LOCUS       Exported                6360 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  AAV expression of ChR2(H134R)-GFP under the Syn promoter.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-Syn-ChR2(H134R)-GFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6360)
  AUTHORS   Boyden ES, Zhang F, Bamberg E, Nagel G, Deisseroth K
  TITLE     Millisecond-timescale, genetically targeted optical control of 
            neural activity.
  JOURNAL   Nat Neurosci. 2005 Sep . 8(9):1263-8.
  PUBMED    16116447
REFERENCE   2  (bases 1 to 6360)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6360
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   562..691
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(753..1341)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(833..852)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1512..2372)
                     /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     2135..2154
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2373..2477)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2545..2563
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2601..2623)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2723..2742
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2759..3214)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2896..2917)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3108..3127
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   3289..3429
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3300..3429
                     /label=AAV2 ITR
     polyA_signal    3475..3951
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     3733..3752
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     misc_feature    3971..4559
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             4077..4088
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     4486..4506
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4587..5300)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(4629..4650)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4978..4997
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5237..5258
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(5319..6245)
                     /codon_start=1
                     /product="H134R mutant of an N-terminal fragment of 
                     channelrhodopsin-2 from the alga Chlamydomonas reinhardtii 
                     (Nagel et al., 2005)"
                     /label=ChR2(H134R)
                     /note="blue light-activated cation channel"
                     /translation="MDYGGALSAVGRELLFVTNPVVVNGSVLVPEDQCYCAGWIESRGT
                     NGAQTASNVLQWLAAGFSILLLMFYAYQTWKSTCGWEEIYVCAIEMVKVILEFFFEFKN
                     PSMLYLATGHRVQWLRYAEWLLTCPVILIRLSNLTGLSNDYSRRTMGLLVSDIGTIVWG
                     ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS
                     WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI
                     RKTTKLNIGGTEIEVETLVEDEAEAGAV"
     regulatory      6242..6251
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     promoter        complement(6264..351)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
ORIGIN
        1 ccccctatct cgcgcctcgc gtggtgcggt ccggctgggc cggcggcggc gcggacgcga
       61 ccaaggtggc cgggaagggg agtttgcggg ggaccggcga gtgacgtcag cgcgccttca
      121 gtgctgaggc ggcggtggcg cgcgccgcca ggcgggggcg aaggcactgt ccgcggtgct
      181 gaagctggca gtgcgcacgc gcctcgccgc atcctgtttc ccctccccct ctctgatagg
      241 ggatgcgcaa tttggggaat gggggttggg tgcttgtcca gtgggtcggg gtcggtcgtc
      301 aggtaggcac ccccaccccg cctcatcctg gtcctaaaac ccacttgcac tcatacgcag
      361 ggccctctgc agtctagatt aattaaccat ttgctgccga cccctccccc caacttctcg
      421 gggactgtgg gcgatgtgcg ctctgcccac tgacgggcac cggagccaat tcccactcct
      481 ttcaagacct agaaggtcca ttagctgcaa agattcctct ctgtttaaaa ctttatccat
      541 ctttgcaacg cgtgcggccg caggaacccc tagtgatgga gttggccact ccctctctgc
      601 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggcggcctca
      661 gtgagcgagc gagcgcgcag ctgcctgcag gacatgtgag caaaaggcca gcaaaaggcc
      721 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
      781 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
      841 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
      901 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
      961 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     1021 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     1081 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     1141 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     1201 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     1261 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     1321 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     1381 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     1441 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     1501 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     1561 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     1621 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     1681 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     1741 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     1801 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     1861 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     1921 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     1981 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     2041 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     2101 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     2161 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     2221 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     2281 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     2341 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     2401 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     2461 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     2521 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt
