LOCUS       Exported                5958 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 E4 regulatory element.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-S5E4-dTom-nlsdTom
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5958)
  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 5958)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5958
                     /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          577..709
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     regulatory      732..741
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             738..1439
                     /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             1461..1517
                     /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             1530..1550
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1551..2249
                     /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    2304..2892
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2357..2377)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2775..2786)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3061..3142
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3098..3117)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3152..3171
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   3221..3361
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3221..3350
                     /label=AAV2 ITR
     rep_origin      3436..3891
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3523..3542)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3733..3754
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3908..3927)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4027..4049
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4087..4105)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4173..4277
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4278..5138
                     /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(4496..4515)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5309..5897
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5798..5817
                     /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 ttttctgaca gagcaagtct
      181 tgacctgctt aacattatgt tatgctagtc attttaaaat gagtctttat ttcccataga
      241 aggtcagttt ttttacatta ttatataatc ttttgacaga ataacaaata acattctgaa
      301 tgtctcattt ctaaatacaa aacatcttag tataaaatta tgcattgttt taaatgcttg
      361 gaagtaggtc cacatgtaga aaacaaagta cgtatgataa aaaatatcaa aattgtatat
      421 tcagaaactt cctgcagccc gggctgggca taaaagtcag ggcagagcca tctattgctt
      481 acatttgctt ctagcctgca agtcgaggag cgcagccttc cagaagcaga gcgcggcgcc
      541 ttaagctgca gaagttggtc gtgaggcact gggcacgtaa gtatcaaggt tacaagacag
      601 gtttaaggag accaatagaa actgggcttg tcgagacaga gaagactctt gcgtttctga
      661 taggcaccta ttggtcttac tgacatccac tttgcctttc tctccacagc gagctcgcta
      721 gcgcgatcgc tgccaccatg gtgagcaagg gcgaggaggt catcaaagag ttcatgcgct
      781 tcaaggtgcg catggagggc tccatgaacg gccacgagtt cgagatcgag ggcgagggcg
      841 agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggcggccccc
      901 tgcccttcgc ctgggacatc ctgtcccccc agttcatgta cggctccaag gcgtacgtga
      961 agcaccccgc cgacatcccc gattacaaga agctgtcctt ccccgagggc ttcaagtggg
     1021 agcgcgtgat gaacttcgag gacggcggtc tggtgaccgt gacccaggac tcctccctgc
     1081 aggacggcac gctgatctac aaggtgaaga tgcgcggcac caacttcccc cccgacggcc
     1141 ccgtaatgca gaagaagacc atgggctggg aggcctccac cgagcgcctg tacccccgcg
     1201 acggcgtgct gaagggcgag atccaccagg ccctgaagct gaaggacggc ggccactacc
     1261 tggtggagtt caagaccatc tacatggcca agaagcccgt gcaactgccc ggctactact
     1321 acgtggacac caagctggac atcacctccc acaacgagga ctacaccatc gtggaacagt
     1381 acgagcgctc cgagggccgc caccacctgt tcctgtacgg catggacgag ctgtacaagt
     1441 ccggagataa cggaagcgga gccactaact tctccctgtt gaaacaagca ggggatgtcg
     1501 aagagaatcc cgggccatct agaatggccc ccaagaagaa gaggaaggtg gtgagcaagg
     1561 gcgaggaggt catcaaagag ttcatgcgct tcaaggtgcg catggagggc tccatgaacg
     1621 gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg
     1681 ccaagctgaa ggtgaccaag ggcggccccc tgcccttcgc ctgggacatc ctgtcccccc
     1741 agttcatgta cggctccaag gcgtacgtga agcaccccgc cgacatcccc gattacaaga
     1801 agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggtc
     1861 tggtgaccgt gacccaggac tcctccctgc aggacggcac gctgatctac aaggtgaaga
     1921 tgcgcggcac caacttcccc cccgacggcc ccgtaatgca gaagaagacc atgggctggg
     1981 aggcctccac cgagcgcctg tacccccgcg acggcgtgct gaagggcgag atccaccagg
     2041 ccctgaagct gaaggacggc ggccactacc tggtggagtt caagaccatc tacatggcca
     2101 agaagcccgt gcaactgccc ggctactact acgtggacac caagctggac atcacctccc
     2161 acaacgagga ctacaccatc gtggaacagt acgagcgctc cgagggccgc caccacctgt
     2221 tcctgtacgg catggacgag ctgtacaagt aagtcgacgg cgcgccccac ccctgcaggg
     2281 aattcgatat caagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga
     2341 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     2401 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     2461 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     2521 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     2581 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     2641 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat
     2701 catcgtcctt tccttggctg ctcgcctatg ttgccacctg gattctgcgc gggacgtcct
     2761 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     2821 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     2881 ccgcctcccc gcatcgatac cgagcgctgc tcgagagatc tgtgatagcg gccatcaagc
     2941 tggccgcgac tctagatcat aatcagccat accacatttg tagaggtttt acttgcttta
     3001 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     3061 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     3121 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatca
     3181 gcttatcgat accgcatgca cgtgcggacc gagcggccgc aggaacccct agtgatggag
     3241 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
     3301 cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag
     3361 gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt
     3421 caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     3481 cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc gctttcttcc
     3541 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     3601 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg
     3661 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     3721 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaactct atctcgggct
     3781 attcttttga tttataaggg attttgccga tttcggtcta ttggttaaaa aatgagctga
     3841 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca
     3901 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
     3961 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     4021 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
     4081 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     4141 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     4201 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     4261 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     4321 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     4381 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     4441 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     4501 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     4561 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     4621 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     4681 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     4741 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     4801 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     4861 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     4921 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     4981 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     5041 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     5101 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     5161 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     5221 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     5281 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     5341 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     5401 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     5461 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     5521 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     5581 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     5641 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     5701 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     5761 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     5821 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     5881 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     5941 ggccttttgc tcacatgt
//
