LOCUS       V009070                 6587 bp    DNA     circular SYN 17-DEC-2018
DEFINITION  Exported.
ACCESSION   V009070
VERSION     V009070
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6587)
  AUTHORS   Tooley AJ, Cai YA, Glazer AN.
  TITLE     Biosynthesis of a fluorescent cyanobacterial C-phycocyanin
            holo-alpha subunit in a heterologous host
  JOURNAL   Proc. Natl. Acad. Sci. U.S.A. 98 (19), 10560-10565 (2001)
   PUBMED   11553806
REFERENCE   2  (bases 1 to 6587)
  AUTHORS   Tooley AJ, Cai YA, Glazer AN.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-MAY-2003) Molecular and Cell Biology, University of
            California, Berkeley, 142 LSA no. 3200, Berkeley, CA 94720-3200, USA
REFERENCE   3  (bases 1 to 6587)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6587)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc. Natl.
            Acad. Sci. U.S.A."; date: "2001"; volume: "98"; issue: "19"; pages:
            "10560-10565"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (04-MAY-2003) Molecular and Cell Biology, University of California,
            Berkeley, 142 LSA no. 3200, Berkeley, CA 94720-3200, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6587
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     gene            10..195
                     /gene="rop"
                     /label="rop"
                     /note="coding region not determined; repressor of primer
                     protein variant; regulation of plasmid replication;
                     maintains medium plasmid copy number"
     misc_feature    297..439
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(625..1213)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             1497..2285
                     /codon_start=1
                     /gene="aadA"
                     /product="AadA"
                     /label="aadA"
                     /note="confers resistance to streptomycin and to
                     spectinomycin via inactivation of the drugs by specific
                     adenylylation"
                     /protein_id="AAP43512.1"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEDRLASRADQLEEFVHYVKGEITKVVGK"
     gene            1497..2285
                     /gene="aadA"
                     /label="aadA"
     CDS             2506..3585
                     /label="lacI"
                     /note="lac repressor"
     regulatory      3820..3825
                     /label="trc promoter"
                     /note="trc promoter"
                     /regulatory_class="minus_35_signal"
     regulatory      3843..3849
                     /label="trc promoter"
                     /note="trc promoter"
                     /regulatory_class="minus_10_signal"
     misc_feature    3855..3856
                     /note="transcription initiation site from the trc promoter"
     protein_bind    3857..3873
                     /label="lac operator"
                     /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)."
     regulatory      3882..3887
                     /gene="cpcA"
                     /regulatory_class="ribosome_binding_site"
     CDS             3900..3917
                     /label="6xHis"
                     /note="6xHis affinity tag"
     CDS             3939..3959
                     /label="TEV site"
                     /note="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
     CDS             3966..4451
                     /gene="cpcA"
                     /label="C-phycocyanin alpha subunit"
                     /note="C-phycocyanin alpha subunit from Synechocystis sp.
                     (strain PCC 6803 / Kazusa). Accession#: Q54715"
     gene            4483..5313
                     /gene="cpcE"
                     /label="cpcE"
     regulatory      4483..4488
                     /gene="cpcE"
                     /regulatory_class="ribosome_binding_site"
     CDS             4495..5313
                     /codon_start=1
                     /gene="cpcE"
                     /product="CpcE"
                     /label="cpcE"
                     /note="required for the covalent attachment of
                     phycocyanobilin to the alpha subunit apoprotein of
                     C-phycocyanin"
                     /protein_id="AAP43515.1"
                     /translation="MSEPNLNPAYTLDQAIANLQQTEDASARYYAAWWIGRFRAAQPET
                     IAALLVALEDETDRSPDGGYPLRRNAAKALGKLGDRQVVPALIKALECEDYYVRESAAQ
                     ALEGLGDARAMAPLMAKLTGGLAAAQLVEGKPHLAQPYEAIIEALGTLQAVESIGLIEP
                     FLEHFSPKVQYAAARALFQLTGDNRYGDLLITALGGTDLQLRRSAMMDLGATGYLPGAQ
                     AIAKAFAENSLKLIALRDLWATHRQRQASSESKALSPASRQILELMDSLL"
