LOCUS       sequence-275998-        7848 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector for phycocyanobilin (PCB) synthesis in mammalian 
            cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7848)
  AUTHORS   Uda Y, Miura H, Goto Y, Yamamoto K, Mii Y, Kondo Y, Takada S, Aoki K
  TITLE     Improvement of Phycocyanobilin Synthesis for Genetically Encoded 
            Phytochrome-Based Optogenetics.
  JOURNAL   ACS Chem Biol. 2020 Nov 20;15(11):2896-2906. doi: 
            10.1021/acschembio.0c00477. Epub 2020 Nov 9.
  PUBMED    33164485
REFERENCE   2  (bases 1 to 7848)
  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"; doi: 
            "10.1021/acschembio.0c00477"; journalName: "ACS Chem Biol"; date: 
            "2020-11-20- 20"; volume: "15"; issue: "11"; pages: "2896-2906"
COMMENT     Sequence Label: pCAGGS-synPCB2.1
FEATURES             Location/Qualifiers
     source          1..7848
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        9..388
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        390..666
                     /label=chicken beta-actin promoter
     intron          667..1687
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1607..1629
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1695..1714
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1741..1815
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             3046..3102
                     /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             3103..3177
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     regulatory      3624..3633
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3913..3936
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3946..4002
                     /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             4006..4080
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             4111..4137
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(4870..4889)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4935..4990
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4989..5008)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5351..5367)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5351..5367)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5364..5386)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5375..5391
                     /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(5399..5405,5406..5423,5424..5429))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5444..5465
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5524..5719
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5570..5705
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5725..5859
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5762..5781)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5816..5835
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5927..5944)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6098..6686)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6178..6197)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6857..7648,7649..7717))
                     /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     7480..7499
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7718..7822)
                     /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 gggggggggg gcgcgcncag gcggggcggg gcggggcgag
      541 gggcggggcg gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg cgcgctccga
      601 aagtttcctt ttatggcgag gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg
      661 cgggcgggag tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc
      721 cgcccgcccc ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct
      781 tctcctccgg gctgtaatta gcgcttggtt taatgacggc ttgtttcttt tctgtggctg
      841 cgtgaaagcc ttgaggggct ccgggagggc cctttgtgcg gggggagcgg ctcggggggt
      901 gcgtgcgtgt gtgtgtgcgt ggggagcgcc gcgtgcggct ccgcgctgcc cggcggctgt
      961 gagcgctgcg ggcgcggcgc ggggctttgt gcgctccgca gtgtgcgcga ggggagcgcg
     1021 gccgggggcg gtgccccgcg gtgcgggggg ggctgcgagg ggaacaaagg ctgcgtgcgg
     1081 ggtgtgtgcg tgggggggtg agcagggggt gtgggcgcgt cggtcgggct gcaacccccc
     1141 ctgcaccccc ctccccgagt tgctgagcac ggcccggctt cgggtgcggg gctccgtacg
     1201 gggcgtggcg cggggctcgc cgtgccgggc ggggggtggc ggcaggtggg ggtgccgggc
     1261 ggggcggggc cgcctcgggc cggggagggc tcgggggagg ggcgcggcgg cccccggagc
     1321 gccggcggct gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag
     1381 agggcgcagg gacttccttt gtcccaaatc tgtgcggagc cgaaatctgg gaggcgccgc
     1441 cgcaccccct ctagcgggcg cggggcgaag cggtgcggcg ccggcaggaa ggaaatgggc
     1501 ggggagggcc ttcgtgcgtc gccgcgccgc cgtccccttc tccctctcca gcctcggggc
     1561 tgtccgcggg gggacggctg ccttcggggg ggacggggca gggcggggtt cggcttctgg
     1621 cgtgtgaccg gcggctctag ctagagcctc tgctaaccat gttcatgcct tcttcttttt
     1681 cctacagctc ctgggcaacg tgctggttat tgtgctgtct catcattttg gcaagaattc
     1741 atgtccgtcc tgacgccgct gctgctgcgg ggcttgacag gctcggcccg gcggctccca
     1801 gtgccgcgcg ccaagatcca ttcgttgctc gagaatattt accgacctaa caatccttgc
     1861 attgggaaag tcattagcaa tgaggagctg gtgcgagagg ggggagaagg gactgtaaaa
     1921 catataatct tcgatatatc aggcacggag cttcggtatc ttgaggggca atctattgga
     1981 atcataccgg cgggtactga tgccaatgga aaaccgcata aattgcgcct ctactctata
     2041 gcttctacta ggcacgggga tttccaggat gataagaccg tctctttgtg tgtccgcagg
     2101 ctggaataca aggataagga aaccggagag acaatatatg gggtatgttc ctcatacctg
     2161 aatcaattgc aaccaggaga tgaagtgaag ataacggggc ctgttggtaa ggagatgctc
     2221 ctgtcagatg acccagaagc tacaattata atgctggcga ctgggactgg catagcgcct
     2281 ttccgagcct tcctctggag aatgttcaag gaaaacaacc ccgattatca attcaagggg
     2341 cttgcctggt tgttttttgg tgtagcatac acggctaaca tcctttacaa agatgaactc
     2401 gaagcgattc aggcgcagta cccggatcat tttcgcttga catacgccat atctagggaa
     2461 cagaagaccc ccgacggggg taaaatgtat atccaagggc gaatagccga acatgcagat
     2521 gaaatctggc aactgctgca aaagaaaaat actcacgtct atatgtgtgg tctcagagga
     2581 atggagccgg gcatagacga agcgatgacc gctgcggcgg caaagaacgg cgccgattgg
