LOCUS       sequence-193720-        7827 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression vector for phycocyanobilin (PCB) synthesis in mammalian 
            cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7827)
  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 7827)
  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-PHFF
FEATURES             Location/Qualifiers
     source          1..7827
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     336..358
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     420..439
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             470..544
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     regulatory      991..1000
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1280..1303
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1313..1369
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1373..1447
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             1478..1504
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2228..2284
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             2285..2359
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             2390..2419
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2720..2776
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             2777..2851
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     primer_bind     complement(4189..4209)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4189..4207)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4228..4244)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4346..4365)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4411..4466
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4465..4484)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4827..4843)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4851..4867
                     /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(4875..4881,4882..4899,4900..4905))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4920..4941
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5000..5195
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5046..5181
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5201..5335
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5238..5257)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5292..5311
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5403..5420)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5574..6162)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5654..5673)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6333..7124,7125..7193))
                     /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     6956..6975
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7194..7298)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        7329..7708
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gcctcgggcc ggggagggct cgggggaggg gcgcggcggc ccccggagcg ccggcggctg
       61 tcgaggcgcg gcgagccgca gccattgcct tttatggtaa tcgtgcgaga gggcgcaggg
      121 acttcctttg tcccaaatct gtgcggagcc gaaatctggg aggcgccgcc gcaccccctc
      181 tagcgggcgc ggggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg gggagggcct
      241 tcgtgcgtcg ccgcgccgcc gtccccttct ccctctccag cctcggggct gtccgcgggg
      301 ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc gtgtgaccgg
      361 cggctctaga gcctctgcta accatgttca tgccttcttc tttttcctac agctcctggg
      421 caacgtgctg gttattgtgc tgtctcatca ttttggcaaa gaattcacca tgtccgtcct
      481 gactccactg ctgctgaggg gcctgactgg gtccgcacgc cgcctgcctg tcccaagagc
      541 caaaatccat agcctgggcg ggtctggagg catgagtctg aggcagcacc agcatcctct
      601 gatccagcgc ctggccgatc gaatcgaggc aatttggcag gccttctttc ccctggcacc
      661 ttacgccctg ccagaagacc tgggctatgt ggagggaaaa ctggagggcg aacgcctgac
      721 aatcgagaac cactgctatc aggccccccc ttttcggaag ctgcatctgg aactggctag
      781 agtgggcgag tctctggaca ttctgcactg cgtcatgttt cccgagccta gatacgatct
      841 gcccatgttc ggatgtgacc tggtgggggg acgcggacag atcagtgccg ctattgtgga
      901 tctgtcacca gtcactggac agctgccagc agcatacacc tgcgctctga atgcactgcc
      961 taagctgacc tttagacagc caagggagct gccaccatgg ggacatatct tctctccctt
     1021 ttgtatcttc attagacctc agggcgaagc cgaggaacag cagttcctgg ataggattgg
     1081 ggagtatctg actctgcact gccagctgag ccagcaggca gtgcctaccg accatccaca
     1141 ggccgtcatc gctggacagc ggcagtactg tcagcagcag cagcagaacg ataaaactcg
     1201 gagagtgctg gaaaaggctt ttggcgtccc atgggcagag agatacatga ccacagtgct
     1261 gttcgatgtg cctccagtgg actataagga cgatgacgat aaaggcagcg gggccaccaa
     1321 cttctccctg ctgaagcagg ctggcgacgt ggaggaaaat ccaggacccg gcatgtcagt
     1381 cctgacacct ctgctgctgc gaggactgac tggaagcgcc aggcgcctgc ctgtgccacg
     1441 agcaaaaatc cactcactgg gcgggagcgg aggcatgtac ccctatgatg tccctgacta
     1501 cgccatgact accagcctgg ctacaaagct gagggaaggc actaagaaag ctcataccat
     1561 ggcagagaac gtggggtttg tccggtgttt cctgaaggga actgtggaga aaagctccta
     1621 tagaaagctg gtggcctctc tgtaccacgt ctatagtgct atggagcagg aaatggagcg
     1681 cctgaaagac catccaatcg tgggcaagat ctactttccc gaactgaaca ggaagtctag
     1741 tctggagcgc gatctgacat actatttcgg gtccaattgg agggaggaaa ttcccccttc
     1801 tcccgcaact caggcctacg tggctcgcat ccacgaagtc gcaaacaccg cccctgagct
     1861 gctggtggcc catagttaca cacggtatct gggagacctg tcagggggac agatcctgaa
     1921 gggcattgct gaacgggcaa tgaatctgca ggatggggag ggaaccgcct tctacagatt
     1981 cgaaagtatc tcagacgaga aggccttcaa acagctgtat cgacagcggc tggatgaact
     2041 gcccgtggac gaggctacag cagatagaat tgtggacgag gctaacgctg catttggcat
     2101 gaatatgaag atcttccagg aactggaggg caacctgatc cgggccattg ggcagctgct
     2161 gtttaatacc ctgacacgac ggaaacagag aggcagcacc gagctggcca cagctgatgg
     2221 ctcaggggca accaacttca gcctgctgaa gcaggccgga gacgtggaag agaatccagg
     2281 acctatgtcc gtcctgaccc cactgctgct gaggggactg acaggatctg caagaaggct
     2341 gccagtgccc agagccaaaa ttcactccct gggaggctct ggaggcatgg agcagaagct
     2401 gatcagcgag gaagatctga tggcctccta tacagtgaaa ctgatcactc ccgacggcga
     2461 atcaagcatt gagtgctccg acgataccta catcctggat gcagctgagg aagcagggct
