LOCUS       sequence-337184-        4947 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV backbone for Cre-dependent transgene expression 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 4947)
  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 4947)
  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-VTKD2
FEATURES             Location/Qualifiers
     source          1..4947
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          872..1004
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     987..1006
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     protein_bind    complement(1053..1086)
                     /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(1137..1170)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    1185..1241
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     protein_bind    1261..1294
                     /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    1345..1378
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    1387..1975
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1440..1460)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1858..1869)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    1983..2207
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2209..2349
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2209..2338
                     /label=AAV2 ITR
     rep_origin      2424..2879
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2511..2530)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2721..2742
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2896..2915)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3015..3037
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3075..3093)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3161..3265
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3266..3334,3335..4126)
                     /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(3484..3503)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4297..4885
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4786..4805
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(4947,1..129)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 ctgcaggcag ctgcgcgctc gctcgctcac tgaggccgcc cgggcgtcgg gcgacctttg
       61 gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg agtggccaac tccatcacta
      121 ggggttcctg ctagactgca gagggcccgc ggccgcacgc gtttattcag aatgttatgc
      181 actcacagtg gtttggcatg catctggtga atttttttta acgaaaaatt agtgttggtt
      241 tcgatgtatg gtagcattct ccctaacgta atttgaataa ttcagcaaag ccccactacc
      301 agctgtactt ctgcagcctc ttccattctt ttcagcatta taattttggt taattttcaa
      361 ttttaggtcc tacgtctctg caatttgtgt atgaataaca gaataatttc cctcttttgt
      421 ttcgcctttc ctgttcctga atctaaataa agatggcttt ttagtattaa aagtggaaga
      481 aaattacagg taattatctt tgacggtaaa aacgctgtaa tcagcgggct acatgaaaaa
      541 ttactctaat tatggctgca tttaagagaa tggaaaaaaa ccttcttgtg gataaaaacc
      601 ttaaattgtc cccaatgtct gcttcaaatt ggatggcact gcagctggag gctttgttca
      661 gaattgatcc tggggagcta cgaacccaaa gtttcacagt aggaagctag atatcgttta
      721 aacttcctgc agcccgggct gggcataaaa gtcagggcag agccatctat tgcttacatt
      781 tgcttctagc ctgcaagtcg aggagcgcag ccttccagaa gcagagcgcg gcgccttaag
      841 ctgcagaagt tggtcgtgag gcactgggca cgtaagtatc aaggttacaa gacaggttta
      901 aggagaccaa tagaaactgg gcttgtcgag acagagaaga ctcttgcgtt tctgataggc
      961 acctattggt cttactgaca tccactttgc ctttctctcc acaggtgtcc actcccagtt
     1021 caattacagc tcttaagaaa gctagctccg gaataacttc gtataggata ctttatacga
     1081 agttatgcag aatggtagct ggattgtagc tgctattagc aatatgaaac ctcttaataa
     1141 cttcgtatag catacattat acgaagttat aacgcgccac tagtgaattc gagctcggta
     1201 cccggggatc ctctagagtc gacctgcagg catgcaagct ttactagtgg tggcgctagc
     1261 ataacttcgt ataaagtatc ctatacgaag ttatttgcct taacccagaa attatcactg
     1321 ttattcttta gaatggtgca aagaataact tcgtataatg tatgctatac gaagttatgg
     1381 cgcgccaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     1441 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     1501 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     1561 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     1621 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     1681 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     1741 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg
     1801 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     1861 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     1921 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga
     1981 tcctgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     2041 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     2101 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     2161 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggcag gaacccctag
     2221 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     2281 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct
     2341 gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     2401 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     2461 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc
     2521 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     2581 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
     2641 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     2701 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat
     2761 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa
     2821 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt
     2881 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     2941 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     3001 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     3061 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     3121 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     3181 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     3241 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     3301 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     3361 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     3421 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     3481 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     3541 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     3601 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     3661 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     3721 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     3781 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     3841 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     3901 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     3961 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     4021 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     4081 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     4141 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     4201 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     4261 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     4321 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     4381 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     4441 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     4501 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     4561 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     4621 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     4681 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     4741 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     4801 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     4861 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     4921 cttttgctgg ccttttgctc acatgtc
//
