LOCUS       Exported                6360 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 E5 regulatory element.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-S5E5-dTom-nlsdTom
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6360)
  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 6360)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6360
                     /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          979..1111
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     regulatory      1134..1143
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1140..1841
                     /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             1863..1919
                     /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             1932..1952
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1953..2651
                     /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    2706..3294
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2759..2779)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3177..3188)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3463..3544
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3500..3519)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3554..3573
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   3623..3763
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3623..3752
                     /label=AAV2 ITR
     rep_origin      3838..4293
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3925..3944)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4135..4156
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4310..4329)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4429..4451
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4489..4507)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4575..4679
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4680..5540
                     /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(4898..4917)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5711..6299
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6200..6219
                     /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 tttaatgttt tgatatttag
      181 gagaaaattg caaaacaaaa tgatgacagt gtttgaaagt gtttgatcag tgccaagcat
      241 cactttatgt acttggcaaa catgacttga ggccttaagc tgtgatttgc aaatgtagat
      301 tggaatcaag atctttatag atgaggaagc aaaaatcaga agacaaaata acattatcaa
      361 cttgatctca tgtgcagcca gggctgaact gcaaatgctg atttgcccca gtctgggctc
      421 ctcaaatcgt tccttggaat cctattagtt ggaactttat ctctgctcgt ggcagggtgc
      481 ctgggaccat gtttataaat atctgctgaa tgaagaataa gtgagtcaat cgaaccagaa
      541 ctcactttgg ttagttaatt tcattcgtgg tatttatgga gagcagaaga aagaattcca
      601 gagacacgat ttgtcaaaac tctctaaaga aaatgatgac actatatatt gatgaaaatg
      661 aatgttcttg ttcttgcttt atttgatttt cttgtccccc cactccccat ctgctagggt
      721 ctcattacag catagttctt gaatatccca ggttgacctg aagttacaat atattcttga
      781 tttagatggc agacattggg aatattttga ctcttaaaat ttaataaaac ttcctgcagc
      841 ccgggctggg cataaaagtc agggcagagc catctattgc ttacatttgc ttctagcctg
      901 caagtcgagg agcgcagcct tccagaagca gagcgcggcg ccttaagctg cagaagttgg
      961 tcgtgaggca ctgggcacgt aagtatcaag gttacaagac aggtttaagg agaccaatag
     1021 aaactgggct tgtcgagaca gagaagactc ttgcgtttct gataggcacc tattggtctt
     1081 actgacatcc actttgcctt tctctccaca gcgagctcgc tagcgcgatc gctgccacca
     1141 tggtgagcaa gggcgaggag gtcatcaaag agttcatgcg cttcaaggtg cgcatggagg
     1201 gctccatgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg
     1261 gcacccagac cgccaagctg aaggtgacca agggcggccc cctgcccttc gcctgggaca
     1321 tcctgtcccc ccagttcatg tacggctcca aggcgtacgt gaagcacccc gccgacatcc
     1381 ccgattacaa gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg
     1441 aggacggcgg tctggtgacc gtgacccagg actcctccct gcaggacggc acgctgatct
     1501 acaaggtgaa gatgcgcggc accaacttcc cccccgacgg ccccgtaatg cagaagaaga
     1561 ccatgggctg ggaggcctcc accgagcgcc tgtacccccg cgacggcgtg ctgaagggcg
     1621 agatccacca ggccctgaag ctgaaggacg gcggccacta cctggtggag ttcaagacca
     1681 tctacatggc caagaagccc gtgcaactgc ccggctacta ctacgtggac accaagctgg
     1741 acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgagcgc tccgagggcc
     1801 gccaccacct gttcctgtac ggcatggacg agctgtacaa gtccggagat aacggaagcg
     1861 gagccactaa cttctccctg ttgaaacaag caggggatgt cgaagagaat cccgggccat
     1921 ctagaatggc ccccaagaag aagaggaagg tggtgagcaa gggcgaggag gtcatcaaag
     1981 agttcatgcg cttcaaggtg cgcatggagg gctccatgaa cggccacgag ttcgagatcg
     2041 agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca
     2101 agggcggccc cctgcccttc gcctgggaca tcctgtcccc ccagttcatg tacggctcca
     2161 aggcgtacgt gaagcacccc gccgacatcc ccgattacaa gaagctgtcc ttccccgagg
     2221 gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg tctggtgacc gtgacccagg
     2281 actcctccct gcaggacggc acgctgatct acaaggtgaa gatgcgcggc accaacttcc
     2341 cccccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc accgagcgcc
     2401 tgtacccccg cgacggcgtg ctgaagggcg agatccacca ggccctgaag ctgaaggacg
     2461 gcggccacta cctggtggag ttcaagacca tctacatggc caagaagccc gtgcaactgc
     2521 ccggctacta ctacgtggac accaagctgg acatcacctc ccacaacgag gactacacca
     2581 tcgtggaaca gtacgagcgc tccgagggcc gccaccacct gttcctgtac ggcatggacg
     2641 agctgtacaa gtaagtcgac ggcgcgcccc acccctgcag ggaattcgat atcaagctta
     2701 tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     2761 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     2821 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     2881 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     2941 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     3001 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     3061 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
     3121 tgctcgccta tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     3181 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     3241 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat
     3301 accgagcgct gctcgagaga tctgtgatag cggccatcaa gctggccgcg actctagatc
     3361 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
     3421 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
     3481 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     3541 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cagcttatcg ataccgcatg
     3601 cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg
     3661 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc
     3721 cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat
     3781 tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg
     3841 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     3901 cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     3961 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     4021 ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat
     4081 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     4141 tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt gatttataag
     4201 ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     4261 cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct
     4321 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     4381 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     4441 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     4501 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     4561 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     4621 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     4681 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     4741 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     4801 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     4861 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     4921 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     4981 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     5041 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     5101 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     5161 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     5221 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     5281 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     5341 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     5401 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     5461 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     5521 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     5581 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     5641 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     5701 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     5761 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     5821 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     5881 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     5941 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     6001 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     6061 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     6121 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     6181 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     6241 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     6301 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