     2521 ggacctgcct tattcttgta gggcaggagc ctgcagtact tgtgcaggca agattaccgc
     2581 cgggagcgtg gatcagagtg accagtcatt tctggacgat gaccagatcg aagctggcta
     2641 cgtgctgaca tgcgtcgcat atcccacttc cgattgtact attgagaccc acaaagagga
     2701 agacctgtac ggatccggcg ctacaaactt ctctctgctg aagcaggcag gggacgtgga
     2761 ggaaaatcct ggaccaatga gtgtcctgac tcccctgctg ctgagaggac tgaccggaag
     2821 cgcccgccga ctgcccgtgc ctagagctaa gatccatagt ctggggggat caggcgggat
     2881 gtacagccca ggatatgtgg ccaccagcag ccggcagtcc gatgcaggaa acaggctgtt
     2941 cgtgtacgag gtcatcggcc tgagccagtc cactatgacc gatgggctgg actatcccat
     3001 tcggagatct ggaagtacat tcatcactgt gcctctgaaa cgaatgaatc aggagatgag
     3061 gcgcatcacc cggatgggag gcaaaatcgt gtcaattaag cctctggaag gcgacagccc
     3121 actgccacac acagagggaa ttgccaagcc atcacagagc gaagggtccg gatctgaggc
     3181 tgtggcaaac cctgctccag aaagcaacaa gactatgaca actaccccta aggagaagaa
     3241 agccgatgac atcccagtga acatctacag gccaaaaacc ccctatattg ggaaggtgct
     3301 ggaaaattac ccactggtcc gcgagggagc tatcggaacc gtgcagcacc tgacatttga
     3361 tctgtctgcc ggcgacctgc ggtatctgga agggcagagt atcggaatca ttccacccgg
     3421 agaggatgac aagggcaaac ctcacaaact gaggctgtac agcattgcct ccacacgcca
     3481 tggggatttc ggagatgaca agactgtgag cctgtgcgtc agacagctgg aataccagaa
     3541 cgaagcagga gagaccgtgc agggagtctg ctccacatac ctgtgcaata tcaaagaggg
     3601 cgatgacatc gccattaccg gcccagtggg gaaggaaatg ctgctgcctc cagatgagga
     3661 cgcaaacatt gtcatgctgg ccacaggcac tgggatcgcc ccctttcggg ctttcctgtg
     3721 gagaatgttc aaagaacagc acgaggacta caagtttaaa ggcctggcct ggctgatctt
     3781 cgggattcca aagtctgaga atatcctgta taaagatgac ctggaaaaga tggcagccga
     3841 gtttcccgat aacttccgcc tgacatacgc tattagccga gaacagcaga atgcagaggg
     3901 gggacgaatg tatatccagc accgggtggc cgagaacgct gaggaactgt ggaacctgat
     3961 gcagaatcca aaaacccata catacatgtg cggactgaag ggcatggaac ccgggatcga
     4021 cgaggccttt accgccctgg ctgaacagaa tggaaaggag tggacaactt tccagagaga
     4081 gatgaagaaa gaacatagat ggcacgtcga gacctacatg gcttcaatga caggaggaca
     4141 gcagatgggg tagcggccgc caccgcggtg gagctccagc ttttgttccc tttagtgagg
     4201 gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     4261 gctaattcac tcctcaggtg caggctgcct atcagaaggt ggtggctggt gtggccaatg
     4321 ccctggctca caaataccac tgagatcttt ttccctctgc caaaaattat ggggacatca
     4381 tgaagcccct tgagcatctg acttctggct aataaaggaa atttattttc attgcaatag
     4441 tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca aatcatttaa
     4501 aacatcagaa tgagtatttg gtttagagtt tggcaacata tgcccatatg ctggctgcca
     4561 tgaacaaagg ttggctataa agaggtcatc agtatatgaa acagccccct gctgtccatt
     4621 ccttattcca tagaaaagcc ttgacttgag gttagatttt ttttatattt tgttttgtgt
     4681 tatttttttc tttaacatcc ctaaaatttt ccttacatgt tttactagcc agatttttcc
     4741 tcctctcctg actactccca gtcatagctg tccctcttct cttatggaga tccctcgacc
     4801 tgcagcccaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     4861 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     4921 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     4981 ctgtcgtgcc agcggatccg catctcaatt agtcagcaac catagtcccg cccctaactc
     5041 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     5101 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     5161 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct aacttgttta ttgcagctta
     5221 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     5281 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatccgctg
     5341 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     5401 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     5461 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     5521 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     5581 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     5641 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     5701 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     5761 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     5821 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     5881 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     5941 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     6001 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     6061 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     6121 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     6181 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     6241 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     6301 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     6361 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     6421 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     6481 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     6541 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     6601 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     6661 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     6721 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     6781 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     6841 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     6901 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     6961 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     7021 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     7081 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     7141 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     7201 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     7261 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     7321 cctgggtcga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
     7381 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
     7441 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
     7501 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     7561 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
     7621 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
     7681 ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc cccacgttct gcttcactct
     7741 ccccatctcc cccccctccc cacccccaat tttgtattta tttatttttt aattattttg
     7801 tgcagcgatg ggggcggggg ggggggg
//
