LOCUS       sequence-451770-        7805 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cation channelrhodopsin ChRmine fused to mRuby2 fluorophore and 
            targeted to the neuronal soma and proximal dendrites for optogenetic
            excitation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7805)
  AUTHORS   Sridharan S, Gajowa MA, Ogando MB, Jagadisan UK, Abdeladim L, 
            Sadahiro M, Bounds HA, Hendricks WD, Turney TS, Tayler I, Gopakumar 
            K, Oldenburg IA, Brohawn SG, Adesnik H
  TITLE     High-performance microbial opsins for spatially and temporally 
            precise perturbations of large neuronal networks.
  JOURNAL   Neuron. 2022 Jan 27. pii: S0896-6273(22)00008-3. doi: 
            10.1016/j.neuron.2022.01.008.
  PUBMED    35120626
REFERENCE   2  (bases 1 to 7805)
  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: "Neuron. 
            2022 Jan 27. pii: S0896-6273(22)00008-3. doi: 
            10.1016/j.neuron.2022.01.008."
COMMENT     Sequence Label: pCAG-ChRmine-mRuby2-ST
FEATURES             Location/Qualifiers
     source          1..7805
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        385..662
                     /label=chicken beta-actin promoter
     intron          663..1679
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1603..1625
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1687..1706
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             2748..3458
                     /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          3700..3832
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     3815..3834
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    3841..4429
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3894..3914)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4312..4323)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4456..4473)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4462..4573
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4669..4689)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4669..4687)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4708..4724)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4826..4845)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4891..4946
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4945..4964)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5307..5323)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5331..5347
                     /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(5355..5361,5362..5379,5380..5385))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5400..5421
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5480..5675
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5526..5661
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5681..5815
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5718..5737)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5772..5791
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5883..5900)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6054..6642)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6134..6153)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6813..7604,7605..7673))
                     /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     7436..7455
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7674..7778)
                     /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 ggggggcgcg cgccaggcgg ggcggggcgg ggcgaggggc
      541 ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
      601 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
      661 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc
      721 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
      781 ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg
      841 aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt
      901 gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc
      961 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg
     1021 ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg
     1081 tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc
     1141 acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc
     1201 gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg
     1261 cggggccgcc tcgggccggg gagggctcgg gggaggggcg cggcggcccc cggagcgccg
     1321 gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg
     1381 cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca
     1441 ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg
     1501 agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc
     1561 cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg
     1621 tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc
     1681 tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttcggtacca
     1741 ccatggcaca cgcaccaggc accgaccaga tgttctacgt gggcacaatg gacggctggt
     1801 atctggatac caagctgaac tccgtggcca tcggcgccca ctggtcttgc tttatcgtgc
     1861 tgacaatcac cacattctac ctgggctatg agtcctggac atctcggggc cctagcaaga
     1921 gaacctcctt ttacgccggc tatcaggagg agcagaacct ggccctgttt gtgaatttct
     1981 ttgccatgct gagctacttc ggcaagatcg tggccgacac actgggccac aacttcggcg
     2041 atgtgggccc cttcatcatc ggcttcggca attacaggta tgccgactac atgctgacct
     2101 gcccaatgct ggtgtacgat ctgctgtatc agctgagggc cccctatcgc gtgtcttgta
     2161 gcgccatcat ctttgccatc ctgatgtctg gcgtgctggc cgagttctac gcagagggcg
     2221 accctaggct gaggaatggc gcctacgcct ggtatggctt tggctgtttc tggtttatct
     2281 tcgcctactc tatcgtgatg agcatcgtgg ccaagcagta tagccggctg gcccagctgg
     2341 cccaggatac aggcgccgag cacagcctgc acgtgctgaa gttcgccgtg tttaccttct
     2401 ccatgctgtg gattctgttt cccctggtgt gggccatctg ccctagaggc ttcggctgga
     2461 tcgacgataa ctggacagag gtggcccact gcgtgtgcga catcgtggcc aagtcctgtt
     2521 acggctttgc cctggcccgg ttcagaaaga cctatgatga ggagctgttt cggctgctgg
