LOCUS       sequence-203780-        6799 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cation channelrhodopsin ChrimsonR fused to mRuby2 fluorophore and 
            targeted to the neuronal soma and proximal dendrites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6799)
  AUTHORS   Pegard NC, Mardinly AR, Oldenburg IA, Sridharan S, Waller L, Adesnik
            H
  TITLE     Three-dimensional scanless holographic optogenetics with temporal 
            focusing (3D-SHOT).
  JOURNAL   Nat Commun. 2017 Oct 31;8(1):1228. doi: 10.1038/s41467-017-01031-3.
  PUBMED    29089483
REFERENCE   2  (bases 1 to 6799)
  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"; journalName: "Nat 
            Commun."; date: "2017-10-31"; volume: "8(1):1228. doi"; pages: " 
            10.1038/s41467-017-01031-3"
COMMENT     Sequence Label: pCAGGS-ChrimsonR-mRuby2-ST
FEATURES             Location/Qualifiers
     source          1..6799
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          169..1185
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1109..1131
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1193..1212
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1246..1255
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1252..2301
                     /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"
     CDS             2314..3024
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
                     MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
                     AKFAGLGGGMDELYK"
     primer_bind     complement(3326..3345)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3391..3446
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3445..3464)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3807..3823)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3807..3823)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3820..3842)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3831..3847
                     /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(3855..3861,3862..3879,3880..3885))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3900..3921
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3980..4175
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4026..4161
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4181..4315
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4218..4237)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4272..4291
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4383..4400)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4554..5142)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4634..4653)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5313..6104,6105..6173))
                     /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     5936..5955
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6174..6278)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        6309..6688
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(6690..6799,1..168)
                     /label=chicken beta-actin promoter
ORIGIN
        1 gggggggggg ggcgcgcgcc aggcggggcg gggcggggcg aggggcgggg cggggcgagg
       61 cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg
      121 aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg agtcgctgcg
      181 cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga
      241 ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc gggctgtaat
      301 tagcgcttgg tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag ccttgagggg
      361 ctccgggagg gccctttgtg cggggggagc ggctcggggg gtgcgtgcgt gtgtgtgtgc
      421 gtggggagcg ccgcgtgcgg ctccgcgctg cccggcggct gtgagcgctg cgggcgcggc
      481 gcggggcttt gtgcgctccg cagtgtgcgc gaggggagcg cggccggggg cggtgccccg
      541 cggtgcgggg ggggctgcga ggggaacaaa ggctgcgtgc ggggtgtgtg cgtggggggg
      601 tgagcagggg gtgtgggcgc gtcggtcggg ctgcaacccc ccctgcaccc ccctccccga
      661 gttgctgagc acggcccggc ttcgggtgcg gggctccgta cggggcgtgg cgcggggctc
      721 gccgtgccgg gcggggggtg gcggcaggtg ggggtgccgg gcggggcggg gccgcctcgg
      781 gccggggagg gctcggggga ggggcgcggc ggcccccgga gcgccggcgg ctgtcgaggc
      841 gcggcgagcc gcagccattg ccttttatgg taatcgtgcg agagggcgca gggacttcct
      901 ttgtcccaaa tctgtgcgga gccgaaatct gggaggcgcc gccgcacccc ctctagcggg
      961 cgcggggcga agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg ccttcgtgcg
     1021 tcgccgcgcc gccgtcccct tctccctctc cagcctcggg gctgtccgcg gggggacggc
     1081 tgccttcggg ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac cggcggctct
     1141 agagcctctg ctaaccatgt tcatgccttc ttctttttcc tacagctcct gggcaacgtg
     1201 ctggttattg tgctgtctca tcattttggc aaagaattct gagccgccac catggctgag
     1261 ctgatcagca gcgccaccag atctctgttt gccgccggag gcatcaaccc ttggcctaac
     1321 ccctaccacc acgaggacat gggctgtgga ggaatgacac ctacaggcga gtgcttcagc
     1381 accgagtggt ggtgtgaccc ttcttacgga ctgagcgacg ccggatacgg atattgcttc
     1441 gtggaggcca caggcggcta cctggtcgtg ggagtggaga agaagcaggc ttggctgcac
     1501 agcagaggca caccaggaga aaagatcggc gcccaggtct gccagtggat tgctttcagc
     1561 atcgccatcg ccctgctgac attctacggc ttcagcgcct ggaaggccac ttgcggttgg
     1621 gaggaggtct acgtctgttg cgtcgaggtg ctgttcgtga ccctggagat cttcaaggag
     1681 ttcagcagcc ccgccacagt gtacctgtct accggcaacc acgcctattg cctgcgctac
     1741 ttcgagtggc tgctgtcttg ccccgtgatc ctgatcagac tgagcaacct gagcggcctg
     1801 aagaacgact acagcaagcg gaccatgggc ctgatcgtgt cttgcgtggg aatgatcgtg
     1861 ttcggcatgg ccgcaggact ggctaccgat tggctcaagt ggctgctgta tatcgtgtct
     1921 tgcatctacg gcggctacat gtacttccag gccgccaagt gctacgtgga agccaaccac
