LOCUS       sequence-193719-        8127 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression vector for phycocyanobilin (PCB) synthesis and shRNA for 
            human BVRA gene in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 8127)
  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 8127)
  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-hBVRA
FEATURES             Location/Qualifiers
     source          1..8127
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     337..359
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     421..440
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             471..545
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     regulatory      992..1001
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1281..1304
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1314..1370
                     /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             1374..1448
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             1479..1505
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2229..2285
                     /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             2286..2360
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             2391..2420
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2721..2777
                     /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             2778..2852
                     /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(4190..4210)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4190..4208)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4229..4245)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4347..4366)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4412..4467
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4466..4485)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4828..4844)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4852..4868
                     /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(4876..4882,4883..4900,4901..4906))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4921..4942
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5001..5196
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5047..5182
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5202..5336
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5239..5258)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5293..5312
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5404..5421)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5575..6163)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5655..5674)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6334..7125,7126..7194))
                     /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     6957..6976
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7195..7299)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        complement(7414..7628)
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     complement(7474..7493)
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     enhancer        7637..8016
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 cgcctcgggc cggggagggc tcgggggagg ggcgcggcgg cccccggagc gccggcggct
       61 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
      121 gacttccttt gtcccaaatc tgtgcggagc cgaaatctgg gaggcgccgc cgcaccccct
      181 ctagcgggcg cggggcgaag cggtgcggcg ccggcaggaa ggaaatgggc ggggagggcc
      241 ttcgtgcgtc gccgcgccgc cgtccccttc tccctctcca gcctcggggc tgtccgcggg
      301 gggacggctg ccttcggggg ggacggggca gggcggggtt cggcttctgg cgtgtgaccg
      361 gcggctctag agcctctgct aaccatgttc atgccttctt ctttttccta cagctcctgg
      421 gcaacgtgct ggttattgtg ctgtctcatc attttggcaa agaattcacc atgtccgtcc
      481 tgactccact gctgctgagg ggcctgactg ggtccgcacg ccgcctgcct gtcccaagag
      541 ccaaaatcca tagcctgggc gggtctggag gcatgagtct gaggcagcac cagcatcctc
      601 tgatccagcg cctggccgat cgaatcgagg caatttggca ggccttcttt cccctggcac
      661 cttacgccct gccagaagac ctgggctatg tggagggaaa actggagggc gaacgcctga
      721 caatcgagaa ccactgctat caggcccccc cttttcggaa gctgcatctg gaactggcta
      781 gagtgggcga gtctctggac attctgcact gcgtcatgtt tcccgagcct agatacgatc
      841 tgcccatgtt cggatgtgac ctggtggggg gacgcggaca gatcagtgcc gctattgtgg
      901 atctgtcacc agtcactgga cagctgccag cagcatacac ctgcgctctg aatgcactgc
      961 ctaagctgac ctttagacag ccaagggagc tgccaccatg gggacatatc ttctctccct
     1021 tttgtatctt cattagacct cagggcgaag ccgaggaaca gcagttcctg gataggattg
     1081 gggagtatct gactctgcac tgccagctga gccagcaggc agtgcctacc gaccatccac
     1141 aggccgtcat cgctggacag cggcagtact gtcagcagca gcagcagaac gataaaactc
     1201 ggagagtgct ggaaaaggct tttggcgtcc catgggcaga gagatacatg accacagtgc
     1261 tgttcgatgt gcctccagtg gactataagg acgatgacga taaaggcagc ggggccacca
     1321 acttctccct gctgaagcag gctggcgacg tggaggaaaa tccaggaccc ggcatgtcag
     1381 tcctgacacc tctgctgctg cgaggactga ctggaagcgc caggcgcctg cctgtgccac
     1441 gagcaaaaat ccactcactg ggcgggagcg gaggcatgta cccctatgat gtccctgact
     1501 acgccatgac taccagcctg gctacaaagc tgagggaagg cactaagaaa gctcatacca
     1561 tggcagagaa cgtggggttt gtccggtgtt tcctgaaggg aactgtggag aaaagctcct
     1621 atagaaagct ggtggcctct ctgtaccacg tctatagtgc tatggagcag gaaatggagc
     1681 gcctgaaaga ccatccaatc gtgggcaaga tctactttcc cgaactgaac aggaagtcta
     1741 gtctggagcg cgatctgaca tactatttcg ggtccaattg gagggaggaa attccccctt
     1801 ctcccgcaac tcaggcctac gtggctcgca tccacgaagt cgcaaacacc gcccctgagc
     1861 tgctggtggc ccatagttac acacggtatc tgggagacct gtcaggggga cagatcctga
     1921 agggcattgc tgaacgggca atgaatctgc aggatgggga gggaaccgcc ttctacagat
     1981 tcgaaagtat ctcagacgag aaggccttca aacagctgta tcgacagcgg ctggatgaac
     2041 tgcccgtgga cgaggctaca gcagatagaa ttgtggacga ggctaacgct gcatttggca
     2101 tgaatatgaa gatcttccag gaactggagg gcaacctgat ccgggccatt gggcagctgc
     2161 tgtttaatac cctgacacga cggaaacaga gaggcagcac cgagctggcc acagctgatg
     2221 gctcaggggc aaccaacttc agcctgctga agcaggccgg agacgtggaa gagaatccag
     2281 gacctatgtc cgtcctgacc ccactgctgc tgaggggact gacaggatct gcaagaaggc
     2341 tgccagtgcc cagagccaaa attcactccc tgggaggctc tggaggcatg gagcagaagc
     2401 tgatcagcga ggaagatctg atggcctcct atacagtgaa actgatcact cccgacggcg
     2461 aatcaagcat tgagtgctcc gacgatacct acatcctgga tgcagctgag gaagcagggc
     2521 tggacctgcc ttattcttgt agggcaggag cctgcagtac ttgtgcaggc aagattaccg
