LOCUS       sequence-213552-        7833 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cation channelrhodopsin CsChR 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 7833)
  AUTHORS   Mardinly AR, Oldenburg IA, Pegard NC, Sridharan S, Lyall EH, Chesnov
            K, Brohawn SG, Waller L, Adesnik H
  TITLE     Precise multimodal optical control of neural ensemble activity.
  JOURNAL   Nat Neurosci. 2018 Apr 30. pii: 10.1038/s41593-018-0139-8. doi: 
            10.1038/s41593-018-0139-8.
  PUBMED    29713079
REFERENCE   2  (bases 1 to 7833)
  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 
            Neurosci. 2018 Apr 30. pii: 10.1038/s41593-018-0139-8. doi: 
            10.1038/s41593-018-0139-8."
COMMENT     Sequence Label: pCAG-CsChR-mRuby2-ST
FEATURES             Location/Qualifiers
     source          1..7833
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1678
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1602..1624
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1686..1705
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             2795..3505
                     /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"
     intron          3729..3861
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     3844..3863
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    3870..4458
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3923..3943)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4341..4352)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4485..4502)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4491..4602
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4698..4718)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4698..4716)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4737..4753)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4855..4874)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4920..4975
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4974..4993)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5336..5352)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5360..5376
                     /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(5384..5390,5391..5408,5409..5414))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5429..5450
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5509..5704
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5555..5690
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5710..5844
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5747..5766)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5801..5820
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5912..5929)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6083..6671)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6163..6182)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6842..7633,7634..7702))
                     /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     7465..7484
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7703..7807)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcgggggggg gcgggggggg
      541 ggggggggcg gggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
      721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
      841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
      901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
      961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
     1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
     1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
     1261 ggggccgcct cgggccgggg agggctcggg ggaggggcgc ggcggccccc ggagcgccgg
     1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
     1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
     1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
     1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1621 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1681 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tcggtaccac
     1741 catgagcaga ctggtcgccg cttcttggct gctggctctc ctcctctgcg gaattaccag
     1801 cacaacaaca gcctctagcg ccccagcagc ttcttctaca gacggaacag ccgccgcagc
     1861 agtgtctcac tacgccatga acggcttcga cgagctggct aaaggagccg tggtgccaga
     1921 agaccacttt gtctgcggac cagccgacaa gtgctattgc tccgcttggc tgcacagcca
     1981 cggaagcaag gaggagaaga ccgccttcac cgtcatgcag tggatcgtgt tcgccgtctg
     2041 catcatcagc ctgctgttct acgcctacca gacttggagg gctacttgcg gttgggagga
     2101 ggtgtacgtg accatcatcg agctggtcca cgtctgcttc ggactctggc acgaggtcga
     2161 tagcccttgt accctgtacc tgagcacagg caacatggtc ctctggctga gatacgccga
     2221 gtggctgctg acttgccccg tgatcctgat ccacctgagc aacctgaccg gcatgaagaa
     2281 cgactacaac aagcggacca tggccctgct ggtgtcagac gtgggctgta tcgtgtgggg
     2341 aacaacagcc gccctgagca ccgatttcgt gaagatcatc ttcttcttcc tgggcctgct
     2401 gtacggcttc tacaccttct acgccgccgc caagatctac atcgaggcct accacaccgt
     2461 gcccaagggc atttgtagac agctcgtgcg gctgcaggcc tacgacttct tcttcacttg
     2521 gagcatgttc cccatcctgt tcatggtcgg cccagaggga ttcggcaaga tcaccgccta
     2581 cagcagcgga atcgcccacg aagtgtgcga tctgctgagc aagaacctct ggggcctgat
     2641 gggccacttc atccgcgtga agatccacga gcacatcctg gtgcacggca acatcaccaa
     2701 gaagaccaag gtcaacgtgg ccggcgacat ggtggaactg gacacctacg tggaccagga
     2761 cgaggaacac gacgagggag cggccgcacc ggtaatggtg tctaagggcg aagagctgat
     2821 caaggaaaat atgcgtatga aggtggtcat ggaaggttcg gtcaacggcc accaattcaa
     2881 atgcacaggt gaaggagaag gcaatccgta catgggaact caaaccatga ggatcaaagt
