LOCUS       sequence-193840-        8122 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression vector for phycocyanobilin (PCB) synthesis and shRNA for 
            mouse BVRA gene in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 8122)
  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 8122)
  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-sh-mBVRA
FEATURES             Location/Qualifiers
     source          1..8122
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     326..348
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     410..429
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             460..534
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     regulatory      981..990
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1270..1293
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1303..1359
                     /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             1363..1437
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             1468..1494
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2218..2274
                     /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             2275..2349
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             2380..2409
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2710..2766
                     /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             2767..2841
                     /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(4179..4199)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4179..4197)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4218..4234)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4336..4355)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4401..4456
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4455..4474)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4817..4833)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4841..4857
                     /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(4865..4871,4872..4889,4890..4895))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4910..4931
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4990..5185
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5036..5171
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5191..5325
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5228..5247)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5282..5301
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5393..5410)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5564..6152)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5644..5663)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6323..7114,7115..7183))
                     /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     6946..6965
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7184..7288)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        complement(7401..7615)
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     complement(7461..7480)
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     enhancer        7624..8003
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ggggagggct cgggggaggg gcgcggcggc ccccggagcg ccggcggctg tcgaggcgcg
       61 gcgagccgca gccattgcct tttatggtaa tcgtgcgaga gggcgcaggg acttcctttg
      121 tcccaaatct gtgcggagcc gaaatctggg aggcgccgcc gcaccccctc tagcgggcgc
      181 ggggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg gggagggcct tcgtgcgtcg
      241 ccgcgccgcc gtccccttct ccctctccag cctcggggct gtccgcgggg ggacggctgc
      301 cttcgggggg gacggggcag ggcggggttc ggcttctggc gtgtgaccgg cggctctaga
      361 gcctctgcta accatgttca tgccttcttc tttttcctac agctcctggg caacgtgctg
      421 gttattgtgc tgtctcatca ttttggcaaa gaattcacca tgtccgtcct gactccactg
      481 ctgctgaggg gcctgactgg gtccgcacgc cgcctgcctg tcccaagagc caaaatccat
      541 agcctgggcg ggtctggagg catgagtctg aggcagcacc agcatcctct gatccagcgc
      601 ctggccgatc gaatcgaggc aatttggcag gccttctttc ccctggcacc ttacgccctg
      661 ccagaagacc tgggctatgt ggagggaaaa ctggagggcg aacgcctgac aatcgagaac
      721 cactgctatc aggccccccc ttttcggaag ctgcatctgg aactggctag agtgggcgag
      781 tctctggaca ttctgcactg cgtcatgttt cccgagccta gatacgatct gcccatgttc
      841 ggatgtgacc tggtgggggg acgcggacag atcagtgccg ctattgtgga tctgtcacca
      901 gtcactggac agctgccagc agcatacacc tgcgctctga atgcactgcc taagctgacc
      961 tttagacagc caagggagct gccaccatgg ggacatatct tctctccctt ttgtatcttc
     1021 attagacctc agggcgaagc cgaggaacag cagttcctgg ataggattgg ggagtatctg
     1081 actctgcact gccagctgag ccagcaggca gtgcctaccg accatccaca ggccgtcatc
     1141 gctggacagc ggcagtactg tcagcagcag cagcagaacg ataaaactcg gagagtgctg
     1201 gaaaaggctt ttggcgtccc atgggcagag agatacatga ccacagtgct gttcgatgtg
     1261 cctccagtgg actataagga cgatgacgat aaaggcagcg gggccaccaa cttctccctg
     1321 ctgaagcagg ctggcgacgt ggaggaaaat ccaggacccg gcatgtcagt cctgacacct
     1381 ctgctgctgc gaggactgac tggaagcgcc aggcgcctgc ctgtgccacg agcaaaaatc
     1441 cactcactgg gcgggagcgg aggcatgtac ccctatgatg tccctgacta cgccatgact
     1501 accagcctgg ctacaaagct gagggaaggc actaagaaag ctcataccat ggcagagaac
     1561 gtggggtttg tccggtgttt cctgaaggga actgtggaga aaagctccta tagaaagctg
     1621 gtggcctctc tgtaccacgt ctatagtgct atggagcagg aaatggagcg cctgaaagac
     1681 catccaatcg tgggcaagat ctactttccc gaactgaaca ggaagtctag tctggagcgc
     1741 gatctgacat actatttcgg gtccaattgg agggaggaaa ttcccccttc tcccgcaact
     1801 caggcctacg tggctcgcat ccacgaagtc gcaaacaccg cccctgagct gctggtggcc
     1861 catagttaca cacggtatct gggagacctg tcagggggac agatcctgaa gggcattgct
     1921 gaacgggcaa tgaatctgca ggatggggag ggaaccgcct tctacagatt cgaaagtatc
     1981 tcagacgaga aggccttcaa acagctgtat cgacagcggc tggatgaact gcccgtggac
     2041 gaggctacag cagatagaat tgtggacgag gctaacgctg catttggcat gaatatgaag
     2101 atcttccagg aactggaggg caacctgatc cgggccattg ggcagctgct gtttaatacc
     2161 ctgacacgac ggaaacagag aggcagcacc gagctggcca cagctgatgg ctcaggggca
     2221 accaacttca gcctgctgaa gcaggccgga gacgtggaag agaatccagg acctatgtcc
     2281 gtcctgaccc cactgctgct gaggggactg acaggatctg caagaaggct gccagtgccc
     2341 agagccaaaa ttcactccct gggaggctct ggaggcatgg agcagaagct gatcagcgag
     2401 gaagatctga tggcctccta tacagtgaaa ctgatcactc ccgacggcga atcaagcatt
     2461 gagtgctccg acgataccta catcctggat gcagctgagg aagcagggct ggacctgcct
     2521 tattcttgta gggcaggagc ctgcagtact tgtgcaggca agattaccgc cgggagcgtg
