LOCUS       sequence-12796-e        7931 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of humanized ChR2(H134R) fused to mCherry, for 
            optogenetic excitation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7931)
  TITLE     hChR2 plasmids
REFERENCE   2  (bases 1 to 7931)
  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)-mCherry
FEATURES             Location/Qualifiers
     source          1..7931
                     /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      2381..2390
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2387..3313
                     /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             3326..4033
                     /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(3468..3486)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3736..3755
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    4058..4646
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4111..4131)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4529..4540)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4678..5154
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4877..4896)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   5194..5334
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5194..5323
                     /label=AAV2 ITR
     rep_origin      5409..5864
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5496..5515)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5706..5727
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5881..5900)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6000..6022
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6060..6078)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6146..6250
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6251..6319,6320..7111)
                     /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      7282..7870
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7771..7790
                     /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 gccaccatgg actatggcgg
     2401 cgctttgtct gccgtcggac gcgaactttt gttcgttact aatcctgtgg tggtgaacgg
     2461 gtccgtcctg gtccctgagg atcaatgtta ctgtgccgga tggattgaat ctcgcggcac
     2521 gaacggcgct cagaccgcgt caaatgtcct gcagtggctt gcagcaggat tcagcatttt
     2581 gctgctgatg ttctatgcct accaaacctg gaaatctaca tgcggctggg aggagatcta
     2641 tgtgtgcgcc attgaaatgg ttaaggtgat tctcgagttc ttttttgagt ttaagaatcc
     2701 ctctatgctc taccttgcca caggacaccg ggtgcagtgg ctgcgctatg cagagtggct
     2761 gctcacttgt cctgtcatcc ttatccgcct gagcaacctc accggcctga gcaacgacta
     2821 cagcaggaga accatgggac tccttgtctc agacatcggg actatcgtgt ggggggctac
     2881 cagcgccatg gcaaccggct atgttaaagt catcttcttt tgtcttggat tgtgctatgg
     2941 cgcgaacaca ttttttcacg ccgccaaagc atatatcgag ggttatcata ctgtgccaaa
     3001 gggtcggtgc cgccaggtcg tgaccggcat ggcatggctg tttttcgtga gctggggtat
     3061 gttcccaatt ctcttcattt tggggcccga aggttttggc gtcctgagcg tctatggctc
     3121 caccgtaggt cacacgatta ttgatctgat gagtaaaaat tgttgggggt tgttgggaca
     3181 ctacctgcgc gtcctgatcc acgagcacat attgattcac ggagatatcc gcaaaaccac
     3241 caaactgaac atcggcggaa cggagatcga ggtcgagact ctcgtcgaag acgaagccga
     3301 ggccggagcc gtgccagcgg ccgccgtgag caagggcgag gaggataaca tggccatcat
     3361 caaggagttc atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga
     3421 gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt
     3481 gaccaagggt ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg
     3541 ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc
     3601 cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac
     3661 ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa
     3721 cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga
     3781 gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa
     3841 ggacggcggc cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca
     3901 gctgcccggc gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta
     3961 caccatcgtg gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga
     4021 gctgtacaag taagaattcg atatcaagct tatcgataat caacctctgg attacaaaat
     4081 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     4141 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     4201 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     4261 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     4321 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     4381 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     4441 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
     4501 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     4561 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     4621 gatctccctt tgggccgcct ccccgcatcg ataccgagcg ctgctcgaga gatctacggg
     4681 tggcatccct gtgacccctc cccagtgcct ctcctggccc tggaagttgc cactccagtg
     4741 cccaccagcc ttgtcctaat aaaattaagt tgcatcattt tgtctgacta ggtgtccttc
     4801 tataatatta tggggtggag gggggtggta tggagcaagg ggcaagttgg gaagacaacc
     4861 tgtagggcct gcggggtcta ttgggaacca agctggagtg cagtggcaca atcttggctc
     4921 actgcaatct ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc cgagttgttg
     4981 ggattccagg catgcatgac caggctcagc taatttttgt ttttttggta gagacggggt
     5041 ttcaccatat tggccaggct ggtctccaac tcctaatctc aggtgatcta cccaccttgg
     5101 cctcccaaat tgctgggatt acaggcgtga accactgctc ccttccctgt ccttctgatt
     5161 ttgtaggtaa ccacgtgcgg accgagcggc cgcaggaacc cctagtgatg gagttggcca
     5221 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
     5281 cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc
     5341 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca
     5401 accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     5461 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     5521 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     5581 ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg
     5641 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     5701 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg gctattcttt
     5761 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     5821 aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag
     5881 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     5941 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     6001 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     6061 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     6121 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     6181 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     6241 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     6301 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     6361 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     6421 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     6481 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     6541 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     6601 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     6661 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     6721 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     6781 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     6841 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     6901 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     6961 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     7021 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     7081 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     7141 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     7201 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     7261 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     7321 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     7381 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     7441 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     7501 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     7561 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     7621 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     7681 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     7741 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     7801 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     7861 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     7921 tgctcacatg t
//
