LOCUS       sequence-183171-        6939 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of humanized ChR2 with H134R mutation fused to 
            mCherry driven by CAG promoter for optogenetic activation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6939)
  TITLE     Deisseroth AAV plasmids
REFERENCE   2  (bases 1 to 6939)
  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.CAG.hChR2(H134R)-mCherry.WPRE.SV40
FEATURES             Location/Qualifiers
     source          1..6939
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     promoter        441..718
                     /label=chicken beta-actin promoter
     primer_bind     997..1019
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1081..1100
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1170..1179
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1176..2102
                     /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             2115..2822
                     /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(2257..2275)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2525..2544
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     intron          2848..2980
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     2963..2982
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    3036..3624
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3089..3109)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3507..3518)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3667..3788
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3678..3697)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3732..3751
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3950..4079
                     /label=AAV2 ITR
     primer_bind     complement(4098..4115)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4098..4114)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4107..4129)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4256..4711
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4343..4362)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4553..4574
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4737..4841
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4842..4910,4911..5702)
                     /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(5060..5079)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5873..6461
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6362..6381
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    6749..6770
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6785..6790,6791..6808,6809..6815)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6823..6839
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     6828..6850
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6847..6863
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6847..6863
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct
      181 aggaagagta ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      241 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      301 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      361 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      421 agtcatcgct attaccatgg tcgaggtgag ccccacgttc tgcttcactc tccccatctc
      481 ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat
      541 gggggcgggg gggggggggg ggcgcgcgcc aggcggggcg gggcggggcg aggggcgggg
      601 cggggcgagg cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc
      661 ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg
      721 agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc
      781 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      841 gggctgtaat tagcgcttgg tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag
      901 ccttgagggg ctccgggagg gccctttgtg cggggggagc ggctcggggc tgtccgcggg
      961 gggacggctg ccttcggggg ggacggggca gggcggggtt cggcttctgg cgtgtgaccg
     1021 gcggctctag agcctctgct aaccatgttc atgccttctt ctttttccta cagctcctgg
     1081 gcaacgtgct ggttattgtg ctgtctcatc attttggcaa agaattggat cgggatccac
     1141 tagtccagtg tggtggaatt gcccttgctg ccaccatgga ctatggcggc gctttgtctg
     1201 ccgtcggacg cgaacttttg ttcgttacta atcctgtggt ggtgaacggg tccgtcctgg
     1261 tccctgagga tcaatgttac tgtgccggat ggattgaatc tcgcggcacg aacggcgctc
     1321 agaccgcgtc aaatgtcctg cagtggcttg cagcaggatt cagcattttg ctgctgatgt
     1381 tctatgccta ccaaacctgg aaatctacat gcggctggga ggagatctat gtgtgcgcca
     1441 ttgaaatggt taaggtgatt ctcgagttct tttttgagtt taagaatccc tctatgctct
     1501 accttgccac aggacaccgg gtgcagtggc tgcgctatgc agagtggctg ctcacttgtc
     1561 ctgtcatcct tatccgcctg agcaacctca ccggcctgag caacgactac agcaggagaa
     1621 ccatgggact ccttgtctca gacatcggga ctatcgtgtg gggggctacc agcgccatgg
     1681 caaccggcta tgttaaagtc atcttctttt gtcttggatt gtgctatggc gcgaacacat
     1741 tttttcacgc cgccaaagca tatatcgagg gttatcatac tgtgccaaag ggtcggtgcc
     1801 gccaggtcgt gaccggcatg gcatggctgt ttttcgtgag ctggggtatg ttcccaattc
     1861 tcttcatttt ggggcccgaa ggttttggcg tcctgagcgt ctatggctcc accgtaggtc
     1921 acacgattat tgatctgatg agtaaaaatt gttgggggtt gttgggacac tacctgcgcg
     1981 tcctgatcca cgagcacata ttgattcacg gagatatccg caaaaccacc aaactgaaca
     2041 tcggcggaac ggagatcgag gtcgagactc tcgtcgaaga cgaagccgag gccggagccg
     2101 tgccagcggc cgccgtgagc aagggcgagg aggataacat ggccatcatc aaggagttca
     2161 tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg
     2221 agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg
     2281 gccccctgcc cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct
     2341 acgtgaagca ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca
     2401 agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct
     2461 ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg
     2521 acggccccgt aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc
     2581 ccgaggacgg cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc
     2641 actacgacgc tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg
     2701 cctacaacgt caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg
     2761 aacagtacga acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagt
     2821 aaagcggccc cggactcgag gccgcaggta agtatcaagg ttacaagaca ggtttaagga
     2881 gaccaataga aactgggctt gtcgagacag agaagactct tgcgtttctg ataggcacct
     2941 attggtctta ctgacatcca ctttgccttt ctctccacag gtgtcgagtg gagctcgcga
     3001 ctagtcgatt cgaattcgat atcaagctta tcgataatca acctctggat tacaaaattt
     3061 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     3121 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     3181 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     3241 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     3301 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     3361 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     3421 tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc
     3481 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     3541 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     3601 tctccctttg ggccgcctcc ccgcatcgat accgtcgacc cgggcggccg cttcgagcag
     3661 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     3721 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     3781 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gagatgtggg
     3841 aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatcgataag gatcttccta
     3901 gagcatggct acgtagataa gtagcatggc gggttaatca ttaactacaa ggaaccccta
     3961 gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca
     4021 aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc
     4081 cttaattaac ctaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     4141 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     4201 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg
     4261 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     4321 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     4381 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     4441 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     4501 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     4561 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     4621 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     4681 aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg
     4741 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     4801 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     4861 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     4921 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     4981 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     5041 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     5101 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     5161 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     5221 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     5281 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     5341 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     5401 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     5461 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     5521 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     5581 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     5641 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     5701 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     5761 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     5821 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     5881 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     5941 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     6001 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     6061 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     6121 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     6181 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     6241 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     6301 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     6361 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     6421 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     6481 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     6541 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     6601 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     6661 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     6721 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     6781 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     6841 ttcacacagg aaacagctat gaccatgatt acgccagatt taattaaggg caactgttgg
     6901 gaagggcgat cggtgcgggc ctcttcgcta ttacgccag
//
