LOCUS       Exported                6599 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  AAV2 transfer vector containing the InSynC (SYP1-miniSOG-citrine) 
            construct.
ACCESSION   .
VERSION     .
KEYWORDS    AAV-SYP1-miniSOG-citrine
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6599)
  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 6599)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6599
                     /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..1037
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             2019..2336
                     /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             2376..3092
                     /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(2421..2442)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2682..2701)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3029..3050
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3108..3696
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3161..3181)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3579..3590)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3699..3715)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3700..3716)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3745..3866
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3756..3775)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3810..3829
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4028..4157
                     /label=AAV2 ITR
     primer_bind     complement(4176..4193)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4176..4192)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4185..4207)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4334..4789
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4421..4440)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4631..4652
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4815..4919
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4920..5780
                     /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(5138..5157)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5951..6539
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6440..6459
                     /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 atccatggac gtggtgaatc
     1081 agctggtggc tgggggtcag ttccgggtgg tcaaggagcc ccttggcttc gtgaaggtgc
     1141 tgcagtgggt ctttgccatc ttcgcctttg ctacgtgtgg cagctacacc ggggagcttc
     1201 ggctgagcgt ggagtgtgcc aacaagacgg agagtgccct caacatcgaa gttgaattcg
     1261 agtacccctt caggctgcac caagtgtact ttgatgcacc ctcctgcgtc aaagggggca
     1321 ctaccaagat cttcctggtt ggggactact cctcgtcggc tgaattcttt gtcaccgtgg
     1381 ctgtgtttgc cttcctctac tccatggggg ccctggccac ctacatcttc ctgcagaaca
     1441 agtaccgaga gaacaacaaa gggcctatga tggactttct ggctacagcc gtgttcgctt
     1501 tcatgtggct agttagttca tcagcctggg ccaaaggcct gtccgatgtg aagatggcca
     1561 cggacccaga gaacattatc aaggagatgc ccatgtgccg ccagacaggg aacacatgca
     1621 aggaactgag ggaccctgtg acttcaggac tcaacacctc agtggtgttt ggcttcctga
     1681 acctggtgct ctgggttggc aacttatggt tcgtgttcaa ggagacaggc tgggcagccc
     1741 cattcatgcg cgcacctcca ggcgccccgg aaaagcaacc agcacctggc gatgcctacg
     1801 gcgatgcggg ctacgggcag ggccccggag gctatgggcc ccaggactcc tacgggcctc
     1861 agggtggtta tcaacccgat tacgggcagc cagccagcgg tggcggtggc tacgggcctc
     1921 agggcgacta tgggcagcaa ggctatggcc aacagggtgc gcccacctcc ttctccaatc
     1981 agatgaaaac cggtggtggc ggcagtggtg gcggcagcat ggagaaaagt ttcgtgataa
     2041 ctgatccacg gctgccagac aatcccatca tcttcgcatc cgatggcttc ctggagctga
     2101 ccgagtattc cagagaggag atcctgggcc gcaatggccg ctttctgcag ggaccagaga
     2161 cagaccaggc cacagtgcag aagattcgcg atgccattag agatcagcgc gagattaccg
     2221 tgcagctgat aaactacaca aaaagcggga agaaattctg gaacctcctg cacctccagc
     2281 ccatgaggga ccagaagggt gagctccagt atttcatcgg agtgcagctg gatggatcag
     2341 gaggaggctc aggaggagga ggcgcgggaa tcgatatggt gagcaagggc gaggagctgt
     2401 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     2461 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct
     2521 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccttc ggctacggcc
     2581 tgatgtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     2641 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     2701 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     2761 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
     2821 acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc
     2881 gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
     2941 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagctaccag tccgccctga
     3001 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     3061 ggatcactct cggcatggac gagctgtaca agtagaagct tatcgataat caacctctgg
     3121 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     3181 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     3241 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
     3301 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
     3361 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
     3421 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     3481 attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca
     3541 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
     3601 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
     3661 agacgagtcg gatctccctt tgggccgcct ccccgcatcg ataccgtcga cctcgacccg
     3721 ggcggccgct tcgagcagac atgataagat acattgatga gtttggacaa accacaacta
     3781 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     3841 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
     3901 ttcaggggga gatgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtaaaa
     3961 tcgataagga tcttcctaga gcatggctac gtagataagt agcatggcgg gttaatcatt
     4021 aactacaagg aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc
     4081 actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg
     4141 agcgagcgag cgcgcagcct taattaacct aattcactgg ccgtcgtttt acaacgtcgt
     4201 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
     4261 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     4321 aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     4381 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     4441 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     4501 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     4561 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     4621 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     4681 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     4741 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt
     4801 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     4861 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     4921 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     4981 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     5041 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     5101 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     5161 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     5221 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     5281 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     5341 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     5401 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     5461 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     5521 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     5581 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     5641 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     5701 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     5761 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     5821 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     5881 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     5941 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     6001 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     6061 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     6121 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     6181 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     6241 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     6301 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     6361 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     6421 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     6481 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     6541 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatg
//
