LOCUS       sequence-337190-        4522 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV backbone for Cre-dependent transgene expression of any transgene
            in neurons under the control of the hSyn1 promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4522)
  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 4522)
  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-VTKS2
FEATURES             Location/Qualifiers
     source          1..4522
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(313..346)
                     /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(397..430)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    445..501
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     protein_bind    521..554
                     /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    605..638
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    647..1235
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(700..720)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1118..1129)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    1243..1467
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1469..1609
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1469..1598
                     /label=AAV2 ITR
     rep_origin      1684..2139
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1771..1790)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1981..2002
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2156..2175)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2275..2297
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2335..2353)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2421..2525
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2526..2594,2595..3386)
                     /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(2744..2763)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3557..4145
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4046..4065
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4207..4336
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        join(4364..4522,1..289)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
ORIGIN
        1 actgccagct tcagcaccgc ggacagtgcc ttcgcccccg cctggcggcg cgcgccaccg
       61 ccgcctcagc actgaaggcg cgctgacgtc actcgccggt cccccgcaaa ctccccttcc
      121 cggccacctt ggtcgcgtcc gcgccgccgc cggcccagcc ggaccgcacc acgcgaggcg
      181 cgagataggg gggcacgggc gcgaccatct gcgctgcggc gccggcgact cagcgctgcc
      241 tcagtctgcg gtgggcagcg gaggagtcgt gtcgtgcctg agagcgcagt cagatatcgt
      301 ttaaactccg gaataacttc gtataggata ctttatacga agttatgcag aatggtagct
      361 ggattgtagc tgctattagc aatatgaaac ctcttaataa cttcgtatag catacattat
      421 acgaagttat aacgcgccac tagtgaattc gagctcggta cccggggatc ctctagagtc
      481 gacctgcagg catgcaagct ttactagtgg tggcgctagc ataacttcgt ataaagtatc
      541 ctatacgaag ttatttgcct taacccagaa attatcactg ttattcttta gaatggtgca
      601 aagaataact tcgtataatg tatgctatac gaagttatgg cgcgccaatc aacctctgga
      661 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
      721 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
      781 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
      841 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
      901 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
      961 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     1021 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac
     1081 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     1141 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     1201 gacgagtcgg atctcccttt gggccgcctc cccgcctgga tcctgtgcct tctagttgcc
     1261 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     1321 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     1381 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     1441 atgctgggga tgcggtgggc tctatggcag gaacccctag tgatggagtt ggccactccc
     1501 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc
     1561 tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg
     1621 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat
     1681 agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     1741 ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg
     1801 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     1861 ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg
     1921 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     1981 gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt
     2041 tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat
     2101 ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa
     2161 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     2221 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     2281 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     2341 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     2401 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     2461 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     2521 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     2581 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     2641 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     2701 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     2761 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     2821 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     2881 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     2941 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     3001 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     3061 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     3121 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     3181 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     3241 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     3301 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     3361 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     3421 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     3481 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     3541 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     3601 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     3661 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     3721 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     3781 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     3841 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     3901 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     3961 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     4021 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     4081 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     4141 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     4201 acatgtcctg caggcagctg cgcgctcgct cgctcactga ggccgcccgg gcgtcgggcg
     4261 acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc
     4321 atcactaggg gttcctgcta gactgcagag ggccctgcgt atgagtgcaa gtgggtttta
     4381 ggaccaggat gaggcggggt gggggtgcct acctgacgac cgaccccgac ccactggaca
     4441 agcacccaac ccccattccc caaattgcgc atcccctatc agagaggggg aggggaaaca
     4501 ggatgcggcg aggcgcgtgc gc
//
