LOCUS       sequence-213546-        7794 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Modified cation channelrhodopsin Chronos 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 7794)
  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 7794)
  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-Chronos(M140T)-mRuby2-ST
FEATURES             Location/Qualifiers
     source          1..7794
                     /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          3690..3822
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     3805..3824
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    3831..4419
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3884..3904)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4302..4313)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4446..4463)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4452..4563
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4659..4679)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4659..4677)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4698..4714)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4816..4835)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4881..4936
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4935..4954)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5297..5313)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5321..5337
                     /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(5345..5351,5352..5369,5370..5375))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5390..5411
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5470..5665
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5516..5651
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5671..5805
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5708..5727)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5762..5781
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5873..5890)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6044..6632)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6124..6143)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6803..7594,7595..7663))
                     /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     7426..7445
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7664..7768)
                     /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 gcacctggct 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 cagcaagaac 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 taaggactcg aggccgcagg taagtatcaa ggttacaaga caggtttaag
     3721 gagaccaata gaaactgggc ttgtcgagac agagaagact cttgcgtttc tgataggcac
     3781 ctattggtct tactgacatc cactttgcct ttctctccac aggtgtcgac aatcaacctc
     3841 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     3901 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     3961 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
     4021 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
     4081 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
     4141 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     4201 acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg
     4261 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
     4321 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
     4381 ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaattcga gctcggtacg
     4441 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     4501 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     4561 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     4621 gggggaggat tgggaagaca atagcccagc ttttgttccc tttagtgagg gttaattgcg
     4681 cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctaattcac
     4741 tcctcaggtg caggctgcct atcagaaggt ggtggctggt gtggccaatg ccctggctca
     4801 caaataccac tgagatcttt ttccctctgc caaaaattat ggggacatca tgaagcccct
     4861 tgagcatctg acttctggct aataaaggaa atttattttc attgcaatag tgtgttggaa
     4921 ttttttgtgt ctctcactcg gaaggacata tgggagggca aatcatttaa aacatcagaa
     4981 tgagtatttg gtttagagtt tggcaacata tgcccatatg ctggctgcca tgaacaaagg
     5041 ttggctataa agaggtcatc agtatatgaa acagccccct gctgtccatt ccttattcca
     5101 tagaaaagcc ttgacttgag gttagatttt ttttatattt tgttttgtgt tatttttttc
     5161 tttaacatcc ctaaaatttt ccttacatgt tttactagcc agatttttcc tcctctcctg
     5221 actactccca gtcatagctg tccctcttct cttatggaga tccctcgacc tgcagcccaa
     5281 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     5341 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     5401 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     5461 agcggatccg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     5521 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     5581 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     5641 ttttggaggc ctaggctttt gcaaaaagct aacttgttta ttgcagctta taatggttac
     5701 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     5761 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatccgctg cattaatgaa
     5821 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     5881 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     5941 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     6001 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     6061 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     6121 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     6181 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     6241 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     6301 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     6361 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     6421 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     6481 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     6541 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     6601 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     6661 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     6721 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     6781 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     6841 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     6901 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     6961 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     7021 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     7081 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     7141 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     7201 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     7261 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     7321 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     7381 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     7441 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     7501 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     7561 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     7621 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     7681 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     7741 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctg
//