     2581 gatcagagtg accagtcatt tctggacgat gaccagatcg aagctggcta cgtgctgaca
     2641 tgcgtcgcat atcccacttc cgattgtact attgagaccc acaaagagga agacctgtac
     2701 ggatccggcg ctacaaactt ctctctgctg aagcaggcag gggacgtgga ggaaaatcct
     2761 ggaccaatga gtgtcctgac tcccctgctg ctgagaggac tgaccggaag cgcccgccga
     2821 ctgcccgtgc ctagagctaa gatccatagt ctggggggat caggcgggat gtacagccca
     2881 ggatatgtgg ccaccagcag ccggcagtcc gatgcaggaa acaggctgtt cgtgtacgag
     2941 gtcatcggcc tgagccagtc cactatgacc gatgggctgg actatcccat tcggagatct
     3001 ggaagtacat tcatcactgt gcctctgaaa cgaatgaatc aggagatgag gcgcatcacc
     3061 cggatgggag gcaaaatcgt gtcaattaag cctctggaag gcgacagccc actgccacac
     3121 acagagggaa ttgccaagcc atcacagagc gaagggtccg gatctgaggc tgtggcaaac
     3181 cctgctccag aaagcaacaa gactatgaca actaccccta aggagaagaa agccgatgac
     3241 atcccagtga acatctacag gccaaaaacc ccctatattg ggaaggtgct ggaaaattac
     3301 ccactggtcc gcgagggagc tatcggaacc gtgcagcacc tgacatttga tctgtctgcc
     3361 ggcgacctgc ggtatctgga agggcagagt atcggaatca ttccacccgg agaggatgac
     3421 aagggcaaac ctcacaaact gaggctgtac agcattgcct ccacacgcca tggggatttc
     3481 ggagatgaca agactgtgag cctgtgcgtc agacagctgg aataccagaa cgaagcagga
     3541 gagaccgtgc agggagtctg ctccacatac ctgtgcaata tcaaagaggg cgatgacatc
     3601 gccattaccg gcccagtggg gaaggaaatg ctgctgcctc cagatgagga cgcaaacatt
     3661 gtcatgctgg ccacaggcac tgggatcgcc ccctttcggg ctttcctgtg gagaatgttc
     3721 aaagaacagc acgaggacta caagtttaaa ggcctggcct ggctgatctt cgggattcca
     3781 aagtctgaga atatcctgta taaagatgac ctggaaaaga tggcagccga gtttcccgat
     3841 aacttccgcc tgacatacgc tattagccga gaacagcaga atgcagaggg gggacgaatg
     3901 tatatccagc accgggtggc cgagaacgct gaggaactgt ggaacctgat gcagaatcca
     3961 aaaacccata catacatgtg cggactgaag ggcatggaac ccgggatcga cgaggccttt
     4021 accgccctgg ctgaacagaa tggaaaggag tggacaactt tccagagaga gatgaagaaa
     4081 gaacatagat ggcacgtcga gacctacatg gcttcaatga caggaggaca gcagatgggg
     4141 tagcggccgc caccgcggtg gagctccagc ttttgttccc tttagtgagg gttaattgcg
     4201 cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctaattcac
     4261 tcctcaggtg caggctgcct atcagaaggt ggtggctggt gtggccaatg ccctggctca
     4321 caaataccac tgagatcttt ttccctctgc caaaaattat ggggacatca tgaagcccct
     4381 tgagcatctg acttctggct aataaaggaa atttattttc attgcaatag tgtgttggaa
     4441 ttttttgtgt ctctcactcg gaaggacata tgggagggca aatcatttaa aacatcagaa
     4501 tgagtatttg gtttagagtt tggcaacata tgcccatatg ctggctgcca tgaacaaagg
     4561 ttggctataa agaggtcatc agtatatgaa acagccccct gctgtccatt ccttattcca
     4621 tagaaaagcc ttgacttgag gttagatttt ttttatattt tgttttgtgt tatttttttc
     4681 tttaacatcc ctaaaatttt ccttacatgt tttactagcc agatttttcc tcctctcctg
     4741 actactccca gtcatagctg tccctcttct cttatggaga tccctcgacc tgcagcccaa
     4801 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     4861 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     4921 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     4981 agcggatccg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     5041 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     5101 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     5161 ttttggaggc ctaggctttt gcaaaaagct aacttgttta ttgcagctta taatggttac
     5221 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     5281 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatccgctg cattaatgaa
     5341 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     5401 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     5461 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     5521 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     5581 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     5641 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     5701 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     5761 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     5821 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     5881 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     5941 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     6001 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     6061 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     6121 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     6181 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     6241 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     6301 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     6361 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     6421 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     6481 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     6541 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     6601 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     6661 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     6721 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     6781 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     6841 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     6901 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     6961 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     7021 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     7081 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     7141 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     7201 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     7261 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctggtcgac
     7321 ggtatcgata agcttaaaaa gccaaatgta ggagtcaata atctcttgaa ttattgactc
     7381 ctacatttgg cgggagatct gtggtctcat acagaactta taagattccc aaatccaaag
     7441 acatttcacg tttatggtga tttcccagaa cacatagcga catgcaaata ttgcagggcg
     7501 ccactcccct gtccctcaca gccatcttcc tgccagggcg cacgcgcgct gggtgttccc
     7561 gcctagtgac actgggcccg cgattccttg gagcgggttg atgacgtcag cgttcgaatt
     7621 ctcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
     7681 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
     7741 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
     7801 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
     7861 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
     7921 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
     7981 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
     8041 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
     8101 cgatgggggc gggggggggg gg
//
