LOCUS       sequence-193839-        7464 bp    DNA     linear   UNA 14-APR-2025
DEFINITION  Expresses Phytochrome B (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 7464)
  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 7464)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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..7464
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     335..357
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     423..442
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(3351..3369)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3619..3638
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(3986..4005)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4051..4106
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4105..4124)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4467..4483)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4467..4483)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4480..4502)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4491..4507
                     /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(4515..4521,4522..4539,4540..4545))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4560..4581
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4640..4835
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4686..4821
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4841..4975
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4878..4897)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4932..4951
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5043..5060)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5214..5802)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5294..5313)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5973..6764,6765..6833))
                     /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     6596..6615
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6834..6938)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        6973..7352
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 cctcgggccg gggagggctc gggggagggg cgcggcggcc cccggagcgc cggcggctgt
       61 cgaggcgcgg cgagccgcag ccattgcctt ttatggtaat cgtgcgagag ggcgcaggga
      121 cttcctttgt cccaaatctg tgcggagccg aaatctggga ggcgccgccg caccccctct
      181 agcgggcgcg gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
      241 cgtgcgtcgc cgcgccgccg tccccttctc cctctccagc ctcggggctg tccgcggggg
      301 gacggctgcc ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc
      361 ggctctagct agagcctctg ctaaccatgt tcatgccttc ttctttttcc tacagctcct
      421 gggcaacgtg ctggttattg tgctgtctca tcattttggc aaagaattca ccatggtgtc
      481 aggagtggga ggaagcggag gcggcagagg aggcgggaga ggaggcgagg aagagccaag
      541 ctcctctcac actcccaaca atcggagagg gggagagcag gctcagagtt caggcactaa
      601 gtccctgagg ccccgctcta acaccgaatc catgtctaaa gccatccagc agtacaccgt
      661 ggatgctagg ctgcatgccg tgtttgaaca gtccggggag tctggaaaga gtttcgacta
      721 cagtcagtca ctgaaaacca caacttatgg gagctccgtg cccgagcagc agattacagc
      781 ttacctgagc aggatccaga ggggaggata tattcagcct tttggatgca tgatcgctgt
      841 ggatgagtct agtttccgga tcattgggta ctctgaaaat gccagagaga tgctgggaat
      901 catgccacag agtgtgccca ctctggaaaa gcctgagatt ctggccatgg gcaccgacgt
      961 gcggtctctg tttacatcaa gctcctctat cctgctggag cgggctttcg tggccagaga
     1021 aatcactctg ctgaacccag tgtggattca cagcaagaat accggcaaac ccttttatgc
     1081 catcctgcat aggattgacg tgggggtggt catcgatctg gaacctgctc gcaccgagga
     1141 tccagccctg agcattgctg gcgccgtgca gtcccagaag ctggctgtga gagccatcag
     1201 ccagctgcag gccctgccag gaggcgacat caagctgctg tgcgatacag tggtggagtc
     1261 cgtgagggac ctgactggat acgatcgcgt gatggtgtat aagttccacg aagacgagca
     1321 tggcgaagtg gtggccgaga gcaaaaggga cgatctggag ccttacatcg gcctgcacta
     1381 tcctgctacc gatattccac aggcctcccg cttcctgttt aagcagaacc gggtgagaat
     1441 gatcgtggac tgtaatgcta caccagtgct ggtggtgcag gacgatcggc tgacccagag
     1501 tatgtgcctg gtgggatcaa cactgagagc cccccacggc tgtcatagcc agtacatggc
     1561 taacatgggg tcaattgcca gcctggctat ggccgtgatc attaacggca atgaggacga
     1621 tgggagcaat gtggcctccg ggaggagttc aatgcgcctg tggggactgg tggtgtgcca
     1681 ccatacaagc tcccggtgta tcccatttcc cctgagatat gcttgcgagt ttctgatgca
     1741 ggccttcggc ctgcagctga acatggagct gcagctggcc ctgcagatga gcgaaaagcg
     1801 ggtgctgaga acccagacac tgctgtgcga catgctgctg agggattccc cagctggaat
     1861 cgtgacccag agcccttcca ttatggacct ggtgaagtgt gatggagccg ctttcctgta
     1921 ccacggcaaa tactatccac tgggagtggc tccttccgag gtgcagatca aggatgtggt
     1981 ggaatggctg ctggctaatc atgccgactc tacaggcctg tccactgact ctctgggcga
     2041 tgctggatac cctggagccg ctgccctggg agatgccgtg tgcggaatgg ctgtggccta
     2101 tattacaaaa agggacttcc tgttttggtt ccgcagccac actgctaagg agatcaaatg
     2161 ggggggagcc aagcaccatc ctgaagataa agacgatggc cagcggatgc atccaagatc
     2221 tagttttcag gccttcctgg aggtggtgaa gtctcggagt cagccatggg aaaccgctga
     2281 gatggatgcc atccacagtc tgcagctgat tctgagagac tcttttaagg agagtgaggc
     2341 tgccatgaac tcaaaagtgg tggacggagt ggtgcagcct tgcagggata tggctggcga
     2401 acagggaatc gacgagctgg gagctgtggc tagggaaatg gtgcgcctga tcgagactgc
     2461 caccgtgcca attttcgctg tggacgccgg cgggtgtatc aacggctgga atgctaagat
     2521 tgccgagctg acaggcctgt ccgtggaaga ggccatgggg aaaagtctgg tgtcagatct
     2581 gatctataag gaaaacgagg ctacagtgaa caagctgctg tctagggccc tgcgcggaga
     2641 cgaagagaag aacgtggagg tgaagctgaa aacttttagt cccgaactgc agggcaaggc
