LOCUS       sequence-271294-        6976 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector to drive the expression of dTomato in PV cortical 
            interneuronsunder the control of the E1 regulatory element.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6976)
  AUTHORS   Vormstein-Schneider D, Lin JD, Pelkey KA, Chittajallu R, Guo B, 
            Arias-Garcia MA, Allaway K, Sakopoulos S, Schneider G, Stevenson O, 
            Vergara J, Sharma J, Zhang Q, Franken TP, Smith J, Ibrahim LA, M 
            Astro KJ, Sabri E, Huang S, Favuzzi E, Burbridge T, Xu Q, Guo L, 
            Vogel I, Sanchez V, Saldi GA, Gorissen BL, Yuan X, Zaghloul KA, 
            Devinsky O, Sabatini BL, Batista-Brito R, Reynolds J, Feng G, Fu Z, 
            McBain CJ, Fishell G, Dimidschstein J
  TITLE     Viral manipulation of functionally distinct interneurons in mice, 
            non-human primates and humans.
  JOURNAL   Nat Neurosci. 2020 Dec;23(12):1629-1636. doi: 
            10.1038/s41593-020-0692-9. Epub 2020 Aug 17.
  PUBMED    32807948
REFERENCE   2  (bases 1 to 6976)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41593-020-0692-9"; journalName: "Nat Neurosci"; date: 
            "2020-12"; volume: "23"; issue: "12"; pages: "1629-1636"
COMMENT     Sequence Label: pAAV-S5E1-dTom-nlsdTom
FEATURES             Location/Qualifiers
     source          1..6976
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     intron          1595..1727
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     regulatory      1750..1759
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1756..2457
                     /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             2479..2535
                     /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             2548..2568
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2569..3267
                     /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    3322..3910
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3375..3395)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3793..3804)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4079..4160
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4116..4135)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4170..4189
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   4239..4379
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4239..4368
                     /label=AAV2 ITR
     rep_origin      4454..4909
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4541..4560)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4751..4772
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4926..4945)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5045..5067
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5105..5123)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5191..5295
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5296..5364,5365..6156)
                     /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(5514..5533)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6327..6915
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6816..6835
                     /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 tttcaaagtg gacagagggg
      181 aggggagggg atgcgagggg aggggagggg aatgatcgcg aaaggcttcc aaaccttgtc
      241 cttgtttttc accatttctg aaatatatgc tgagtgcaac tatgggaaga ccattttcat
      301 aatctataat actcctcctt ttaaaggact ttcgttaacc gcttgcaaag gtgagtgccg
      361 ggtagaggac attagctcgc taagtcccta gaaatcacac ttggagacta agcaggcttt
      421 cccaggagaa gtccaaagcc aacataagca ggaggctggg ggctggccgt taaccgcaag
      481 gcagtggttg agccctcggg atcatcccgg cggggggcgc agcatctccg ccaaggccgc
      541 aggctctcac catcagctgc ccgagccacc ctgtacctcg cagtccactc gccctgccca
      601 cgccccgcgc cgcccgctca ccttcagccc ctgggagtcc atggccgccg gctacccgga
      661 gggtgcccac cgctgcccgc cgcagggtta ggggttcaga cccacttccc gggccctcaa
      721 accctaaggg acgcggcgtg cagcacgagg ggcgtggccc gatctccatt ggttgtcggc
      781 gtgagggggc ggagcttagt tgtaggacta ggaaggaggg ggccaccgga gcaggcgagg
      841 agggaacccc gagggaggac cgcgagggcg actggggctg gaatccgctg agcattgagt
      901 gctgccgagt tgtggggcta gaggagggag gtccagcctg gaaacggcgc gaggaggagg
      961 gattgggtgg agcaagagat atgagattaa agaataaaga tgatgaagca gcaaatagga
     1021 gggagagcca tgccgctttt catccctgca aacaaaggcc gactccattt tctcagcatt
     1081 ttttgtggaa gccgatttgc gcaatgcggc ttagtacttg accagggaaa atgatttacc
     1141 tgacacgtgt agtaatcgtg tctgggccac aaggtggcgc agaaaaatca cagttcggca
     1201 aaaaccttga agcctggctt gggcttgttc taaatctttt caggcgctgc tgtaattttg
     1261 ctattcgagt gcttattaaa ctgctccgcc agatttccac ccccaaagtc ttatttaaaa
     1321 atatgtggtt acctctttta gatttctatt tcttaagtgt ttgctgtagt ttggatctaa
     1381 actgtccctc aaagacacac gtgctgaatg ttccccagcc cgtgtgctgt tggagtggtg
     1441 gaaaacttcc tgcagcccgg gctgggcata aaagtcaggg cagagccatc tattgcttac
     1501 atttgcttct agcctgcaag tcgaggagcg cagccttcca gaagcagagc gcggcgcctt
     1561 aagctgcaga agttggtcgt gaggcactgg gcacgtaagt atcaaggtta caagacaggt
     1621 ttaaggagac caatagaaac tgggcttgtc gagacagaga agactcttgc gtttctgata
     1681 ggcacctatt ggtcttactg acatccactt tgcctttctc tccacagcga gctcgctagc
     1741 gcgatcgctg ccaccatggt gagcaagggc gaggaggtca tcaaagagtt catgcgcttc
     1801 aaggtgcgca tggagggctc catgaacggc cacgagttcg agatcgaggg cgagggcgag
     1861 ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg cggccccctg
     1921 cccttcgcct gggacatcct gtccccccag ttcatgtacg gctccaaggc gtacgtgaag
     1981 caccccgccg acatccccga ttacaagaag ctgtccttcc ccgagggctt caagtgggag
