LOCUS       sequence-197245-        8231 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Phytochrome B (1-908 aa) fused with mCherry and Hras 
            C-terminus in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8231)
  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 8231)
  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-hPhyB-mCherry-HRasCT
FEATURES             Location/Qualifiers
     source          1..8231
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        9..388
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        390..665
                     /label=chicken beta-actin promoter
     intron          666..1682
                     /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     1690..1709
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(4618..4636)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4886..4905
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(5253..5272)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5318..5373
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5372..5391)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5734..5750)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5734..5750)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5747..5769)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5758..5774
                     /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(5782..5788,5789..5806,5807..5812))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5827..5848
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5907..6102
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5953..6088
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6108..6242
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6145..6164)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6199..6218
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6310..6327)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6481..7069)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6561..6580)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7240..8031,8032..8100))
                     /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     7863..7882
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8101..8205)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgatcga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
       61 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
      121 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      181 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      241 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      301 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      361 ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc cccacgttct gcttcactct
      421 ccccatctcc cccccctccc cacccccaat tttgtattta tttatttttt aattattttg
      481 tgcagcgatg ggggcggggg gggggggggc gcgcgccagg cggggcgggg cgggggcgag
      541 ggcggggcgg ggcgaggcgg agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa
      601 agtttccttt tatggcgagg cggcggcggc ggcggcccta taaaaagcga agcgcgcggc
      661 gggcgggagt cgctgcgttg ccttcgcccc gtgccccgct ccgcgccgcc tcgcgccgcc
      721 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
      781 ctccgggctg taattagcgc ttggtttaat gacggctcgt ttcttttctg tggctgcgtg
      841 aaagccttaa agggctccgg gagggccctt tgtgcggggg ggagcggctc ggggggtgcg
      901 tgcgtgtgtg tgtgcgtggg gagcgccgcg tgcggcccgc gctgcccggc ggctgtgagc
      961 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcgtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg
     1081 tgcgtggggg ggtgagcagg gggtgtgggc gcggcggtcg ggctgtaacc cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtgcggggcg
     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 tctagctaga gcctctgcta accatgttca tgccttcttc tttttcctac
     1681 agctcctggg caacgtgctg gttattgtgc tgtctcatca ttttggcaaa gaattcacca
     1741 tggtgtcagg agtgggagga agcggaggcg gcagaggagg cgggagagga ggcgaggaag
     1801 agccaagctc ctctcacact cccaacaatc ggagaggggg agagcaggct cagagttcag
     1861 gcactaagtc cctgaggccc cgctctaaca ccgaatccat gtctaaagcc atccagcagt
     1921 acaccgtgga tgctaggctg catgccgtgt ttgaacagtc cggggagtct ggaaagagtt
     1981 tcgactacag tcagtcactg aaaaccacaa cttatgggag ctccgtgccc gagcagcaga
     2041 ttacagctta cctgagcagg atccagaggg gaggatatat tcagcctttt ggatgcatga
     2101 tcgctgtgga tgagtctagt ttccggatca ttgggtactc tgaaaatgcc agagagatgc
     2161 tgggaatcat gccacagagt gtgcccactc tggaaaagcc tgagattctg gccatgggca
     2221 ccgacgtgcg gtctctgttt acatcaagct cctctatcct gctggagcgg gctttcgtgg
     2281 ccagagaaat cactctgctg aacccagtgt ggattcacag caagaatacc ggcaaaccct
     2341 tttatgccat cctgcatagg attgacgtgg gggtggtcat cgatctggaa cctgctcgca
     2401 ccgaggatcc agccctgagc attgctggcg ccgtgcagtc ccagaagctg gctgtgagag
     2461 ccatcagcca gctgcaggcc ctgccaggag gcgacatcaa gctgctgtgc gatacagtgg
     2521 tggagtccgt gagggacctg actggatacg atcgcgtgat ggtgtataag ttccacgaag
     2581 acgagcatgg cgaagtggtg gccgagagca aaagggacga tctggagcct tacatcggcc
     2641 tgcactatcc tgctaccgat attccacagg cctcccgctt cctgtttaag cagaaccggg
     2701 tgagaatgat cgtggactgt aatgctacac cagtgctggt ggtgcaggac gatcggctga
     2761 cccagagtat gtgcctggtg ggatcaacac tgagagcccc ccacggctgt catagccagt
     2821 acatggctaa catggggtca attgccagcc tggctatggc cgtgatcatt aacggcaatg
     2881 aggacgatgg gagcaatgtg gcctccggga ggagttcaat gcgcctgtgg ggactggtgg
     2941 tgtgccacca tacaagctcc cggtgtatcc catttcccct gagatatgct tgcgagtttc
     3001 tgatgcaggc cttcggcctg cagctgaaca tggagctgca gctggccctg cagatgagcg
     3061 aaaagcgggt gctgagaacc cagacactgc tgtgcgacat gctgctgagg gattccccag
     3121 ctggaatcgt gacccagagc ccttccatta tggacctggt gaagtgtgat ggagccgctt
     3181 tcctgtacca cggcaaatac tatccactgg gagtggctcc ttccgaggtg cagatcaagg
