LOCUS       pAAV_hSyn_ChRger        6413 bp ds-DNA     circular     22-JAN-2019
DEFINITION  AAV expression of humanized ChR2 with H134R mutation fused to EYFP
            driven by human Synapsin I promoter for optogenetic activation.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-hSyn-hChR2(H134R)-EYFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6413)
  TITLE     hChR2 plasmids
REFERENCE   2  (bases 1 to 6413)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sunday, Aug 12, 2018 from SnapGene 3.3.4
            http://www.snapgene.com
COMMENT     
COMMENT     ApEinfo:methylated:1
FEATURES             Location/Qualifiers
     source          1..6413
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /label=source:synthetic DNA construct
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     repeat_region   1..141
                     /note="inverted terminal repeat of adeno-associated virus
                     serotype 2"
                     /label=AAV2 ITR
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     repeat_region   12..141
                     /label=AAV2 ITR(1)
                     /ApEinfo_label=AAV2 ITR
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     promoter        186..633
                     /note="human synapsin I promoter; confers neuron-specific
                     expression (Kugler et al., 2003)"
                     /label=hSyn promoter
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     CDS             659..1726
                     /codon_start=1
                     /translation="MSRLVAASWLLALLLCGITSTTTASAHIVMVDAYKPTKSAPAAS
                     S
                     TDGTAAAAVSHYAMNGFDELAKGAVVPEDHFVCGPADKCYCSAWLHSRGALWEQETAR
                     G
                     FQWFAVFLSALFLAFYGWHAYKASVGWEEVYVCSVELIKVILEIYFEFTSPAMLFLYG
                     G
                     NITPWLRYAEWLLTCPVILIHLSNITGLSEAYNKRTMALLVSDLGTICMGVTAALATG
                     W
                     VKWLFYCIGLVYGTQTFYNAGIIYVEAYHTVPKGRCRQVVTGMAWLFFVSWGMFPILF
                     I
                     LGPEGFGVLSVAGSTIGHTIADLLSKNIWGLLGHFLRIKIHEHIIMYGDIRRPVSSQF
                     L GRKVDVLAFVTEEDKV"
                     /label=ChRger2
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     misc_feature    1736..1795
                     /note="ApEinfo_fwdcolor=#00ff00
                     ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0 ApEinfo_revcolor=green"
                     /label=TS
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     CDS             1796..2512
                     /codon_start=1
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     K
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     G
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     V
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     E FVTAAGITLGMDELYK"
                     /label=EYFP (no start)
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     misc_feature    2540..3128
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
                     /label=WPRE
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     CDS             complement(3011..3022)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /translation="IEGR"
                     /label=Factor Xa site
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     polyA_signal    3160..3636
                     /note="human growth hormone polyadenylation signal"
                     /label=hGH poly(A) signal
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     repeat_region   3676..3816
                     /note="inverted terminal repeat of adeno-associated virus
                     serotype 2"
                     /label=AAV2 ITR(2)
                     /ApEinfo_label=AAV2 ITR
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     repeat_region   3676..3805
                     /label=AAV2 ITR(3)
                     /ApEinfo_label=AAV2 ITR
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     rep_origin      3891..4346
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label=f1 ori
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     promoter        4628..4732
                     /gene="bla"
                     /label=AmpR promoter
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     CDS             4733..5593
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="confers resistance to ampicillin, carbenicillin,
                     and related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     I
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     S
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     W
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     A
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     S LIKHW"
                     /label=AmpR
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     rep_origin      5764..6352
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label=ori
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtgtgtc tagactgcag agggccctgc
      181 gtatgagtgc aagtgggttt taggaccagg atgaggcggg gtgggggtgc ctacctgacg
      241 accgaccccg acccactgga caagcaccca acccccattc cccaaattgc gcatccccta
      301 tcagagaggg ggaggggaaa caggatgcgg cgaggcgcgt gcgcactgcc agcttcagca
      361 ccgcggacag tgccttcgcc cccgcctggc ggcgcgcgcc accgccgcct cagcactgaa
      421 ggcgcgctga cgtcactcgc cggtcccccg caaactcccc ttcccggcca ccttggtcgc
      481 gtccgcgccg ccgccggccc agccggaccg caccacgcga ggcgcgagat aggggggcac
      541 gggcgcgacc atctgcgctg cggcgccggc gactcagcgc tgcctcagtc tgcggtgggc
      601 agcggaggag tcgtgtcgtg cctgagagcg cagtcgagaa ggtaccggat ccgccaccat
      661 gagcagactg gtcgccgctt cttggctgct ggctctcctc ctctgcggaa ttaccagcac
      721 aacaacagcc tctgcccaca tcgtgatggt ggacgcctac aagcccacca agagcgcccc
      781 agcagcttct tctacagacg gaacagccgc cgcagcagtg tctcactacg ccatgaacgg
      841 cttcgacgag ctggctaaag gagccgtggt gccagaagac cactttgtct gcggaccagc
      901 cgacaagtgc tattgctccg cttggctgca cagcagagga gccctgtggg agcaggaaac
      961 agccaggggc ttccagtggt ttgccgtgtt cctgtctgcc ctgtttctgg ctttctacgg
