LOCUS       sequence-269825-        7838 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 7838)
  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 7838)
  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..7838
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        8..387
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        390..665
                     /label=chicken beta-actin promoter
     intron          666..1678
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1598..1620
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     CDS             1733..1807
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             3038..3094
                     /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             3095..3169
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     regulatory      3616..3625
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3905..3928
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3938..3994
                     /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             3998..4072
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             4103..4129
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(4862..4881)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4927..4982
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4981..5000)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5341..5357)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5341..5357)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5354..5376)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5365..5381
                     /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(5389..5395,5396..5413,5414..5419))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5434..5455
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5514..5709
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5560..5695
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5715..5849
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5752..5771)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5806..5825
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5917..5934)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6088..6676)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6168..6187)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6847..7638,7639..7707))
                     /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     7470..7489
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7708..7812)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgatcgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
       61 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
      121 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
      181 atagggactt tccattgacg tcaatgggtg gactatttac ggtaaactgc ccacttggca
      241 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
      301 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
      361 tacgtattag tcatcgctat taccatgggt 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 ggcggccccg gagcgccggc
     1321 ggctgtcgag gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg
     1381 cagggacttc ctttgtccca aatctggcgg agccgaaatc tgggaggcgc cgccgcaccc
     1441 cctctagcgg gcgcgggcga agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg
     1501 ccttcgtgcg tcgccgcgcc gccgtcccct tctccatctc cagcctcggg gctgccgcag
     1561 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
     1621 ggcggctcta gctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1681 cctgggcaac gtgctggttg ttgtgctgtc tcatcatttt ggcaagaatt caatgtccgt
     1741 cctgacgccg ctgctgctgc ggggcttgac aggctcggcc cggcggctcc cagtgccgcg
     1801 cgccaagatc cattcgttgc tcgagaatat ttaccgacct aacaatcctt gcattgggaa
     1861 agtcattagc aatgaggagc tggtgcgaga ggggggagaa gggactgtaa aacatataat
     1921 cttcgatata tcaggcacgg agcttcggta tcttgagggg caatctattg gaatcatacc
     1981 ggcgggtact gatgccaatg gaaaaccgca taaattgcgc ctctactcta tagcttctac
     2041 taggcacggg gatttccagg atgataagac cgtctctttg tgtgtccgca ggctggaata
     2101 caaggataag gaaaccggag agacaatata tggggtatgt tcctcatacc tgaatcaatt
     2161 gcaaccagga gatgaagtga agataacggg gcctgttggt aaggagatgc tcctgtcaga
     2221 tgacccagaa gctacaatta taatgctggc gactgggact ggcatagcgc ctttccgagc
     2281 cttcctctgg agaatgttca aggaaaacaa ccccgattat caattcaagg ggcttgcctg
     2341 gttgtttttt ggtgtagcat acacggctaa catcctttac aaagatgaac tcgaagcgat
     2401 tcaggcgcag tacccggatc attttcgctt gacatacgcc atatctaggg aacagaagac
     2461 ccccgacggg ggtaaaatgt atatccaagg gcgaatagcc gaacatgcag atgaaatctg
     2521 gcaactgctg caaaagaaaa atactcacgt ctatatgtgt ggtctcagag gaatggagcc
     2581 gggcatagac gaagcgatga ccgctgcggc ggcaaagaac ggcgccgatt ggcaagaatt
     2641 tctcaagggt acacttaaga aggaggggcg ctggcacgtc gagacgtacg gcggatccgg
     2701 aggttccggc ggctcaggag gttctatggc tacctataag gtcacgctgg ttcgaccaga
     2761 tggtagcgag actaccattg atgtccctga agacgaatat attctggacg tagctgagga
     2821 gcaaggcttg gatcttcctt tcagctgccg ggctggtgct tgcagcacgt gcgcgggaaa
     2881 acttctggag ggagaggtcg atcagagcga tcagtccttc ctcgacgatg accaaataga
     2941 aaagggtttc gttctcactt gcgttgccta cccaagaagc gattgcaaaa tactcacaaa
     3001 ccaggaggaa gagctgtacg gcggccgcgg ctcaggtgcg acgaatttta gtctactgaa
     3061 acaagcggga gacgtggagg aaaaccctgg acctatgtca gtgctgactc cactgctgct
     3121 gaggggcctg actgggtccg cacgccgcct gcctgtccca agagccaaaa tccatagcct
