LOCUS       40924_5979        8193 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pBBR1-accBCD1, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8193)
  AUTHORS   Yang D, Kim WJ, Yoo SM, Choi JH, Ha SH, Lee MH, Lee SY.
  TITLE     Repurposing type III polyketide synthase as a malonyl-CoA biosensor 
            for metabolic engineering in bacteria
  JOURNAL   Proc. Natl. Acad. Sci. U.S.A. (2018) In press
  PUBMED    30232266
REFERENCE   2  (bases 1 to 8193)
  AUTHORS   Yang D, Kim WJ, Yoo SM, Choi JH, Ha SH, Lee MH, Lee SY.
  TITLE     Novel Enzyme-coupled Malonyl-CoA Biosensor based on Type III 
            Polyketide Synthase and Use of the same (KR 10-2018-0066323)
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 8193)
  AUTHORS   Yang D, Kim WJ, Yoo SM, Choi JH, Ha SH, Lee MH, Lee SY.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-JUL-2018) Dept. Chemical and Biomolecular Engineering,
            Korea Advanced Institute of Science and Technology, Daehak-ro 291, 
            Daejeon 34141, South Korea
REFERENCE   4  (bases 1 to 8193)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 8193)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc. Natl.
            Acad. Sci. U.S.A. (2018) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (19-JUL-2018) Dept. Chemical and Biomolecular Engineering, Korea 
            Advanced Institute of Science and Technology, Daehak-ro 291, Daejeon
            34141, South Korea"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8193
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..30
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    38..54
                     /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)."
     CDS             77..1852
                     /codon_start=1
                     /gene="accBC"
                     /product="acetyl-CoA carboxylase subunit alpha"
                     /label=accBC
                     /protein_id="AYD60196.1"
                     /translation="MSVETRKITKVLVANRGEIAIRVFRAARDEGIGSVAVYAEPDADA
                     PFVSYADEAFALGGQTSAESYLVIDKIIDAARKSGADAIHPGYGFLAENADFAEAVINE
                     GLIWIGPSPESIRSLGDKVTARHIADTAKAPMAPGTKEPVKDAAEVVAFAEEFGLPIAI
                     KAAFGGGGRGMKVAYKMEEVADLFESATREATAAFGRGECFVERYLDKARHVEAQVIAD
                     KHGNVVVAGTRDCSLQRRFQKLVEEAPAPFLTDDQRERLHSSAKAICKEAGYYGAGTVE
                     YLVGSDGLISFLEVNTRLQVEHPVTEETTGIDLVREMFRIAEGHELSIKEDPAPRGHAF
                     EFRINGEDAGSNFMPAPGKITSYREPQGPGVRMDSGVVEGSEISGQFDSMLAKLIVWGD
                     TREQALQRSRRALAEYVVEGMPTVIPFHQHIVENPAFVGNDEGFEIYTKWIEEVWDNPI
                     APYVDASELDEDEDKTPAQKVVVEINGRRVEVALPGDLALGGTAGPKKKAKKRRAGGAK
                     AGVSGDAVAAPMQGTVIKVNVEEGAEVNEGDTVVVLEAMKMENPVKAHKSGTVTGLTVA
                     AGEGVNKGVVLLEIK"
     gene            77..1852
                     /gene="accBC"
                     /label=accBC
     regulatory      1894..1922
                     /label=Tac promoter
                     /note="Tac promoter"
                     /regulatory_class="promoter"
     promoter        1894..1922
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    1930..1946
                     /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)."
