LOCUS       sequence-193838-        6602 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses Phytochrome B (1-621 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 6602)
  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 6602)
  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-hPhyB621-mCherry-HRasCT
FEATURES             Location/Qualifiers
     source          1..6602
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     331..353
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     419..438
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(2498..2516)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2766..2785
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(3133..3152)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3198..3253
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3252..3271)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3614..3630)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3614..3630)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3627..3649)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3638..3654
                     /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(3662..3668,3669..3686,3687..3692))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3707..3728
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3787..3982
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3833..3968
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3988..4122
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4025..4044)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4079..4098
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4190..4207)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4361..4949)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4441..4460)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5120..5911,5912..5980))
                     /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     5743..5762
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5981..6085)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        6120..6499
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gggccgggga gggctcgggg gaggggcgcg gcggcccccg gagcgccggc ggctgtcgag
       61 gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg cagggacttc
      121 ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc ccctctagcg
      181 ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag ggccttcgtg
      241 cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg gggctgtccg cggggggacg
      301 gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct
      361 ctagctagag cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc
      421 aacgtgctgg ttattgtgct gtctcatcat tttggcaaag aattcaccat ggtgtcagga
      481 gtgggaggaa gcggaggcgg cagaggaggc gggagaggag gcgaggaaga gccaagctcc
      541 tctcacactc ccaacaatcg gagaggggga gagcaggctc agagttcagg cactaagtcc
      601 ctgaggcccc gctctaacac cgaatccatg tctaaagcca tccagcagta caccgtggat
      661 gctaggctgc atgccgtgtt tgaacagtcc ggggagtctg gaaagagttt cgactacagt
      721 cagtcactga aaaccacaac ttatgggagc tccgtgcccg agcagcagat tacagcttac
      781 ctgagcagga tccagagggg aggatatatt cagccttttg gatgcatgat cgctgtggat
      841 gagtctagtt tccggatcat tgggtactct gaaaatgcca gagagatgct gggaatcatg
      901 ccacagagtg tgcccactct ggaaaagcct gagattctgg ccatgggcac cgacgtgcgg
      961 tctctgttta catcaagctc ctctatcctg ctggagcggg ctttcgtggc cagagaaatc
     1021 actctgctga acccagtgtg gattcacagc aagaataccg gcaaaccctt ttatgccatc
     1081 ctgcatagga ttgacgtggg ggtggtcatc gatctggaac ctgctcgcac cgaggatcca
     1141 gccctgagca ttgctggcgc cgtgcagtcc cagaagctgg ctgtgagagc catcagccag
     1201 ctgcaggccc tgccaggagg cgacatcaag ctgctgtgcg atacagtggt ggagtccgtg
     1261 agggacctga ctggatacga tcgcgtgatg gtgtataagt tccacgaaga cgagcatggc
     1321 gaagtggtgg ccgagagcaa aagggacgat ctggagcctt acatcggcct gcactatcct
     1381 gctaccgata ttccacaggc ctcccgcttc ctgtttaagc agaaccgggt gagaatgatc
     1441 gtggactgta atgctacacc agtgctggtg gtgcaggacg atcggctgac ccagagtatg
     1501 tgcctggtgg gatcaacact gagagccccc cacggctgtc atagccagta catggctaac
     1561 atggggtcaa ttgccagcct ggctatggcc gtgatcatta acggcaatga ggacgatggg
     1621 agcaatgtgg cctccgggag gagttcaatg cgcctgtggg gactggtggt gtgccaccat
     1681 acaagctccc ggtgtatccc atttcccctg agatatgctt gcgagtttct gatgcaggcc
     1741 ttcggcctgc agctgaacat ggagctgcag ctggccctgc agatgagcga aaagcgggtg
     1801 ctgagaaccc agacactgct gtgcgacatg ctgctgaggg attccccagc tggaatcgtg
     1861 acccagagcc cttccattat ggacctggtg aagtgtgatg gagccgcttt cctgtaccac
     1921 ggcaaatact atccactggg agtggctcct tccgaggtgc agatcaagga tgtggtggaa
     1981 tggctgctgg ctaatcatgc cgactctaca ggcctgtcca ctgactctct gggcgatgct
     2041 ggataccctg gagccgctgc cctgggagat gccgtgtgcg gaatggctgt ggcctatatt
     2101 acaaaaaggg acttcctgtt ttggttccgc agccacactg ctaaggagat caaatggggg
     2161 ggagccaagc accatcctga agataaagac gatggccagc ggatgcatcc aagatctagt
     2221 tttcaggcct tcctggaggt ggtgaagtct cggagtcagc catgggaaac cgctgagatg
     2281 gatgccatcc acagtctgca gctgattctg agagactctt ttaaggagag tggaggtagt
     2341 ggaggcggcc gcatggtgag caagggcgag gaggataaca tggccatcat caaggagttc
     2401 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
     2461 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt
     2521 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc
     2581 tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc
     2641 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc
     2701 tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc
     2761 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac
     2821 cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc
     2881 cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc
     2941 gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg
     3001 gaacagtacg aacgcgccga gggccgccac tccactctcg gcatggacga gctgtctaga
     3061 aagctgaacc ctcctgatga gagtggcccc ggctgcatga gctgcaagtg tgtgctctcc
     3121 tgactcgacg gatctttttc cctctgccaa aaattatggg gacatcatga agccccttga
     3181 gcatctgact tctggctaat aaaggaaatt tattttcatt gcaatagtgt gttggaattt
     3241 tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttaaaac atcagaatga
     3301 gtatttggtt tagagtttgg caacatatgc ccatatgctg gctgccatga acaaaggttg
     3361 gctataaaga ggtcatcagt atatgaaaca gccccctgct gtccattcct tattccatag
     3421 aaaagccttg acttgaggtt agattttttt tatattttgt tttgtgttat ttttttcttt
     3481 aacatcccta aaattttcct tacatgtttt actagccaga tttttcctcc tctcctgact
     3541 actcccagtc atagctgtcc ctcttctctt atggagatcc ctcgacctgc agcccaagct
     3601 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     3661 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     3721 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     3781 ggatccgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     3841 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     3901 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     3961 tggaggccta ggcttttgca aaaagctaac ttgtttattg cagcttataa tggttacaaa
     4021 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     4081 ggtttgtcca aactcatcaa tgtatcttat catgtctgga tccgctgcat taatgaatcg
     4141 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     4201 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     4261 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     4321 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     4381 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     4441 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     4501 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     4561 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     4621 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     4681 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     4741 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     4801 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     4861 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     4921 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     4981 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     5041 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     5101 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     5161 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     5221 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     5281 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     5341 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     5401 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     5461 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     5521 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     5581 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     5641 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     5701 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     5761 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     5821 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     5881 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     5941 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     6001 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     6061 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gggtcgatcg
     6121 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
     6181 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     6241 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     6301 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     6361 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     6421 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     6481 agtcatcgct attaccatgg tcgaggtgag ccccacgttc tgcttcactc tccccatctc
     6541 ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat
     6601 gg
//
