LOCUS       sequence-12776-e        6236 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of humanized ChR2 with H134R mutation fused to EYFP 
            driven by human Synapsin I promoter for optogenetic excitation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6236)
  TITLE     hChR2 plasmids
REFERENCE   2  (bases 1 to 6236)
  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)-EYFP
FEATURES             Location/Qualifiers
     source          1..6236
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     promoter        186..633
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      671..680
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             677..1603
                     /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      1613..1622
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1619..1621,1622..1624,1625..2338)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1664..1685)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1925..1944)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2272..2293
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2363..2951
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2416..2436)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2834..2845)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2983..3459
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3182..3201)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3499..3639
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3499..3628
                     /label=AAV2 ITR
     rep_origin      3714..4169
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3801..3820)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4011..4032
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4186..4205)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4305..4327
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4365..4383)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4451..4555
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4556..4624,4625..5416)
                     /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(4774..4793)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5587..6175
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6076..6095
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
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 cctctagagt
      661 cgactccgga gccaccatgg actatggcgg cgctttgtct gccgtcggac gcgaactttt
      721 gttcgttact aatcctgtgg tggtgaacgg gtccgtcctg gtccctgagg atcaatgtta
      781 ctgtgccgga tggattgaat ctcgcggcac gaacggcgct cagaccgcgt caaatgtcct
      841 gcagtggctt gcagcaggat tcagcatttt gctgctgatg ttctatgcct accaaacctg
      901 gaaatctaca tgcggctggg aggagatcta tgtgtgcgcc attgaaatgg ttaaggtgat
      961 tctcgagttc ttttttgagt ttaagaatcc ctctatgctc taccttgcca caggacaccg
     1021 ggtgcagtgg ctgcgctatg cagagtggct gctcacttgt cctgtcatcc ttatccgcct
     1081 gagcaacctc accggcctga gcaacgacta cagcaggaga accatgggac tccttgtctc
     1141 agacatcggg actatcgtgt ggggggctac cagcgccatg gcaaccggct atgttaaagt
     1201 catcttcttt tgtcttggat tgtgctatgg cgcgaacaca ttttttcacg ccgccaaagc
     1261 atatatcgag ggttatcata ctgtgccaaa gggtcggtgc cgccaggtcg tgaccggcat
     1321 ggcatggctg tttttcgtga gctggggtat gttcccaatt ctcttcattt tggggcccga
     1381 aggttttggc gtcctgagcg tctatggctc caccgtaggt cacacgatta ttgatctgat
     1441 gagtaaaaat tgttgggggt tgttgggaca ctacctgcgc gtcctgatcc acgagcacat
     1501 attgattcac ggagatatcc gcaaaaccac caaactgaac atcggcggaa cggagatcga
     1561 ggtcgagact ctcgtcgaag acgaagccga ggccggagcc gtgccagcgg ccgccaccat
     1621 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     1681 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     1741 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     1801 cgtgaccacc ttcggctacg gcctgcagtg cttcgcccgc taccccgacc acatgaagca
     1861 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     1921 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     1981 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     2041 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     2101 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     2161 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     2221 cctgagctac cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     2281 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaaga
     2341 attcgatatc aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac
     2401 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     2461 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     2521 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     2581 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     2641 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     2701 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     2761 atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     2821 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     2881 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     2941 cgcctccccg catcgatacc gagcgctgct cgagagatct acgggtggca tccctgtgac
     3001 ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac cagccttgtc
     3061 ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa tattatgggg
     3121 tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag ggcctgcggg
     3181 gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc aatctccgcc
     3241 tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt ccaggcatgc
     3301 atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac catattggcc
     3361 aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc caaattgctg
     3421 ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgattttgta ggtaaccacg
     3481 tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg
     3541 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg
     3601 cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc
     3661 tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc
     3721 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     3781 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     3841 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     3901 acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc
     3961 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     4021 gttccaaact ggaacaacac tcaaccctat ctcgggctat tcttttgatt tataagggat
     4081 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     4141 ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga
     4201 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     4261 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     4321 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     4381 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     4441 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     4501 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     4561 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     4621 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     4681 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     4741 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     4801 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     4861 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     4921 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     4981 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     5041 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     5101 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     5161 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     5221 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     5281 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     5341 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     5401 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     5461 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     5521 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     5581 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     5641 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5701 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     5761 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     5821 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     5881 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     5941 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     6001 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     6061 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     6121 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     6181 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgt
//