     3181 gggcgggtct ggaggcatga gtctgaggca gcaccagcat cctctgatcc agcgcctggc
     3241 cgatcgaatc gaggcaattt ggcaggcctt ctttcccctg gcaccttacg ccctgccaga
     3301 agacctgggc tatgtggagg gaaaactgga gggcgaacgc ctgacaatcg agaaccactg
     3361 ctatcaggcc cccccttttc ggaagctgca tctggaactg gctagagtgg gcgagtctct
     3421 ggacattctg cactgcgtca tgtttcccga gcctagatac gatctgccca tgttcggatg
     3481 tgacctggtg gggggacgcg gacagatcag tgccgctatt gtggatctgt caccagtcac
     3541 tggacagctg ccagcagcat acacctgcgc tctgaatgca ctgcctaagc tgacctttag
     3601 acagccaagg gagctgccac catggggaca tatcttctct cccttttgta tcttcattag
     3661 acctcagggc gaagccgagg aacagcagtt cctggatagg attggggagt atctgactct
     3721 gcactgccag ctgagccagc aggcagtgcc taccgaccat ccacaggccg tcatcgctgg
     3781 acagcggcag tattgtcagc agcagcagca gaacgataaa actcggagag tgctggaaaa
     3841 ggcttttggc gtcccatggg cagagagata catgaccaca gtgctgttcg atgtgcctcc
     3901 agtggactat aaagacgatg acgataaagg cagcggagcg acgaatttta gtctactaaa
     3961 acaagcgggt gatgtagaag aaaatccggg tccgggcatg tcagttctga cacctctgct
     4021 gctgcgtgga ctgactggaa gcgccaggcg cctgcctgtg ccacgagcaa aaatccactc
     4081 actgggcggg agcggaggca tgtaccccta tgatgtccct gactacgcct ctagaatgac
     4141 taccagcctg gctacaaagc tgagggaagg cactaagaaa gctcatacca tggcagagaa
     4201 cgtggggttt gtccggtgtt tcctgaaggg aactgtggag aaaagctcct atagaaagct
     4261 ggtggcctct ctgtaccacg tctatagtgc tatggagcag gaaatggagc gcctgaaaga
     4321 ccatccaatc ttgggcaaaa tctactttcc cgaactgaac aggaagtcta gtctggagcg
     4381 cgatctgaca tactatttcg ggtccaattg gagggaggaa attccccctt ctcccgcaac
     4441 tcaggcctac gtggctcgca tccacgaagt cgcaaacacc gcccctgagc tgctggtggc
     4501 ccatagttac acacggtatc tgggagacct gtcaggggga cagatcctga agggcattgc
     4561 tgaacgggca atgaatctgc aggatgggga gggaaccgcc ttctacagat tcgaaagtat
     4621 ctcagacgag aaggccttca aacagctgta tcgacagcgg ctggatgaac tgcccgtgga
     4681 cgaggctaca gcagatagaa ttgtggacga ggctaacgct gcatttggca tgaatatgaa
     4741 gattttccag gaactggagg gcaacctgat ccgggccatt gggcagctgc tgtttaatac
     4801 cctgacacga cggaaacaga gaggcagcac cgagctggcc acagctgatt aagtcgacgg
     4861 atctttttcc ctctgccaaa aattatgggg acatcatgaa gccccttgag catctgactt
     4921 ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct
     4981 cactcggaag gacatatggg agggcaaatc atttaaaaca tcagaatgag tatttggttt
     5041 agagtttggc aacatatgcc atatgctggc tgccatgaac aaaggtggct ataaagaggt
     5101 catcagtata tgaaacagcc ccctgctgtc cattccttat tccatagaaa agccttgact
     5161 tgaggttaga ttttttttta tattttgttt gtgttatttt tttctttaac atccctaaaa
     5221 ttttccttac atgttttact agccagattt ttcctcctct cctgactact cccagtcata
     5281 gctgtccctc ttctcttatg aagatccctc gacctgcagc ccaagcttgg cgtaatcatg
     5341 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     5401 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     5461 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagcgga tccgcatctc
     5521 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     5581 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     5641 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     5701 ttttgcaaaa agctaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     5761 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     5821 tcatcaatgt atcttatcat gtctggatcc gctgcattaa tgaatcggcc aacgcgcggg
     5881 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     5941 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     6001 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     6061 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     6121 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     6181 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     6241 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta
     6301 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     6361 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     6421 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     6481 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     6541 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6601 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     6661 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6721 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6781 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     6841 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     6901 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     6961 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     7021 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     7081 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     7141 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     7201 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     7261 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     7321 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     7381 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     7441 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     7501 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     7561 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     7621 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     7681 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     7741 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     7801 aggggttccg cgcacatttc cccgaaaagt gccacctg
//
