LOCUS       sequence-193843-        7472 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses Phytochrome B-Y276H (1-908 aa) fused with mCherry and Hras
            C-terminus in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7472)
  AUTHORS   Uda Y, Goto Y, Oda S, Kohchi T, Matsuda M, Aoki K
  TITLE     Efficient synthesis of phycocyanobilin in mammalian cells for 
            optogenetic control of cell signaling.
  JOURNAL   Proc Natl Acad Sci U S A. 2017 Oct 24. pii: 201707190. doi: 
            10.1073/pnas.1707190114.
  PUBMED    29078307
REFERENCE   2  (bases 1 to 7472)
  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: "Proc Natl 
            Acad Sci U S A. 2017 Oct 24. pii: 201707190. doi: 
            10.1073/pnas.1707190114."
COMMENT     Sequence Label: pCAGGS-hPhyBY276H-mCherry-HRasCT
FEATURES             Location/Qualifiers
     source          1..7472
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     338..360
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     426..445
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(3354..3372)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3622..3641
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(3989..4008)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4054..4109
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4108..4127)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4470..4486)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4470..4486)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4483..4505)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4494..4510
                     /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(4518..4524,4525..4542,4543..4548))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4563..4584
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4643..4838
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4689..4824
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4844..4978
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4881..4900)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4935..4954
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5046..5063)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5217..5805)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5297..5316)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5976..6767,6768..6836))
                     /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     6599..6618
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6837..6941)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        6976..7355
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 cagcctcggg ccggggaggg ctcgggggag gggcgcggcg gcccccggag cgccggcggc
       61 tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag
      121 ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc
      181 tctagcgggc gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc
      241 cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg
      301 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
      361 ggcggctcta gctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
      421 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tcaccatggt
      481 gtcaggagtg ggaggaagcg gaggcggcag aggaggcggg agaggaggcg aggaagagcc
      541 aagctcctct cacactccca acaatcggag agggggagag caggctcaga gttcaggcac
      601 taagtccctg aggccccgct ctaacaccga atccatgtct aaagccatcc agcagtacac
      661 cgtggatgct aggctgcatg ccgtgtttga acagtccggg gagtctggaa agagtttcga
      721 ctacagtcag tcactgaaaa ccacaactta tgggagctcc gtgcccgagc agcagattac
      781 agcttacctg agcaggatcc agaggggagg atatattcag ccttttggat gcatgatcgc
      841 tgtggatgag tctagtttcc ggatcattgg gtactctgaa aatgccagag agatgctggg
      901 aatcatgcca cagagtgtgc ccactctgga aaagcctgag attctggcca tgggcaccga
      961 cgtgcggtct ctgtttacat caagctcctc tatcctgctg gagcgggctt tcgtggccag
     1021 agaaatcact ctgctgaacc cagtgtggat tcacagcaag aataccggca aaccctttta
     1081 tgccatcctg cataggattg acgtgggggt ggtcatcgat ctggaacctg ctcgcaccga
     1141 ggatccagcc ctgagcattg ctggcgccgt gcagtcccag aagctggctg tgagagccat
     1201 cagccagctg caggccctgc caggaggcga catcaagctg ctgtgcgata cagtggtgga
     1261 gtccgtgagg gacctgactg gatacgatcg cgtgatggtg cataagttcc acgaagacga
     1321 gcatggcgaa gtggtggccg agagcaaaag ggacgatctg gagccttaca tcggcctgca
     1381 ctatcctgct accgatattc cacaggcctc ccgcttcctg tttaagcaga accgggtgag
     1441 aatgatcgtg gactgtaatg ctacaccagt gctggtggtg caggacgatc ggctgaccca
     1501 gagtatgtgc ctggtgggat caacactgag agccccccac ggctgtcata gccagtacat
     1561 ggctaacatg gggtcaattg ccagcctggc tatggccgtg atcattaacg gcaatgagga
     1621 cgatgggagc aatgtggcct ccgggaggag ttcaatgcgc ctgtggggac tggtggtgtg
     1681 ccaccataca agctcccggt gtatcccatt tcccctgaga tatgcttgcg agtttctgat
     1741 gcaggccttc ggcctgcagc tgaacatgga gctgcagctg gccctgcaga tgagcgaaaa
     1801 gcgggtgctg agaacccaga cactgctgtg cgacatgctg ctgagggatt ccccagctgg
     1861 aatcgtgacc cagagccctt ccattatgga cctggtgaag tgtgatggag ccgctttcct
     1921 gtaccacggc aaatactatc cactgggagt ggctccttcc gaggtgcaga tcaaggatgt
     1981 ggtggaatgg ctgctggcta atcatgccga ctctacaggc ctgtccactg actctctggg
     2041 cgatgctgga taccctggag ccgctgccct gggagatgcc gtgtgcggaa tggctgtggc
     2101 ctatattaca aaaagggact tcctgttttg gttccgcagc cacactgcta aggagatcaa
     2161 atggggggga gccaagcacc atcctgaaga taaagacgat ggccagcgga tgcatccaag
     2221 atctagtttt caggccttcc tggaggtggt gaagtctcgg agtcagccat gggaaaccgc
     2281 tgagatggat gccatccaca gtctgcagct gattctgaga gactctttta aggagagtga
     2341 ggctgccatg aactcaaaag tggtggacgg agtggtgcag ccttgcaggg atatggctgg
     2401 cgaacaggga atcgacgagc tgggagctgt ggctagggaa atggtgcgcc tgatcgagac
