LOCUS       sequence-355040-        6600 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector for Cre- and Flp-dependent transgene expression (CRE-ON /
            FLP-ON) of dTom in cortical interneurons under the control of the 
            Dlx enhancer (Dimidschstein et al 2016).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6600)
  AUTHORS   Pouchelon G, Vergara J, McMahon J, Gorissen BL, Lin JD, 
            Vormstein-Schneider D, Niehaus JL, Burbridge TJ, Wester JC, Sherer 
            M, Fernandez-Otero M, Allaway KC, Pelkey K, Chittajallu R, McBain 
            CJ, Fan M, Nasse JS, Wildenberg GA, Fishell G, Dimidschstein J
  TITLE     A versatile viral toolkit for functional discovery in the nervous 
            system.
  JOURNAL   Cell Rep Methods. 2022 May 26;2(6):100225. doi: 
            10.1016/j.crmeth.2022.100225. eCollection 2022 Jun 20.
  PUBMED    35784651
REFERENCE   2  (bases 1 to 6600)
  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"; journalName: "Cell Rep 
            Methods."; date: "2022-05-26"; volume: "2(6):100225. doi"; pages: " 
            10.1016/j.crmeth.2022.100225. eCollection 2022 Jun 20"
COMMENT     Sequence Label: pAAV-VTKD4-dTom
FEATURES             Location/Qualifiers
     source          1..6600
                     /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          873..1005
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     988..1007
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     protein_bind    1048..1081
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     polyA_signal    1127..1175
                     /note="synthetic polyadenylation signal"
     polyA_signal    1184..1660
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1383..1402)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(1695..1714)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1739..1787
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     protein_bind    1947..1980
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     protein_bind    complement(1987..2020)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(2071..2104)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(2161..2865)
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="MAVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQT
                     AKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDG
                     GLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEI
                     HQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGR
                     HHLFLYGMDELYK"
     primer_bind     complement(2244..2267)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(2442..2461)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2711..2729
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      2862..2871
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2915..2948
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    2999..3032
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3041..3629
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3094..3114)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3512..3523)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3637..3861
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3863..4003
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3863..3992
                     /label=AAV2 ITR
     rep_origin      4078..4533
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4165..4184)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4375..4396
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4550..4569)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4669..4691
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4729..4747)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4815..4919
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4920..4988,4989..5780)
                     /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(5138..5157)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5951..6539
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6440..6459
                     /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 gctagactgc agagggcccg cggccgcacg cgtttattca gaatgttatg
      181 cactcacagt ggtttggcat gcatctggtg aatttttttt aacgaaaaat tagtgttggt
      241 ttcgatgtat ggtagcattc tccctaacgt aatttgaata attcagcaaa gccccactac
      301 cagctgtact tctgcagcct cttccattct tttcagcatt ataattttgg ttaattttca
      361 attttaggtc ctacgtctct gcaatttgtg tatgaataac agaataattt ccctcttttg
      421 tttcgccttt cctgttcctg aatctaaata aagatggctt tttagtatta aaagtggaag
      481 aaaattacag gtaattatct ttgacggtaa aaacgctgta atcagcgggc tacatgaaaa
      541 attactctaa ttatggctgc atttaagaga atggaaaaaa accttcttgt ggataaaaac
      601 cttaaattgt ccccaatgtc tgcttcaaat tggatggcac tgcagctgga ggctttgttc
      661 agaattgatc ctggggagct acgaacccaa agtttcacag taggaagcta gatatcgttt
      721 aaacttcctg cagcccgggc tgggcataaa agtcagggca gagccatcta ttgcttacat
      781 ttgcttctag cctgcaagtc gaggagcgca gccttccaga agcagagcgc ggcgccttaa
      841 gctgcagaag ttggtcgtga ggcactgggc acgtaagtat caaggttaca agacaggttt
      901 aaggagacca atagaaactg ggcttgtcga gacagagaag actcttgcgt ttctgatagg
      961 cacctattgg tcttactgac atccactttg cctttctctc cacaggtgtc cactcccagt
     1021 tcaattacag ctcttaagaa agctagcgaa gttcctattc tctagaaagt ataggaactt
     1081 ccctcagcac catggctagc ggcagcctcg gagtttgaat agatagaata aaatatcttt
     1141 attttcattc catctgtgtg ttggtttttt gtgtgagatc tacgggtggc atccctgtga
     1201 cccctcccca gtgcctctcc tggccctgga agttgccact ccagtgccca ccagccttgt
     1261 cctaataaaa ttaagttgca tcattttgtc tgactaggtg tccttctata atattatggg
     1321 gtggaggggg gtggtatgga gcaaggggca agttgggaag acaacctgta gggcctgcgg
     1381 ggtctattgg gaaccaagct ggagtgcagt ggcacaatct tggctcactg caatctccgc
