LOCUS       sequence-313556-        6204 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of humanized ChR2 with H134R mutation fused to 
            mCherry, for optogenetic excitation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6204)
  TITLE     hChR2 plasmids
REFERENCE   2  (bases 1 to 6204)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pAAV-hSyn-hChR2(H134R)-mCherry
FEATURES             Location/Qualifiers
     source          1..6204
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..448
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      480..489
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             486..1412
                     /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             1425..2132
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(1567..1585)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1835..1854
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2157..2745
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2210..2230)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2628..2639)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2777..3253
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2976..2995)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3293..3433
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3293..3422
                     /label=AAV2 ITR
     rep_origin      3508..3963
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3595..3614)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3805..3826
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3980..3999)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4099..4121
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4159..4177)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4245..4349
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4350..4418,4419..5210)
                     /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(4568..4587)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5381..5969
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5870..5889
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   6031..6160
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 agtgcaagtg ggttttagga ccaggatgag gcggggtggg ggtgcctacc tgacgaccga
       61 ccccgaccca ctggacaagc acccaacccc cattccccaa attgcgcatc ccctatcaga
      121 gagggggagg ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg
      181 gacagtgcct tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc
      241 gctgacgtca ctcgccggtc ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg
      301 cgccgccgcc ggcccagccg gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg
      361 cgaccatctg cgctgcggcg ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg
      421 aggagtcgtg tcgtgcctga gagcgcagtc gagaaggtac cggatcctct agagtcgacg
      481 ccaccatgga ctatggcggc gctttgtctg ccgtcggacg cgaacttttg ttcgttacta
      541 atcctgtggt ggtgaacggg tccgtcctgg tccctgagga tcaatgttac tgtgccggat
      601 ggattgaatc tcgcggcacg aacggcgctc agaccgcgtc aaatgtcctg cagtggcttg
      661 cagcaggatt cagcattttg ctgctgatgt tctatgccta ccaaacctgg aaatctacat
      721 gcggctggga ggagatctat gtgtgcgcca ttgaaatggt taaggtgatt ctcgagttct
      781 tttttgagtt taagaatccc tctatgctct accttgccac aggacaccgg gtgcagtggc
      841 tgcgctatgc agagtggctg ctcacttgtc ctgtcatcct tatccgcctg agcaacctca
      901 ccggcctgag caacgactac agcaggagaa ccatgggact ccttgtctca gacatcggga
      961 ctatcgtgtg gggggctacc agcgccatgg caaccggcta tgttaaagtc atcttctttt
     1021 gtcttggatt gtgctatggc gcgaacacat tttttcacgc cgccaaagca tatatcgagg
     1081 gttatcatac tgtgccaaag ggtcggtgcc gccaggtcgt gaccggcatg gcatggctgt
     1141 ttttcgtgag ctggggtatg ttcccaattc tcttcatttt ggggcccgaa ggttttggcg
     1201 tcctgagcgt ctatggctcc accgtaggtc acacgattat tgatctgatg agtaaaaatt
     1261 gttgggggtt gttgggacac tacctgcgcg tcctgatcca cgagcacata ttgattcacg
     1321 gagatatccg caaaaccacc aaactgaaca tcggcggaac ggagatcgag gtcgagactc
     1381 tcgtcgaaga cgaagccgag gccggagccg tgccagcggc cgccgtgagc aagggcgagg
     1441 aggataacat ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg gagggctccg
     1501 tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc
     1561 agaccgccaa gctgaaggtg accaagggtg gccccctgcc cttcgcctgg gacatcctgt
     1621 cccctcagtt catgtacggc tccaaggcct acgtgaagca ccccgccgac atccccgact
     1681 acttgaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg
     1741 gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggcgagttc atctacaagg
     1801 tgaagctgcg cggcaccaac ttcccctccg acggccccgt aatgcagaag aagaccatgg
     1861 gctgggaggc ctcctccgag cggatgtacc ccgaggacgg cgccctgaag ggcgagatca
     1921 agcagaggct gaagctgaag gacggcggcc actacgacgc tgaggtcaag accacctaca
     1981 aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt caacatcaag ttggacatca
     2041 cctcccacaa cgaggactac accatcgtgg aacagtacga acgcgccgag ggccgccact
     2101 ccaccggcgg catggacgag ctgtacaagt aagaattcga tatcaagctt atcgataatc
     2161 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     2221 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     2281 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     2341 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     2401 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     2461 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     2521 gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct
     2581 atgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     2641 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     2701 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcga taccgagcgc
     2761 tgctcgagag atctacgggt ggcatccctg tgacccctcc ccagtgcctc tcctggccct
     2821 ggaagttgcc actccagtgc ccaccagcct tgtcctaata aaattaagtt gcatcatttt
     2881 gtctgactag gtgtccttct ataatattat ggggtggagg ggggtggtat ggagcaaggg
     2941 gcaagttggg aagacaacct gtagggcctg cggggtctat tgggaaccaa gctggagtgc
     3001 agtggcacaa tcttggctca ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc
     3061 tcagcctccc gagttgttgg gattccaggc atgcatgacc aggctcagct aatttttgtt
     3121 tttttggtag agacggggtt tcaccatatt ggccaggctg gtctccaact cctaatctca
     3181 ggtgatctac ccaccttggc ctcccaaatt gctgggatta caggcgtgaa ccactgctcc
     3241 cttccctgtc cttctgattt tgtaggtaac cacgtgcgga ccgagcggcc gcaggaaccc
     3301 ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga
     3361 ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc
     3421 agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     3481 caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg
     3541 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt
     3601 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     3661 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     3721 atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     3781 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaact
     3841 ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc tattggttaa
     3901 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa
     3961 ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
     4021 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     4081 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     4141 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     4201 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     4261 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     4321 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     4381 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     4441 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     4501 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     4561 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     4621 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     4681 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     4741 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     4801 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     4861 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     4921 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     4981 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     5041 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     5101 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     5161 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     5221 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     5281 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     5341 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     5401 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     5461 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     5521 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     5581 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     5641 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     5701 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     5761 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     5821 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     5881 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     5941 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     6001 tggccttttg ctggcctttt gctcacatgt cctgcaggca gctgcgcgct cgctcgctca
     6061 ctgaggccgc ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg
     6121 cgcagagagg gagtggccaa ctccatcact aggggttcct gcggccgcac gcgtgtgtct
     6181 agactgcaga gggccctgcg tatg
//