     2581 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     2641 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     2701 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccataaa
     2761 attgtaaacg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     2821 tttaaccaat agaccgaaat cggcaaaatc ccttataaat caaaagaata gcccgagata
     2881 gagttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     2941 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcacccaaa
     3001 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     3061 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     3121 aaaggagcgg gcgctaaggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     3181 cccgccgcgc ttaatgcgcc gctacagggc gcgtactatg gttgctttga cgtatgcggt
     3241 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgcccc tgcaggcagc
     3301 tgcgcgctcg ctcgctcact gaggccgccc gggcaaagcc cgggcgtcgg gcgacctttg
     3361 gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg agtggccaac tccatcacta
     3421 ggggttcctg cggccgctcg gtccgcacgt ggttacctac aaaatcagct cgagaaggac
     3481 agggaaggga gcagtggttc acgcctgtaa tcccagcaat ttgggaggcc aaggtgggta
     3541 gatcacctga gattaggagt tggagaccag cctggccaat atggtgaaac cccgtctcta
     3601 ccaaaaaaac aaaaattagc tgagcctggt catgcatgcc tggaatccca acaactcggg
     3661 aggctgaggc aggagaatcg cttgaaccca ggaggcggag attgcagtga gccaagattg
     3721 tgccactgca ctccagcttg gttcccaata gaccccgcag gccctacagg ttgtcttccc
     3781 aacttgcccc ttgctccata ccacccccct ccaccccata atattataga aggacaccta
     3841 gtcagacaaa atgatgcaac ttaattttat taggacaagg ctggtgggca ctggagtggc
     3901 aacttccagg gccaggagag gcactgggga ggggtcacag ggatgccacc cgtctcgagc
     3961 cggtatcgat gcggggaggc ggcccaaagg gagatccgac tcgtctgagg gcgaaggcga
     4021 agacgcggaa gaggccgcag agccggcagc aggccgcggg aaggaaggtc cgctggattg
     4081 agggccgaag ggacgtagca gaaggacgtc ccgcgcagaa tccaggtggc aacataggcg
     4141 agcagccaag gaaaggacga tgatttcccc gacaacacca cggaattgtc agtgcccaac
     4201 agccgagccc ctgtccagca gcgggcaagg caggcggcga tgagttccgc cgtggcaata
     4261 gggaggggga aagcgaaagt cccggaaagg agctgacagg tggtggcaat gccccaacca
     4321 gtgggggttg cgtcagcaaa cacagtgcac accacgccac gttgcctgac aacgggccac
     4381 aactcctcat aaagagacag caaccaggat ttatacaagg aggagaaaat gaaagccata
     4441 cgggaagcaa tagcatgata caaaggcatt aaagcagcgt atccacatag cgtaaaagga
     4501 gcaacatagt taagaatacc agtcaatctt tcacaaattt tgtaatccag aggttgatta
     4561 tcgataagct ttgtacggaa ttcttacttg tacagctcgt ccatgccgag agtgatcccg
     4621 gcggcggtca cgaactccag caggaccatg tgatcgcgct tctcgttggg gtctttgctc
     4681 agggcggact gggtgctcag gtagtggttg tcgggcagca gcacggggcc gtcgccgatg
     4741 ggggtgttct gctggtagtg gtcggcgagc tgcacgctgc cgtcctcgat gttgtggcgg
     4801 atcttgaagt tcaccttgat gccgttcttc tgcttgtcgg ccatgatata gacgttgtgg
     4861 ctgttgtagt tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg
     4921 cccttcagct cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc
     4981 ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg
     5041 gcggacttga agaagtcgtg ctgcttcatg tggtcggggt agcggctgaa gcactgcacg
     5101 ccgtaggtca gggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcaa
     5161 atgaacttca gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg
     5221 aacttgtggc cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac
     5281 agctcctcgc ccttgctcac tgctactacc ggtgccgcca cggctccggc ctcggcttcg
     5341 tcttcgacga gagtctcgac ctcgatctcc gttccgccga tgttcagttt ggtggttttg
     5401 cggatatctc cgtgaatcaa tatgtgctcg tggatcagga cgcgcaggta gtgtcccaac
     5461 aacccccaac aatttttact catcagatca ataatcgtgt gacctacggt ggagccatag
     5521 acgctcagga cgccaaaacc ttcgggcccc aaaatgaaga gaattgggaa cataccccag
     5581 ctcacgaaaa acagccatgc catgccggtc acgacctggc ggcaccgacc ctttggcaca
     5641 gtatgataac cctcgatata tgctttggcg gcgtgaaaaa atgtgttcgc gccatagcac
     5701 aatccaagac aaaagaagat gactttaaca tagccggttg ccatggcgct ggtagccccc
     5761 cacacgatag tcccgatgtc tgagacaagg agtcccatgg ttctcctgct gtagtcgttg
     5821 ctcaggccgg tgaggttgct cagtctgata aggatgacag gacaagtgag cagccactct
     5881 gcatagcgca gccactgcac ccggtgtcct gtggcaaggt agagcataga gggattctta
     5941 aactcaaaaa agaactcgag aatcacctta accatttcaa tggcgcacac atagatctcc
     6001 tcccagccgc atgtagattt ccaggtttgg taggcataga acatcagcag caaaatgctg
     6061 aatcctgctg caagccactg caggacattt gacgcggtct gagcgccgtt cgtgccgcga
     6121 gattcaatcc atccggcaca gtaacattga tcctcaggga ccaggacgga cccgttcacc
     6181 accacaggat tagtaacgaa caaaagttcg cgtccgacgg cagacaaagc gccgccatag
     6241 tccatggtgg cggatcctct cgactgcgct ctcaggcacg acacgactcc tccgctgccc
     6301 accgcagact gaggcagcgc tgagtcgccg gcgccgcagc gcagatggtc gcgcccgtgc
//
