LOCUS       V009065                 6399 bp    DNA     circular SYN 17-DEC-2018
DEFINITION  Exported.
ACCESSION   V009065
VERSION     V009065
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6399)
  AUTHORS   Tooley AJ, Glazer AN.
  TITLE     Biosynthesis of the cyanobacterial light-harvesting polypeptide
            phycoerythrocyanin holo-alpha subunit in a heterologous host
  JOURNAL   J. Bacteriol. 184 (17), 4666-4671 (2002)
   PUBMED   12169589
REFERENCE   2  (bases 1 to 6399)
  AUTHORS   Tooley AJ, Glazer AN.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-MAY-2003) Molecular and Cell Biology, University of
            California, Berkeley, 142 LSA no. 3200, Berkeley, CA 94720-3200, USA
REFERENCE   3  (bases 1 to 6399)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6399)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J.
            Bacteriol."; date: "2002"; volume: "184"; issue: "17"; pages:
            "4666-4671"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (18-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..6399
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     gene            10..195
                     /gene="rop"
                     /label="rop"
                     /note="coding region not determined; ROP'; repressor of
                     primer protein variant; regulation of plasmid replication;
                     helps maintain 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="AAP45701.1"
                     /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
                     /label="transcription initiation site from trc promoter"
                     /note="transcription initiation site from 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="pecA"
                     /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="pecA"
                     /label="Phycoerythrocyanin alpha chain"
                     /note="Phycoerythrocyanin alpha chain from Nostoc sp.
                     (strain PCC 7120 / SAG 25.82 / UTEX 2576). Accession#:
                     P35796"
     gene            4487..5259
                     /gene="pecE"
                     /label="pecE"
     regulatory      4487..4492
                     /gene="pecE"
                     /regulatory_class="ribosome_binding_site"
     CDS             4498..5259
                     /codon_start=1
                     /gene="pecE"
                     /product="PecE"
                     /label="pecE"
                     /note="catalyzes both the covalent attachment of
                     phycocyanobilin to the phycoerythrocyanin alpha subunit and
                     its concurrent isomerization to phycobiliviolin"
                     /protein_id="AAP45704.1"
                     /translation="MTAEPILSPETAIAALSGEDNQIRYYAAWWLGKHNVQAGFTALCV
                     ALFDERYRIPSGGYPLRRQAARALGLLKNPQAVPALIAALECDEDLRLREAVICSLAAI
                     GDKRAVTPLLNLLQSSQAQPYEALIEALATLQVWSARPQIEPFLQHYSERVQCAAARYM
                     YLLTQESEYIERIVKNLNHDNMYLRWAAVFDLGAVAHQQAVQAILTAQVPNSLKLLNLK
                     RILEAMLNNNSVNHEKAALLFGAIDDLLIQL"
     gene            5268..5800
                     /gene="pecF"
                     /label="pecF"
     regulatory      5268..5273
                     /gene="pecF"
                     /regulatory_class="ribosome_binding_site"
     CDS             5279..5800
                     /codon_start=1
                     /gene="pecF"
                     /product="PecF"
                     /label="pecF"
                     /note="catalyzes both the covalent attachment of
                     phycocyanobilin to the phycoerythrocyanin alpha subunit and
