LOCUS       sequence-91634-e        7301 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector for ChrimsonR-tdTomato.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7301)
  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 7301)
  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: pCAG-ChrimsonR-tdT
FEATURES             Location/Qualifiers
     source          1..7301
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        385..660
                     /label=chicken beta-actin promoter
     intron          661..1674
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1598..1620
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1682..1701
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1735..1744
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1741..2790
                     /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"
     primer_bind     complement(4323..4342)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4388..4443
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4442..4461)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4804..4820)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4804..4820)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4817..4839)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4828..4844
                     /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)."
     promoter        complement(join(4852..4858,4859..4876,4877..4882))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4897..4918
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4977..5172
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5023..5158
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5178..5312
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5215..5234)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5269..5288
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5380..5397)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5551..6139)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5631..5650)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6310..7101,7102..7170))
                     /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     6933..6952
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7171..7275)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
      421 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
      481 cgatgggggc gggggggggg ggggcgcgcg ccaggcgggg cggggcgggg cgaggggcgg
      541 ggcggggcga ggcggagagg tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt
      601 ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg
      661 ggagtcgctg cgttgccttc gccccgtgcc ccgctccgcg ccgcctcgcg ccgcccgccc
      721 cggctctgac tgaccgcgtt actcccacag gtgagcgggc gggacggccc ttctcctccg
      781 ggctgtaatt agcgcttggt ttaatgacgg ctcgtttctt ttctgtggct gcgtgaaagc
      841 cttgaggggc tccgggaggg ccctttgtgc ggggggagcg gctcgggggg tgcgtgcgtg
      901 tgtgtgtgcg tggggagcgc cgcgtgcggc tccgcgctgc ccggcggctg tgagcgctgc
      961 gggcgcggcg cggggctttg tgcgctccgc agtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc
     1081 gtgggggggt gagcaggggg tgtgggcgcg tcggtcgggc tgcaaccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtac ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gcccccggag cgccggcggc
     1321 tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag
     1381 ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc
     1441 tctagcgggc gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc
     1501 cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg
     1561 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
     1621 ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg
     1681 ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattctg agccgccacc
     1741 atggctgagc tgatcagcag cgccaccaga tctctgtttg ccgccggagg catcaaccct
     1801 tggcctaacc cctaccacca cgaggacatg ggctgtggag gaatgacacc tacaggcgag
     1861 tgcttcagca ccgagtggtg gtgtgaccct tcttacggac tgagcgacgc cggatacgga
     1921 tattgcttcg tggaggccac aggcggctac ctggtcgtgg gagtggagaa gaagcaggct
     1981 tggctgcaca gcagaggcac accaggagaa aagatcggcg cccaggtctg ccagtggatt
     2041 gctttcagca tcgccatcgc cctgctgaca ttctacggct tcagcgcctg gaaggccact
     2101 tgcggttggg aggaggtcta cgtctgttgc gtcgaggtgc tgttcgtgac cctggagatc
     2161 ttcaaggagt tcagcagccc cgccacagtg tacctgtcta ccggcaacca cgcctattgc
     2221 ctgcgctact tcgagtggct gctgtcttgc cccgtgatcc tgatcagact gagcaacctg
     2281 agcggcctga agaacgacta cagcaagcgg accatgggcc tgatcgtgtc ttgcgtggga
     2341 atgatcgtgt tcggcatggc cgcaggactg gctaccgatt ggctcaagtg gctgctgtat
     2401 atcgtgtctt gcatctacgg cggctacatg tacttccagg ccgccaagtg ctacgtggaa
     2461 gccaaccaca gcgtgcctaa aggccattgc cgcatggtcg tgaagctgat ggcctacgct
     2521 tacttcgcct cttggggcag ctacccaatc ctctgggcag tgggaccaga aggactgctg
     2581 aagctgagcc cttacgccaa cagcatcggc cacagcatct gcgacatcat cgccaaggag
     2641 ttttggacct tcctggccca ccacctgagg atcaagatcc acgagcacat cctgatccac
     2701 ggcgacatcc ggaagaccac caagatggag atcggaggcg aggaggtgga agtggaagag
     2761 ttcgtggagg aggaggacga ggacacagtg gcggcaccgg tagtagcagt gagtaagggc
