LOCUS       sequence-12795-e        7952 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of humanized ChR2(H134R) fused to EYFP, for 
            optogenetic excitation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7952)
  TITLE     hChR2 plasmids
REFERENCE   2  (bases 1 to 7952)
  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-GFAP-hChR2(H134R)-EYFP
FEATURES             Location/Qualifiers
     source          1..7952
                     /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
     regulatory      2387..2396
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2393..3319
                     /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      3329..3338
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3335..3337,3338..3340,3341..4054)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3380..3401)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3641..3660)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3988..4009
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4079..4667
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4132..4152)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4550..4561)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4699..5175
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4898..4917)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   5215..5355
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5215..5344
                     /label=AAV2 ITR
     rep_origin      5430..5885
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5517..5536)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5727..5748
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5902..5921)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6021..6043
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6081..6099)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6167..6271
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6272..6340,6341..7132)
                     /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"
     rep_origin      7303..7891
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7792..7811
                     /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 cgcgtcccac ctccctctct gtgctgggac
      181 tcacagaggg agacctcagg aggcagtctg tccatcacat gtccaaatgc agagcatacc
      241 ctgggctggg cgcagtggcg cacaactgta attccagcac tttgggaggc tgatgtggaa
      301 ggatcacttg agcccagaag ttctagacca gcctgggcaa catggcaaga ccctatctct
      361 acaaaaaaag ttaaaaaatc agccacgtgt ggtgacacac acctgtagtc ccagctattc
      421 aggaggctga ggtgagggga tcacttaagg ctgggaggtt gaggctgcag tgagtcgtgg
      481 ttgcgccact gcactccagc ctgggcaaca gtgagaccct gtctcaaaag acaaaaaaaa
      541 aaaaaaaaaa aaaaagaaca tatcctggtg tggagtaggg gacgctgctc tgacagaggc
      601 tcgggggcct gagctggctc tgtgagctgg ggaggaggca gacagccagg ccttgtctgc
      661 aagcagacct ggcagcattg ggctggccgc cccccagggc ctcctcttca tgcccagtga
      721 atgactcacc ttggcacaga cacaatgttc ggggtgggca cagtgcctgc ttcccgccgc
      781 accccagccc ccctcaaatg ccttccgaga agcccattga gcagggggct tgcattgcac
      841 cccagcctga cagcctggca tcttgggata aaagcagcac agccccctag gggctgccct
      901 tgctgtgtgg cgccaccggc ggtggagaac aaggctctat tcagcctgtg cccaggaaag
      961 gggatcaggg gatgcccagg catggacagt gggtggcagg gggggagagg agggctgtct
     1021 gcttcccaga agtccaagga cacaaatggg tgaggggact gggcagggtt ctgaccctgt
     1081 gggaccagag tggagggcgt agatggacct gaagtctcca gggacaacag ggcccaggtc
     1141 tcaggctcct agttgggccc agtggctcca gcgtttccaa acccatccat ccccagaggt
     1201 tcttcccatc tctccaggct gatgtgtggg aactcgagga aataaatctc cagtgggaga
     1261 cggaggggtg gccagggaaa cggggcgctg caggaataaa gacgagccag cacagccagc
     1321 tcatgtgtaa cggctttgtg gagctgtcaa ggcctggtct ctgggagaga ggcacaggga
     1381 ggccagacaa ggaaggggtg acctggaggg acagatccag gggctaaagt cctgataagg
     1441 caagagagtg ccggccccct cttgccctat caggacctcc actgccacat agaggccatg
     1501 attgaccctt agacaaaggg ctggtgtcca atcccagccc ccagccccag aactccaggg
     1561 aatgaatggg cagagagcag gaatgtggga catctgtgtt caagggaagg actccaggag
     1621 tctgctggga atgaggccta gtaggaaatg aggtggccct tgagggtaca gaacaggttc
     1681 attcttcgcc aaattcccag caccttgcag gcacttacag ctgagtgaga taatgcctgg
     1741 gttatgaaat caaaaagttg gaaagcaggt cagaggtcat ctggtacagc ccttccttcc
     1801 cttttttttt tttttttttt gtgagacaag gtctctctct gttgcccagg ctggagtggc
     1861 gcaaacacag ctcactgcag cctcaaccta ctgggctcaa gcaatcctcc agcctcagcc
     1921 tcccaaagtg ctgggattac aagcatgagc caccccactc agccctttcc ttccttttta
     1981 attgatgcat aataattgta agtattcatc atggtccaac caaccctttc ttgacccacc
     2041 ttcctagaga gagggtcctc ttgcttcagc ggtcagggcc ccagacccat ggtctggctc
     2101 caggtaccac ctgcctcatg caggagttgg cgtgcccagg aagctctgcc tctgggcaca
     2161 gtgacctcag tggggtgagg ggagctctcc ccatagctgg gctgcggccc aaccccaccc
     2221 cctcaggcta tgccaggggg tgttgccagg ggcacccggg catcgccagt ctagcccact
     2281 ccttcataaa gccctcgcat cccaggagcg agcagagcca gagcaggttg gagaggagac
     2341 gcatcacctc cgctgctcgc cgggatcctc tagagtcgac tccggagcca ccatggacta
     2401 tggcggcgct ttgtctgccg tcggacgcga acttttgttc gttactaatc ctgtggtggt
     2461 gaacgggtcc gtcctggtcc ctgaggatca atgttactgt gccggatgga ttgaatctcg
     2521 cggcacgaac ggcgctcaga ccgcgtcaaa tgtcctgcag tggcttgcag caggattcag
     2581 cattttgctg ctgatgttct atgcctacca aacctggaaa tctacatgcg gctgggagga
     2641 gatctatgtg tgcgccattg aaatggttaa ggtgattctc gagttctttt ttgagtttaa
     2701 gaatccctct atgctctacc ttgccacagg acaccgggtg cagtggctgc gctatgcaga
     2761 gtggctgctc acttgtcctg tcatccttat ccgcctgagc aacctcaccg gcctgagcaa
     2821 cgactacagc aggagaacca tgggactcct tgtctcagac atcgggacta tcgtgtgggg
