LOCUS       sequence-219823-        7675 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-mediated expression of ChrimsonR-tdTomato under the CaMKII 
            promoter. tdTomato has codons varied between the first and second 
            tandem repeats to reduce recombination. Using SV40 signal. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7675)
  AUTHORS   Klapoetke NC, Murata Y, Kim SS, Pulver SR, Birdsey-Benson A, Cho YK,
            Morimoto TK, Chuong AS, Carpenter EJ, Tian Z, Wang J, Xie Y, Yan Z, 
            Zhang Y, Chow BY, Surek B, Melkonian M, Jayaraman V, 
            Constantine-Paton M, Wong GK, Boyden ES
  TITLE     Independent optical excitation of distinct neural populations.
  JOURNAL   Nat Methods. 2014 Mar;11(3):338-46. doi: 10.1038/nmeth.2836. Epub 
            2014 Feb 9.
  PUBMED    24509633
REFERENCE   2  (bases 1 to 7675)
  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"; doi: 
            "10.1038/nmeth.2836"; journalName: "Nat Methods"; date: "2014-03"; 
            volume: "11"; issue: "3"; pages: "338-46"
COMMENT     Sequence Label: pAAV-CaMKII-ChrimsonR-tdTomato
FEATURES             Location/Qualifiers
     source          1..7675
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(182..770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(262..281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(941..1732,1733..1801))
                     /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     1564..1583
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1802..1906)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1932..2387)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2069..2090)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2281..2300
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2459..2588
                     /label=AAV2 ITR
     promoter        2661..3949
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     regulatory      3959..3968
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3965..5014
                     /codon_start=1
                     /product="red light-drivable channelrhodopsin from 
                     Chlamydomonas noctigama (Klapoetke et al., 2014)"
                     /label=ChrimsonR
                     /note="K176R mutant with accelerated closing kinetics"
                     /translation="MAELISSATRSLFAAGGINPWPNPYHHEDMGCGGMTPTGECFSTE
                     WWCDPSYGLSDAGYGYCFVEATGGYLVVGVEKKQAWLHSRGTPGEKIGAQVCQWIAFSI
                     AIALLTFYGFSAWKATCGWEEVYVCCVEVLFVTLEIFKEFSSPATVYLSTGNHAYCLRY
                     FEWLLSCPVILIRLSNLSGLKNDYSKRTMGLIVSCVGMIVFGMAAGLATDWLKWLLYIV
                     SCIYGGYMYFQAAKCYVEANHSVPKGHCRMVVKLMAYAYFASWGSYPILWAVGPEGLLK
                     LSPYANSIGHSICDIIAKEFWTFLAHHLRIKIHEHILIHGDIRKTTKMEIGGEEVEVEE
                     FVEEEDEDTV"
     misc_feature    6485..7073
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6538..6558)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6956..6967)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    7114..7321
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   7491..7620
                     /label=AAV2 ITR
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1921 aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     1981 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     2041 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     2101 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     2161 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     2221 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     2281 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     2341 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt
     2401 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     2461 gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg
     2521 tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag
     2581 gggttccttg tagttaatga ttaacccgcc atgctactta tctacgtagc catgctctag
     2641 gaagatcgta ggttaattaa cattatggcc ttaggtcact tcatctccat ggggttcttc
     2701 ttctgatttt ctagaaaatg agatgggggt gcagagagct tcctcagtga cctgcccagg
     2761 gtcacatcag aaatgtcaga gctagaactt gaactcagat tactaatctt aaattccatg
     2821 ccttgggggc atgcaagtac gatatacaga aggagtgaac tcattagggc agatgaccaa
     2881 tgagtttagg aaagaagagt ccagggcagg gtacatctac accacccgcc cagccctggg
     2941 tgagtccagc cacgttcacc tcattatagt tgcctctctc cagtcctacc ttgacgggaa
     3001 gcacaagcag aaactgggac aggagcccca ggagaccaaa tcttcatggt ccctctggga
     3061 ggatgggtgg ggagagctgt ggcagaggcc tcaggagggg ccctgctgct cagtggtgac
     3121 agataggggt gagaaagcag acagagtcat tccgtcagca ttctgggtct gtttggtact
     3181 tcttctcacg ctaaggtggc ggtgtgatat gcacaatggc taaaaagcag ggagagctgg
     3241 aaagaaacaa ggacagagac agaggccaag tcaaccagac caattcccag aggaagcaaa
     3301 gaaaccatta cagagactac aagggggaag ggaaggagag atgaattagc ttcccctgta
     3361 aaccttagaa cccagctgtt gccagggcaa cggggcaata cctgtctctt cagaggagat
     3421 gaagttgcca gggtaactac atcctgtctt tctcaaggac catcccagaa tgtggcaccc
