LOCUS       sequence-76818-e        6027 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV2 transfer vector containing the InSynC (miniSOG-VAMP2-citrine) 
            construct.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6027)
  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 6027)
  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-miniSOG-VAMP2-citrine
FEATURES             Location/Qualifiers
     source          1..6027
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    228..249
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(264..269,270..287,288..294)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    302..318
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     307..329
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     326..342
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     326..342
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   380..509
                     /label=AAV2 ITR
     promoter        591..1037
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             1067..1384
                     /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             join(1799..1801,1802..1804,1805..2515)
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1844..1865)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2105..2124)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2452..2473
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2536..3124
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2589..2609)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3007..3018)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3127..3143)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3128..3144)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3173..3294
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3184..3203)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3238..3257
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3456..3585
                     /label=AAV2 ITR
     primer_bind     complement(3604..3621)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3604..3620)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3613..3635)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3762..4217
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3849..3868)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4059..4080
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4243..4347
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4348..4416,4417..5208)
                     /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(4566..4585)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5379..5967
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5868..5887
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
       61 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
      121 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
      181 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
      241 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
      301 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccagattt
      361 aattaaggcc ttaattaggc tgcgcgctcg ctcgctcact gaggccgccc gggcaaagcc
      421 cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg
      481 agtggccaac tccatcacta ggggttcctt gtagttaatg attaacccgc catgctactt
      541 atctacgtag ccatgctcta ggaagatctc tgcagagggc cctgcgtatg agtgcaagtg
      601 ggttttagga ccaggatgag gcggggtggg ggtgcctacc tgacgaccga ccccacccac
      661 tggacaagca cccaaccccc attccccaaa ttgcgcatcc cctatcagag agggggaggg
      721 gaaacaggat gcggcgaggc gcgtgcgcac tgccagcttc agcaccgcgg acagtgcctt
      781 cgcccccgcc tggcggcgcg cgccaccgcc gcctcagcac tgaaggcgcg ctgacgtcac
      841 tcgccggtcc cccgcaaact ccccttcccg gccaccttgg tcgcgtccgc gccgccgccg
      901 gcccagccgg accgcaccac gcgaggcgcg agataggggg gcacgggcgc gaccatctgc
      961 gctgcggcgc cggcgactca gcgctgcctc agtctgcggt gggcagcgga ggagtcgtgt
     1021 cgtgcctgag agcgcagtcg aattcaagct gctagcaagg atcctcatgg agaaaagttt
     1081 cgtgataact gatccacggc tgccagacaa tcccatcatc ttcgcatccg atggcttcct
     1141 ggagctgacc gagtattcca gagaggagat cctgggccgc aatggccgct ttctgcaggg
     1201 accagagaca gaccaggcca cagtgcagaa gattcgcgat gccattagag atcagcgcga
     1261 gattaccgtg cagctgataa actacacaaa aagcgggaag aaattctgga acctcctgca
     1321 cctccagccc atgagggacc agaagggtga gctccagtat ttcatcggag tgcagctgga
     1381 tggatcagga ggaggctcag gaggaggagg cgcgcctacc ggtatgtcgg ctaccgctgc
     1441 caccgtcccg cctgccgccc cggccggcga gggtggcccc cctgcacctc ctccaaacct
     1501 tactagtaac aggagactgc agcagaccca ggcccaggtg gatgaggtgg tggacatcat
     1561 gagggtgaat gtggacaagg tcctggagcg ggaccagaag ttgtcggagc tggatgaccg
     1621 tgcagatgcc ctccaggcag gggcctccca gtttgaaaca agtgcagcca agctcaagcg
     1681 caaatactgg tggaaaaacc tcaagatgat gatcatcttg ggagtgatct gcgccatcat
     1741 cctcatcatc atcatcgttt acttcagcac tagcggcgga agcggcgggg gaatcgatat
     1801 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     1861 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     1921 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     1981 cgtgaccacc ttcggctacg gcctgatgtg cttcgcccgc taccccgacc acatgaagca
     2041 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     2101 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     2161 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     2221 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     2281 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     2341 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     2401 cctgagctac cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     2461 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtagaa
     2521 ttcaagctta tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     2581 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     2641 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     2701 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     2761 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     2821 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     2881 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc
     2941 tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     3001 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     3061 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     3121 ccgcatcgat accgtcgacc tcgacccggg cggccgcttc gagcagacat gataagatac
     3181 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     3241 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     3301 aacaattgca ttcattttat gtttcaggtt cagggggaga tgtgggaggt tttttaaagc
     3361 aagtaaaacc tctacaaatg tggtaaaatc gataaggatc ttcctagagc atggctacgt
     3421 agataagtag catggcgggt taatcattaa ctacaaggaa cccctagtga tggagttggc
     3481 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
     3541 cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagcctta attaacctaa
     3601 ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
     3661 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
     3721 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc
     3781 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     3841 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     3901 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     3961 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     4021 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4081 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     4141 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     4201 attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     4261 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     4321 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     4381 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     4441 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     4501 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     4561 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     4621 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     4681 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     4741 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     4801 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     4861 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     4921 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     4981 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     5041 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     5101 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     5161 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     5221 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     5281 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     5341 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     5401 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     5461 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     5521 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     5581 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     5641 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     5701 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     5761 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     5821 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     5881 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     5941 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     6001 gccttttgct ggccttttgc tcacatg
//
