LOCUS       sequence-421884-        6358 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV construct expressing dTomato driven by PV+ basket cell-targeting
            enhancer. Alias: pAAV_WP0003_dTomato_nlsdTomato.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6358)
  AUTHORS   Furlanis E, Dai M, Leyva Garcia B, Tran T, Vergara J, Pereira A, 
            Gorissen BL, Wills S, Vlachos A, Hairston A, Dwivedi D, Du S, 
            McMahon J, Huang S, Morabito A, Vazquez A, Kim S, Lee AT, Chang EF, 
            Razzaq T, Qazi A, Vargish G, Yuan X, Caccavano A, Hunt S, 
            Chittajallu R, McLean N, Hewitt L, Paranzino E, Rice H, Cummins AC, 
            Plotnikova A, Mohanty A, Tangen AC, Shin JH, Azadi R, Eldridge MAG, 
            Alvarez VA, Averbeck BB, Alyahyay M, Vallejo TR, Soheib M, Vattino 
            LG, MacGregor CP, Chatain CP, Banks E, Olah VJ, Naskar S, Hill S, 
            Liebergall S, Badiani R, Hyde L, Hanley E, Xu Q, Allaway KC, 
            Goldberg EM, Rowan MJM, Nowakowski TJ, Lee S, Favuzzi E, Kaeser PS, 
            Sjulson L, Batista-Brito R, Takesian AE, Ibrahim LA, Iqbal A, Pelkey
            KA, McBain CJ, Dimidschstein J, Fishell G, Wang Y
  TITLE     An enhancer-AAV toolbox to target and manipulate distinct 
            interneuron subtypes.
  JOURNAL   Neuron. 2025 May 21;113(10):1525-1547.e15. doi: 
            10.1016/j.neuron.2025.05.002.
  PUBMED    40403705
REFERENCE   2  (bases 1 to 6358)
  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: 
            "10.1016/j.neuron.2025.05"; journalName: "Neuron"; date: 
            "2025-05-21- 21"; volume: "113"; issue: "10"; pages: "1525-1547.e15"
COMMENT     Sequence Label: pAAV_BiPVe3_dTomato_nlsdTomato
FEATURES             Location/Qualifiers
     source          1..6358
                     /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          977..1109
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     regulatory      1132..1141
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1138..1839
                     /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             1861..1917
                     /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             1930..1950
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1951..2649
                     /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    2704..3292
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2757..2777)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3175..3186)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3461..3542
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3498..3517)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3552..3571
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   3621..3761
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3621..3750
                     /label=AAV2 ITR
     rep_origin      3836..4291
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3923..3942)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4133..4154
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4308..4327)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4427..4449
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4487..4505)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4573..4677
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4678..4746,4747..5538)
                     /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(4896..4915)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5709..6297
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6198..6217
                     /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 gcgtatcgat agaccgctgt tgcagtattt cagggaacaa
      181 gatcagaatg gtatagccta cctgaaaaat aggcaatcct ttcaaaatta aatgcacttt
      241 ctcttgggta ggagacaaga taatctatga taagggaaga gagatgtctc cagcttttat
      301 tttttttctt gtaatttgca tttaagccct ggtaacagtt taatatgaag gtgactatgg
      361 cagctccatt tattcaatct atcaatactc tgttgtcaca ggaggagcca tgaggacttg
      421 agtgcagagg aggaatcccc acaaagcagt cttcactctt atcatagctt ccatgtgccc
      481 ttcttattca gcagctctca ttaaaggaac actatcttta gcacagggat tctacttgac
      541 attgacaagg acaggctcag tagcaaatgg gccaaaacta tgaataggcg attcacagca
      601 gagtaaaatc caatggcagc agacataaaa agaagctcaa aatgacaaac aagtcagaca
      661 aacacaaatt caagtaacaa taaaaaatgg agcactggtt tgactcgtac acaagcaaac
      721 agacacaaac aaaacaaaag agtcattaca cctggggcta aagacttctc ttatgtattg
      781 caaagtggaa ataagacctc caggcaaaat gtgaatcgat gtttaaactt cctgcagccc
      841 gggctgggca taaaagtcag ggcagagcca tctattgctt acatttgctt ctagcctgca
      901 agtcgaggag cgcagccttc cagaagcaga gcgcggcgcc ttaagctgca gaagttggtc
      961 gtgaggcact gggcacgtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
     1021 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
     1081 tgacatccac tttgcctttc tctccacagc gagctcgcta gcgcgatcgc tgccaccatg
     1141 gtgagcaagg gcgaggaggt catcaaagag ttcatgcgct tcaaggtgcg catggagggc
     1201 tccatgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc
     1261 acccagaccg ccaagctgaa ggtgaccaag ggcggccccc tgcccttcgc ctgggacatc
     1321 ctgtcccccc agttcatgta cggctccaag gcgtacgtga agcaccccgc cgacatcccc
     1381 gattacaaga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag
     1441 gacggcggtc tggtgaccgt gacccaggac tcctccctgc aggacggcac gctgatctac
