LOCUS       sequence-476707-        6361 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV2 transfer vector containing the InSynC 
            (SYP1-miniSOG-T2A-mCherry) construct.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6361)
  AUTHORS   Lin JY, Sann SB, Zhou K, Nabavi S, Proulx CD, Malinow R, Jin Y, 
            Tsien RY
  TITLE     Optogenetic inhibition of synaptic release with chromophore-assisted
            light inactivation (CALI).
  JOURNAL   Neuron. 2013 Jul 24;79(2):241-53. doi: 10.1016/j.neuron.2013.05.022.
  PUBMED    23889931
REFERENCE   2  (bases 1 to 6361)
  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"; doi: 
            "10.1016/j.neuron.2013.05"; journalName: "Neuron"; date: 
            "2013-07-24- 24"; volume: "79"; issue: "2"; pages: "241-53"
COMMENT     Sequence Label: AAV-SYP1-miniSOG-T2A-mCherry
FEATURES             Location/Qualifiers
     source          1..6361
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   120..249
                     /label=AAV2 ITR
     promoter        331..778
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             1778..2095
                     /codon_start=1
                     /product="mini singlet oxygen generator, a fluorescent 
                     flavoprotein suitable for correlated light and electron 
                     microscopy (Shu et al., 2011)"
                     /label=miniSOG
                     /translation="MEKSFVITDPRLPDNPIIFASDGFLELTEYSREEILGRNGRFLQG
                     PETDQATVQKIRDAIRDQREITVQLINYTKSGKKFWNLLHLQPMRDQKGELQYFIGVQL
                     DG"
     CDS             2123..2176
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             2183..2893
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2328..2346)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2596..2615
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2918..3506
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2971..2991)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3389..3400)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3509..3525)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3510..3526)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3555..3676
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3566..3585)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3620..3639
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3838..3967
                     /label=AAV2 ITR
     rep_origin      complement(4204..4792)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4284..4303)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4963..5754,5755..5823))
                     /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     5586..5605
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5824..5928)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(join(5954..6361,1..48))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6091..6112)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6303..6322
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat
       61 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
      121 tgcgcgctcg ctcgctcact gaggccgccc gggcaaagcc cgggcgtcgg gcgacctttg
      181 gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg agtggccaac tccatcacta
      241 ggggttcctt gtagttaatg attaacccgc catgctactt atctacgtag ccatgctcta
      301 ggaagatctc tgcagagggc cctgcgtatg agtgcaagtg ggttttagga ccaggatgag
      361 gcggggtggg ggtgcctacc tgacgaccga ccccgaccca ctggacaagc acccaacccc
      421 cattccccaa attgcgcatc ccctatcaga gagggggagg ggaaacagga tgcggcgagg
      481 cgcgtgcgca ctgccagctt cagcaccgcg gacagtgcct tcgcccccgc ctggcggcgc
      541 gcgccaccgc cgcctcagca ctgaaggcgc gctgacgtca ctcgccggtc ccccgcaaac
      601 tccccttccc ggccaccttg gtcgcgtccg cgccgccgcc ggcccagccg gaccgcacca
      661 cgcgaggcgc gagatagggg ggcacgggcg cgaccatctg cgctgcggcg ccggcgactc
      721 agcgctgcct cagtctgcgg tgggcagcgg aggagtcgtg tcgtgcctga gagcgcagtc
      781 gaattcaagc tgctagcaag gatccgtgac cggcggctct agaatggacg tggtgaatca
      841 gctggtggct gggggtcagt tccgggtggt caaggagccc cttggcttcg tgaaggtgct
      901 gcagtgggtc tttgccatct tcgcctttgc tacgtgtggc agctacaccg gggagcttcg
      961 gctgagcgtg gagtgtgcca acaagacgga gagtgccctc aacatcgaag ttgaattcga
     1021 gtaccccttc aggctgcacc aagtgtactt tgatgcaccc tcctgcgtca aagggggcac
     1081 taccaagatc ttcctggttg gggactactc ctcgtcggct gaattctttg tcaccgtggc
     1141 tgtgtttgcc ttcctctact ccatgggggc cctggccacc tacatcttcc tgcagaacaa
     1201 gtaccgagag aacaacaaag ggcctatgat ggactttctg gctacagccg tgttcgcttt
     1261 catgtggcta gttagttcat cagcctgggc caaaggcctg tccgatgtga agatggccac
     1321 ggacccagag aacattatca aggagatgcc catgtgccgc cagacaggga acacatgcaa
     1381 ggaactgagg gaccctgtga cttcaggact caacacctca gtggtgtttg gcttcctgaa
     1441 cctggtgctc tgggttggca acttatggtt cgtgttcaag gagacaggct gggcagcccc
     1501 attcatgcgc gcacctccag gcgccccgga aaagcaacca gcacctggcg atgcctacgg
     1561 cgatgcgggc tacgggcagg gccccggagg ctatgggccc caggactcct acgggcctca
     1621 gggtggttat caacccgatt acgggcagcc agccagcggt ggcggtggct acgggcctca
     1681 gggcgactat gggcagcaag gctatggcca acagggtgcg cccacctcct tctccaatca
     1741 gatgaaaacc ggtggtggcg gcagtggtgg cggcagcatg gagaaaagtt tcgtgataac
     1801 tgatccacgg ctgccagaca atcccatcat cttcgcatcc gatggcttcc tggagctgac
     1861 cgagtattcc agagaggaga tcctgggccg caatggccgc tttctgcagg gaccagagac
     1921 agaccaggcc acagtgcaga agattcgcga tgccattaga gatcagcgcg agattaccgt
     1981 gcagctgata aactacacaa aaagcgggaa gaaattctgg aacctcctgc acctccagcc
     2041 catgagggac cagaagggtg agctccagta tttcatcgga gtgcagctgg atggagctag
     2101 cggaggtgga tctggtggcg gtgagggcag aggaagtctt ctaacatgcg gtgacgtgga
     2161 ggagaatccc ggccctcgta cgatggtgag caagggcgag gaggataaca tggccatcat
     2221 caaggagttc atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga
     2281 gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt
     2341 gaccaagggt ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg
     2401 ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc
     2461 cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac
     2521 ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa
     2581 cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga
     2641 gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa
     2701 ggacggcggc cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca
     2761 gctgcccggc gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta
     2821 caccatcgtg gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga
     2881 gctgtacaag taagaattcg atatcaagct tatcgataat caacctctgg attacaaaat
     2941 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     3001 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     3061 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     3121 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     3181 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     3241 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     3301 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
     3361 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     3421 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     3481 gatctccctt tgggccgcct ccccgcatcg ataccgtcga cctcgacccg ggcggccgct
     3541 tcgagcagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     3601 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     3661 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     3721 gatgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtaaaa tcgataagga
     3781 tcttcctaga gcatggctac gtagataagt agcatggcgg gttaatcatt aactacaagg
     3841 aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg
     3901 ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag
     3961 cgcgcagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     4021 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     4081 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     4141 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     4201 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     4261 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     4321 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     4381 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     4441 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     4501 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     4561 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     4621 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     4681 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     4741 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     4801 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     4861 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     4921 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     4981 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     5041 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     5101 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     5161 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     5221 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     5281 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     5341 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     5401 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     5461 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     5521 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     5581 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     5641 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     5701 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     5761 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     5821 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     5881 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     5941 aaaagtgcca cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     6001 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     6061 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     6121 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     6181 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     6241 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     6301 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     6361 c
//
