LOCUS       Exported                6072 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  AAV2 transfer vector containing the InSynC 
            (miniSOG-VAMP2-T2A-mCherry) construct.
ACCESSION   .
VERSION     .
KEYWORDS    AAV2-miniSOG-VAMP2-T2A-mCherry
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6072)
  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 6072)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6072
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     94..111
                     /label=L4440
                     /note="L4440 vector, forward primer"
     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        264..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..1038
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             1069..1386
                     /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             1786..1839
                     /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             1846..2556
                     /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(1991..2009)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2259..2278
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2581..3169
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2634..2654)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3052..3063)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3172..3188)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3173..3189)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3218..3339
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3229..3248)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3283..3302
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3501..3630
                     /label=AAV2 ITR
     primer_bind     complement(3649..3666)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3649..3665)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3658..3680)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3807..4262
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3894..3913)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4104..4125
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4288..4392
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4393..5253
                     /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(4611..4630)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5424..6012
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5913..5932
                     /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 ccccgaccca
      661 ctggacaagc acccaacccc cattccccaa attgcgcatc ccctatcaga gagggggagg
      721 ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg gacagtgcct
      781 tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc gctgacgtca
      841 ctcgccggtc ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg cgccgccgcc
      901 ggcccagccg gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg cgaccatctg
      961 cgctgcggcg ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg aggagtcgtg
     1021 tcgtgcctga gagcgcagtc gaattcaagc tgctagcaag gatccggtat ggagaaaagt
     1081 ttcgtgataa ctgatccacg gctgccagac aatcccatca tcttcgcatc cgatggcttc
     1141 ctggagctga ccgagtattc cagagaggag atcctgggcc gcaatggccg ctttctgcag
     1201 ggaccagaga cagaccaggc cacagtgcag aagattcgcg atgccattag agatcagcgc
     1261 gagattaccg tgcagctgat aaactacaca aaaagcggga agaaattctg gaacctcctg
     1321 cacctccagc ccatgaggga ccagaagggt gagctccagt atttcatcgg agtgcagctg
     1381 gatggatcag gaggaggctc aggaggagga atgtcggcta ccgctgccac cgtcccgcct
     1441 gccgccccgg ccggcgaggg tggcccccct gcacctcctc caaaccttac tagtaacagg
     1501 agactgcagc agacccaggc ccaggtggat gaggtggtgg acatcatgag ggtgaatgtg
     1561 gacaaggtcc tggagcggga ccagaagttg tcggagctgg atgaccgtgc agatgccctc
     1621 caggcagggg cctcccagtt tgaaacaagt gcagccaagc tcaagcgcaa atactggtgg
     1681 aaaaacctca agatgatgat catcttggga gtgatctgcg ccatcatcct catcatcatc
     1741 atcgtttact tcagcactgc tagcggaggt ggatctggtg gcggtgaggg cagaggaagt
     1801 cttctaacat gcggtgacgt ggaggagaat cccggccctc gtacgatggt gagcaagggc
     1861 gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc
     1921 tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc
     1981 acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc
     2041 ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc
     2101 gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag
     2161 gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac
     2221 aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc
     2281 atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag
     2341 atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc
     2401 tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac
     2461 atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc
     2521 cactccaccg gcggcatgga cgagctgtac aagtaagaat tcgatatcaa gcttatcgat
     2581 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     2641 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     2701 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     2761 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     2821 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     2881 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     2941 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc
     3001 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     3061 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     3121 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgataccgt
     3181 cgacctcgac ccgggcggcc gcttcgagca gacatgataa gatacattga tgagtttgga
     3241 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     3301 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     3361 tttatgtttc aggttcaggg ggagatgtgg gaggtttttt aaagcaagta aaacctctac
     3421 aaatgtggta aaatcgataa ggatcttcct agagcatggc tacgtagata agtagcatgg
     3481 cgggttaatc attaactaca aggaacccct agtgatggag ttggccactc cctctctgcg
     3541 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg
     3601 ggcggcctca gtgagcgagc gagcgcgcag ccttaattaa cctaattcac tggccgtcgt
     3661 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
     3721 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     3781 gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg
     3841 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     3901 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     3961 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     4021 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     4081 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     4141 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     4201 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat
     4261 ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     4321 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     4381 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     4441 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     4501 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     4561 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     4621 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     4681 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     4741 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     4801 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     4861 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     4921 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     4981 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     5041 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     5101 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     5161 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     5221 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     5281 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     5341 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     5401 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     5461 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     5521 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     5581 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     5641 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     5701 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     5761 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     5821 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     5881 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     5941 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     6001 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     6061 tttgctcaca tg
//
