LOCUS       sequence-354847-        5862 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector for Cre- and Flp-dependent transgene expression (CRE-ON /
            FLP-OFF) of V5-APEX2-NES 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 5862)
  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 5862)
  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-VTKD5-V5-APEX2-NES
FEATURES             Location/Qualifiers
     source          1..5862
                     /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)"
     protein_bind    complement(1088..1121)
                     /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(1172..1205)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1243..1269)
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             complement(2029..2070)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     regulatory      2070..2079
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2143..2176
                     /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    2227..2260
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    2261..2294
                     /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)"
     misc_feature    2303..2891
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2356..2376)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2774..2785)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2899..3123
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3125..3265
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3125..3254
                     /label=AAV2 ITR
     rep_origin      3340..3795
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3427..3446)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3637..3658
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3812..3831)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3931..3953
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3991..4009)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4077..4181
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4182..4250,4251..5042)
                     /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(4400..4419)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5213..5801
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5702..5721
                     /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 ctccggaata acttcgtata ggatacttta tacgaagtta tgcagaatgg tagctggatt
     1141 gtagctgcta ttagcaatat gaaacctctt aataacttcg tatagcatac attatacgaa
     1201 gttataacgc gccactagtg aattcgagct cggtacctag tccagggtca ggcgctccag
     1261 gggtggcagc tgcagggcgt cggcaaatcc cagttctgac agcttctgat gtgcttctgc
     1321 gtaatctgcg aaaaaagcat cctcgtcggc tgcatactta tcgaccagag gtctgaacac
     1381 gggatcggac agcagggctt tgtcgctggg cagctgcagc agcccttcct tctctccact
     1441 cagcagttca gtaaagtaag aattgtcgaa aatcaggggg ttgcttgtcc agggtccctc
     1501 gaagcctgac ctttccttat gagcggcgcc aatggtgtgc cctccagaca gtgcgacgat
     1561 atcctggtca gtcaggccca tagctttccc aaacacatct cgcagatggt cggagccctt
     1621 tgtcggatct ggcaggcgcc cctctggagg tggttctggt ttgtcctccc gtcctgggtg
     1681 gaaaggcacc ttggggcccc cagtgacctc cacagcgacc actcctgcca gctgataaaa
     1741 atcggcgtag gacagaattg ggaactctgc tttcagaggt tccagcagcc gcacggcgat
     1801 gtccagccca ttgttggcac tatgtgccag ttcagcaggg tgcttgattg ttccaaaggg
     1861 tccgcctgtt ttggttccct tgtcaaaggt gcctgcgctg tggaaagcca gtctcagcat
     1921 cagaggtgcg cacctctttt ctgcgataaa ccctctcagc ttcttcttgg ctttctccac
     1981 ggcatcctgg tagtctgcgg acactgttgg gtatgatttc ccaccggtgg tgctgtccag
     2041 gcccagcagg gggttgggga tgggcttgcc catggtggcg tacccgggga tcctctagag
     2101 tcgacctgca ggcatgcaag ctttactagt ggtggcgcta gcataacttc gtataaagta
     2161 tcctatacga agttatttgc cttaacccag aaattatcac tgttattctt tagaatggtg
     2221 caaagaataa cttcgtataa tgtatgctat acgaagttat gaagttccta ttctctagaa
     2281 agtataggaa cttcggcgcg ccaatcaacc tctggattac aaaatttgtg aaagattgac
     2341 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     2401 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     2461 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     2521 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     2581 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     2641 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct
     2701 gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     2761 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     2821 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     2881 cgcctccccg cctggatcct gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     2941 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     3001 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     3061 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     3121 tggcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact
     3181 gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc
     3241 gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg
     3301 tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat
     3361 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccttag
     3421 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     3481 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     3541 ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     3601 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     3661 caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg
     3721 tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     3781 taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     3841 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     3901 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     3961 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     4021 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     4081 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     4141 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     4201 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     4261 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     4321 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     4381 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     4441 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     4501 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     4561 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     4621 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     4681 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     4741 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     4801 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     4861 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     4921 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     4981 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     5041 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     5101 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     5161 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     5221 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     5281 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     5341 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     5401 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     5461 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     5521 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     5581 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     5641 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     5701 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     5761 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     5821 ttttacggtt cctggccttt tgctggcctt ttgctcacat gt
//
