LOCUS       sequence-337185-        4929 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV backbone for Flp-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 4929)
  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 4929)
  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-VTKD3
FEATURES             Location/Qualifiers
     source          1..4929
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   29..158
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     intron          901..1033
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1016..1035
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     protein_bind    1076..1109
                     /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    1205..1261
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     protein_bind    complement(1272..1305)
                     /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    1398..1986
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1451..1471)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1869..1880)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    1994..2218
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2220..2360
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2220..2349
                     /label=AAV2 ITR
     rep_origin      2435..2890
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2522..2541)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2732..2753
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2907..2926)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3026..3048
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3086..3104)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3172..3276
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3277..3345,3346..4137)
                     /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(3495..3514)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4308..4896
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4797..4816
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gccttttgct ggccttttgc tcacatgtcc tgcaggcagc tgcgcgctcg ctcgctcact
       61 gaggccgccc gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg
      121 cagagaggga gtggccaact ccatcactag gggttcctgc tagactgcag agggcccgcg
      181 gccgcacgcg tttattcaga atgttatgca ctcacagtgg tttggcatgc atctggtgaa
      241 ttttttttaa cgaaaaatta gtgttggttt cgatgtatgg tagcattctc cctaacgtaa
      301 tttgaataat tcagcaaagc cccactacca gctgtacttc tgcagcctct tccattcttt
      361 tcagcattat aattttggtt aattttcaat tttaggtcct acgtctctgc aatttgtgta
      421 tgaataacag aataatttcc ctcttttgtt tcgcctttcc tgttcctgaa tctaaataaa
      481 gatggctttt tagtattaaa agtggaagaa aattacaggt aattatcttt gacggtaaaa
      541 acgctgtaat cagcgggcta catgaaaaat tactctaatt atggctgcat ttaagagaat
      601 ggaaaaaaac cttcttgtgg ataaaaacct taaattgtcc ccaatgtctg cttcaaattg
      661 gatggcactg cagctggagg ctttgttcag aattgatcct ggggagctac gaacccaaag
      721 tttcacagta ggaagctaga tatcgtttaa acttcctgca gcccgggctg ggcataaaag
      781 tcagggcaga gccatctatt gcttacattt gcttctagcc tgcaagtcga ggagcgcagc
      841 cttccagaag cagagcgcgg cgccttaagc tgcagaagtt ggtcgtgagg cactgggcac
      901 gtaagtatca aggttacaag acaggtttaa ggagaccaat agaaactggg cttgtcgaga
      961 cagagaagac tcttgcgttt ctgataggca cctattggtc ttactgacat ccactttgcc
     1021 tttctctcca caggtgtcca ctcccagttc aattacagct cttaagaaag ctagcgaagt
     1081 tcctattctc tagaaagtat aggaacttcg cagaatggta gctggattgt agctgctatt
     1141 agcaatatga aacctcttag aagttcctat tcttcaaaag gtataggaac ttcaacgcgc
     1201 cctagaattc gagctcggta cccggggatc ctctagagtc gacctgcagg catgcaagct
     1261 tggtggctag cgaagttcct atactttcta gagaatagga acttcttgcc ttaacccaga
     1321 aattatcact gttattcttt agaatggtgc aaagagaagt tcctatacct tttgaagaat
     1381 aggaacttcg gcgcgccaat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     1441 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     1501 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     1561 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     1621 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     1681 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     1741 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt
     1801 cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct
     1861 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     1921 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     1981 ccccgcctgg atcctgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     2041 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     2101 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     2161 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggca
     2221 ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc
     2281 cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg
     2341 agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     2401 tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc
     2461 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cttagcgccc
     2521 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     2581 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     2641 aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     2701 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     2761 ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggtctat
     2821 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     2881 tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     2941 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     3001 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     3061 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     3121 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     3181 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     3241 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     3301 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     3361 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     3421 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     3481 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     3541 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     3601 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     3661 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     3721 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     3781 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     3841 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     3901 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     3961 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     4021 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     4081 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     4141 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     4201 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     4261 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     4321 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     4381 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     4441 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     4501 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     4561 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     4621 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     4681 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     4741 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     4801 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     4861 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     4921 cggttcctg
//
