LOCUS       sequence-337188-        4997 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV backbone for Cre- and Flp-dependent transgene expression 
            (CRE-OFF / FLP-ON) of any transgene in cortical interneurons under 
            the control of the Dlx enhancer..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4997)
  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 4997)
  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
FEATURES             Location/Qualifiers
     source          1..4997
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          853..985
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     968..987
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     protein_bind    complement(1028..1061)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    1062..1095
                     /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    1191..1247
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     protein_bind    complement(1258..1291)
                     /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(1376..1409)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    1418..2006
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1471..1491)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1889..1900)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2014..2238
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2240..2380
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2240..2369
                     /label=AAV2 ITR
     rep_origin      2455..2910
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2542..2561)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2752..2773
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2927..2946)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3046..3068
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3106..3124)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3192..3296
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3297..3365,3366..4157)
                     /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(3515..3534)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4328..4916
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4817..4836
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(4978..4997,1..110)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag
       61 cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct gctagactgc
      121 agagggcccg cggccgcacg cgtttattca gaatgttatg cactcacagt ggtttggcat
      181 gcatctggtg aatttttttt aacgaaaaat tagtgttggt ttcgatgtat ggtagcattc
      241 tccctaacgt aatttgaata attcagcaaa gccccactac cagctgtact tctgcagcct
      301 cttccattct tttcagcatt ataattttgg ttaattttca attttaggtc ctacgtctct
      361 gcaatttgtg tatgaataac agaataattt ccctcttttg tttcgccttt cctgttcctg
      421 aatctaaata aagatggctt tttagtatta aaagtggaag aaaattacag gtaattatct
      481 ttgacggtaa aaacgctgta atcagcgggc tacatgaaaa attactctaa ttatggctgc
      541 atttaagaga atggaaaaaa accttcttgt ggataaaaac cttaaattgt ccccaatgtc
      601 tgcttcaaat tggatggcac tgcagctgga ggctttgttc agaattgatc ctggggagct
      661 acgaacccaa agtttcacag taggaagcta gatatcgttt aaacttcctg cagcccgggc
      721 tgggcataaa agtcagggca gagccatcta ttgcttacat ttgcttctag cctgcaagtc
      781 gaggagcgca gccttccaga agcagagcgc ggcgccttaa gctgcagaag ttggtcgtga
      841 ggcactgggc acgtaagtat caaggttaca agacaggttt aaggagacca atagaaactg
      901 ggcttgtcga gacagagaag actcttgcgt ttctgatagg cacctattgg tcttactgac
      961 atccactttg cctttctctc cacaggtgtc cactcccagt tcaattacag ctcttaagaa
     1021 agctagcata acttcgtata gcatacatta tacgaagtta tgaagttcct attctctaga
     1081 aagtatagga acttcgcaga atggtagctg gattgtagct gctattagca atatgaaacc
     1141 tcttagaagt tcctattctt caaaaggtat aggaacttca acgcgcccta gaattcgagc
     1201 tcggtacccg gggatcctct agagtcgacc tgcaggcatg caagcttggt ggctagcgaa
     1261 gttcctatac tttctagaga ataggaactt cttgccttaa cccagaaatt atcactgtta
     1321 ttctttagaa tggtgcaaag agaagttcct ataccttttg aagaatagga acttcataac
     1381 ttcgtatagc atacattata cgaagttatg gcgcgccaat caacctctgg attacaaaat
     1441 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     1501 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     1561 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     1621 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     1681 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     1741 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     1801 gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct
     1861 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     1921 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     1981 gatctccctt tgggccgcct ccccgcctgg atcctgtgcc ttctagttgc cagccatctg
     2041 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     2101 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     2161 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     2221 atgcggtggg ctctatggca ggaaccccta gtgatggagt tggccactcc ctctctgcgc
     2281 gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg
     2341 gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt
     2401 ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc
     2461 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     2521 ttgccagcgc cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     2581 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     2641 tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc
     2701 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     2761 tgttccaaac tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga
     2821 ttttgccgat ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     2881 attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg
     2941 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     3001 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     3061 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     3121 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     3181 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     3241 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     3301 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     3361 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     3421 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     3481 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     3541 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     3601 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     3661 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     3721 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     3781 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     3841 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     3901 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     3961 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     4021 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     4081 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     4141 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     4201 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     4261 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     4321 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     4381 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     4441 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     4501 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4561 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     4621 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     4681 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     4741 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     4801 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     4861 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     4921 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtcct
     4981 gcaggcagct gcgcgct
//