     2461 tgccaccgtg ccaattttcg ctgtggacgc cggcgggtgt atcaacggct ggaatgctaa
     2521 gattgccgag ctgacaggcc tgtccgtgga agaggccatg gggaaaagtc tggtgtcaga
     2581 tctgatctat aaggaaaacg aggctacagt gaacaagctg ctgtctaggg ccctgcgcgg
     2641 agacgaagag aagaacgtgg aggtgaagct gaaaactttt agtcccgaac tgcagggcaa
     2701 ggccgtgttc gtggtggtga atgcctgctc aagcaaagat tacctgaaca atatcgtggg
     2761 cgtgtgcttt gtgggacagg acgtgacctc tcagaagatc gtgatggata aattcatcaa
     2821 catccagggc gactataagg ccatcgtgca cagccctaat ccactgatcc cccctatttt
     2881 cgctgccgac gaaaacacct gctgtctgga gtggaatatg gccatggaaa agctgacagg
     2941 atggtcaaga agcgaagtga tcggcaaaat gattgtggga gaagtgtttg gcccctgctg
     3001 tatgctgaag ggcccagatg ctctgactaa attcatgatc gtgctgcata acgccattgg
     3061 aggccaggac accgataagt ttcccttccc tttctttgac aggaacggga aattcgtgca
     3121 ggctccgctg accgccaata agcgcgtgtc actggagggg aaagtgatcg gagccttttg
     3181 cttcctgcag attcccagcg gcggccgcat ggtgagcaag ggcgaggagg ataacatggc
     3241 catcatcaag gagttcatgc gcttcaaggt gcacatggag ggctccgtga acggccacga
     3301 gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct
     3361 gaaggtgacc aagggtggcc ccctgccctt cgcctgggac atcctgtccc ctcagttcat
     3421 gtacggctcc aaggcctacg tgaagcaccc cgccgacatc cccgactact tgaagctgtc
     3481 cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac
     3541 cgtgacccag gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg
     3601 caccaacttc ccctccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc
     3661 ctccgagcgg atgtaccccg aggacggcgc cctgaagggc gagatcaagc agaggctgaa
     3721 gctgaaggac ggcggccact acgacgctga ggtcaagacc acctacaagg ccaagaagcc
     3781 cgtgcagctg cccggcgcct acaacgtcaa catcaagttg gacatcacct cccacaacga
     3841 ggactacacc atcgtggaac agtacgaacg cgccgagggc cgccactcca ctctcggcat
     3901 ggacgagctg tctagaaagc tgaaccctcc tgatgagagt ggccccggct gcatgagctg
     3961 caagtgtgtg ctctcctgac tcgacggatc tttttccctc tgccaaaaat tatggggaca
     4021 tcatgaagcc ccttgagcat ctgacttctg gctaataaag gaaatttatt ttcattgcaa
     4081 tagtgtgttg gaattttttg tgtctctcac tcggaaggac atatgggagg gcaaatcatt
     4141 taaaacatca gaatgagtat ttggtttaga gtttggcaac atatgcccat atgctggctg
     4201 ccatgaacaa aggttggcta taaagaggtc atcagtatat gaaacagccc cctgctgtcc
     4261 attccttatt ccatagaaaa gccttgactt gaggttagat tttttttata ttttgttttg
     4321 tgttattttt ttctttaaca tccctaaaat tttccttaca tgttttacta gccagatttt
     4381 tcctcctctc ctgactactc ccagtcatag ctgtccctct tctcttatgg agatccctcg
     4441 acctgcagcc caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     4501 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     4561 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     4621 aacctgtcgt gccagcggat ccgcatctca attagtcagc aaccatagtc ccgcccctaa
     4681 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4741 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     4801 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctaacttgt ttattgcagc
     4861 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     4921 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatccg
     4981 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     5041 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     5101 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     5161 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5221 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     5281 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     5341 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     5401 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     5461 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     5521 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     5581 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     5641 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     5701 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     5761 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     5821 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     5881 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     5941 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     6001 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     6061 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     6121 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     6181 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     6241 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     6301 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     6361 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     6421 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     6481 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     6541 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     6601 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     6661 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     6721 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     6781 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     6841 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     6901 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     6961 ccacctgggt cgatcgacat tgattattga ctagttatta atagtaatca attacggggt
     7021 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
     7081 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
     7141 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
     7201 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
     7261 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
     7321 agtacatcta cgtattagtc atcgctatta ccatggtcga ggtgagcccc acgttctgct
     7381 tcactctccc catctccccc ccctccccac ccccaatttt gtatttattt attttttaat
     7441 tattttgtgc agcgatgggg gcgggggggg gg
//