                     its concurrent isomerization to phycobiliviolin"
                     /protein_id="AAP45705.1"
                     /translation="MNQASLSVDAITNLIEAFHHHHPAVRSAAVDELIKLGSITVNLLI
                     AAYDDSQDQGFQAQIIQVLAQIGDAKALKLLAEVVGTSVANHCQGNVRRIAARGLGKIA
                     STTSNTEIINNAQEKLIWALLTPEDWGLRYAAAVSLQEIATPKAKAALQQAIAQETDPV
                     VRSRMAIALS"
     terminator      6136..6222
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      6314..6341
                     /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 aacacctttg accgaagcga tctctgctgc tgatgttcgt ggttcctacc
     4021 tcagcaacac tgaaatgcaa gcagtttttg gtcgttttaa ccgcgctcgc gctggtttag
     4081 cggctgctca agcattttct aataatggca aaaaatgggc agaagcagca gcaaatcatg
     4141 tttatcaaaa attcccttac accactcaaa tgagtgggcc tcaatatgct tccactccag
     4201 aaggtaaatc caagtgtgtt cgtgatattg accactatct tcgtactatt agctattgct
     4261 gcgtagtcgg tggtactggc cccttagatg aatatgtagt atctggttta agcgaactca
     4321 acagcgccct tggtttatct cctagctggt atgtagctgc actagaattt gtccgggata
     4381 atcacggttt aaatggtgat gttgcaggtg aagctaatat ctacctgaac tatgcaatta
     4441 acgctttaag ttaagagact cgaattcaaa ggcctacgtc gacaggagga aagccatatg
     4501 actgctgaac caattctttc tccagaaaca gcgatcgcag ctttatctgg tgaggataac
     4561 caaattcgtt attatgctgc ttggtggttg ggtaaacata acgtacaggc tggttttacg
     4621 gcgctatgtg tagccttatt tgatgaacgc tatcgcatcc catcaggagg gtatcctctg
     4681 cgtcgtcagg ctgcacgggc tttaggattg ttgaaaaatc cccaagctgt accagcctta
     4741 attgcagcat tggaatgtga tgaggatttg cgtctgaggg aagcagtaat ttgctcttta
     4801 gcggcgatcg gtgacaaacg tgctgtgact cctttactca atcttttgca atcaagtcaa
     4861 gcacaacctt atgaagcttt aatcgaagct ttagctactc tacaagtttg gtcagcccgt
     4921 ccacaaattg aaccttttct tcagcattat tctgagcgtg tgcaatgtgc agccgctaga
     4981 tatatgtatc ttctcaccca agaatctgaa tacattgaac gtattgtcaa aaatctcaat
     5041 catgacaata tgtatttacg ttgggcggcg gtatttgatt tgggtgcagt tgctcatcaa
     5101 caagccgtac aggcaatctt aacggctcag gttcctaata gcttaaagct cttaaattta
     5161 aagcgtattc ttgaagctat gttgaataat aatagtgtta atcatgaaaa agcagcttta
     5221 ttatttggag caattgatga tttattaatt caactttaaa gcggccgagg aggaacatat
     5281 gaatcaagct tcattgagcg tagacgctat taccaattta atcgaagcat ttcatcatca
     5341 ccatcctgct gttagaagtg cggctgtaga tgaattaatc aaattgggaa gcatcacagt
     5401 taatttgtta attgctgctt atgatgattc tcaagatcaa ggttttcaag cccagatcat
     5461 tcaggtttta gctcaaattg gtgatgcaaa ggcactaaaa ttattagccg aggtggtggg
     5521 gacaagtgta gccaatcact gtcaaggtaa tgtccgccgc attgctgcac gagggttagg
     5581 aaaaatcgcc agcaccacta gtaatacaga aataatcaac aatgctcaag aaaagttaat
     5641 ttgggcatta cttacccccg aagattgggg tctacgttac gctgctgctg tctctttaca
     5701 agaaattgcc acgccaaaag ctaaagctgc tttacaacaa gcgatcgccc aagaaaccga
     5761 tccagttgtt cgctcacgta tggcgatcgc cttgagttaa gggactcgga tccggagaca
     5821 tgtctagaat gcatgatatc tcgagatctg gtacccgggc gcgcctgcag gcatgcaagc
     5881 ttggctgttt tggcggatga gagaagagca gcctgaatgg cgaatggcag cttggctgtt
     5941 ttggcggatg agataagatt ttcagcctga tacagattaa atcagaacgc agaagcggtc
     6001 tgataaaaca gaatttgcct ggcggcagta gcgcggtggt cccacctgac cccatgccga
     6061 actcagaagt gaaacgccgt agcgccgatg gtagtgtggg gtctccccat gcgagagtag
     6121 ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt
     6181 atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccggg agcggatttg
     6241 aacgttgcga agcaacggcc cggagggtgg cgggcaggac gcccgccata aactgccagg
     6301 catcaaatta agcagaaggc catcctgacg gatggccttt ttgcgtttct acaaactctt
     6361 tttgtttatt tttctaaata cattcaaata tgtatccgc
//