     2581 ccgggagcgt ggatcagagt gaccagtcat ttctggacga tgaccagatc gaagctggct
     2641 acgtgctgac atgcgtcgca tatcccactt ccgattgtac tattgagacc cacaaagagg
     2701 aagacctgta cggatccggc gctacaaact tctctctgct gaagcaggca ggggacgtgg
     2761 aggaaaatcc tggaccaatg agtgtcctga ctcccctgct gctgagagga ctgaccggaa
     2821 gcgcccgccg actgcccgtg cctagagcta agatccatag tctgggggga tcaggcggga
     2881 tgtacagccc aggatatgtg gccaccagca gccggcagtc cgatgcagga aacaggctgt
     2941 tcgtgtacga ggtcatcggc ctgagccagt ccactatgac cgatgggctg gactatccca
     3001 ttcggagatc tggaagtaca ttcatcactg tgcctctgaa acgaatgaat caggagatga
     3061 ggcgcatcac ccggatggga ggcaaaatcg tgtcaattaa gcctctggaa ggcgacagcc
     3121 cactgccaca cacagaggga attgccaagc catcacagag cgaagggtcc ggatctgagg
     3181 ctgtggcaaa ccctgctcca gaaagcaaca agactatgac aactacccct aaggagaaga
     3241 aagccgatga catcccagtg aacatctaca ggccaaaaac cccctatatt gggaaggtgc
     3301 tggaaaatta cccactggtc cgcgagggag ctatcggaac cgtgcagcac ctgacatttg
     3361 atctgtctgc cggcgacctg cggtatctgg aagggcagag tatcggaatc attccacccg
     3421 gagaggatga caagggcaaa cctcacaaac tgaggctgta cagcattgcc tccacacgcc
     3481 atggggattt cggagatgac aagactgtga gcctgtgcgt cagacagctg gaataccaga
     3541 acgaagcagg agagaccgtg cagggagtct gctccacata cctgtgcaat atcaaagagg
     3601 gcgatgacat cgccattacc ggcccagtgg ggaaggaaat gctgctgcct ccagatgagg
     3661 acgcaaacat tgtcatgctg gccacaggca ctgggatcgc cccctttcgg gctttcctgt
     3721 ggagaatgtt caaagaacag cacgaggact acaagtttaa aggcctggcc tggctgatct
     3781 tcgggattcc aaagtctgag aatatcctgt ataaagatga cctggaaaag atggcagccg
     3841 agtttcccga taacttccgc ctgacatacg ctattagccg agaacagcag aatgcagagg
     3901 ggggacgaat gtatatccag caccgggtgg ccgagaacgc tgaggaactg tggaacctga
     3961 tgcagaatcc aaaaacccat acatacatgt gcggactgaa gggcatggaa cccgggatcg
     4021 acgaggcctt taccgccctg gctgaacaga atggaaagga gtggacaact ttccagagag
     4081 agatgaagaa agaacataga tggcacgtcg agacctacat ggcttcaatg acaggaggac
     4141 agcagatggg gtagcggccg ccaccgcggt ggagctccag cttttgttcc ctttagtgag
     4201 ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     4261 cgctaattca ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg tgtggccaat
     4321 gccctggctc acaaatacca ctgagatctt tttccctctg ccaaaaatta tggggacatc
     4381 atgaagcccc ttgagcatct gacttctggc taataaagga aatttatttt cattgcaata
     4441 gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc aaatcattta
     4501 aaacatcaga atgagtattt ggtttagagt ttggcaacat atgcccatat gctggctgcc
     4561 atgaacaaag gttggctata aagaggtcat cagtatatga aacagccccc tgctgtccat
     4621 tccttattcc atagaaaagc cttgacttga ggttagattt tttttatatt ttgttttgtg
     4681 ttattttttt ctttaacatc cctaaaattt tccttacatg ttttactagc cagatttttc
     4741 ctcctctcct gactactccc agtcatagct gtccctcttc tcttatggag atccctcgac
     4801 ctgcagccca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     4861 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     4921 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     4981 cctgtcgtgc cagcggatcc gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     5041 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     5101 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     5161 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc taacttgttt attgcagctt
     5221 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     5281 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatccgct
     5341 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     5401 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     5461 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     5521 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     5581 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     5641 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     5701 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     5761 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     5821 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     5881 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     5941 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     6001 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     6061 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     6121 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     6181 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     6241 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     6301 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     6361 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     6421 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     6481 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     6541 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     6601 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     6661 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     6721 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     6781 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     6841 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     6901 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     6961 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     7021 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     7081 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     7141 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     7201 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     7261 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     7321 acctggtcga cggtatcgat aagcttaaaa aagcgaaagg aagatcagta tatgatctct
     7381 tgaatcatat actgatcttc ctttcgcaga tctgtggtct catacagaac ttataagatt
     7441 cccaaatcca aagacatttc acgtttatgg tgatttccca gaacacatag cgacatgcaa
     7501 atattgcagg gcgccactcc cctgtccctc acagccatct tcctgccagg gcgcacgcgc
     7561 gctgggtgtt cccgcctagt gacactgggc ccgcgattcc ttggagcggg ttgatgacgt
     7621 cagcgttcga attctcgaca ttgattattg actagttatt aatagtaatc aattacgggg
     7681 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
     7741 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
     7801 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
     7861 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
     7921 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
     7981 cagtacatct acgtattagt catcgctatt accatggtcg aggtgagccc cacgttctgc
     8041 ttcactctcc ccatctcccc cccctcccca cccccaattt tgtatttatt tattttttaa
     8101 ttattttgtg cagcgatggg ggcgggg
//