     2701 cgtgttcgtg gtggtgaatg cctgctcaag caaagattac ctgaacaata tcgtgggcgt
     2761 gtgctttgtg ggacaggacg tgacctctca gaagatcgtg atggataaat tcatcaacat
     2821 ccagggcgac tataaggcca tcgtgcacag ccctaatcca ctgatccccc ctattttcgc
     2881 tgccgacgaa aacacctgct gtctggagtg gaatatggcc atggaaaagc tgacaggatg
     2941 gtcaagaagc gaagtgatcg gcaaaatgat tgtgggagaa gtgtttggct cctgctgtat
     3001 gctgaagggc ccagatgctc tgactaaatt catgatcgtg ctgcataacg ccattggagg
     3061 ccaggacacc gataagtttc ccttcccttt ctttgacagg aacgggaaat tcgtgcaggc
     3121 tctgctgacc gccaataagc gcgtgtcact ggaggggaaa gtgatcggag ccttttgctt
     3181 cctgcagatt cccagcggcg gccgcatggt gagcaagggc gaggaggata acatggccat
     3241 catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg gccacgagtt
     3301 cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa
     3361 ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc agttcatgta
     3421 cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga agctgtcctt
     3481 ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt
     3541 gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc tgcgcggcac
     3601 caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctcctc
     3661 cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagcaga ggctgaagct
     3721 gaaggacggc ggccactacg acgctgaggt caagaccacc tacaaggcca agaagcccgt
     3781 gcagctgccc ggcgcctaca acgtcaacat caagttggac atcacctccc acaacgagga
     3841 ctacaccatc gtggaacagt acgaacgcgc cgagggccgc cactccactc tcggcatgga
     3901 cgagctgtct agaaagctga accctcctga tgagagtggc cccggctgca tgagctgcaa
     3961 gtgtgtgctc tcctgactcg acggatcttt ttccctctgc caaaaattat ggggacatca
     4021 tgaagcccct tgagcatctg acttctggct aataaaggaa atttattttc attgcaatag
     4081 tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca aatcatttaa
     4141 aacatcagaa tgagtatttg gtttagagtt tggcaacata tgcccatatg ctggctgcca
     4201 tgaacaaagg ttggctataa agaggtcatc agtatatgaa acagccccct gctgtccatt
     4261 ccttattcca tagaaaagcc ttgacttgag gttagatttt ttttatattt tgttttgtgt
     4321 tatttttttc tttaacatcc ctaaaatttt ccttacatgt tttactagcc agatttttcc
     4381 tcctctcctg actactccca gtcatagctg tccctcttct cttatggaga tccctcgacc
     4441 tgcagcccaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     4501 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     4561 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     4621 ctgtcgtgcc agcggatccg catctcaatt agtcagcaac catagtcccg cccctaactc
     4681 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     4741 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     4801 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct aacttgttta ttgcagctta
     4861 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     4921 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatccgctg
     4981 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     5041 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     5101 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     5161 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     5221 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     5281 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     5341 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     5401 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     5461 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     5521 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     5581 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     5641 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     5701 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     5761 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     5821 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     5881 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     5941 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     6001 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     6061 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     6121 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     6181 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     6241 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     6301 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     6361 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     6421 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     6481 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     6541 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     6601 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     6661 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     6721 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     6781 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     6841 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     6901 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     6961 cctgggtcga tcgacattga ttattgacta gttattaata gtaatcaatt acggggtcat
     7021 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
     7081 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
     7141 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
     7201 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
     7261 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
     7321 acatctacgt attagtcatc gctattacca tggtcgaggt gagccccacg ttctgcttca
     7381 ctctccccat ctcccccccc tccccacccc caattttgta tttatttatt ttttaattat
     7441 tttgtgcagc gatgggggcg gggg
//