     1501 aaggtgaaga tgcgcggcac caacttcccc cccgacggcc ccgtaatgca gaagaagacc
     1561 atgggctggg aggcctccac cgagcgcctg tacccccgcg acggcgtgct gaagggcgag
     1621 atccaccagg ccctgaagct gaaggacggc ggccactacc tggtggagtt caagaccatc
     1681 tacatggcca agaagcccgt gcaactgccc ggctactact acgtggacac caagctggac
     1741 atcacctccc acaacgagga ctacaccatc gtggaacagt acgagcgctc cgagggccgc
     1801 caccacctgt tcctgtacgg catggacgag ctgtacaagt ccggagataa cggaagcgga
     1861 gccactaact tctccctgtt gaaacaagca ggggatgtcg aagagaatcc cgggccatct
     1921 agaatggccc ccaagaagaa gaggaaggtg gtgagcaagg gcgaggaggt catcaaagag
     1981 ttcatgcgct tcaaggtgcg catggagggc tccatgaacg gccacgagtt cgagatcgag
     2041 ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag
     2101 ggcggccccc tgcccttcgc ctgggacatc ctgtcccccc agttcatgta cggctccaag
     2161 gcgtacgtga agcaccccgc cgacatcccc gattacaaga agctgtcctt ccccgagggc
     2221 ttcaagtggg agcgcgtgat gaacttcgag gacggcggtc tggtgaccgt gacccaggac
     2281 tcctccctgc aggacggcac gctgatctac aaggtgaaga tgcgcggcac caacttcccc
     2341 cccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctccac cgagcgcctg
     2401 tacccccgcg acggcgtgct gaagggcgag atccaccagg ccctgaagct gaaggacggc
     2461 ggccactacc tggtggagtt caagaccatc tacatggcca agaagcccgt gcaactgccc
     2521 ggctactact acgtggacac caagctggac atcacctccc acaacgagga ctacaccatc
     2581 gtggaacagt acgagcgctc cgagggccgc caccacctgt tcctgtacgg catggacgag
     2641 ctgtacaagt aagtcgacgg cgcgccccac ccctgcaggg aattcgatat caagcttatc
     2701 gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     2761 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     2821 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     2881 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     2941 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     3001 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     3061 ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg
     3121 ctcgcctatg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     3181 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     3241 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgatac
     3301 cgagcgctgc tcgagagatc tgtgatagcg gccatcaagc tggccgcgac tctagatcat
     3361 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     3421 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     3481 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     3541 gcattctagt tgtggtttgt ccaaactcat caatgtatca gcttatcgat accgcatgca
     3601 cgtgcggacc gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg
     3661 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg
     3721 ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt
     3781 tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg
     3841 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     3901 cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     3961 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     4021 ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg
     4081 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     4141 ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga tttataaggg
     4201 attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     4261 aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct
     4321 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     4381 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     4441 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     4501 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     4561 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4621 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     4681 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     4741 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     4801 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     4861 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     4921 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     4981 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     5041 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     5101 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     5161 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     5221 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     5281 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     5341 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     5401 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     5461 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     5521 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     5581 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     5641 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     5701 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     5761 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     5821 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     5881 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     5941 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     6001 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     6061 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     6121 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     6181 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     6241 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     6301 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgt
//
