LOCUS       sequence-238426-        7213 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV plasmid with human synapsin promoter driving synaptophysin fused
            to GFP, iLID(V416I) and BoNT/B amino acids 1-146. Co-express with 
            SSPB-BoNT(147-441, Y365A) for vPA-BoNT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7213)
  AUTHORS   Liu Q, Sinnen BL, Boxer EE, Schneider MW, Grybko MJ, Buchta WC, 
            Gibson ES, Wysoczynski CL, Ford CP, Gottschalk A, Aoto J, Tucker CL,
            Kennedy MJ
  TITLE     A Photoactivatable Botulinum Neurotoxin for Inducible Control of 
            Neurotransmission.
  JOURNAL   Neuron. 2019 Jan 15. pii: S0896-6273(19)30003-0. doi: 
            10.1016/j.neuron.2019.01.002.
  PUBMED    30704911
REFERENCE   2  (bases 1 to 7213)
  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: "Neuron. 
            2019 Jan 15. pii: S0896-6273(19)30003-0. doi: 
            10.1016/j.neuron.2019.01.002."
COMMENT     Sequence Label: pAAV hSyn syph-GFP-myc-BoNT/B(1-146)-iLID
FEATURES             Location/Qualifiers
     source          1..7213
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   48..177
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        222..669
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      695..704
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1667..1669,1670..1672,1673..2383)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1667..1669,1670..1672,1673..2383)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1712..1733)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1973..1992)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2320..2341
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2384..2413
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2924..3361
                     /codon_start=1
                     /product="improved light-inducible dimer when used with 
                     SspB nano or SspB micro (Guntas et al., 2015)"
                     /label=iLID
                     /translation="LATTLERIEKNFIITDPRLPDNPIIFASDSFLQLTEYSREEILGR
                     NCRFLQGPETDRATVRKIRDAIDNQTEVTVQLINYTKSGKKFWNVFHLQPMRDYKGDVQ
                     YFIGVQLDGTERLHGAAEREAVCLIKKTAFQIAEAANDENYF"
     misc_feature    3387..3975
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3440..3460)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3858..3869)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4007..4483
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4206..4225)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4523..4663
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4523..4652
                     /label=AAV2 ITR
     rep_origin      4738..5193
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4825..4844)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5035..5056
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5210..5229)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5329..5351
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5389..5407)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5475..5579
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5580..5648,5649..6440)
                     /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(5798..5817)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6611..7199
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7100..7119
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtcct gcaggcagct
       61 gcgcgctcgc tcgctcactg aggccgcccg ggcgtcgggc gacctttggt cgcccggcct
      121 cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg ggttcctgcg
      181 gccgcacgcg tgtgtctaga ctgcagaggg ccctgcgtat gagtgcaagt gggttttagg
      241 accaggatga ggcggggtgg gggtgcctac ctgacgaccg accccgaccc actggacaag
      301 cacccaaccc ccattcccca aattgcgcat cccctatcag agagggggag gggaaacagg
      361 atgcggcgag gcgcgtgcgc actgccagct tcagcaccgc ggacagtgcc ttcgcccccg
      421 cctggcggcg cgcgccaccg ccgcctcagc actgaaggcg cgctgacgtc actcgccggt
      481 cccccgcaaa ctccccttcc cggccacctt ggtcgcgtcc gcgccgccgc cggcccagcc
      541 ggaccgcacc acgcgaggcg cgagataggg gggcacgggc gcgaccatct gcgctgcggc
      601 gccggcgact cagcgctgcc tcagtctgcg gtgggcagcg gaggagtcgt gtcgtgcctg
      661 agagcgcagt cgagaaggta ccggatctgc tagcgccacc atggacgtgg tgaatcagct
      721 ggtggctggg ggtcagttcc gggtggtcaa ggagcccctt ggcttcgtga aggtgctgca
      781 gtgggtcttt gccatcttcg cctttgctac gtgtggcagc tacaccgggg agcttcggct
      841 gagcgtggag tgtgccaaca agacggagag tgccctcaac atcgaagttg aattcgagta
      901 ccccttcagg ctgcaccaag tgtactttga tgcaccctcc tgcgtcaaag ggggcactac
      961 caagatcttc ctggttgggg actactcctc gtcggctgaa ttctttgtca ccgtggctgt
     1021 gtttgccttc ctctactcca tgggggccct ggccacctac atcttcctgc agaacaagta
     1081 ccgagagaac aacaaagggc ctatgatgga ctttctggct acagccgtgt tcgctttcat
     1141 gtggctagtt agttcatcag cctgggccaa aggcctgtcc gatgtgaaga tggccacgga
     1201 cccagagaac attatcaagg agatgcccat gtgccgccag acagggaaca catgcaagga
     1261 actgagggac cctgtgactt caggactcaa cacctcagtg gtgtttggct tcctgaacct
     1321 ggtgctctgg gttggcaact tatggttcgt gttcaaggag acaggctggg cagccccatt
     1381 catgcgcgca cctccaggcg ccccggaaaa gcaaccagca cctggcgatg cctacggcga
     1441 tgcgggctac gggcagggcc ccggaggcta tgggccccag gactcctacg ggcctcaggg
     1501 tggttatcaa cccgattacg ggcagccagc cagcggtggc ggtggctacg ggcctcaggg
     1561 cgactatggg cagcaaggct atggccaaca gggtgcgccc acctccttct ccaatcagat
     1621 gggatccaaa ggggttgagc ctaaaactta ttgttattat tcttctatgg tgagcaaggg
     1681 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     1741 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     1801 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     1861 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