     gene            5330..5988
                     /gene="cpcF"
                     /label="cpcF"
     regulatory      5330..5335
                     /gene="cpcF"
                     /regulatory_class="ribosome_binding_site"
     CDS             5344..5988
                     /codon_start=1
                     /gene="cpcF"
                     /product="CpcF"
                     /label="cpcF"
                     /note="required for the covalent attachment of
                     phycocyanobilin to the alpha subunit apoprotein of
                     C-phycocyanin"
                     /protein_id="AAP43516.1"
                     /translation="MEGNSVVTPEIERLIQAVETADSAAKLVGAVRALAATRSPLAVPQ
                     LTTVLRYNNPGAAVAAVDGLIQIGDAAMTHLLANMDGYNYGARAWATRACAGIGDPRAL
                     ALLQEAALTDFALSVRRAAAKGLGFLRWQSLPQEEQETVQKAIYDTLIQVCEDPEWVVR
                     YGAIAGLENLAKQAQHYRQPLKDFLQSFVEQEPEAIVGERILWTLENIGPI"
     terminator      6324..6410
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      6502..6529
                     /label="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
ORIGIN
        1 tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc gctttatcag aagccagaca
       61 ttaacgcttc tggagaaact caacgagctg gacgcggatg aacaggcaga catctgtgaa
      121 tcgcttcacg accacgctga tgagctttac cgcagctgcc tcgcgcgttt cggtgatgac
      181 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
      241 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca
      301 gccatgaccc agtcacgtag cgatagcgga gtgtatactg gcttaactat gcggcatcag
      361 agcagattgt actgagagtg caccatatat gcggtgtgaa ataccgcaca gatgcgtaag
      421 gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
      481 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
      541 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
      601 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
      661 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
      721 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
      781 gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct
      841 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
      901 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
      961 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     1021 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     1081 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     1141 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     1201 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     1261 aaactcacgt taagggattt tggtcatggt tcagcgatcg ctcctcatct tcaataagca
     1321 gggcatgagc cagcgttaag caaatcaaat caaatctcgc ttctgggctt caataaatgg
     1381 ttccgattga tgataggttg attcatgcaa gctagcttgg agcacaggat gacgcctaac
     1441 aattcattca agccgacacc gcttcgcggc gcggcttaat tcaggagtta aacatcatga
     1501 gggaagcggt gatcgccgaa gtatcgactc aactatcaga ggtagttggc gtcatcgagc
     1561 gccatctcga accgacgttg ctggccgtac atttgtacgg ctccgcagtg gatggcggcc
     1621 tgaagccaca cagtgatatt gatttgctgg ttacggtgac cgtaaggctt gatgaaacaa
     1681 cgcggcgagc tttgatcaac gaccttttgg aaacttcggc ttcccctgga gagagcgaga
     1741 ttctccgcgc tgtagaagtc accattgttg tgcacgacga catcattccg tggcgttatc
     1801 cagctaagcg cgaactgcaa tttggagaat ggcagcgcaa tgacattctt gcaggtatct
     1861 tcgagccagc cacgatcgac attgatctgg ctatcttgct gacaaaagca agagaacata
     1921 gcgttgcctt ggtaggtcca gcggcggagg aactctttga tccggttcct gaacaggatc
     1981 tatttgaggc gctaaatgaa accttaacgc tatggaactc gccgcccgac tgggctggcg
     2041 atgagcgaaa tgtagtgctt acgttgtccc gcatttggta cagcgcagta accggcaaaa
     2101 tcgcgccgaa ggatgtcgct gccgactggg caatggagcg cctgccggcc cagtatcagc
     2161 ccgtcatact tgaagctagg caggcttatc ttggacaaga agatcgcttg gcctcgcgcg
     2221 cagatcagtt ggaagaattt gttcactacg tgaaaggcga gatcaccaag gtagtcggca
     2281 aataatgtct aacaattcgt tcaagccgac gccgcttcgc ggcgcggctt aactcaagcg
     2341 ttagagagct ggggaagact atgcgcgatc tgttgaaggt ggttctaagc ctcgtacttg
     2401 cgatggcatc ggggcaggca cttgctgacc tgccaattat ggtgcaaaac ctttcgcggt
     2461 atggcatgat agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa accagtaacg
     2521 ttatacgatg tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg cgtggtgaac
     2581 caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat ggcggagctg
     2641 aattacattc ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt gctgattggc
     2701 gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc gattaaatct
     2761 cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag cggcgtcgaa
     2821 gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg tcagtgggct gatcattaac
