LOCUS       sequence-375693-        5717 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector for Cre- and Flp-dependent transgene expression (CRE-OFF 
            / 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 5717)
  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 5717)
  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-VTKD6-dTom
FEATURES             Location/Qualifiers
     source          1..5717
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   2..142
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2..131
                     /label=AAV2 ITR
     rep_origin      217..672
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(304..323)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     514..535
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(689..708)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     808..830
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(868..886)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        954..1058
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1059..1127,1128..1919)
                     /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(1277..1296)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2090..2678
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2579..2598
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2740..2869
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     intron          3612..3744
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     3727..3746
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     protein_bind    complement(3787..3820)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    3821..3854
                     /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)"
     CDS             complement(3992..4696)
                     /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(4075..4098)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(4273..4292)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     4542..4560
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      4693..4702
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(4737..4770)
                     /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(4855..4888)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    4897..5485
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4950..4970)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5368..5379)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    5493..5717
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
       61 gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag
      121 cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc
      181 ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa
      241 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccttagcgc
      301 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
      361 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
      421 aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
      481 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
      541 cactcaactc tatctcgggc tattcttttg atttataagg gattttgccg atttcggtct
      601 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
      661 cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
      721 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
      781 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
      841 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
      901 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
      961 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     1021 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     1081 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     1141 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     1201 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     1261 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     1321 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     1381 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     1441 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     1501 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     1561 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     1621 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     1681 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     1741 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     1801 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     1861 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     1921 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     1981 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     2041 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     2101 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     2161 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     2221 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     2281 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     2341 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     2401 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     2461 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     2521 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     2581 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     2641 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     2701 tacggttcct ggccttttgc tggccttttg ctcacatgtc ctgcaggcag ctgcgcgctc
     2761 gctcgctcac tgaggccgcc cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc
     2821 gagcgagcgc gcagagaggg agtggccaac tccatcacta ggggttcctg ctagactgca
     2881 gagggcccgc ggccgcacgc gtttattcag aatgttatgc actcacagtg gtttggcatg
     2941 catctggtga atttttttta acgaaaaatt agtgttggtt tcgatgtatg gtagcattct
     3001 ccctaacgta atttgaataa ttcagcaaag ccccactacc agctgtactt ctgcagcctc
     3061 ttccattctt ttcagcatta taattttggt taattttcaa ttttaggtcc tacgtctctg
     3121 caatttgtgt atgaataaca gaataatttc cctcttttgt ttcgcctttc ctgttcctga
     3181 atctaaataa agatggcttt ttagtattaa aagtggaaga aaattacagg taattatctt
     3241 tgacggtaaa aacgctgtaa tcagcgggct acatgaaaaa ttactctaat tatggctgca
     3301 tttaagagaa tggaaaaaaa ccttcttgtg gataaaaacc ttaaattgtc cccaatgtct
     3361 gcttcaaatt ggatggcact gcagctggag gctttgttca gaattgatcc tggggagcta
     3421 cgaacccaaa gtttcacagt aggaagctag atatcgttta aacttcctgc agcccgggct
     3481 gggcataaaa gtcagggcag agccatctat tgcttacatt tgcttctagc ctgcaagtcg
     3541 aggagcgcag ccttccagaa gcagagcgcg gcgccttaag ctgcagaagt tggtcgtgag
     3601 gcactgggca cgtaagtatc aaggttacaa gacaggttta aggagaccaa tagaaactgg
     3661 gcttgtcgag acagagaaga ctcttgcgtt tctgataggc acctattggt cttactgaca
     3721 tccactttgc ctttctctcc acaggtgtcc actcccagtt caattacagc tcttaagaaa
     3781 gctagcataa cttcgtatag catacattat acgaagttat gaagttccta ttctctagaa
     3841 agtataggaa cttcgcagaa tggtagctgg attgtagctg ctattagcaa tatgaaacct
     3901 cttagaagtt cctattcttc aaaaggtata ggaacttcaa cgcgccctag aattcgagct
     3961 cggtacccgg ggatcctcta gagtcgactt acttgtacag ctcgtccatg ccgtacagga
     4021 acaggtggtg gcggccctcg gagcgctcgt actgttccac gatggtgtag tcctcgttgt
     4081 gggaggtgat gtccagcttg gtgtccacgt agtagtagcc gggcagttgc acgggcttct
     4141 tggccatgta gatggtcttg aactccacca ggtagtggcc gccgtccttc agcttcaggg
     4201 cctggtggat ctcgcccttc agcacgccgt cgcgggggta caggcgctcg gtggaggcct
     4261 cccagcccat ggtcttcttc tgcattacgg ggccgtcggg ggggaagttg gtgccgcgca
     4321 tcttcacctt gtagatcagc gtgccgtcct gcagggagga gtcctgggtc acggtcacca
     4381 gaccgccgtc ctcgaagttc atcacgcgct cccacttgaa gccctcgggg aaggacagct
     4441 tcttgtaatc ggggatgtcg gcggggtgct tcacgtacgc cttggagccg tacatgaact
     4501 ggggggacag gatgtcccag gcgaagggca gggggccgcc cttggtcacc ttcagcttgg
     4561 cggtctgggt gccctcgtag gggcggccct cgccctcgcc ctcgatctcg aactcgtggc
     4621 cgttcatgga gccctccatg cgcaccttga agcgcatgaa ctctttgatg acctcctcgc
     4681 ccttgctcac ggccatggtg gcgtcgacct gcaggcatgc aagcttggtg gctagcgaag
     4741 ttcctatact ttctagagaa taggaacttc ttgccttaac ccagaaatta tcactgttat
     4801 tctttagaat ggtgcaaaga gaagttccta taccttttga agaataggaa cttcataact
     4861 tcgtatagca tacattatac gaagttatgg cgcgccaatc aacctctgga ttacaaaatt
     4921 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     4981 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     5041 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     5101 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     5161 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     5221 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     5281 ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg
     5341 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     5401 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     5461 atctcccttt gggccgcctc cccgcctgga tcctgtgcct tctagttgcc agccatctgt
     5521 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     5581 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     5641 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     5701 tgcggtgggc tctatgg
//
