LOCUS       sequence-355038-        6308 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector for Cre- and Flp-dependent transgene expression (CRE-ON /
            FLP-ON) of V5-APEX2-NES in neurons under the control of the hSyn1 
            promoter (Kugler et al 2003).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6308)
  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 6308)
  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-VTKS4-V5-APEX2-NES
FEATURES             Location/Qualifiers
     source          1..6308
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        158..605
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    629..662
                     /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    708..756
                     /note="synthetic polyadenylation signal"
     polyA_signal    765..1241
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(964..983)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(1276..1295)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1320..1368
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     protein_bind    1528..1561
                     /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(1568..1601)
                     /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(1652..1685)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1723..1749)
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             complement(2509..2550)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     regulatory      2550..2559
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2623..2656
                     /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    2707..2740
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2749..3337
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2802..2822)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3220..3231)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3345..3569
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3571..3711
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3571..3700
                     /label=AAV2 ITR
     rep_origin      3786..4241
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3873..3892)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4083..4104
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4258..4277)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4377..4399
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4437..4455)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4523..4627
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4628..4696,4697..5488)
                     /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(4846..4865)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5659..6247
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6148..6167
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gctagactgc agagggccct gcgtatgagt gcaagtgggt tttaggacca
      181 ggatgaggcg gggtgggggt gcctacctga cgaccgaccc cgacccactg gacaagcacc
      241 caacccccat tccccaaatt gcgcatcccc tatcagagag ggggagggga aacaggatgc
      301 ggcgaggcgc gtgcgcactg ccagcttcag caccgcggac agtgccttcg cccccgcctg
      361 gcggcgcgcg ccaccgccgc ctcagcactg aaggcgcgct gacgtcactc gccggtcccc
      421 cgcaaactcc ccttcccggc caccttggtc gcgtccgcgc cgccgccggc ccagccggac
      481 cgcaccacgc gaggcgcgag ataggggggc acgggcgcga ccatctgcgc tgcggcgccg
      541 gcgactcagc gctgcctcag tctgcggtgg gcagcggagg agtcgtgtcg tgcctgagag
      601 cgcagtcaga tatcgtttaa actccggaga agttcctatt ctctagaaag tataggaact
      661 tccctcagca ccatggctag cggcagcctc ggagtttgaa tagatagaat aaaatatctt
      721 tattttcatt ccatctgtgt gttggttttt tgtgtgagat ctacgggtgg catccctgtg
      781 acccctcccc agtgcctctc ctggccctgg aagttgccac tccagtgccc accagccttg
      841 tcctaataaa attaagttgc atcattttgt ctgactaggt gtccttctat aatattatgg
      901 ggtggagggg ggtggtatgg agcaaggggc aagttgggaa gacaacctgt agggcctgcg
      961 gggtctattg ggaaccaagc tggagtgcag tggcacaatc ttggctcact gcaatctccg
     1021 cctcctgggt tcaagcgatt ctcctgcctc aggctcccga gttgttggga ttccaggcat
     1081 gcatgaccag gctcagctaa tttttgtttt tttggtagag acggggtttc accatattgg
     1141 ccaggctggt ctccaactcc taatctcagg tgatctaccc accttggcct cccaaattgc
     1201 tgggattaca ggcgtgaacc actgctccct tccctgtcct tctgcctcag ctaattgagt
     1261 aggggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac
     1321 ggcaataaaa agacagaata aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg
     1381 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     1441 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     1501 cagccaacgt cggggcggca ggccgtggaa gttcctattc tctagaaagt ataggaactt
     1561 caccggaata acttcgtata ggatacttta tacgaagtta tgcagaatgg tagctggatt
     1621 gtagctgcta ttagcaatat gaaacctctt aataacttcg tatagcatac attatacgaa
     1681 gttataacgc gccactagtg aattcgagct cggtacctag tccagggtca ggcgctccag
     1741 gggtggcagc tgcagggcgt cggcaaatcc cagttctgac agcttctgat gtgcttctgc
     1801 gtaatctgcg aaaaaagcat cctcgtcggc tgcatactta tcgaccagag gtctgaacac
     1861 gggatcggac agcagggctt tgtcgctggg cagctgcagc agcccttcct tctctccact
     1921 cagcagttca gtaaagtaag aattgtcgaa aatcaggggg ttgcttgtcc agggtccctc
     1981 gaagcctgac ctttccttat gagcggcgcc aatggtgtgc cctccagaca gtgcgacgat
     2041 atcctggtca gtcaggccca tagctttccc aaacacatct cgcagatggt cggagccctt
     2101 tgtcggatct ggcaggcgcc cctctggagg tggttctggt ttgtcctccc gtcctgggtg
     2161 gaaaggcacc ttggggcccc cagtgacctc cacagcgacc actcctgcca gctgataaaa
     2221 atcggcgtag gacagaattg ggaactctgc tttcagaggt tccagcagcc gcacggcgat
     2281 gtccagccca ttgttggcac tatgtgccag ttcagcaggg tgcttgattg ttccaaaggg
     2341 tccgcctgtt ttggttccct tgtcaaaggt gcctgcgctg tggaaagcca gtctcagcat
     2401 cagaggtgcg cacctctttt ctgcgataaa ccctctcagc ttcttcttgg ctttctccac
     2461 ggcatcctgg tagtctgcgg acactgttgg gtatgatttc ccaccggtgg tgctgtccag
     2521 gcccagcagg gggttgggga tgggcttgcc catggtggcg tacccgggga tcctctagag
     2581 tcgacctgca ggcatgcaag ctttactagt ggtggcgcta gcataacttc gtataaagta
     2641 tcctatacga agttatttgc cttaacccag aaattatcac tgttattctt tagaatggtg
     2701 caaagaataa cttcgtataa tgtatgctat acgaagttat ggcgcgccaa tcaacctctg
     2761 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     2821 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     2881 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     2941 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     3001 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     3061 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     3121 aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc
     3181 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     3241 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     3301 cagacgagtc ggatctccct ttgggccgcc tccccgcctg gatcctgtgc cttctagttg
     3361 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     3421 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     3481 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     3541 gcatgctggg gatgcggtgg gctctatggc aggaacccct agtgatggag ttggccactc
     3601 cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg
     3661 gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga
     3721 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc
     3781 atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     3841 gaccgctaca cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct
     3901 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     3961 atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag
     4021 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     4081 tagtggactc ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga
     4141 tttataaggg attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa
     4201 atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac
     4261 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     4321 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     4381 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
     4441 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     4501 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     4561 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     4621 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     4681 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     4741 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     4801 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     4861 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     4921 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     4981 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     5041 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     5101 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     5161 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     5221 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     5281 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     5341 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     5401 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     5461 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     5521 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     5581 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     5641 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     5701 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     5761 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     5821 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     5881 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     5941 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     6001 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     6061 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     6121 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     6181 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     6241 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     6301 tcacatgt
//
