LOCUS       sequence-354815-        6727 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector for Cre- and Flp-dependent transgene expression (CRE-ON /
            FLP-ON) 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 6727)
  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 6727)
  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-V5-APEX2-NES
FEATURES             Location/Qualifiers
     source          1..6727
                     /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(2142..2168)
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             complement(2928..2969)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     regulatory      2969..2978
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    3042..3075
                     /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    3126..3159
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3168..3756
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3221..3241)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3639..3650)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3764..3988
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3990..4130
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3990..4119
                     /label=AAV2 ITR
     rep_origin      4205..4660
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4292..4311)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4502..4523
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4677..4696)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4796..4818
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4856..4874)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4942..5046
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5047..5115,5116..5907)
                     /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(5265..5284)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6078..6666
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6567..6586
                     /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 ggtacctagt ccagggtcag gcgctccagg
     2161 ggtggcagct gcagggcgtc ggcaaatccc agttctgaca gcttctgatg tgcttctgcg
     2221 taatctgcga aaaaagcatc ctcgtcggct gcatacttat cgaccagagg tctgaacacg
     2281 ggatcggaca gcagggcttt gtcgctgggc agctgcagca gcccttcctt ctctccactc
     2341 agcagttcag taaagtaaga attgtcgaaa atcagggggt tgcttgtcca gggtccctcg
     2401 aagcctgacc tttccttatg agcggcgcca atggtgtgcc ctccagacag tgcgacgata
     2461 tcctggtcag tcaggcccat agctttccca aacacatctc gcagatggtc ggagcccttt
     2521 gtcggatctg gcaggcgccc ctctggaggt ggttctggtt tgtcctcccg tcctgggtgg
     2581 aaaggcacct tggggccccc agtgacctcc acagcgacca ctcctgccag ctgataaaaa
     2641 tcggcgtagg acagaattgg gaactctgct ttcagaggtt ccagcagccg cacggcgatg
     2701 tccagcccat tgttggcact atgtgccagt tcagcagggt gcttgattgt tccaaagggt
     2761 ccgcctgttt tggttccctt gtcaaaggtg cctgcgctgt ggaaagccag tctcagcatc
     2821 agaggtgcgc acctcttttc tgcgataaac cctctcagct tcttcttggc tttctccacg
     2881 gcatcctggt agtctgcgga cactgttggg tatgatttcc caccggtggt gctgtccagg
     2941 cccagcaggg ggttggggat gggcttgccc atggtggcgt acccggggat cctctagagt
     3001 cgacctgcag gcatgcaagc tttactagtg gtggcgctag cataacttcg tataaagtat
     3061 cctatacgaa gttatttgcc ttaacccaga aattatcact gttattcttt agaatggtgc
     3121 aaagaataac ttcgtataat gtatgctata cgaagttatg gcgcgccaat caacctctgg
     3181 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     3241 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     3301 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
     3361 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
     3421 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
     3481 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     3541 attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca
     3601 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
     3661 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
     3721 agacgagtcg gatctccctt tgggccgcct ccccgcctgg atcctgtgcc ttctagttgc
     3781 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     3841 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     3901 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     3961 catgctgggg atgcggtggg ctctatggca ggaaccccta gtgatggagt tggccactcc
     4021 ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg
     4081 ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat
     4141 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca
     4201 tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     4261 accgctacac ttgccagcgc cttagcgccc gctcctttcg ctttcttccc ttcctttctc
     4321 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     4381 tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt
     4441 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     4501 agtggactct tgttccaaac tggaacaaca ctcaactcta tctcgggcta ttcttttgat
     4561 ttataaggga ttttgccgat ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa
     4621 tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca
     4681 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
     4741 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     4801 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc
     4861 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     4921 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     4981 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     5041 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     5101 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     5161 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     5221 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     5281 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     5341 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     5401 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     5461 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     5521 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     5581 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     5641 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     5701 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     5761 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     5821 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     5881 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     5941 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     6001 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     6061 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     6121 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     6181 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     6241 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     6301 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     6361 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     6421 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     6481 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     6541 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     6601 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     6661 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     6721 cacatgt
//
