttgaaacaa gcaggggatg
     2881 tcgaagagaa tcccgggcca tctagaatgg cccccaagaa gaagaggaag gtggtgagca
     2941 agggcgagga ggtcatcaaa gagttcatgc gcttcaaggt gcgcatggag ggctccatga
     3001 acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga
     3061 ccgccaagct gaaggtgacc aagggcggcc ccctgccctt cgcctgggac atcctgtccc
     3121 cccagttcat gtacggctcc aaggcgtacg tgaagcaccc cgccgacatc cccgattaca
     3181 agaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg
     3241 gtctggtgac cgtgacccag gactcctccc tgcaggacgg cacgctgatc tacaaggtga
     3301 agatgcgcgg caccaacttc ccccccgacg gccccgtaat gcagaagaag accatgggct
     3361 gggaggcctc caccgagcgc ctgtaccccc gcgacggcgt gctgaagggc gagatccacc
     3421 aggccctgaa gctgaaggac ggcggccact acctggtgga gttcaagacc atctacatgg
     3481 ccaagaagcc cgtgcaactg cccggctact actacgtgga caccaagctg gacatcacct
     3541 cccacaacga ggactacacc atcgtggaac agtacgagcg ctccgagggc cgccaccacc
     3601 tgttcctgta cggcatggac gagctgtaca agtaagtcga cggcgcgccc cacccctgca
     3661 gggaattcga tatcaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat
     3721 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     3781 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     3841 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     3901 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     3961 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     4021 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     4081 aatcatcgtc ctttccttgg ctgctcgcct atgttgccac ctggattctg cgcgggacgt
     4141 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     4201 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     4261 gggccgcctc cccgcatcga taccgagcgc tgctcgagag atctgtgata gcggccatca
     4321 agctggccgc gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct
     4381 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     4441 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     4501 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     4561 tcagcttatc gataccgcat gcacgtgcgg accgagcggc cgcaggaacc cctagtgatg
     4621 gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc
     4681 gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg
     4741 caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     4801 cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     4861 ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct
     4921 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     4981 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg
     5041 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     5101 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg
     5161 gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc
     5221 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt
     5281 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
     5341 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     5401 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     5461 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     5521 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     5581 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     5641 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     5701 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     5761 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     5821 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     5881 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     5941 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     6001 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     6061 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     6121 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     6181 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     6241 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     6301 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     6361 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     6421 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     6481 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     6541 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     6601 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     6661 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     6721 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     6781 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     6841 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     6901 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     6961 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     7021 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     7081 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     7141 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     7201 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     7261 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     7321 gctggccttt tgctcacatg t
//