     CDS             1969..3600
                     /codon_start=1
                     /gene="accD1"
                     /product="acetyl-CoA carboxylase subunit beta"
                     /label=accD1
                     /protein_id="AYD60197.1"
                     /translation="MTISSPLIDVANLPDINTTAGKIADLKARRAEAHFPMGEKAVEKV
                     HAAGRLTARERLDYLLDEGSFIETDQLARHRTTAFGLGAKRPATDGIVTGWGTIDGREV
                     CIFSQDGTVFGGALGEVYGEKMIKIMELAIDTGRPLIGLYEGAGARIQDGAVSLDFISQ
                     TFYQNIQASGVIPQISVIMGACAGGNAYGPALTDFVVMVDKTSKMFVTGPDVIKTVTGE
                     EITQEELGGATTHMVTAGNSHYTAATDEEALDWVQDLVSFLPSNNRSYTPLEDFDEEEG
                     GVEENITADDLKLDEIIPDSATVPYDVRDVIECLTDDGEYLEIQADRAENVVIAFGRIE
                     GQSVGFVANQPTQFAGCLDIDSSEKAARFVRTCDAFNIPIVMLVDVPGFLPGAGQEYGG
                     ILRRGAKLLYAYGEATVPKITVTMRKAYGGAYCVMGSKGLGSDINLAWPTAQIAVMGAA
                     GAVGFIYRKELMAADAKGLDTVALAKSFEREYEDHMLNPYHAAERGLIDAVILPSETRG
                     QISRNLRLLKHKNVTRPARKHGNMPL"
     gene            1969..3600
                     /gene="accD1"
                     /label=accD1
     terminator      3849..3935
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      4027..4054
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     protein_bind    complement(4075..4096)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(4210..4842)
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     promoter        complement(4843..4945)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             complement(5095..6114)
                     /codon_start=1
                     /gene="mob"
                     /product="Mob"
                     /function="required for plasmid mobilization"
                     /label=mob
                     /protein_id="AYD60199.1"
                     /translation="MAAYAIMRCKKLAKMGNVAASLKHAYRERETPNADASRTPENEHW
                     AASSTDEAMGRLRELLPEKRRKDAVLAVEYVMTASPEWWKSASQEQQAAFFEKAHKWLA
                     DKYGADRIVTASIHRDETSPHMTAFVVPLTQDGRLSAKEFIGNKAQMTRDQTTFAAAVA
                     DLGLQRGIEGSKARHTRIQAFYEALERPPVGHVTISPQAVEPRAYAPQGLAEKLGISKR
                     VETPEAVADRLTKAVRQGYEPALQAAAGAREMRKKADQAQETARDLRERLKPVLDALGP
                     LNRDMQAKAAAIIKAVGEKLLTEQREVQRQKQAQRQQERGRAHFPEKCHLTSKKPLLS"
     gene            complement(5095..6114)
                     /gene="mob"
                     /label=mob
     rep_origin      6338..7109
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             7110..7769
                     /label=pBBR1 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica"
ORIGIN
        1 gttgacaatt aatcatcggc tcgtataatg tgtggaattg tgagcggata acaatttcac
       61 acaggaaaca gaattcgtgt cagtcgagac taggaagatc accaaggttc ttgtcgctaa
      121 ccgtggtgag attgcaatcc gcgtgttccg tgcagctcga gatgaaggca tcggatctgt
      181 cgccgtctac gcagagccag atgcagatgc accattcgtg tcatatgcag acgaggcttt
      241 tgccctcggt ggccaaacat ccgctgagtc ctaccttgtc attgacaaga tcatcgatgc
      301 ggcccgcaag tccggcgccg acgccatcca ccccggctac ggcttcctcg cagaaaacgc
      361 tgacttcgca gaagcagtca tcaacgaagg cctgatctgg attggacctt cacctgagtc
      421 catccgctcc ctcggcgaca aggtcaccgc tcgccacatc gcagataccg ccaaggctcc
