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   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        341..788
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      801..810
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             807..1733
                     /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"
     CDS             1752..2465
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1794..1815)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2055..2074)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2402..2423
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2493..3081
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2546..2566)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2964..2975)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3101..3577
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3300..3319)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3623..3763
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3623..3752
                     /label=AAV2 ITR
     rep_origin      3838..4293
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3925..3944)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4135..4156
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4310..4329)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4429..4451
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4489..4507)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4575..4679
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4680..5540
                     /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(4898..4917)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5711..6299
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6200..6219
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgttgcaaa gatggataaa gttttaaaca gagaggaatc
      181 tttgcagcta atggaccttc taggtcttga aaggagtggg aattggctcc ggtgcccgtc
      241 agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcagca
      301 aatggttaat taatctagac tgcagagggc cctgcgtatg agtgcaagtg ggttttagga
      361 ccaggatgag gcggggtggg ggtgcctacc tgacgaccga ccccgaccca ctggacaagc
      421 acccaacccc cattccccaa attgcgcatc ccctatcaga gagggggagg ggaaacagga
      481 tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg gacagtgcct tcgcccccgc
      541 ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc gctgacgtca ctcgccggtc
      601 ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg cgccgccgcc ggcccagccg
      661 gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg cgaccatctg cgctgcggcg
      721 ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg aggagtcgtg tcgtgcctga
      781 gagcgcagtc gagaggatcc gccaccatgg actatggcgg cgctttgtct gccgtcggac
      841 gcgaactttt gttcgttact aatcctgtgg tggtgaacgg gtccgtcctg gtccctgagg
      901 atcaatgtta ctgtgccgga tggattgaat ctcgcggcac gaacggcgct cagaccgcgt
      961 caaatgtcct gcagtggctt gcagcaggat tcagcatttt gctgctgatg ttctatgcct
     1021 accaaacctg gaaatctaca tgcggctggg aggagatcta tgtgtgcgcc attgaaatgg
     1081 ttaaggtgat tctcgagttc ttttttgagt ttaagaatcc ctctatgctc taccttgcca
     1141 caggacaccg ggtgcagtgg ctgcgctatg cagagtggct gctcacttgt cctgtcatcc
     1201 ttatcagact gagcaacctc accggcctga gcaacgacta cagcaggaga accatgggac
     1261 tccttgtctc agacatcggg actatcgtgt ggggggctac cagcgccatg gcaaccggct
     1321 atgttaaagt catcttcttt tgtcttggat tgtgctatgg cgcgaacaca ttttttcacg
     1381 ccgccaaagc atatatcgag ggttatcata ctgtgccaaa gggtcggtgc cgccaggtcg
     1441 tgaccggcat ggcatggctg tttttcgtga gctggggtat gttcccaatt ctcttcattt
     1501 tggggcccga aggttttggc gtcctgagcg tctatggctc caccgtaggt cacacgatta
     1561 ttgatctgat gagtaaaaat tgttgggggt tgttgggaca ctacctgcgc gtcctgatcc
     1621 acgagcacat attgattcac ggagatatcc gcaaaaccac caaactgaac atcggcggaa
     1681 cggagatcga ggtcgagact ctcgtcgaag acgaagccga ggccggagcc gtggcggcac
     1741 cggtagtagc agtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     1801 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
     1861 ccacctacgg caagctgacc ctgaagttca tttgcaccac cggcaagctg cccgtgccct
     1921 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
     1981 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     2041 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     2101 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     2161 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
     2221 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
     2281 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     2341 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     2401 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     2461 acaagtaaga attccgtaca aagcttatcg ataatcaacc tctggattac aaaatttgtg
     2521 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     2581 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     2641 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     2701 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     2761 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     2821 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     2881 cggggaaatc atcgtccttt ccttggctgc tcgcctatgt tgccacctgg attctgcgcg
     2941 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     3001 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     3061 ccctttgggc cgcctccccg catcgatacc ggctcgagac gggtggcatc cctgtgaccc
     3121 ctccccagtg cctctcctgg ccctggaagt tgccactcca gtgcccacca gccttgtcct
     3181 aataaaatta agttgcatca ttttgtctga ctaggtgtcc ttctataata ttatggggtg
     3241 gaggggggtg gtatggagca aggggcaagt tgggaagaca acctgtaggg cctgcggggt
     3301 ctattgggaa ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc
     3361 ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg ttgggattcc aggcatgcat
     3421 gaccaggctc agctaatttt tgtttttttg gtagagacgg ggtttcacca tattggccag
     3481 gctggtctcc aactcctaat ctcaggtgat ctacccacct tggcctccca aattgctggg
     3541 attacaggcg tgaaccactg ctcccttccc tgtccttctc gagctgattt tgtaggtaac
     3601 cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg
     3661 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc
     3721 cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat
     3781 tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg
     3841 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     3901 cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     3961 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     4021 ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat
     4081 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     4141 tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt gatttataag
     4201 ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     4261 cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct
     4321 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     4381 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     4441 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     4501 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     4561 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     4621 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     4681 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     4741 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     4801 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     4861 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     4921 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     4981 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     5041 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     5101 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     5161 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     5221 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     5281 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     5341 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     5401 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     5461 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     5521 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     5581 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     5641 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     5701 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     5761 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     5821 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     5881 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     5941 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     6001 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     6061 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     6121 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     6181 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     6241 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     6301 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