     1441 ctcctgggtt caagcgattc tcctgcctca ggctcccgag ttgttgggat tccaggcatg
     1501 catgaccagg ctcagctaat ttttgttttt ttggtagaga cggggtttca ccatattggc
     1561 caggctggtc tccaactcct aatctcaggt gatctaccca ccttggcctc ccaaattgct
     1621 gggattacag gcgtgaacca ctgctccctt ccctgtcctt ctgcctcagc taattgagta
     1681 ggggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     1741 gcaataaaaa gacagaataa aacgcacggg tgttgggtcg tttgttcata aacgcggggt
     1801 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     1861 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     1921 agccaacgtc ggggcggcag gccgtggaag ttcctattct ctagaaagta taggaacttc
     1981 tccggaataa cttcgtatag gatactttat acgaagttat gcagaatggt agctggattg
     2041 tagctgctat tagcaatatg aaacctctta ataacttcgt atagcataca ttatacgaag
     2101 ttataacgcg ccactagtga attcgagctc ggtacccggg gatcctctag agtcgactta
     2161 cttgtacagc tcgtccatgc cgtacaggaa caggtggtgg cggccctcgg agcgctcgta
     2221 ctgttccacg atggtgtagt cctcgttgtg ggaggtgatg tccagcttgg tgtccacgta
     2281 gtagtagccg ggcagttgca cgggcttctt ggccatgtag atggtcttga actccaccag
     2341 gtagtggccg ccgtccttca gcttcagggc ctggtggatc tcgcccttca gcacgccgtc
     2401 gcgggggtac aggcgctcgg tggaggcctc ccagcccatg gtcttcttct gcattacggg
     2461 gccgtcgggg gggaagttgg tgccgcgcat cttcaccttg tagatcagcg tgccgtcctg
     2521 cagggaggag tcctgggtca cggtcaccag accgccgtcc tcgaagttca tcacgcgctc
     2581 ccacttgaag ccctcgggga aggacagctt cttgtaatcg gggatgtcgg cggggtgctt
     2641 cacgtacgcc ttggagccgt acatgaactg gggggacagg atgtcccagg cgaagggcag
     2701 ggggccgccc ttggtcacct tcagcttggc ggtctgggtg ccctcgtagg ggcggccctc
     2761 gccctcgccc tcgatctcga actcgtggcc gttcatggag ccctccatgc gcaccttgaa
     2821 gcgcatgaac tctttgatga cctcctcgcc cttgctcacg gccatggtgg cgtcgacctg
     2881 caggcatgca agctttacta gtggtggcgc tagcataact tcgtataaag tatcctatac
     2941 gaagttattt gccttaaccc agaaattatc actgttattc tttagaatgg tgcaaagaat
     3001 aacttcgtat aatgtatgct atacgaagtt atggcgcgcc aatcaacctc tggattacaa
     3061 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     3121 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     3181 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     3241 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     3301 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     3361 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     3421 ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat
     3481 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     3541 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     3601 tcggatctcc ctttgggccg cctccccgcc tggatcctgt gccttctagt tgccagccat
     3661 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     3721 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     3781 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     3841 gggatgcggt gggctctatg gcaggaaccc ctagtgatgg agttggccac tccctctctg
     3901 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc
     3961 cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat
     4021 tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg
     4081 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     4141 cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     4201 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     4261 ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat
     4321 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     4381 tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt gatttataag
     4441 ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     4501 cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct
     4561 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     4621 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     4681 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     4741 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     4801 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     4861 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     4921 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     4981 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     5041 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     5101 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     5161 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     5221 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     5281 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     5341 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     5401 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     5461 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     5521 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     5581 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     5641 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     5701 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     5761 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     5821 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     5881 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     5941 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     6001 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     6061 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     6121 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     6181 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     6241 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     6301 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     6361 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     6421 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     6481 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     6541 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