      481 aatggctcct ggcaccaagg aaccagtaaa agacgcagca gaagttgtgg ctttcgctga
      541 agaattcggt ctcccaatcg ccatcaaggc agctttcggt ggcggcggac gtggcatgaa
      601 ggttgcctac aagatggaag aagtcgctga cctcttcgag tccgcaaccc gtgaagcaac
      661 cgcagcgttc ggccgcggcg agtgcttcgt ggagcgctac ctggacaagg cacgccacgt
      721 tgaggctcag gtcatcgccg ataagcacgg caacgttgtt gtcgccggaa cccgtgactg
      781 ctccctgcag cgccgtttcc agaagctcgt cgaagaagca ccagcaccat tcctcaccga
      841 tgaccagcgc gagcgtctcc actcctccgc gaaggctatc tgtaaggaag ctggctacta
      901 cggtgcaggc accgttgagt acctcgttgg ctccgacggc ctgatctcct tcctcgaggt
      961 caacacccgc ctccaggtgg aacacccagt caccgaagag accaccggca tcgacctggt
     1021 ccgcgaaatg ttccgcatcg cagaaggcca cgagctctcc atcaaggaag atccagctcc
     1081 acgcggccac gcattcgagt tccgcatcaa cggcgaagac gctggctcca acttcatgcc
     1141 tgcaccaggc aagatcacca gctaccgcga gccacagggc ccaggcgtcc gcatggactc
     1201 cggtgtcgtt gaaggttccg aaatctccgg acagttcgac tccatgctgg caaagctgat
     1261 cgtttggggc gacacccgcg agcaggctct ccagcgctcc cgccgtgcac ttgcagagta
     1321 cgttgtcgag ggcatgccaa ccgttatccc attccaccag cacatcgtgg aaaacccagc
     1381 attcgtgggc aacgacgaag gcttcgagat ctacaccaag tggatcgaag aggtttggga
     1441 taacccaatc gcaccttacg ttgacgcttc cgagctcgac gaagatgagg acaagacccc
     1501 agcacagaag gttgttgtgg agatcaacgg ccgtcgcgtt gaggttgcac tcccaggcga
     1561 tctggcactc ggtggcaccg ctggtcctaa gaagaaggcc aagaagcgtc gcgcaggtgg
     1621 tgcaaaggct ggcgtatccg gcgatgcagt ggcagctcca atgcagggca ctgtcatcaa
     1681 ggtcaacgtc gaagaaggcg ctgaagtcaa cgaaggcgac accgttgttg tcctcgaggc
     1741 tatgaagatg gaaaaccctg tgaaggctca taagtccgga accgtaaccg gccttactgt
     1801 cgctgcaggc gagggtgtca acaagggcgt tgttctcctc gagatcaagt aatggtaccc
     1861 ggggatcctc tagagtcgac ctgcaggcat gcattgacaa ttaatcatcg gctcgtataa
     1921 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagaattcat gaccatttcc
     1981 tcacctttga ttgacgtcgc caaccttcca gacatcaaca ccactgccgg caagatcgcc
     2041 gaccttaagg ctcgccgcgc ggaagcccat ttccccatgg gtgaaaaggc agtagagaag
     2101 gtccacgctg ctggacgcct cactgcccgt gagcgcttgg attacttact cgatgagggc
     2161 tccttcatcg agaccgatca gctggctcgc caccgcacca ccgctttcgg cctgggcgct
     2221 aagcgtcctg caaccgacgg catcgtgacc ggctggggca ccattgatgg acgcgaagtc
     2281 tgcatcttct cgcaggacgg caccgtattc ggtggcgcgc ttggtgaggt gtacggcgaa
     2341 aagatgatca agatcatgga gctggcaatc gacaccggcc gcccattgat cggtctttac
     2401 gaaggcgctg gcgctcgcat tcaggacggc gctgtctccc tggacttcat ttcccagacc
     2461 ttctaccaaa acattcaggc ttctggcgtt atcccacaga tctccgtcat catgggcgca
     2521 tgtgcaggtg gcaacgctta cggcccagcc ctgaccgact tcgtggtcat ggtggacaag
     2581 acctccaaga tgttcgttac cggcccagac gtgatcaaga ccgtcaccgg cgaggaaatc
     2641 acccaggaag agcttggcgg agcaaccacc cacatggtga ccgctggcaa ctcccactac
     2701 accgctgcga ccgatgagga agcactggat tgggtacagg acctggtgtc cttcctccca
     2761 tccaacaatc gctcttacac accactggaa gacttcgacg aggaagaagg cggcgttgaa
     2821 gaaaacatca ccgctgacga tctgaagctc gacgagatca tcccagattc cgcgaccgtt
     2881 ccttacgacg tccgcgatgt catcgaatgc ctcaccgacg atggcgaata cctggaaatc
     2941 caggcagacc gcgcagaaaa cgttgttatt gcattcggcc gcatcgaagg ccagtccgtt
     3001 ggatttgttg ccaaccagcc aacccagttc gctggctgcc tggacatcga ctcctctgag