     2881 ggctaccagc gccatggcaa ccggctatgt taaagtcatc ttcttttgtc ttggattgtg
     2941 ctatggcgcg aacacatttt ttcacgccgc caaagcatat atcgagggtt atcatactgt
     3001 gccaaagggt cggtgccgcc aggtcgtgac cggcatggca tggctgtttt tcgtgagctg
     3061 gggtatgttc ccaattctct tcattttggg gcccgaaggt tttggcgtcc tgagcgtcta
     3121 tggctccacc gtaggtcaca cgattattga tctgatgagt aaaaattgtt gggggttgtt
     3181 gggacactac ctgcgcgtcc tgatccacga gcacatattg attcacggag atatccgcaa
     3241 aaccaccaaa ctgaacatcg gcggaacgga gatcgaggtc gagactctcg tcgaagacga
     3301 agccgaggcc ggagccgtgc cagcggccgc caccatggtg agcaagggcg aggagctgtt
     3361 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     3421 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     3481 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccttcg gctacggcct
     3541 gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     3601 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     3661 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     3721 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     3781 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg
     3841 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     3901 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agctaccagt ccgccctgag
     3961 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     4021 gatcactctc ggcatggacg agctgtacaa gtaagaattc gatatcaagc ttatcgataa
     4081 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     4141 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     4201 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     4261 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     4321 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     4381 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     4441 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc
     4501 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     4561 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     4621 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgagc
     4681 gctgctcgag agatctacgg gtggcatccc tgtgacccct ccccagtgcc tctcctggcc
     4741 ctggaagttg ccactccagt gcccaccagc cttgtcctaa taaaattaag ttgcatcatt
     4801 ttgtctgact aggtgtcctt ctataatatt atggggtgga ggggggtggt atggagcaag
     4861 gggcaagttg ggaagacaac ctgtagggcc tgcggggtct attgggaacc aagctggagt
     4921 gcagtggcac aatcttggct cactgcaatc tccgcctcct gggttcaagc gattctcctg
     4981 cctcagcctc ccgagttgtt gggattccag gcatgcatga ccaggctcag ctaatttttg
     5041 tttttttggt agagacgggg tttcaccata ttggccaggc tggtctccaa ctcctaatct
     5101 caggtgatct acccaccttg gcctcccaaa ttgctgggat tacaggcgtg aaccactgct
     5161 cccttccctg tccttctgat tttgtaggta accacgtgcg gaccgagcgg ccgcaggaac
     5221 ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc
     5281 gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc
     5341 gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     5401 cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc
     5461 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     5521 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     5581 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     5641 tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     5701 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     5761 ccctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     5821 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac
     5881 aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
     5941 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     6001 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     6061 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     6121 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     6181 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     6241 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     6301 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     6361 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     6421 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     6481 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     6541 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     6601 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     6661 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     6721 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     6781 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     6841 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     6901 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     6961 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     7021 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     7081 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     7141 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     7201 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     7261 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     7321 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     7381 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     7441 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     7501 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     7561 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     7621 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     7681 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     7741 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     7801 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     7861 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     7921 cctggccttt tgctggcctt ttgctcacat gt
//