     3481 actagccgtt accatagcaa ctgcctcttt gccccactta atcccatccc gtctgttaaa
     3541 agggccctat agttggaggt gggggaggta ggaagagcga tgatcacttg tggactaagt
     3601 ttgttcgcat ccccttctcc aaccccctca gtacatcacc ctgggggaac agggtccact
     3661 tgctcctggg cccacacagt cctgcagtat tgtgtatata aggccagggc aaagaggagc
     3721 aggttttaaa gtgaaaggca ggcaggtgtt ggggaggcag ttaccggggc aacgggaaca
     3781 gggcgtttcg gaggtggttg ccatggggac ctggatgctg acgaaggctc gcgaggctgt
     3841 gagcagccac agtgccctgc tcagaagccc caagctcgtc agtcaagccg gttctccgtt
     3901 tgcactcagg agcacgggca ggcgagtggc ccctagttct gggggcagcg ggggatccgc
     3961 caccatggct gagctgatca gcagcgccac cagatctctg tttgccgccg gaggcatcaa
     4021 cccttggcct aacccctacc accacgagga catgggctgt ggaggaatga cacctacagg
     4081 cgagtgcttc agcaccgagt ggtggtgtga cccttcttac ggactgagcg acgccggata
     4141 cggatattgc ttcgtggagg ccacaggcgg ctacctggtc gtgggagtgg agaagaagca
     4201 ggcttggctg cacagcagag gcacaccagg agaaaagatc ggcgcccagg tctgccagtg
     4261 gattgctttc agcatcgcca tcgccctgct gacattctac ggcttcagcg cctggaaggc
     4321 cacttgcggt tgggaggagg tctacgtctg ttgcgtcgag gtgctgttcg tgaccctgga
     4381 gatcttcaag gagttcagca gccccgccac agtgtacctg tctaccggca accacgccta
     4441 ttgcctgcgc tacttcgagt ggctgctgtc ttgccccgtg atcctgatca gactgagcaa
     4501 cctgagcggc ctgaagaacg actacagcaa gcggaccatg ggcctgatcg tgtcttgcgt
     4561 gggaatgatc gtgttcggca tggccgcagg actggctacc gattggctca agtggctgct
     4621 gtatatcgtg tcttgcatct acggcggcta catgtacttc caggccgcca agtgctacgt
     4681 ggaagccaac cacagcgtgc ctaaaggcca ttgccgcatg gtcgtgaagc tgatggccta
     4741 cgcttacttc gcctcttggg gcagctaccc aatcctctgg gcagtgggac cagaaggact
     4801 gctgaagctg agcccttacg ccaacagcat cggccacagc atctgcgaca tcatcgccaa
     4861 ggagttttgg accttcctgg cccaccacct gaggatcaag atccacgagc acatcctgat
     4921 ccacggcgac atccggaaga ccaccaagat ggagatcgga ggcgaggagg tggaagtgga
     4981 agagttcgtg gaggaggagg acgaggacac agtggcggca ccggtagtag cagtgagtaa
     5041 gggcgaggaa gtgatcaaag agttcatgcg gtttaaggtg agaatggaag gaagcatgaa
     5101 cggccacgag ttcgaaattg agggagaagg agagggacgg ccctacgagg gcacccagac
     5161 agccaagctg aaagtgacaa agggcgggcc tctgccattc gcttgggaca tcctgagccc
     5221 acagtttatg tacggctcca aggcctatgt gaaacatcca gctgacattc ccgattataa
     5281 gaaactgagc ttccccgagg ggtttaagtg ggaaagagtg atgaacttcg aggacggagg
     5341 cctggtgact gtgacccagg acagctccct gcaggatggg accctgatct acaaggtgaa
     5401 aatgagaggg acaaattttc cccctgatgg acctgtgatg cagaagaaaa ctatgggatg
     5461 ggaggcctcc accgaaaggc tgtatccacg cgacggggtg ctgaaaggag aaatccacca
     5521 ggctctgaag ctgaaagatg ggggacatta cctggtggag ttcaagacaa tctacatggc
     5581 caagaaacct gtgcagctgc caggctacta ttacgtggac acaaaactgg atatcacttc
     5641 acacaacgag gactacacta ttgtggagca gtatgaacgg agcgagggga gacaccatct
     5701 gttcctgggc catgggactg gaagtaccgg ctcagggtct agtggaaccg cctcaagcga
     5761 ggataacaat atggctgtga tcaaagagtt catgaggttt aaggtgcgca tggagggcag
     5821 catgaatggg cacgaatttg agattgaagg agagggcgaa gggaggcctt acgagggcac
     5881 acagactgcc aagctgaaag tgaccaaggg aggaccactg cctttcgctt gggatatcct
     5941 gtctcctcag tttatgtacg gaagtaaggc ctatgtcaag catcccgctg acattcctga
     6001 ttacaagaaa ctgtctttcc cagagggctt taagtgggag agagtgatga attttgaaga
     6061 tggaggcctg gtgaccgtga cacaggactc ctctctgcag gatggcactc tgatctacaa
     6121 agtcaaaatg cgcggcacca attttccacc cgatgggccc gtgatgcaga agaaaacaat
     6181 ggggtgggag gccagcactg aacggctgta tcctagagac ggagtgctga agggcgaaat
     6241 ccaccaggcc ctgaagctga aagacggcgg ccactacctg gtggagttca aaaccatcta
     6301 catggccaag aaaccagtgc agctgcccgg ctattactat gtggacacca agctggatat
     6361 cacatcccac aatgaagact acaccattgt ggaacagtat gagaggtctg aaggacgcca
     6421 ccatctgttt ctgtacggca tggatgagct gtataagtaa gaattcgata tcaagcttat
     6481 cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     6541 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     6601 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     6661 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     6721 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     6781 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     6841 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct
     6901 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     6961 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     7021 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata
     7081 ccgtcgaccc gggcggccgc tcataaagct cgactgtgcc ttctagttgc cagccatctg
     7141 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     7201 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     7261 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     7321 atgcggtccg gactgtacaa ttgcattcat tttatgtttc aggttcaggg ggagatgtgg
     7381 gaggtttttt aaagcaagta aaacctctac aaatgtggta aaatcgataa ggatcttcct
     7441 agagcatggc tacgtagata agtagcatgg cgggttaatc attaactaca aggaacccct
     7501 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc
     7561 aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag
     7621 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgct
//