     1921 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     1981 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     2041 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     2101 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
     2161 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     2221 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
     2281 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     2341 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aaggaacaaa aactcatctc
     2401 agaagaggat ctgggtcgac ctcgagccac catgccagtt acaataaata attttaatta
     2461 taatgatcct attgataata ataatattat tatgatggag cctccatttg cgagaggtac
     2521 ggggagatat tataaagctt ttaaaatcac agatcgtatt tggataatac cggaaagata
     2581 tacttttgga tataaacctg aggattttaa taaaagttcc ggtattttta atagagatgt
     2641 ttgtgaatat tatgatccag attacttaaa tactaatgat aaaaagaata tatttttaca
     2701 aacaatgatc aagttattta atagaatcaa atcaaaacca ttgggtgaaa agttattaga
     2761 gatgattata aatggtatac cttatcttgg agatagacgt gttccactcg aagagtttaa
     2821 cacaaacatt gctagtgtaa ctgttaataa attaatcagt aatccaggag gcggccgcag
     2881 cggtggcggt ggctctggag gtgggtccga gctcggggag tttctggcaa ccacactgga
     2941 acggatcgag aaaaatttca tcattactga tccgagactg cctgacaacc caatcatttt
     3001 tgcgagcgat tccttcctgc agctgacaga atattctcgg gaagagatcc tggggcgcaa
     3061 ttgccgtttt ctgcagggac ccgagacaga ccgtgccact gttcggaaaa tcagagatgc
     3121 tattgacaac cagactgaag tgaccgttca gctgatcaat tataccaaga gcggcaagaa
     3181 gttctggaac gtgttccacc tgcagccgat gcgcgattat aagggcgacg tccagtactt
     3241 cattggcgtg cagctggatg gcaccgaacg tcttcatggc gccgctgagc gtgaggcggt
     3301 ctgcctgatc aaaaagacag cctttcagat tgctgaggca gcgaacgacg aaaattactt
     3361 ttgaattcga tatcaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat
     3421 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     3481 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     3541 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     3601 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     3661 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     3721 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     3781 aatcatcgtc ctttccttgg ctgctcgcct atgttgccac ctggattctg cgcgggacgt
     3841 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     3901 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     3961 gggccgcctc cccgcatcga taccgagcgc tgctcgagag atctacgggt ggcatccctg
     4021 tgacccctcc ccagtgcctc tcctggccct ggaagttgcc actccagtgc ccaccagcct
     4081 tgtcctaata aaattaagtt gcatcatttt gtctgactag gtgtccttct ataatattat
     4141 ggggtggagg ggggtggtat ggagcaaggg gcaagttggg aagacaacct gtagggcctg
     4201 cggggtctat tgggaaccaa gctggagtgc agtggcacaa tcttggctca ctgcaatctc
     4261 cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc gagttgttgg gattccaggc
     4321 atgcatgacc aggctcagct aatttttgtt tttttggtag agacggggtt tcaccatatt
     4381 ggccaggctg gtctccaact cctaatctca ggtgatctac ccaccttggc ctcccaaatt
     4441 gctgggatta caggcgtgaa ccactgctcc cttccctgtc cttctgattt tgtaggtaac
     4501 cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg
     4561 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc
     4621 cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat
     4681 tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg
     4741 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     4801 cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     4861 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     4921 ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat
     4981 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     5041 tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt gatttataag
     5101 ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     5161 cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct
     5221 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     5281 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     5341 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     5401 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     5461 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     5521 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     5581 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     5641 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     5701 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     5761 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     5821 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     5881 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     5941 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     6001 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     6061 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     6121 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     6181 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     6241 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     6301 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     6361 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     6421 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     6481 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     6541 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     6601 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     6661 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     6721 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     6781 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     6841 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     6901 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     6961 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     7021 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     7081 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     7141 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     7201 acgccagcaa cgc
//
