LOCUS       sequence-271297-        6244 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector to drive the expression of dTomato in PV cortical 
            interneuronsunder the control of the E10 regulatory element.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6244)
  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 6244)
  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-S5E10-dTom-nlsdTom
FEATURES             Location/Qualifiers
     source          1..6244
                     /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          863..995
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     regulatory      1018..1027
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1024..1725
                     /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             1747..1803
                     /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             1816..1836
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1837..2535
                     /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    2590..3178
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2643..2663)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3061..3072)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3347..3428
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3384..3403)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3438..3457
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   3507..3647
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3507..3636
                     /label=AAV2 ITR
     rep_origin      3722..4177
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3809..3828)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4019..4040
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4194..4213)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4313..4335
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4373..4391)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4459..4563
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4564..4632,4633..5424)
                     /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(4782..4801)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5595..6183
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6084..6103
                     /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 ttttattgca aaaggaagga
      181 atgagacagt ttatgcagag ctaagggttt gtgcgttatt atgattaatc acaaggacag
      241 ctgccaagct tccatcatga caatattctc tgggagaatt catcaggttc tactgtctat
      301 taatttctgt tgatgtatct tatctggcat cttcaatgac agaggacact tgttagtttt
      361 tttttttaag tgaaggttaa aagacaaagt tcattaaaga aatgatttat atatgacatt
      421 taagaactag caatgtcatt gcttcaagaa aattatgaga atttagtctt ggtaggagtt
      481 tacaccatgt ccttgaagtg tctaattatg tgacttgata gttttactta gtacatatcg
      541 attaggctgt atctattatt tatcaagaaa ttatggaagg aggcaatgtg gcataggcat
      601 acacattctg attttaaaat aatcctgctt ttaccattaa ctccttctca gataattctg
      661 aatacatatc ttgtctatga atctgtgtaa tcatggaaaa agaaaaaatc aaacttcctg
      721 cagcccgggc tgggcataaa agtcagggca gagccatcta ttgcttacat ttgcttctag
      781 cctgcaagtc gaggagcgca gccttccaga agcagagcgc ggcgccttaa gctgcagaag
      841 ttggtcgtga ggcactgggc acgtaagtat caaggttaca agacaggttt aaggagacca
      901 atagaaactg ggcttgtcga gacagagaag actcttgcgt ttctgatagg cacctattgg
      961 tcttactgac atccactttg cctttctctc cacagcgagc tcgctagcgc gatcgctgcc
     1021 accatggtga gcaagggcga ggaggtcatc aaagagttca tgcgcttcaa ggtgcgcatg
     1081 gagggctcca tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac
     1141 gagggcaccc agaccgccaa gctgaaggtg accaagggcg gccccctgcc cttcgcctgg
     1201 gacatcctgt ccccccagtt catgtacggc tccaaggcgt acgtgaagca ccccgccgac
     1261 atccccgatt acaagaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac
     1321 ttcgaggacg gcggtctggt gaccgtgacc caggactcct ccctgcagga cggcacgctg
     1381 atctacaagg tgaagatgcg cggcaccaac ttcccccccg acggccccgt aatgcagaag
     1441 aagaccatgg gctgggaggc ctccaccgag cgcctgtacc cccgcgacgg cgtgctgaag
     1501 ggcgagatcc accaggccct gaagctgaag gacggcggcc actacctggt ggagttcaag
     1561 accatctaca tggccaagaa gcccgtgcaa ctgcccggct actactacgt ggacaccaag
     1621 ctggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga gcgctccgag
     1681 ggccgccacc acctgttcct gtacggcatg gacgagctgt acaagtccgg agataacgga
     1741 agcggagcca ctaacttctc cctgttgaaa caagcagggg atgtcgaaga gaatcccggg
     1801 ccatctagaa tggcccccaa gaagaagagg aaggtggtga gcaagggcga ggaggtcatc
     1861 aaagagttca tgcgcttcaa ggtgcgcatg gagggctcca tgaacggcca cgagttcgag
     1921 atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg
     1981 accaagggcg gccccctgcc cttcgcctgg gacatcctgt ccccccagtt catgtacggc
     2041 tccaaggcgt acgtgaagca ccccgccgac atccccgatt acaagaagct gtccttcccc
     2101 gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggtctggt gaccgtgacc
     2161 caggactcct ccctgcagga cggcacgctg atctacaagg tgaagatgcg cggcaccaac
     2221 ttcccccccg acggccccgt aatgcagaag aagaccatgg gctgggaggc ctccaccgag
     2281 cgcctgtacc cccgcgacgg cgtgctgaag ggcgagatcc accaggccct gaagctgaag
     2341 gacggcggcc actacctggt ggagttcaag accatctaca tggccaagaa gcccgtgcaa
     2401 ctgcccggct actactacgt ggacaccaag ctggacatca cctcccacaa cgaggactac
     2461 accatcgtgg aacagtacga gcgctccgag ggccgccacc acctgttcct gtacggcatg
     2521 gacgagctgt acaagtaagt cgacggcgcg ccccacccct gcagggaatt cgatatcaag
     2581 cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     2641 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     2701 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     2761 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     2821 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     2881 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     2941 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct
     3001 tggctgctcg cctatgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     3061 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     3121 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat
     3181 cgataccgag cgctgctcga gagatctgtg atagcggcca tcaagctggc cgcgactcta
     3241 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca
     3301 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
     3361 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     3421 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcagctt atcgataccg
     3481 catgcacgtg cggaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc
     3541 tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt
     3601 tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg
     3661 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag
     3721 tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     3781 gctacacttg ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     3841 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     3901 agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg
     3961 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     4021 ggactcttgt tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta
     4081 taagggattt tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt
     4141 aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc
     4201 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
     4261 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     4321 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg
     4381 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     4441 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     4501 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     4561 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     4621 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     4681 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     4741 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     4801 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     4861 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     4921 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     4981 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     5041 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     5101 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     5161 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     5221 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     5281 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     5341 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     5401 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     5461 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     5521 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     5581 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     5641 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     5701 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     5761 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     5821 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     5881 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     5941 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     6001 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     6061 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     6121 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     6181 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     6241 atgt
//