     3061 aaggcagctc gcttcgtccg cacctgcgac gcgtttaaca tcccaatcgt catgcttgtc
     3121 gacgtccccg gcttccttcc aggcgcaggc caggagtatg gtggcatcct gcgtcgtggc
     3181 gcaaagctgc tctacgcata cggcgaagca accgttccaa agattaccgt caccatgcgt
     3241 aaggcttacg gcggagcgta ctgcgtgatg ggttccaagg gcttgggctc tgacatcaac
     3301 cttgcatggc caaccgcaca gatcgccgtc atgggcgctg ctggcgcagt cggattcatc
     3361 taccgcaagg agctcatggc agctgatgcc aagggcctcg ataccgtagc tctggctaag
     3421 tccttcgagc gcgagtacga agaccacatg ctcaacccgt accacgctgc agaacgtggc
     3481 ctgatcgacg ccgtgatcct gccaagcgaa acccgcggac agatttcccg caaccttcgc
     3541 ctgctcaagc acaagaacgt cactcgccct gctcgcaagc acggcaacat gccactgtaa
     3601 tggtacccgg ggatcctcta gagtcgacct gcaggcatgc aagcttggct gttttggcgg
     3661 atgagagaag attttcagcc tgatacagat taaatcagaa cgcagaagcg gtctgataaa
     3721 acagaatttg cctggcggca gtagcgcggt ggtcccacct gaccccatgc cgaactcaga
     3781 agtgaaacgc cgtagcgccg atggtagtgt ggggtctccc catgcgagag tagggaactg
     3841 ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt
     3901 gtttgtcggt gaacgctctc ctgagtagga caaatccgtc gggagcggat ttgaacgttg
     3961 cgaagcaacg gcccggaggg tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa
     4021 ttaagcagaa ggccatcctg acggatggcc ttttactagt gcctggggtg cctaatgagt
     4081 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     4141 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     4201 catgcataaa aactgttgta attcattaag cattctgccg acatggaagc catcacaaac
     4261 ggcatgatga acctgaatcg ccagcggcat cagcaccttg tcgccttgcg tataatattt
     4321 gcccatggtg aaaacggggg cgaagaagtt gtccatattg gccacgttta aatcaaaact
     4381 ggtgaaactc acccagggat tggctgagac gaaaaacata ttctcaataa accctttagg
     4441 gaaataggcc aggttttcac cgtaacacgc cacatcttgc gaatatatgt gtagaaactg
     4501 ccggaaatcg tcgtggtatt cactccagag cgatgaaaac gtttcagttt gctcatggaa
     4561 aacggtgtaa caagggtgaa cactatccca tatcaccagc tcaccgtctt tcattgccat
     4621 acggaattcc ggatgagcat tcatcaggcg ggcaagaatg tgaataaagg ccggataaaa
     4681 cttgtgctta tttttcttta cggtctttaa aaaggccgta atatccagct gaacggtctg
     4741 gttataggta cattgagcaa ctgactgaaa tgcctcaaaa tgttctttac gatgccattg
     4801 ggatatatca acggtggtat atccagtgat ttttttctcc attttagctt ccttagctcc
     4861 tgaaaatctc gataactcaa aaaatacgcc cggtagtgat cttatttcat tatggtgaaa
     4921 gttggaacct cttacgtgcc gatcaacgtc tcattttcgc caaaagttgg cccagggctt
     4981 cccggtatca acagggacac caggatttat ttattctgcg aagtgatctt ccgtcacagg
     5041 tatttattcg aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     5101 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc gcccgcgttc
     5161 ctgctggcgc tgggcctgtt tctggcgctg gacttcccgc tgttccgtca gcagcttttc
     5221 gcccacggcc ttgatgatcg cggcggcctt ggcctgcata tcccgattca acggccccag
     5281 ggcgtccaga acgggcttca ggcgctcccg aaggtctcgg gccgtctctt gggcttgatc
     5341 ggccttcttg cgcatctcac gcgctcctgc ggcggcctgt agggcaggct catacccctg
     5401 ccgaaccgct tttgtcagcc ggtcggccac ggcttccggc gtctcaacgc gctttgagat
     5461 tcccagcttt tcggccaatc cctgcggtgc ataggcgcgt ggctcgaccg cttgcgggct
     5521 gatggtgacg tggcccactg gtggccgctc cagggcctcg tagaacgcct gaatgcgcgt
     5581 gtgacgtgcc ttgctgccct cgatgccccg ttgcagccct agatcggcca cagcggccgc