     2821 gaggaagtga tcaaagagtt catgcggttt aaggtgagaa tggaaggaag catgaacggc
     2881 cacgagttcg aaattgaggg agaaggagag ggacggccct acgagggcac ccagacagcc
     2941 aagctgaaag tgacaaaggg cgggcctctg ccattcgctt gggacatcct gagcccacag
     3001 tttatgtacg gctccaaggc ctatgtgaaa catccagctg acattcccga ttataagaaa
     3061 ctgagcttcc ccgaggggtt taagtgggaa agagtgatga acttcgagga cggaggcctg
     3121 gtgactgtga cccaggacag ctccctgcag gatgggaccc tgatctacaa ggtgaaaatg
     3181 agagggacaa attttccccc tgatggacct gtgatgcaga agaaaactat gggatgggag
     3241 gcctccaccg aaaggctgta tccacgcgac ggggtgctga aaggagaaat ccaccaggct
     3301 ctgaagctga aagatggggg acattacctg gtggagttca agacaatcta catggccaag
     3361 aaacctgtgc agctgccagg ctactattac gtggacacaa aactggatat cacttcacac
     3421 aacgaggact acactattgt ggagcagtat gaacggagcg aggggagaca ccatctgttc
     3481 ctgggccatg ggactggaag taccggctca gggtctagtg gaaccgcctc aagcgaggat
     3541 aacaatatgg ctgtgatcaa agagttcatg aggtttaagg tgcgcatgga gggcagcatg
     3601 aatgggcacg aatttgagat tgaaggagag ggcgaaggga ggccttacga gggcacacag
     3661 actgccaagc tgaaagtgac caagggagga ccactgcctt tcgcttggga tatcctgtct
     3721 cctcagttta tgtacggaag taaggcctat gtcaagcatc ccgctgacat tcctgattac
     3781 aagaaactgt ctttcccaga gggctttaag tgggagagag tgatgaattt tgaagatgga
     3841 ggcctggtga ccgtgacaca ggactcctct ctgcaggatg gcactctgat ctacaaagtc
     3901 aaaatgcgcg gcaccaattt tccacccgat gggcccgtga tgcagaagaa aacaatgggg
     3961 tgggaggcca gcactgaacg gctgtatcct agagacggag tgctgaaggg cgaaatccac
     4021 caggccctga agctgaaaga cggcggccac tacctggtgg agttcaaaac catctacatg
     4081 gccaagaaac cagtgcagct gcccggctat tactatgtgg acaccaagct ggatatcaca
     4141 tcccacaatg aagactacac cattgtggaa cagtatgaga ggtctgaagg acgccaccat
     4201 ctgtttctgt acggcatgga tgagctgtat aagtaaagcg gccgcactcc tcaggtgcag
     4261 gctgcctatc agaaggtggt ggctggtgtg gccaatgccc tggctcacaa ataccactga
     4321 gatctttttc cctctgccaa aaattatggg gacatcatga agccccttga gcatctgact
     4381 tctggctaat aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc
     4441 tcactcggaa ggacatatgg gagggcaaat catttaaaac atcagaatga gtatttggtt
     4501 tagagtttgg caacatatgc ccatatgctg gctgccatga acaaaggttg gctataaaga
     4561 ggtcatcagt atatgaaaca gccccctgct gtccattcct tattccatag aaaagccttg
     4621 acttgaggtt agattttttt tatattttgt tttgtgttat ttttttcttt aacatcccta
     4681 aaattttcct tacatgtttt actagccaga tttttcctcc tctcctgact actcccagtc
     4741 atagctgtcc ctcttctctt atggagatcc ctcgacctgc agcccaagct tggcgtaatc
     4801 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     4861 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     4921 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc ggatccgcat
     4981 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     5041 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     5101 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     5161 ggcttttgca aaaagctaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     5221 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     5281 aactcatcaa tgtatcttat catgtctgga tccgctgcat taatgaatcg gccaacgcgc
     5341 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     5401 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5461 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5521 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5581 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5641 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5701 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     5761 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5821 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5881 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5941 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     6001 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     6061 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     6121 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     6181 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     6241 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     6301 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6361 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     6421 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6481 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6541 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6601 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6661 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6721 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     6781 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     6841 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6901 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6961 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     7021 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     7081 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     7141 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     7201 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     7261 aataggggtt ccgcgcacat ttccccgaaa agtgccacct g
//