     1981 agcgtgccta aaggccattg ccgcatggtc gtgaagctga tggcctacgc ttacttcgcc
     2041 tcttggggca gctacccaat cctctgggca gtgggaccag aaggactgct gaagctgagc
     2101 ccttacgcca acagcatcgg ccacagcatc tgcgacatca tcgccaagga gttttggacc
     2161 ttcctggccc accacctgag gatcaagatc cacgagcaca tcctgatcca cggcgacatc
     2221 cggaagacca ccaagatgga gatcggaggc gaggaggtgg aagtggaaga gttcgtggag
     2281 gaggaggacg aggacacagt ggcggcaccg gtaatggtgt ctaagggcga agagctgatc
     2341 aaggaaaata tgcgtatgaa ggtggtcatg gaaggttcgg tcaacggcca ccaattcaaa
     2401 tgcacaggtg aaggagaagg caatccgtac atgggaactc aaaccatgag gatcaaagtc
     2461 atcgagggag gacccctgcc atttgccttt gacattcttg ccacgtcgtt catgtatggc
     2521 agccgtactt ttatcaagta cccgaaaggc attcctgatt tctttaaaca gtcctttcct
     2581 gagggtttta cttgggaaag agttacgaga tacgaagatg gtggagtcgt caccgtcatg
     2641 caggacacca gccttgagga tggctgtctc gtttaccacg tccaagtcag aggggtaaac
     2701 tttccctcca atggtcccgt gatgcagaag aagaccaagg gttgggagcc taatacagag
     2761 atgatgtatc cagcagatgg tggtctgagg ggatacactc atatggcact gaaagttgat
     2821 ggtggtggcc atctgtcttg ctctttcgta acaacttaca ggtcaaaaaa gaccgtcggg
     2881 aacatcaaga tgcccggtat ccatgccgtt gatcaccgcc tggaaaggtt agaggaaagt
     2941 gacaatgaaa tgttcgtagt acaacgcgaa cacgcagttg ccaagttcgc cgggcttggt
     3001 ggtgggatgg acgagctgta caaggcccta gagcagtccc agcccatcct caacaccaag
     3061 gagatggccc cgcagagcaa gcctccagag gagctggaga tgagcagcat gcccagcccc
     3121 gtggcccctc tgcccgcacg cacggagggc gtcatcgaca tgcggagcat gtccagcatt
     3181 gacagcttca tcagctgtgc cacggacttc cctgaagcca ccagattcta aggactcgag
     3241 gcggccgcac tcctcaggtg caggctgcct atcagaaggt ggtggctggt gtggccaatg
     3301 ccctggctca caaataccac tgagatcttt ttccctctgc caaaaattat ggggacatca
     3361 tgaagcccct tgagcatctg acttctggct aataaaggaa atttattttc attgcaatag
     3421 tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca aatcatttaa
     3481 aacatcagaa tgagtatttg gtttagagtt tggcaacata tgcccatatg ctggctgcca
     3541 tgaacaaagg ttggctataa agaggtcatc agtatatgaa acagccccct gctgtccatt
     3601 ccttattcca tagaaaagcc ttgacttgag gttagatttt ttttatattt tgttttgtgt
     3661 tatttttttc tttaacatcc ctaaaatttt ccttacatgt tttactagcc agatttttcc
     3721 tcctctcctg actactccca gtcatagctg tccctcttct cttatggaga tccctcgacc
     3781 tgcagcccaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     3841 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     3901 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     3961 ctgtcgtgcc agcggatccg catctcaatt agtcagcaac catagtcccg cccctaactc
     4021 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     4081 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     4141 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct aacttgttta ttgcagctta
     4201 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     4261 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatccgctg
     4321 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     4381 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     4441 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     4501 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4561 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     4621 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     4681 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     4741 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     4801 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     4861 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     4921 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     4981 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     5041 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     5101 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     5161 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     5221 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     5281 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     5341 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     5401 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     5461 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     5521 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     5581 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     5641 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     5701 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     5761 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     5821 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     5881 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     5941 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     6001 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     6061 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     6121 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     6181 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     6241 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     6301 cctgggtcga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
     6361 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
     6421 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
     6481 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     6541 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
     6601 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
     6661 ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc cccacgttct gcttcactct
     6721 ccccatctcc cccccctccc cacccccaat tttgtattta tttatttttt aattattttg
     6781 tgcagcgatg ggggcgggg
//
