LOCUS       sequence-194451-        7037 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of humanized ChR2 with E123T/H134R mutations fused to
            EYFP driven by CaMKIIa promoter for optogenetic activation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7037)
  TITLE     Deisseroth AAV plasmids
REFERENCE   2  (bases 1 to 7037)
  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.CaMKIIa.ChETA(E123T/H134R)-eYFP.WPRE.hGH
FEATURES             Location/Qualifiers
     source          1..7037
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   52..181
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        200..1488
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     regulatory      1523..1532
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1529..2455
                     /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
                     PSMLYLATGHRVQWLRYATWLLTCPVILIRLSNLTGLSNDYSRRTMGLLVSDIGTIVWG
                     ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS
                     WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI
                     RKTTKLNIGGTEIEVETLVEDEAEAGAV"
     regulatory      2465..2474
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2471..2473,2474..2476,2477..3190)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2516..2537)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2777..2796)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3124..3145
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3215..3803
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3268..3288)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3686..3697)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3835..4311
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4034..4053)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4351..4491
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4351..4480
                     /label=AAV2 ITR
     rep_origin      4566..5021
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4653..4672)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4863..4884
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5038..5057)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5157..5179
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5217..5235)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5303..5407
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5408..5476,5477..6268)
                     /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(5626..5645)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6439..7027
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6928..6947
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg tcctgcaggc
       61 agctgcgcgc tcgctcgctc actgaggccg cccgggcgtc gggcgacctt tggtcgcccg
      121 gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc
      181 tgcggccgca cgcgtttaac attatggcct taggtcactt catctccatg gggttcttct
      241 tctgattttc tagaaaatga gatgggggtg cagagagctt cctcagtgac ctgcccaggg
      301 tcacatcaga aatgtcagag ctagaacttg aactcagatt actaatctta aattccatgc
      361 cttgggggca tgcaagtacg atatacagaa ggagtgaact cattagggca gatgaccaat
      421 gagtttagga aagaagagtc cagggcaggg tacatctaca ccacccgccc agccctgggt
      481 gagtccagcc acgttcacct cattatagtt gcctctctcc agtcctacct tgacgggaag
      541 cacaagcaga aactgggaca ggagccccag gagaccaaat cttcatggtc cctctgggag
      601 gatgggtggg gagagctgtg gcagaggcct caggaggggc cctgctgctc agtggtgaca
      661 gataggggtg agaaagcaga cagagtcatt ccgtcagcat tctgggtctg tttggtactt
      721 cttctcacgc taaggtggcg gtgtgatatg cacaatggct aaaaagcagg gagagctgga
      781 aagaaacaag gacagagaca gaggccaagt caaccagacc aattcccaga ggaagcaaag
      841 aaaccattac agagactaca agggggaagg gaaggagaga tgaattagct tcccctgtaa
      901 accttagaac ccagctgttg ccagggcaac ggggcaatac ctgtctcttc agaggagatg
      961 aagttgccag ggtaactaca tcctgtcttt ctcaaggacc atcccagaat gtggcaccca
     1021 ctagccgtta ccatagcaac tgcctctttg ccccacttaa tcccatcccg tctgttaaaa
     1081 gggccctata gttggaggtg ggggaggtag gaagagcgat gatcacttgt ggactaagtt
     1141 tgttcgcatc cccttctcca accccctcag tacatcaccc tgggggaaca gggtccactt
     1201 gctcctgggc ccacacagtc ctgcagtatt gtgtatataa ggccagggca aagaggagca
     1261 ggttttaaag tgaaaggcag gcaggtgttg gggaggcagt taccggggca acgggaacag
     1321 ggcgtttcgg aggtggttgc catggggacc tggatgctga cgaaggctcg cgaggctgtg
     1381 agcagccaca gtgccctgct cagaagcccc aagctcgtca gtcaagccgg ttctccgttt
     1441 gcactcagga gcacgggcag gcgagtggcc cctagttctg ggggcagctc tagagcggta
     1501 ccggatcctc tagagtcgac tagccaccat ggactatggc ggcgctttgt ctgccgtcgg
     1561 acgcgaactt ttgttcgtta ctaatcctgt ggtggtgaac gggtccgtcc tggtccctga
     1621 ggatcaatgt tactgtgccg gatggattga atctcgcggc acgaacggcg ctcagaccgc
     1681 gtcaaatgtc ctgcagtggc ttgcagcagg attcagcatt ttgctgctga tgttctatgc
     1741 ctaccaaacc tggaaatcta catgcggctg ggaggagatc tatgtgtgcg ccattgaaat
     1801 ggttaaggtg attctcgagt tcttttttga gtttaagaat ccctctatgc tctaccttgc
     1861 cacaggacac cgggtgcagt ggctgcgcta tgcaacctgg ctgctcactt gtcctgtcat
     1921 ccttatccgc ctgagcaacc tcaccggcct gagcaacgac tacagcagga gaaccatggg
     1981 actccttgtc tcagacatcg ggactatcgt gtggggggct accagcgcca tggcaaccgg
     2041 ctatgttaaa gtcatcttct tttgtcttgg attgtgctat ggcgcgaaca cattttttca
     2101 cgccgccaaa gcatatatcg agggttatca tactgtgcca aagggtcggt gccgccaggt
     2161 cgtgaccggc atggcatggc tgtttttcgt gagctggggt atgttcccaa ttctcttcat
