LOCUS       sequence-197258-        7382 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Phytochrome B (1-621 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 7382)
  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 7382)
  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..7382
                     /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(3769..3787)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4037..4056
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(4404..4423)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4469..4524
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4523..4542)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4885..4901)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4885..4901)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4898..4920)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4909..4925
                     /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(4933..4939,4940..4957,4958..4963))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4978..4999
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5058..5253
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5104..5239
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5259..5393
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5296..5315)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5350..5369
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5461..5478)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5632..6220)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5712..5731)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6391..7182,7183..7251))
                     /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     7014..7033
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7252..7356)
                     /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 gtggaggtag tggaggcggc cgcatggtga gcaagggcga ggaggataac atggccatca
     3661 tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg
     3721 agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg
     3781 tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg
     3841 gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc
     3901 ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga
     3961 cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca
     4021 acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg
     4081 agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga
     4141 aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc
     4201 agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact
     4261 acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccactctc ggcatggacg
     4321 agctgtctag aaagctgaac cctcctgatg agagtggccc cggctgcatg agctgcaagt
     4381 gtgtgctctc ctgactcgac ggatcttttt ccctctgcca aaaattatgg ggacatcatg
     4441 aagccccttg agcatctgac ttctggctaa taaaggaaat ttattttcat tgcaatagtg
     4501 tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa tcatttaaaa
     4561 catcagaatg agtatttggt ttagagtttg gcaacatatg cccatatgct ggctgccatg
     4621 aacaaaggtt ggctataaag aggtcatcag tatatgaaac agccccctgc tgtccattcc
     4681 ttattccata gaaaagcctt gacttgaggt tagatttttt ttatattttg ttttgtgtta
     4741 tttttttctt taacatccct aaaattttcc ttacatgttt tactagccag atttttcctc
     4801 ctctcctgac tactcccagt catagctgtc cctcttctct tatggagatc cctcgacctg
     4861 cagcccaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     4921 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     4981 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     5041 gtcgtgccag cggatccgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     5101 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     5161 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     5221 ggaggctttt ttggaggcct aggcttttgc aaaaagctaa cttgtttatt gcagcttata
     5281 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     5341 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg atccgctgca
     5401 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     5461 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     5521 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     5581 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     5641 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5701 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     5761 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     5821 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     5881 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5941 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     6001 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     6061 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     6121 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     6181 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     6241 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     6301 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     6361 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     6421 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     6481 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     6541 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     6601 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     6661 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     6721 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     6781 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     6841 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     6901 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     6961 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     7021 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     7081 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     7141 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     7201 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     7261 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     7321 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     7381 tg
//