     2581 agcagctggg acacgacgag gatgagttcc agaagctgga gctggatatg aggctgagca
     2641 gcaatggcga gcgcctgcgg agactgtctg cggccgccaa gagcaggatc accagcgagg
     2701 gcgagtacat ccccctggac cagatcgaca tcaacgtggt accggtaatg gtgtctaagg
     2761 gcgaagagct gatcaaggaa aatatgcgta tgaaggtggt catggaaggt tcggtcaacg
     2821 gccaccaatt caaatgcaca ggtgaaggag aaggcaatcc gtacatggga actcaaacca
     2881 tgaggatcaa agtcatcgag ggaggacccc tgccatttgc ctttgacatt cttgccacgt
     2941 cgttcatgta tggcagccgt acttttatca agtacccgaa aggcattcct gatttcttta
     3001 aacagtcctt tcctgagggt tttacttggg aaagagttac gagatacgaa gatggtggag
     3061 tcgtcaccgt catgcaggac accagccttg aggatggctg tctcgtttac cacgtccaag
     3121 tcagaggggt aaactttccc tccaatggtc ccgtgatgca gaagaagacc aagggttggg
     3181 agcctaatac agagatgatg tatccagcag atggtggtct gaggggatac actcatatgg
     3241 cactgaaagt tgatggtggt ggccatctgt cttgctcttt cgtaacaact tacaggtcaa
     3301 aaaagaccgt cgggaacatc aagatgcccg gtatccatgc cgttgatcac cgcctggaaa
     3361 ggttagagga aagtgacaat gaaatgttcg tagtacaacg cgaacacgca gttgccaagt
     3421 tcgccgggct tggtggtggg atggacgagc tgtacaaggc cctagagcag tcccagccca
     3481 tcctcaacac caaggagatg gccccgcaga gcaagcctcc agaggagctg gagatgagca
     3541 gcatgcccag ccccgtggcc cctctgcccg cacgcacgga gggcgtcatc gacatgcgga
     3601 gcatgtccag cattgacagc ttcatcagct gtgccacgga cttccctgaa gccaccagat
     3661 tcttctgcta cgagaacgag gtgtaactcg aggccgcagg taagtatcaa ggttacaaga
     3721 caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact cttgcgtttc
     3781 tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac aggtgtcgac
     3841 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     3901 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     3961 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     4021 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     4081 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     4141 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     4201 ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc
     4261 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     4321 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     4381 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaattcga
     4441 gctcggtacg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     4501 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     4561 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     4621 aggacagcaa gggggaggat tgggaagaca atagcccagc ttttgttccc tttagtgagg
     4681 gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     4741 gctaattcac tcctcaggtg caggctgcct atcagaaggt ggtggctggt gtggccaatg
     4801 ccctggctca caaataccac tgagatcttt ttccctctgc caaaaattat ggggacatca
     4861 tgaagcccct tgagcatctg acttctggct aataaaggaa atttattttc attgcaatag
     4921 tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca aatcatttaa
     4981 aacatcagaa tgagtatttg gtttagagtt tggcaacata tgcccatatg ctggctgcca
     5041 tgaacaaagg ttggctataa agaggtcatc agtatatgaa acagccccct gctgtccatt
     5101 ccttattcca tagaaaagcc ttgacttgag gttagatttt ttttatattt tgttttgtgt
     5161 tatttttttc tttaacatcc ctaaaatttt ccttacatgt tttactagcc agatttttcc
     5221 tcctctcctg actactccca gtcatagctg tccctcttct cttatggaga tccctcgacc
     5281 tgcagcccaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     5341 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     5401 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     5461 ctgtcgtgcc agcggatccg catctcaatt agtcagcaac catagtcccg cccctaactc
     5521 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     5581 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     5641 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct aacttgttta ttgcagctta
     5701 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     5761 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatccgctg
     5821 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     5881 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     5941 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     6001 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     6061 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     6121 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     6181 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     6241 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     6301 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     6361 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     6421 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     6481 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     6541 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     6601 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     6661 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     6721 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     6781 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     6841 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     6901 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     6961 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     7021 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     7081 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     7141 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     7201 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     7261 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     7321 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     7381 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     7441 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     7501 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     7561 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     7621 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     7681 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     7741 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     7801 cctgg
//