     2221 tttggggccc gaaggttttg gcgtcctgag cgtctatggc tccaccgtag gtcacacgat
     2281 tattgatctg atgagtaaaa attgttgggg gttgttggga cactacctgc gcgtcctgat
     2341 ccacgagcac atattgattc acggagatat ccgcaaaacc accaaactga acatcggcgg
     2401 aacggagatc gaggtcgaga ctctcgtcga agacgaagcc gaggccggag ccgtgccagc
     2461 ggccgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     2521 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
     2581 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     2641 ctggcccacc ctcgtgacca ccttcggcta cggcctgcag tgcttcgccc gctaccccga
     2701 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     2761 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     2821 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     2881 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
     2941 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
     3001 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     3061 cgacaaccac tacctgagct accagtccgc cctgagcaaa gaccccaacg agaagcgcga
     3121 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     3181 gtacaagtaa gaattcgata tcaagcttat cgataatcaa cctctggatt acaaaatttg
     3241 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     3301 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     3361 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     3421 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     3481 gctcctttcc ggaactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     3541 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     3601 gtcggggaaa tcatcgtcct ttccttggct gctcgcctat gttgccacct ggattctgcg
     3661 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     3721 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     3781 ctccctttgg gccgcctccc cgcatcgata ccgagcgctg ctcgagagat ctacgggtgg
     3841 catccctgtg acccctcccc agtgcctctc ctggccctgg aagttgccac tccagtgccc
     3901 accagccttg tcctaataaa attaagttgc atcattttgt ctgactaggt gtccttctat
     3961 aatattatgg ggtggagggg ggtggtatgg agcaaggggc aagttgggaa gacaacctgt
     4021 agggcctgcg gggtctattg ggaaccaagc tggagtgcag tggcacaatc ttggctcact
     4081 gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc agcctcccga gttgttggga
     4141 ttccaggcat gcatgaccag gctcagctaa tttttgtttt tttggtagag acggggtttc
     4201 accatattgg ccaggctggt ctccaactcc taatctcagg tgatctaccc accttggcct
     4261 cccaaattgc tgggattaca ggcgtgaacc actgctccct tccctgtcct tctgattttg
     4321 taggtaacca cgtgcggacc gagcggccgc aggaacccct agtgatggag ttggccactc
     4381 cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg
     4441 gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga
     4501 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc
     4561 atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     4621 gaccgctaca cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct
     4681 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     4741 atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag
     4801 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     4861 tagtggactc ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga
     4921 tttataaggg attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa
     4981 atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac
     5041 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     5101 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     5161 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
     5221 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     5281 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     5341 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     5401 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     5461 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     5521 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     5581 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     5641 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     5701 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     5761 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     5821 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     5881 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     5941 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     6001 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     6061 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     6121 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     6181 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     6241 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     6301 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     6361 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     6421 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     6481 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     6541 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     6601 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     6661 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     6721 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     6781 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     6841 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     6901 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     6961 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     7021 atggaaaaac gccagca
//