     5641 aaacgtggtc tggtcgcggg tcatctgcgc tttgttgccg atgaactcct tggccgacag
     5701 cctgccgtcc tgcgtcagcg gcaccacgaa cgcggtcatg tgcgggctgg tttcgtcacg
     5761 gtggatgctg gccgtcacga tgcgatccgc cccgtacttg tccgccagcc acttgtgcgc
     5821 cttctcgaag aacgccgcct gctgttcttg gctggccgac ttccaccatt ccgggctggc
     5881 cgtcatgacg tactcgaccg ccaacacagc gtccttgcgc cgcttctctg gcagcaactc
     5941 gcgcagtcgg cccatcgctt catcggtgct gctggccgcc cagtgctcgt tctctggcgt
     6001 cctgctggcg tcagcgttgg gcgtctcgcg ctcgcggtag gcgtgcttga gactggccgc
     6061 cacgttgccc attttcgcca gcttcttgca tcgcatgatc gcgtatgccg ccatgcctgc
     6121 ccctcccttt tggtgtccaa ccggctcgac gggggcagcg caaggcggtg cctccggcgg
     6181 gccactcaat gcttgagtat actcactaga ctttgcttcg caaagtcgtg accgcctacg
     6241 gcggctgcgg cgccctacgg gcttgctctc cgggcttcgc cctgcgcggt cgctgcgctc
     6301 ccttgccagc ccgtggatat gtggacgatg gccgcgagcg gccaccggct ggctcgcttc
     6361 gctcggcccg tggacaaccc tgctggacaa gctgatggac aggctgcgcc tgcccacgag
     6421 cttgaccaca gggattgccc accggctacc cagccttcga ccacataccc accggctcca
     6481 actgcgcggc ctgcggcctt gccccatcaa tttttttaat tttctctggg gaaaagcctc
     6541 cggcctgcgg cctgcgcgct tcgcttgccg gttggacacc aagtggaagg cgggtcaagg
     6601 ctcgcgcagc gaccgcgcag cggcttggcc ttgacgcgcc tggaacgacc caagcctatg
     6661 cgagtggggg cagtcgaagg gcgaagcccg cccgcctgcc ccccgagcct cacggcggcg
     6721 agtgcggggg ttccaagggg gcagcgccac cttgggcaag gccgaaggcc gcgcagtcga
     6781 tcaacaagcc ccggaggggc cactttttgc cggaggggga gccgcgccga aggcgtgggg
     6841 gaaccccgca ggggtgccct tctttgggca ccaaagaact agatataggg cgaaatgcga
     6901 aagacttaaa aatcaacaac ttaaaaaagg ggggtacgca acagctcatt gcggcacccc
     6961 ccgcaatagc tcattgcgta ggttaaagaa aatctgtaat tgactgccac ttttacgcaa
     7021 cgcataattg ttgtcgcgct gccgaaaagt tgcagctgat tgcgcatggt gccgcaaccg
     7081 tgcggcaccc ctaccgcatg gagataagca tggccacgca gtccagagaa atcggcattc
     7141 aagccaagaa caagcccggt cactgggtgc aaacggaacg caaagcgcat gaggcgtggg
     7201 ccgggcttat tgcgaggaaa cccacggcgg caatgctgct gcatcacctc gtggcgcaga
     7261 tgggccacca gaacgccgtg gtggtcagcc agaagacact ttccaagctc atcggacgtt
     7321 ctttgcggac ggtccaatac gcagtcaagg acttggtggc cgagcgctgg atctccgtcg
     7381 tgaagctcaa cggccccggc accgtgtcgg cctacgtggt caatgaccgc gtggcgtggg
     7441 gccagccccg cgaccagttg cgcctgtcgg tgttcagtgc cgccgtggtg gttgatcacg
     7501 acgaccagga cgaatcgctg ttggggcatg gcgacctgcg ccgcatcccg accctgtatc
     7561 cgggcgagca gcaactaccg accggccccg gcgaggagcc gcccagccag cccggcattc
     7621 cgggcatgga accagacctg ccagccttga ccgaaacgga ggaatgggaa cggcgcgggc
     7681 agcagcgcct gccgatgccc gatgagccgt gttttctgga cgatggcgag ccgttggagc
     7741 cgccgacacg ggtcacgctg ccgcgccggt agcacttggg ttgcgcagca acccgtaagt
     7801 gcgctgttcc agactatcgg ctgtagccgc ctcgccgccc tataccttgt ctgcctcccc
     7861 gcgttgcgtc gcggtgcatg gagccgggcc acctcgacct gaatggaagc cggcggcacc
     7921 tcgctaacgg attcaccgtt tttatcaggc tctgggaggc agaataaatg atcatatcgt
     7981 caattattac ctccacgggg agagcctgag caaactggcc tcaggcattt gagaagcaca
     8041 cggtcacact gcttccggta gtcaataaac cggtaaacca gcaatagaca taagcggcta
     8101 tttaacgacc ctgccctgaa ccgacgaccg ggtcgaattt gctttcgaat ttctgccatt
     8161 catccgctta ttatcactta ttcaggcgta gca
//