     2941 catcgaggga ggacccctgc catttgcctt tgacattctt gccacgtcgt tcatgtatgg
     3001 cagccgtact tttatcaagt acccgaaagg cattcctgat ttctttaaac agtcctttcc
     3061 tgagggtttt acttgggaaa gagttacgag atacgaagat ggtggagtcg tcaccgtcat
     3121 gcaggacacc agccttgagg atggctgtct cgtttaccac gtccaagtca gaggggtaaa
     3181 ctttccctcc aatggtcccg tgatgcagaa gaagaccaag ggttgggagc ctaatacaga
     3241 gatgatgtat ccagcagatg gtggtctgag gggatacact catatggcac tgaaagttga
     3301 tggtggtggc catctgtctt gctctttcgt aacaacttac aggtcaaaaa agaccgtcgg
     3361 gaacatcaag atgcccggta tccatgccgt tgatcaccgc ctggaaaggt tagaggaaag
     3421 tgacaatgaa atgttcgtag tacaacgcga acacgcagtt gccaagttcg ccgggcttgg
     3481 tggtgggatg gacgagctgt acaaggccct agagcagtcc cagcccatcc tcaacaccaa
     3541 ggagatggcc ccgcagagca agcctccaga ggagctggag atgagcagca tgcccagccc
     3601 cgtggcccct ctgcccgcac gcacggaggg cgtcatcgac atgcggagca tgtccagcat
     3661 tgacagcttc atcagctgtg ccacggactt ccctgaagcc accagattct aaggactcga
     3721 ggccgcaggt aagtatcaag gttacaagac aggtttaagg agaccaatag aaactgggct
     3781 tgtcgagaca gagaagactc ttgcgtttct gataggcacc tattggtctt actgacatcc
     3841 actttgcctt tctctccaca ggtgtcgaca atcaacctct ggattacaaa atttgtgaaa
     3901 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     3961 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     4021 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     4081 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     4141 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     4201 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     4261 ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     4321 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     4381 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     4441 tttgggccgc ctccccgcct ggaattcgag ctcggtacga tcagcctcga ctgtgccttc
     4501 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     4561 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     4621 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     4681 tagcccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta atcatggtca
     4741 tagctgtttc ctgtgtgaaa ttgttatccg ctaattcact cctcaggtgc aggctgccta
     4801 tcagaaggtg gtggctggtg tggccaatgc cctggctcac aaataccact gagatctttt
     4861 tccctctgcc aaaaattatg gggacatcat gaagcccctt gagcatctga cttctggcta
     4921 ataaaggaaa tttattttca ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg
     4981 aaggacatat gggagggcaa atcatttaaa acatcagaat gagtatttgg tttagagttt
     5041 ggcaacatat gcccatatgc tggctgccat gaacaaaggt tggctataaa gaggtcatca
     5101 gtatatgaaa cagccccctg ctgtccattc cttattccat agaaaagcct tgacttgagg
     5161 ttagattttt tttatatttt gttttgtgtt atttttttct ttaacatccc taaaattttc
     5221 cttacatgtt ttactagcca gatttttcct cctctcctga ctactcccag tcatagctgt
     5281 ccctcttctc ttatggagat ccctcgacct gcagcccaag cttggcgtaa tcatggtcat
     5341 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     5401 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     5461 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gcggatccgc atctcaatta
     5521 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     5581 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     5641 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     5701 caaaaagcta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     5761 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     5821 aatgtatctt atcatgtctg gatccgctgc attaatgaat cggccaacgc gcggggagag
     5881 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     5941 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     6001 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     6061 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     6121 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     6181 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     6241 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     6301 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     6361 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     6421 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     6481 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     6541 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     6601 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     6661 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     6721 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     6781 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     6841 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     6901 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     6961 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     7021 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     7081 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     7141 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     7201 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     7261 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     7321 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     7381 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     7441 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     7501 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     7561 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     7621 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     7681 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     7741 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     7801 ttccgcgcac atttccccga aaagtgccac ctg
//