     3241 atgtggtgga atggctgctg gctaatcatg ccgactctac aggcctgtcc actgactctc
     3301 tgggcgatgc tggataccct ggagccgctg ccctgggaga tgccgtgtgc ggaatggctg
     3361 tggcctatat tacaaaaagg gacttcctgt tttggttccg cagccacact gctaaggaga
     3421 tcaaatgggg gggagccaag caccatcctg aagataaaga cgatggccag cggatgcatc
     3481 caagatctag ttttcaggcc ttcctggagg tggtgaagtc tcggagtcag ccatgggaaa
     3541 ccgctgagat ggatgccatc cacagtctgc agctgattct gagagactct tttaaggaga
     3601 gtgaggctgc catgaactca aaagtggtgg acggagtggt gcagccttgc agggatatgg
     3661 ctggcgaaca gggaatcgac gagctgggag ctgtggctag ggaaatggtg cgcctgatcg
     3721 agactgccac cgtgccaatt ttcgctgtgg acgccggcgg gtgtatcaac ggctggaatg
     3781 ctaagattgc cgagctgaca ggcctgtccg tggaagaggc catggggaaa agtctggtgt
     3841 cagatctgat ctataaggaa aacgaggcta cagtgaacaa gctgctgtct agggccctgc
     3901 gcggagacga agagaagaac gtggaggtga agctgaaaac ttttagtccc gaactgcagg
     3961 gcaaggccgt gttcgtggtg gtgaatgcct gctcaagcaa agattacctg aacaatatcg
     4021 tgggcgtgtg ctttgtggga caggacgtga cctctcagaa gatcgtgatg gataaattca
     4081 tcaacatcca gggcgactat aaggccatcg tgcacagccc taatccactg atccccccta
     4141 ttttcgctgc cgacgaaaac acctgctgtc tggagtggaa tatggccatg gaaaagctga
     4201 caggatggtc aagaagcgaa gtgatcggca aaatgattgt gggagaagtg tttggctcct
     4261 gctgtatgct gaagggccca gatgctctga ctaaattcat gatcgtgctg cataacgcca
     4321 ttggaggcca ggacaccgat aagtttccct tccctttctt tgacaggaac gggaaattcg
     4381 tgcaggctct gctgaccgcc aataagcgcg tgtcactgga ggggaaagtg atcggagcct
     4441 tttgcttcct gcagattccc agcggcggcc gcatggtgag caagggcgag gaggataaca
     4501 tggccatcat caaggagttc atgcgcttca aggtgcacat ggagggctcc gtgaacggcc
     4561 acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca
     4621 agctgaaggt gaccaagggt ggccccctgc ccttcgcctg ggacatcctg tcccctcagt
     4681 tcatgtacgg ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc
     4741 tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg
     4801 tgaccgtgac ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc
     4861 gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg
     4921 cctcctccga gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagcagaggc
     4981 tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa gaccacctac aaggccaaga
     5041 agcccgtgca gctgcccggc gcctacaacg tcaacatcaa gttggacatc acctcccaca
     5101 acgaggacta caccatcgtg gaacagtacg aacgcgccga gggccgccac tccactctcg
     5161 gcatggacga gctgtctaga aagctgaacc ctcctgatga gagtggcccc ggctgcatga
     5221 gctgcaagtg tgtgctctcc tgactcgacg gatctttttc cctctgccaa aaattatggg
     5281 gacatcatga agccccttga gcatctgact tctggctaat aaaggaaatt tattttcatt
     5341 gcaatagtgt gttggaattt tttgtgtctc tcactcggaa ggacatatgg gagggcaaat
     5401 catttaaaac atcagaatga gtatttggtt tagagtttgg caacatatgc ccatatgctg
     5461 gctgccatga acaaaggttg gctataaaga ggtcatcagt atatgaaaca gccccctgct
     5521 gtccattcct tattccatag aaaagccttg acttgaggtt agattttttt tatattttgt
     5581 tttgtgttat ttttttcttt aacatcccta aaattttcct tacatgtttt actagccaga
     5641 tttttcctcc tctcctgact actcccagtc atagctgtcc ctcttctctt atggagatcc
     5701 ctcgacctgc agcccaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     5761 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     5821 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     5881 gggaaacctg tcgtgccagc ggatccgcat ctcaattagt cagcaaccat agtcccgccc
     5941 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     6001 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     6061 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctaac ttgtttattg
     6121 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     6181 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga
     6241 tccgctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     6301 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     6361 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     6421 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     6481 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     6541 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     6601 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     6661 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     6721 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     6781 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     6841 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     6901 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     6961 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     7021 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     7081 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     7141 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     7201 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     7261 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     7321 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     7381 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     7441 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     7501 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     7561 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     7621 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     7681 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     7741 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     7801 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     7861 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     7921 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     7981 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     8041 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     8101 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     8161 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     8221 agtgccacct g
//