     2881 tatccgctgg atgaccagga tgccattgct gtggaagctg cctgcactaa tgttccggcg
     2941 ttatttcttg atgtctctga ccagacaccc atcaacagta ttattttctc ccatgaagac
     3001 ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc accagcaaat cgcgctgtta
     3061 gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca taaatatctc
     3121 actcgcaatc aaattcagcc gatagcggaa cgggaaggcg actggagtgc catgtccggt
     3181 tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat gctggttgcc
     3241 aacgatcaga tggcgctggg cgcaatgcgc gccattaccg agtccgggct gcgcgttggt
     3301 gcggatatct cggtagtggg atacgacgat accgaagaca gctcatgtta tatcccgccg
     3361 tcaaccacca tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga ccgcttgctg
     3421 caactctctc agggccaggc ggtgaagggc aatcagctgt tgcccgtctc actggtgaaa
     3481 agaaaaacca ccctggcacc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     3541 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     3601 taatgtgagt tagcgcgaat tgatctggtt tgacagctta tcatcgactg cacggtgcac
     3661 caatgcttct ggcgtcaggc agccatcgga agctgtggta tggctgtgca ggtcgtaaat
     3721 cactgcataa ttcgtgtcgc tcaaggcgca ctcccgttct ggataatgtt ttttgcgccg
     3781 acatcataac ggttctggca aatattctga aatgagctgt tgacaattaa tcatccggtc
     3841 cgtataatct gtggaattgt gagcggataa caatttcaca caggaaacag accatgggtc
     3901 atcatcatca tcatcacgat tacgatatcc caacgaccga aaacctgtat tttcagggcg
     3961 cccatatgaa aaccccttta actgaagccg tttccaccgc tgactctcaa ggtcgctttc
     4021 tgagcagcac cgaattgcaa attgctttcg gtcgtctacg tcaagctaat gctggtttgc
     4081 aagccgctaa agctctgacc gacaatgccc agagcttggt aaatggtgct gcccaagccg
     4141 tttataacaa attcccctac accacccaaa cccaaggcaa caactttgct gcggatcaac
     4201 ggggtaaaga caagtgtgcc cgggacatcg gctactacct ccgcatcgtt acctactgct
     4261 tagttgctgg tggtaccggt cctttggatg agtacttgat cgccggtatt gatgaaatca
     4321 accgcacctt tgacctctcc cccagctggt atgttgaagc tctgaaatac atcaaagcta
     4381 accacggctt gagtggcgat gcccgtgacg aagctaattc ctacctcgat tacgccatca
     4441 atgctctgag ctagtcttcg aattcaaagg cctacgtcga caaggagctt gcatatgagt
     4501 gaacctaacc tcaaccccgc ctataccctc gaccaggcga tcgccaattt gcaacaaaca
     4561 gaagatgcca gtgcccgtta ctatgcggcc tggtggattg gtcgttttcg cgctgctcaa
     4621 ccggaaacca ttgctgcttt gttggtcgcc ttggaagatg aaactgatcg ctccccggac
     4681 ggtggttatc ccctgcggcg taatgcggct aaagcgttgg gaaaattagg cgatcgccaa
     4741 gtggtgccgg ccctaatcaa agccttggaa tgtgaagatt actatgtgcg agagtcagcg
     4801 gcccaggcgc tggaaggttt gggggatgct agagcgatgg cacctctaat ggccaagtta
     4861 acggggggat tagcagcggc ccagcttgtg gagggtaaac cccatttagc ccaaccctac
     4921 gaagcgatta ttgaagccct gggtactctc caagctgtgg agtccattgg tttaattgaa
     4981 ccgtttctag agcatttttc cccgaaagtc caatacgccg ccgccagggc cctatttcag
     5041 ttaactgggg ataaccgtta cggcgacctt ctcattactg ctctgggggg gacagactta
     5101 caattacgcc gttccgccat gatggatttg ggagccactg gctacttgcc cggcgcccag
     5161 gcgatcgcca aagcctttgc agaaaatagc ctcaagttga tcgccctgcg agatttgtgg
     5221 gccacccatc gccaaaggca agcaagttct gaatcgaagg cgctcagtcc agctagccga
     5281 caaatattgg aattaatgga tagtttactc taaaagcgtc gcggccgcta ggagggctaa
     5341 catatggagg gtaatagcgt cgtaactcca gaaatcgaaa gattgatcca agcggtggaa
     5401 accgcagact cggcagccaa attagtgggg gctgtgcggg ctttggcggc cacccgatcg
     5461 ccgttggcag tgccccaatt aaccacggtg ttgcgctaca acaatccagg ggcagcggtg
     5521 gcggcggtgg atgggctgat ccaaatcggc gatgcggcca tgactcactt gctggccaat
     5581 atggatggct acaattatgg cgctagggcc tgggccacca gagcctgtgc tggcatcggc
     5641 gaccccaggg cattggccct attgcaggaa gcggccctga cggattttgc cttgagtgtg
     5701 cggcgggcgg cggccaaggg actgggtttt ctccgttggc aatccctacc ccaggaagaa
     5761 caagagacag tacaaaaagc catttacgac actctgatcc aagtttgtga agatcctgaa
     5821 tgggtggtac gctacggggc gatcgccggt ctggaaaatt tagccaagca ggcccagcat
     5881 taccgccaac ccctaaagga ttttctccag tcttttgtag aacaagaacc agaggcgatc
     5941 gtgggggaac gcatcctttg gaccctggaa aacatcggcc caatctagtc ttctaggatc
     6001 cggagacatg tctagaatgc atgatatctc gagatctggt acccgggcgc gcctgcaggc
     6061 atgcaagctt ggctgttttg gcggatgaga gaagagcagc ctgaatggcg aatggcagct
     6121 tggctgtttt ggcggatgag ataagatttt cagcctgata cagattaaat cagaacgcag
     6181 aagcggtctg ataaaacaga atttgcctgg cggcagtagc gcggtggtcc cacctgaccc
     6241 catgccgaac tcagaagtga aacgccgtag cgccgatggt agtgtggggt ctccccatgc
     6301 gagagtaggg aactgccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct
     6361 ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccgggag
     6421 cggatttgaa cgttgcgaag caacggcccg gagggtggcg ggcaggacgc ccgccataaa
     6481 ctgccaggca tcaaattaag cagaaggcca tcctgacgga tggccttttt gcgtttctac
     6541 aaactctttt tgtttatttt tctaaataca ttcaaatatg tatccgc
//