     2641 caagaatttc tcaagggtac acttaagaag gaggggcgct ggcacgtcga gacgtacggc
     2701 ggatccggag gttccggcgg ctcaggaggt tctatggcta cctataaggt cacgctggtt
     2761 cgaccagatg gtagcgagac taccattgat gtccctgaag acgaatatat tctggacgta
     2821 gctgaggagc aaggcttgga tcttcctttc agctgccggg ctggtgcttg cagcacgtgc
     2881 gcgggaaaac ttctggaggg agaggtcgat cagagcgatc agtccttcct cgacgatgac
     2941 caaatagaaa agggtttcgt tctcacttgc gttgcctacc caagaagcga ttgcaaaata
     3001 ctcacaaacc aggaggaaga gctgtacggc ggccgcggct caggtgcgac gaattttagt
     3061 ctactgaaac aagcgggaga cgtggaggaa aaccctggac ctatgtcagt gctgactcca
     3121 ctgctgctga ggggcctgac tgggtccgca cgccgcctgc ctgtcccaag agccaaaatc
     3181 catagcctgg gcgggtctgg aggcatgagt ctgaggcagc accagcatcc tctgatccag
     3241 cgcctggccg atcgaatcga ggcaatttgg caggccttct ttcccctggc accttacgcc
     3301 ctgccagaag acctgggcta tgtggaggga aaactggagg gcgaacgcct gacaatcgag
     3361 aaccactgct atcaggcccc cccttttcgg aagctgcatc tggaactggc tagagtgggc
     3421 gagtctctgg acattctgca ctgcgtcatg tttcccgagc ctagatacga tctgcccatg
     3481 ttcggatgtg acctggtggg gggacgcgga cagatcagtg ccgctattgt ggatctgtca
     3541 ccagtcactg gacagctgcc agcagcatac acctgcgctc tgaatgcact gcctaagctg
     3601 acctttagac agccaaggga gctgccacca tggggacata tcttctctcc cttttgtatc
     3661 ttcattagac ctcagggcga agccgaggaa cagcagttcc tggataggat tggggagtat
     3721 ctgactctgc actgccagct gagccagcag gcagtgccta ccgaccatcc acaggccgtc
     3781 atcgctggac agcggcagta ttgtcagcag cagcagcaga acgataaaac tcggagagtg
     3841 ctggaaaagg cttttggcgt cccatgggca gagagataca tgaccacagt gctgttcgat
     3901 gtgcctccag tggactataa agacgatgac gataaaggca gcggagcgac gaattttagt
     3961 ctactaaaac aagcgggtga tgtagaagaa aatccgggtc cgggcatgtc agttctgaca
     4021 cctctgctgc tgcgtggact gactggaagc gccaggcgcc tgcctgtgcc acgagcaaaa
     4081 atccactcac tgggcgggag cggaggcatg tacccctatg atgtccctga ctacgcctct
     4141 agaatgacta ccagcctggc tacaaagctg agggaaggca ctaagaaagc tcataccatg
     4201 gcagagaacg tggggtttgt ccggtgtttc ctgaagggaa ctgtggagaa aagctcctat
     4261 agaaagctgg tggcctctct gtaccacgtc tatagtgcta tggagcagga aatggagcgc
     4321 ctgaaagacc atccaatctt gggcaaaatc tactttcccg aactgaacag gaagtctagt
     4381 ctggagcgcg atctgacata ctatttcggg tccaattgga gggaggaaat tcccccttct
     4441 cccgcaactc aggcctacgt ggctcgcatc cacgaagtcg caaacaccgc ccctgagctg
     4501 ctggtggccc atagttacac acggtatctg ggagacctgt cagggggaca gatcctgaag
     4561 ggcattgctg aacgggcaat gaatctgcag gatggggagg gaaccgcctt ctacagattc
     4621 gaaagtatct cagacgagaa ggccttcaaa cagctgtatc gacagcggct ggatgaactg
     4681 cccgtggacg aggctacagc agatagaatt gtggacgagg ctaacgctgc atttggcatg
     4741 aatatgaaga ttttccagga actggagggc aacctgatcc gggccattgg gcagctgctg
     4801 tttaataccc tgacacgacg gaaacagaga ggcagcaccg agctggccac agctgattaa
     4861 gtcgacggat ctttttccct ctgccaaaaa ttatggggac atcatgaagc cccttgagca
     4921 tctgacttct ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt
     4981 gtgtctctca ctcggaagga catatgggag ggcaaatcat ttaaaacatc agaatgagta
     5041 tttggtttag agtttggcaa catatgccca tatgctggct gccatgaaca aaggttggct
     5101 ataaagaggt catcagtata tgaaacagcc ccctgctgtc cattccttat tccatagaaa
     5161 agccttgact tgaggttaga ttttttttat attttgtttt gtgttatttt tttctttaac
     5221 atccctaaaa ttttccttac atgttttact agccagattt ttcctcctct cctgactact
     5281 cccagtcata gctgtccctc ttctcttatg gagatccctc gacctgcagc ccaagcttgg
     5341 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     5401 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     5461 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagcgga
     5521 tccgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     5581 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     5641 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     5701 aggcctaggc ttttgcaaaa agctaacttg tttattgcag cttataatgg ttacaaataa
     5761 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     5821 ttgtccaaac tcatcaatgt atcttatcat gtctggatcc gctgcattaa tgaatcggcc
     5881 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     5941 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     6001 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     6061 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     6121 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     6181 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     6241 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     6301 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     6361 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     6421 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     6481 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     6541 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     6601 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     6661 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     6721 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     6781 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     6841 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     6901 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     6961 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     7021 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     7081 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     7141 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     7201 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     7261 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     7321 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     7381 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     7441 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     7501 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     7561 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     7621 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     7681 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     7741 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     7801 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctg
//
