LOCUS       sequence-130463-        6872 bp    DNA     circular SYN 14-APR-2025
DEFINITION  AAV-mediated expression of Chrimson-GFP under the CaMKII promoter. 
            Using SV40pA signal..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6872)
  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 6872)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-Chrimson-GFP
FEATURES             Location/Qualifiers
     source          1..6872
                     /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=Chrimson
                     /translation="MAELISSATRSLFAAGGINPWPNPYHHEDMGCGGMTPTGECFSTE
                     WWCDPSYGLSDAGYGYCFVEATGGYLVVGVEKKQAWLHSRGTPGEKIGAQVCQWIAFSI
                     AIALLTFYGFSAWKATCGWEEVYVCCVEVLFVTLEIFKEFSSPATVYLSTGNHAYCLRY
                     FEWLLSCPVILIKLSNLSGLKNDYSKRTMGLIVSCVGMIVFGMAAGLATDWLKWLLYIV
                     SCIYGGYMYFQAAKCYVEANHSVPKGHCRMVVKLMAYAYFASWGSYPILWAVGPEGLLK
                     LSPYANSIGHSICDIIAKEFWTFLAHHLRIKIHEHILIHGDIRKTTKMEIGGEEVEVEE
                     FVEEEDEDTV"
     CDS             join(5033..5035,5036..5749)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(5033..5035,5036..5746)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(5075..5096)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5336..5355)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5683..5704
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    5774..6362
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5827..5847)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6245..6256)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    6405..6526
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6416..6435)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6470..6489
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   6688..6817
                     /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 agctgagcaa
     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 cagtgagcaa
     5041 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     5101 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     5161 cctgaagttc atttgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     5221 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
     5281 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     5341 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     5401 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     5461 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt
     5521 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     5581 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     5641 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     5701 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaag aattcgatat
     5761 caagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     5821 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     5881 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     5941 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     6001 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     6061 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     6121 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt
     6181 tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     6241 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     6301 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     6361 gcatcgatac cgtcgacccg ggcggccgct tcgagcagac atgataagat acattgatga
     6421 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     6481 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     6541 cattcatttt atgtttcagg ttcaggggga gatgtgggag gttttttaaa gcaagtaaaa
     6601 cctctacaaa tgtggtaaaa tcgataagga tcttcctaga gcatggctac gtagataagt
     6661 agcatggcgg gttaatcatt aactacaagg aacccctagt gatggagttg gccactccct
     6721 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct
     6781 ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cattaatgaa tcggccaacg
     6841 cgcggggaga ggcggtttgc gtattgggcg ct
//