     1021 ctggcacgcc tataaggcca gcgtgggctg ggaggaggtg tacgtgtgct ccgtggagct
     1081 gatcaaagtg attctggaga tctatttcga gtttaccagt cctgcgatgc tgttcctgta
     1141 cggaggcaac attaccccat ggcttcgtta cgccgagtgg ctgctgacat gccctgtcat
     1201 tcttatccat ctgagcaaca tcaccggtct gagtgaggca tataataagc gtaccatggc
     1261 tctgctggtg tcagacctgg gcactatctg catgggcgtg acagccgctc tggccactgg
     1321 gtgggtgaag tggctgtttt actgtatcgg cctggtgtac ggcacccaga cattctacaa
     1381 cgcaggaatc atctacgtgg aggcgtacca caccgtgccg aagggccggt gtcgccaggt
     1441 ggtgactggc atggcttggc tcttcttcgt atcatggggt atgttcccca tcctgttcat
     1501 cctcggcccc gagggcttcg gcgtcctgag cgtggctggg tccactattg gccacaccat
     1561 cgccgacctg ctgtccaaga atatttgggg actgctcggg cacttcctgc gcatcaaaat
     1621 ccacgagcat atcattatgt acggcgacat tcgcagacca gtgagctccc agtttctggg
     1681 ccgcaaggtg gacgtcctgg ccttcgtgac agaggaggat aaagtggcgg ccgccaagag
     1741 caggatcacc agcgagggcg agtacatccc cctggaccag atcgacatca acgtggtgag
     1801 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     1861 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     1921 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     1981 caccttcggc tacggcctgc agtgcttcgc ccgctacccc gaccacatga agcagcacga
     2041 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     2101 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     2161 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     2221 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
     2281 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     2341 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     2401 ctaccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     2461 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt aataagaatt
     2521 cgatatcaag cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg
     2581 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     2641 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     2701 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     2761 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     2821 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     2881 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc
     2941 gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     3001 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     3061 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     3121 ctccccgcat cgataccgag cgctgctcga gagatctacg ggtggcatcc ctgtgacccc
     3181 tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag ccttgtccta
     3241 ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat tatggggtgg
     3301 aggggggtgg tatggagcaa ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc
     3361 tattgggaac caagctggag tgcagtggca caatcttggc tcactgcaat ctccgcctcc
     3421 tgggttcaag cgattctcct gcctcagcct cccgagttgt tgggattcca ggcatgcatg
     3481 accaggctca gctaattttt gtttttttgg tagagacggg gtttcaccat attggccagg
     3541 ctggtctcca actcctaatc tcaggtgatc tacccacctt ggcctcccaa attgctggga
     3601 ttacaggcgt gaaccactgc tcccttccct gtccttctga ttttgtaggt aaccacgtgc
     3661 ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc
     3721 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg
     3781 cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc
     3841 ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg
     3901 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     3961 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     4021 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     4081 gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg
     4141 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     4201 ccaaactgga acaacactca accctatctc gggctattct tttgatttat aagggatttt
     4261 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     4321 taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc
     4381 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     4441 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     4501 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     4561 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     4621 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     4681 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     4741 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     4801 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     4861 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     4921 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     4981 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     5041 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     5101 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     5161 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     5221 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     5281 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     5341 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     5401 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     5461 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     5521 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     5581 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     5641 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     5701 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     5761 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     5821 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     5881 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     5941 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     6001 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     6061 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     6121 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     6181 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     6241 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     6301 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     6361 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgt
//
