LOCUS       sequence-213543-        7791 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Modified cation channelrhodopsin ChroME 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 7791)
  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 7791)
  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-ChroME(S273A)-mRuby2-ST
FEATURES             Location/Qualifiers
     source          1..7791
                     /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             2756..3466
                     /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          3687..3819
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     3802..3821
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    3828..4416
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3881..3901)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4299..4310)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4443..4460)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4449..4560
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4656..4676)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4656..4674)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4695..4711)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4813..4832)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4878..4933
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4932..4951)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5294..5310)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5318..5334
                     /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(5342..5348,5349..5366,5367..5372))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5387..5408
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5467..5662
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5513..5648
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5668..5802
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5705..5724)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5759..5778
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5870..5887)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6041..6629)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6121..6140)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6800..7591,7592..7660))
                     /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     7423..7442
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7661..7765)
                     /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 gccggggggg gcgggggggg gggggggggg
      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 tcggtaccgc
     1741 gggcccggga tccaccggtg ctagtatgga aacagccgcc acaatgaccc acgcctttat
     1801 ctcagccgtg cctagcgccg aagccacaat tagaggcctg ctgagcgccg cagcagtggt
     1861 gacaccagca gcagacgctc acggagaaac ctctaacgcc acaacagccg gagccgatca
     1921 cggttgcttc ccccacatca accacggaac cgagctgcag cacaagatcg cagtgggact
     1981 ccagtggttc accgtgatcg tggctatcgt gcagctcatc ttctacggtt ggcacagctt
     2041 caaggccaca accggctggg aggaggtcta cgtctgcgtg atcgagctcg tcaagtgctt
     2101 catcgagctg ttccacgagg tcgacagccc agccacagtg taccagacca acggaggagc
     2161 cgtgatttgg ctgcggtaca gcgagtggct cctgacttgc cccgtgatcc tgatccacct
     2221 gagcaacctg accggactgc acgaagagta cagcaagcgg accatgacca tcctggtgac
     2281 cgacatcggc aacatcgtgt gggggatcac agccgccttt acaaagggcc ccctgaagat
     2341 cctgttcttc atgatcggcc tgttctacgg cgtgacttgc ttcttccaga tcgccaaggt
     2401 gtatatcgag agctaccaca ccctgcccaa aggcgtctgc cggaagattt gcaagatcat
     2461 ggcctacgtc ttcttctgct cttggctgat gttccccgtg atgttcatcg ccggacacga
     2521 gggactgggc ctgatcacac cttacaccag cggaatcggc cacctgatcc tggatctgat
     2581 cgccaagaac acttggggct tcctgggcca ccacctgaga gtgaagatcc acgagcacat
     2641 cctgatccac ggcgacatcc ggaagacaac caccatcaac gtggccggcg agaacatgga
     2701 gatcgagacc ttcgtcgacg aggaggagga gggaggagtg gcggccgcac cggtaatggt
     2761 gtctaagggc gaagagctga tcaaggaaaa tatgcgtatg aaggtggtca tggaaggttc
     2821 ggtcaacggc caccaattca aatgcacagg tgaaggagaa ggcaatccgt acatgggaac
     2881 tcaaaccatg aggatcaaag tcatcgaggg aggacccctg ccatttgcct ttgacattct
     2941 tgccacgtcg ttcatgtatg gcagccgtac ttttatcaag tacccgaaag gcattcctga
     3001 tttctttaaa cagtcctttc ctgagggttt tacttgggaa agagttacga gatacgaaga
     3061 tggtggagtc gtcaccgtca tgcaggacac cagccttgag gatggctgtc tcgtttacca
     3121 cgtccaagtc agaggggtaa actttccctc caatggtccc gtgatgcaga agaagaccaa
     3181 gggttgggag cctaatacag agatgatgta tccagcagat ggtggtctga ggggatacac
     3241 tcatatggca ctgaaagttg atggtggtgg ccatctgtct tgctctttcg taacaactta
     3301 caggtcaaaa aagaccgtcg ggaacatcaa gatgcccggt atccatgccg ttgatcaccg
     3361 cctggaaagg ttagaggaaa gtgacaatga aatgttcgta gtacaacgcg aacacgcagt
     3421 tgccaagttc gccgggcttg gtggtgggat ggacgagctg tacaaggccc tagagcagtc
     3481 ccagcccatc ctcaacacca aggagatggc cccgcagagc aagcctccag aggagctgga
     3541 gatgagcagc atgcccagcc ccgtggcccc tctgcccgca cgcacggagg gcgtcatcga
     3601 catgcggagc atgtccagca ttgacagctt catcagctgt gccacggact tccctgaagc
     3661 caccagattc taactcgagg ccgcaggtaa gtatcaaggt tacaagacag gtttaaggag
     3721 accaatagaa actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta
     3781 ttggtcttac tgacatccac tttgcctttc tctccacagg tgtcgacaat caacctctgg
     3841 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     3901 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     3961 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
     4021 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
     4081 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
     4141 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     4201 attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca
     4261 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
     4321 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
     4381 agacgagtcg gatctccctt tgggccgcct ccccgcctgg aattcgagct cggtacgatc
     4441 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     4501 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     4561 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     4621 ggaggattgg gaagacaata gcccagcttt tgttcccttt agtgagggtt aattgcgcgc
     4681 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct aattcactcc
     4741 tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg gccaatgccc tggctcacaa
     4801 ataccactga gatctttttc cctctgccaa aaattatggg gacatcatga agccccttga
     4861 gcatctgact tctggctaat aaaggaaatt tattttcatt gcaatagtgt gttggaattt
     4921 tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttaaaac atcagaatga
     4981 gtatttggtt tagagtttgg caacatatgc ccatatgctg gctgccatga acaaaggttg
     5041 gctataaaga ggtcatcagt atatgaaaca gccccctgct gtccattcct tattccatag
     5101 aaaagccttg acttgaggtt agattttttt tatattttgt tttgtgttat ttttttcttt
     5161 aacatcccta aaattttcct tacatgtttt actagccaga tttttcctcc tctcctgact
     5221 actcccagtc atagctgtcc ctcttctctt atggagatcc ctcgacctgc agcccaagct
     5281 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     5341 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     5401 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     5461 ggatccgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     5521 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     5581 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     5641 tggaggccta ggcttttgca aaaagctaac ttgtttattg cagcttataa tggttacaaa
     5701 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     5761 ggtttgtcca aactcatcaa tgtatcttat catgtctgga tccgctgcat taatgaatcg
     5821 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     5881 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     5941 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     6001 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     6061 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     6121 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     6181 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     6241 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     6301 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     6361 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     6421 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     6481 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     6541 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     6601 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     6661 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     6721 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     6781 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     6841 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     6901 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     6961 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     7021 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     7081 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     7141 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     7201 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     7261 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     7321 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     7381 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     7441 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     7501 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     7561 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     7621 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     7681 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     7741 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct g
//