     2041 cgcgtgatga acttcgagga cggcggtctg gtgaccgtga cccaggactc ctccctgcag
     2101 gacggcacgc tgatctacaa ggtgaagatg cgcggcacca acttcccccc cgacggcccc
     2161 gtaatgcaga agaagaccat gggctgggag gcctccaccg agcgcctgta cccccgcgac
     2221 ggcgtgctga agggcgagat ccaccaggcc ctgaagctga aggacggcgg ccactacctg
     2281 gtggagttca agaccatcta catggccaag aagcccgtgc aactgcccgg ctactactac
     2341 gtggacacca agctggacat cacctcccac aacgaggact acaccatcgt ggaacagtac
     2401 gagcgctccg agggccgcca ccacctgttc ctgtacggca tggacgagct gtacaagtcc
     2461 ggagataacg gaagcggagc cactaacttc tccctgttga aacaagcagg ggatgtcgaa
     2521 gagaatcccg ggccatctag aatggccccc aagaagaaga ggaaggtggt gagcaagggc
     2581 gaggaggtca tcaaagagtt catgcgcttc aaggtgcgca tggagggctc catgaacggc
     2641 cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc
     2701 aagctgaagg tgaccaaggg cggccccctg cccttcgcct gggacatcct gtccccccag
     2761 ttcatgtacg gctccaaggc gtacgtgaag caccccgccg acatccccga ttacaagaag
     2821 ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggtctg
     2881 gtgaccgtga cccaggactc ctccctgcag gacggcacgc tgatctacaa ggtgaagatg
     2941 cgcggcacca acttcccccc cgacggcccc gtaatgcaga agaagaccat gggctgggag
     3001 gcctccaccg agcgcctgta cccccgcgac ggcgtgctga agggcgagat ccaccaggcc
     3061 ctgaagctga aggacggcgg ccactacctg gtggagttca agaccatcta catggccaag
     3121 aagcccgtgc aactgcccgg ctactactac gtggacacca agctggacat cacctcccac
     3181 aacgaggact acaccatcgt ggaacagtac gagcgctccg agggccgcca ccacctgttc
     3241 ctgtacggca tggacgagct gtacaagtaa gtcgacggcg cgccccaccc ctgcagggaa
     3301 ttcgatatca agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact
     3361 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     3421 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     3481 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     3541 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     3601 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     3661 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     3721 tcgtcctttc cttggctgct cgcctatgtt gccacctgga ttctgcgcgg gacgtccttc
     3781 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     3841 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     3901 gcctccccgc atcgataccg agcgctgctc gagagatctg tgatagcggc catcaagctg
     3961 gccgcgactc tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     4021 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     4081 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     4141 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatcagc
     4201 ttatcgatac cgcatgcacg tgcggaccga gcggccgcag gaacccctag tgatggagtt
     4261 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg
     4321 acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg
     4381 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca
     4441 aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     4501 cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct
     4561 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     4621 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt
     4681 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     4741 ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat
     4801 tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt
     4861 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact
     4921 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     4981 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     5041 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     5101 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     5161 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     5221 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     5281 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     5341 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     5401 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     5461 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     5521 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     5581 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     5641 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     5701 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     5761 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     5821 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     5881 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     5941 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     6001 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     6061 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     6121 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     6181 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     6241 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     6301 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     6361 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     6421 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     6481 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     6541 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     6601 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     6661 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     6721 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     6781 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     6841 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     6901 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     6961 ccttttgctc acatgt
//
