LOCUS       sequence-272255-        6303 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector to drive the expression of dTomato in VIP cortical 
            interneurons under the control of the E6 regulatory element.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6303)
  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 6303)
  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-S5E6-dTom-nlsdTom
FEATURES             Location/Qualifiers
     source          1..6303
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             complement(255..266)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     RBS             677..688
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     intron          922..1054
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     regulatory      1077..1086
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1083..1784
                     /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             1806..1862
                     /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             1875..1895
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1896..2594
                     /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    2649..3237
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2702..2722)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3120..3131)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3406..3487
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3443..3462)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3497..3516
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   3566..3706
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3566..3695
                     /label=AAV2 ITR
     rep_origin      3781..4236
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3868..3887)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4078..4099
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4253..4272)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4372..4394
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4432..4450)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4518..4622
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4623..4691,4692..5483)
                     /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(4841..4860)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5654..6242
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6143..6162
                     /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 tttttgtcac tttgttactc
      181 tacagtgttg cctggagttc gatacttcat tactctatag tggggtgaag aagttcacct
      241 tcttattttc atttccttcc ctcaatgatt tcttcagagc tagctcttac cagctagaaa
      301 ttcttcaaac gacactcgtg ccttccttca cacaggttga actatttgtc tctaatgccc
      361 taaagtactg gtgttcaatc ttccaggcac ttccaatgat ctgaaatctg acctgcttag
      421 gtcagctggc tctgagatta tggtattcta gtcctcaaac caacctgttg gctcgttggt
      481 tttgtaccaa acacactgac ttacatagct caaaatacca ctggcctttt aaaaatggca
      541 tatcacattc caggggagga tcaaaactgc tggctggtga tatttgtcaa gtctctcaaa
      601 gttgcacttt ccaggatttt caattcactg aattcttaga cagacatgtt tatgtgaaag
      661 aattctttat atattttttc tcctctttga gtgggcaaat gaaaatcttg acctctgggt
      721 tccttatttt atttgactct ctgtagtatt taaatcttaa aattttccta aacttcctgc
      781 agcccgggct gggcataaaa gtcagggcag agccatctat tgcttacatt tgcttctagc
      841 ctgcaagtcg aggagcgcag ccttccagaa gcagagcgcg gcgccttaag ctgcagaagt
      901 tggtcgtgag gcactgggca cgtaagtatc aaggttacaa gacaggttta aggagaccaa
      961 tagaaactgg gcttgtcgag acagagaaga ctcttgcgtt tctgataggc acctattggt
     1021 cttactgaca tccactttgc ctttctctcc acagcgagct cgctagcgcg atcgctgcca
     1081 ccatggtgag caagggcgag gaggtcatca aagagttcat gcgcttcaag gtgcgcatgg
     1141 agggctccat gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg
     1201 agggcaccca gaccgccaag ctgaaggtga ccaagggcgg ccccctgccc ttcgcctggg
     1261 acatcctgtc cccccagttc atgtacggct ccaaggcgta cgtgaagcac cccgccgaca
     1321 tccccgatta caagaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact
     1381 tcgaggacgg cggtctggtg accgtgaccc aggactcctc cctgcaggac ggcacgctga
     1441 tctacaaggt gaagatgcgc ggcaccaact tcccccccga cggccccgta atgcagaaga
     1501 agaccatggg ctgggaggcc tccaccgagc gcctgtaccc ccgcgacggc gtgctgaagg
     1561 gcgagatcca ccaggccctg aagctgaagg acggcggcca ctacctggtg gagttcaaga
     1621 ccatctacat ggccaagaag cccgtgcaac tgcccggcta ctactacgtg gacaccaagc
     1681 tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgag cgctccgagg
     1741 gccgccacca cctgttcctg tacggcatgg acgagctgta caagtccgga gataacggaa
     1801 gcggagccac taacttctcc ctgttgaaac aagcagggga tgtcgaagag aatcccgggc
     1861 catctagaat ggcccccaag aagaagagga aggtggtgag caagggcgag gaggtcatca
     1921 aagagttcat gcgcttcaag gtgcgcatgg agggctccat gaacggccac gagttcgaga
     1981 tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga
     2041 ccaagggcgg ccccctgccc ttcgcctggg acatcctgtc cccccagttc atgtacggct
     2101 ccaaggcgta cgtgaagcac cccgccgaca tccccgatta caagaagctg tccttccccg
     2161 agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggtctggtg accgtgaccc
     2221 aggactcctc cctgcaggac ggcacgctga tctacaaggt gaagatgcgc ggcaccaact
     2281 tcccccccga cggccccgta atgcagaaga agaccatggg ctgggaggcc tccaccgagc
     2341 gcctgtaccc ccgcgacggc gtgctgaagg gcgagatcca ccaggccctg aagctgaagg
     2401 acggcggcca ctacctggtg gagttcaaga ccatctacat ggccaagaag cccgtgcaac
     2461 tgcccggcta ctactacgtg gacaccaagc tggacatcac ctcccacaac gaggactaca
     2521 ccatcgtgga acagtacgag cgctccgagg gccgccacca cctgttcctg tacggcatgg
     2581 acgagctgta caagtaagtc gacggcgcgc cccacccctg cagggaattc gatatcaagc
     2641 ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     2701 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     2761 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     2821 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     2881 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
     2941 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     3001 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt
     3061 ggctgctcgc ctatgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     3121 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     3181 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc
     3241 gataccgagc gctgctcgag agatctgtga tagcggccat caagctggcc gcgactctag
     3301 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     3361 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     3421 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     3481 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcagctta tcgataccgc
     3541 atgcacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct
     3601 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     3661 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg
     3721 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt
     3781 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     3841 ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     3901 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     3961 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc
     4021 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     4081 gactcttgtt ccaaactgga acaacactca actctatctc gggctattct tttgatttat
     4141 aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta
     4201 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct
     4261 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     4321 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     4381 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     4441 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     4501 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     4561 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     4621 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     4681 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     4741 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     4801 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     4861 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     4921 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     4981 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     5041 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     5101 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     5161 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     5221 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     5281 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     5341 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     5401 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     5461 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     5521 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     5581 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     5641 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     5701 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     5761 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     5821 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     5881 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     5941 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     6001 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     6061 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     6121 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     6181 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     6241 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     6301 tgt
//
