LOCUS       sequence-337186-        5880 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV backbone for Cre- and Flp-dependent transgene expression (CRE-ON
            / 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 5880)
  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 5880)
  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
FEATURES             Location/Qualifiers
     source          1..5880
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     191..210
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   352..481
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     intron          1224..1356
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1339..1358
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     protein_bind    1399..1432
                     /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    1478..1526
                     /note="synthetic polyadenylation signal"
     polyA_signal    1535..2011
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1734..1753)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2046..2065)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2090..2138
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     protein_bind    2298..2331
                     /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(2338..2371)
                     /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(2422..2455)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2470..2526
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     protein_bind    2546..2579
                     /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    2630..2663
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2672..3260
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2725..2745)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3143..3154)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3268..3492
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3494..3634
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3494..3623
                     /label=AAV2 ITR
     rep_origin      3709..4164
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3796..3815)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4006..4027
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4181..4200)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4300..4322
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4360..4378)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4446..4550
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4551..4619,4620..5411)
                     /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(4769..4788)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(5582..5880,1..290)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
       61 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
      121 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
      181 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
      241 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
      301 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg tcctgcaggc
      361 agctgcgcgc tcgctcgctc actgaggccg cccgggcgtc gggcgacctt tggtcgcccg
      421 gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc
      481 tgctagactg cagagggccc gcggccgcac gcgtttattc agaatgttat gcactcacag
      541 tggtttggca tgcatctggt gaattttttt taacgaaaaa ttagtgttgg tttcgatgta
      601 tggtagcatt ctccctaacg taatttgaat aattcagcaa agccccacta ccagctgtac
      661 ttctgcagcc tcttccattc ttttcagcat tataattttg gttaattttc aattttaggt
      721 cctacgtctc tgcaatttgt gtatgaataa cagaataatt tccctctttt gtttcgcctt
      781 tcctgttcct gaatctaaat aaagatggct ttttagtatt aaaagtggaa gaaaattaca
      841 ggtaattatc tttgacggta aaaacgctgt aatcagcggg ctacatgaaa aattactcta
      901 attatggctg catttaagag aatggaaaaa aaccttcttg tggataaaaa ccttaaattg
      961 tccccaatgt ctgcttcaaa ttggatggca ctgcagctgg aggctttgtt cagaattgat
     1021 cctggggagc tacgaaccca aagtttcaca gtaggaagct agatatcgtt taaacttcct
     1081 gcagcccggg ctgggcataa aagtcagggc agagccatct attgcttaca tttgcttcta
     1141 gcctgcaagt cgaggagcgc agccttccag aagcagagcg cggcgcctta agctgcagaa
     1201 gttggtcgtg aggcactggg cacgtaagta tcaaggttac aagacaggtt taaggagacc
     1261 aatagaaact gggcttgtcg agacagagaa gactcttgcg tttctgatag gcacctattg
     1321 gtcttactga catccacttt gcctttctct ccacaggtgt ccactcccag ttcaattaca
     1381 gctcttaaga aagctagcga agttcctatt ctctagaaag tataggaact tccctcagca
     1441 ccatggctag cggcagcctc ggagtttgaa tagatagaat aaaatatctt tattttcatt
     1501 ccatctgtgt gttggttttt tgtgtgagat ctacgggtgg catccctgtg acccctcccc
     1561 agtgcctctc ctggccctgg aagttgccac tccagtgccc accagccttg tcctaataaa
     1621 attaagttgc atcattttgt ctgactaggt gtccttctat aatattatgg ggtggagggg
     1681 ggtggtatgg agcaaggggc aagttgggaa gacaacctgt agggcctgcg gggtctattg
     1741 ggaaccaagc tggagtgcag tggcacaatc ttggctcact gcaatctccg cctcctgggt
     1801 tcaagcgatt ctcctgcctc aggctcccga gttgttggga ttccaggcat gcatgaccag
     1861 gctcagctaa tttttgtttt tttggtagag acggggtttc accatattgg ccaggctggt
     1921 ctccaactcc taatctcagg tgatctaccc accttggcct cccaaattgc tgggattaca
     1981 ggcgtgaacc actgctccct tccctgtcct tctgcctcag ctaattgagt aggggggagg
     2041 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     2101 agacagaata aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     2161 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     2221 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     2281 cggggcggca ggccgtggaa gttcctattc tctagaaagt ataggaactt ctccggaata
     2341 acttcgtata ggatacttta tacgaagtta tgcagaatgg tagctggatt gtagctgcta
     2401 ttagcaatat gaaacctctt aataacttcg tatagcatac attatacgaa gttataacgc
     2461 gccactagtg aattcgagct cggtacccgg ggatcctcta gagtcgacct gcaggcatgc
     2521 aagctttact agtggtggcg ctagcataac ttcgtataaa gtatcctata cgaagttatt
     2581 tgccttaacc cagaaattat cactgttatt ctttagaatg gtgcaaagaa taacttcgta
     2641 taatgtatgc tatacgaagt tatggcgcgc caatcaacct ctggattaca aaatttgtga
     2701 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     2761 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     2821 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     2881 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     2941 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     3001 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     3061 ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     3121 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     3181 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     3241 cctttgggcc gcctccccgc ctggatcctg tgccttctag ttgccagcca tctgttgttt
     3301 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     3361 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     3421 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     3481 tgggctctat ggcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     3541 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
     3601 tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt
     3661 acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta
     3721 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     3781 gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     3841 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     3901 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
     3961 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     4021 aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc
     4081 cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     4141 acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg
     4201 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     4261 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     4321 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     4381 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     4441 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     4501 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     4561 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     4621 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     4681 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     4741 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     4801 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     4861 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     4921 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     4981 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     5041 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     5101 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     5161 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     5221 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     5281 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     5341 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     5401 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     5461 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     5521 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     5581 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     5641 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     5701 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     5761 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     5821 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
//
