LOCUS       sequence-354854-        5369 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector for Cre-dependent transgene expression 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 5369)
  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 5369)
  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-V5-APEX2-NES
FEATURES             Location/Qualifiers
     source          1..5369
                     /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    complement(629..662)
                     /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(713..746)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(784..810)
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             complement(1570..1611)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     regulatory      1611..1620
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    1684..1717
                     /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    1768..1801
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    1810..2398
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1863..1883)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2281..2292)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2406..2630
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2632..2772
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2632..2761
                     /label=AAV2 ITR
     rep_origin      2847..3302
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2934..2953)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3144..3165
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3319..3338)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3438..3460
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3498..3516)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3584..3688
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3689..3757,3758..4549)
                     /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(3907..3926)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4720..5308
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5209..5228
                     /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 actccggaat aacttcgtat aggatacttt atacgaagtt
      661 atgcagaatg gtagctggat tgtagctgct attagcaata tgaaacctct taataacttc
      721 gtatagcata cattatacga agttataacg cgccactagt gaattcgagc tcggtaccta
      781 gtccagggtc aggcgctcca ggggtggcag ctgcagggcg tcggcaaatc ccagttctga
      841 cagcttctga tgtgcttctg cgtaatctgc gaaaaaagca tcctcgtcgg ctgcatactt
      901 atcgaccaga ggtctgaaca cgggatcgga cagcagggct ttgtcgctgg gcagctgcag
      961 cagcccttcc ttctctccac tcagcagttc agtaaagtaa gaattgtcga aaatcagggg
     1021 gttgcttgtc cagggtccct cgaagcctga cctttcctta tgagcggcgc caatggtgtg
     1081 ccctccagac agtgcgacga tatcctggtc agtcaggccc atagctttcc caaacacatc
     1141 tcgcagatgg tcggagccct ttgtcggatc tggcaggcgc ccctctggag gtggttctgg
     1201 tttgtcctcc cgtcctgggt ggaaaggcac cttggggccc ccagtgacct ccacagcgac
     1261 cactcctgcc agctgataaa aatcggcgta ggacagaatt gggaactctg ctttcagagg
     1321 ttccagcagc cgcacggcga tgtccagccc attgttggca ctatgtgcca gttcagcagg
     1381 gtgcttgatt gttccaaagg gtccgcctgt tttggttccc ttgtcaaagg tgcctgcgct
     1441 gtggaaagcc agtctcagca tcagaggtgc gcacctcttt tctgcgataa accctctcag
     1501 cttcttcttg gctttctcca cggcatcctg gtagtctgcg gacactgttg ggtatgattt
     1561 cccaccggtg gtgctgtcca ggcccagcag ggggttgggg atgggcttgc ccatggtggc
     1621 gtacccgggg atcctctaga gtcgacctgc aggcatgcaa gctttactag tggtggcgct
     1681 agcataactt cgtataaagt atcctatacg aagttatttg ccttaaccca gaaattatca
     1741 ctgttattct ttagaatggt gcaaagaata acttcgtata atgtatgcta tacgaagtta
     1801 tggcgcgcca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     1861 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     1921 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     1981 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     2041 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     2101 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     2161 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca
     2221 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     2281 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     2341 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct
     2401 ggatcctgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     2461 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     2521 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     2581 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg caggaacccc
     2641 tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac
     2701 caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca
     2761 gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     2821 accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg
     2881 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccttagcgc ccgctccttt
     2941 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     3001 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     3061 tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     3121 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaactc
     3181 tatctcgggc tattcttttg atttataagg gattttgccg atttcggtct attggttaaa
     3241 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat
     3301 tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
     3361 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     3421 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     3481 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     3541 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     3601 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     3661 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     3721 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     3781 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     3841 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     3901 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     3961 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     4021 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     4081 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     4141 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     4201 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     4261 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     4321 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     4381 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     4441 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     4501 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     4561 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     4621 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     4681 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     4741 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     4801 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     4861 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     4921 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     4981 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     5041 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     5101 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     5161 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     5221 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     5281 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     5341 ggccttttgc tggccttttg ctcacatgt
//
