LOCUS       V009262                 5619 bp    DNA     circular SYN 17-DEC-2018
DEFINITION  Exported.
ACCESSION   V009262
VERSION     V009262
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 5619)
  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 5619)
  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  JOURNAL   Unpublished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 5619)
  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 5619)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 5619)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc. Natl.
            Acad. Sci. U.S.A. (2018) In press"
            SGRef: number: 2; type: "Journal Article"; journalName:
            "Unpublished"
            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"
            SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5619
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..31
                     /label="trc promoter"
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    39..55
                     /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             76..1011
                     /gene="mdh"
                     /label="Malate dehydrogenase"
                     /note="Malate dehydrogenase from Escherichia coli O6:H1
                     (strain CFT073 / ATCC 700928 / UPEC). Accession#: P61890"
     misc_feature    1017..1072
                     /label="MCS"
                     /note="pUC18/19 multiple cloning site"
     terminator      1275..1361
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      1453..1480
                     /label="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     protein_bind    complement(1501..1522)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(1636..2268)
                     /label="CmR"
                     /note="chloramphenicol acetyltransferase"
     promoter        complement(2269..2371)
                     /label="cat promoter"
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             complement(2521..3540)
                     /codon_start=1
                     /gene="mob"
                     /product="Mob"
                     /function="required for plasmid mobilization"
                     /label="mob"
                     /protein_id="AYD60189.1"
                     /translation="MAAYAIMRCKKLAKMGNVAASLKHAYRERETPNADASRTPENEHW
                     AASSTDEAMGRLRELLPEKRRKDAVLAVEYVMTASPEWWKSASQEQQAAFFEKAHKWLA
                     DKYGADRIVTASIHRDETSPHMTAFVVPLTQDGRLSAKEFIGNKAQMTRDQTTFAAAVA
                     DLGLQRGIEGSKARHTRIQAFYEALERPPVGHVTISPQAVEPRAYAPQGLAEKLGISKR
                     VETPEAVADRLTKAVRQGYEPALQAAAGAREMRKKADQAQETARDLRERLKPVLDALGP
                     LNRDMQAKAAAIIKAVGEKLLTEQREVQRQKQAQRQQERGRAHFPEKCHLTSKKPLLS"
     gene            complement(2521..3540)
                     /gene="mob"
                     /label="mob"
     rep_origin      3764..4535
                     /label="pBBR1 oriV"
                     /note="replication origin of the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1
                     Rep protein for replication"
     CDS             4536..5195
                     /label="pBBR1 Rep"
                     /note="replication protein for the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica"
ORIGIN
        1 gttgacaatt aatcatccgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca
       61 cacaggaaac agaccatgaa agtcgcagtc ctcggcgctg ctggcggtat tggccaggcg
      121 cttgcactac tgttaaaaac ccaactgcct tcaggttcag aactctctct gtatgatatc
      181 gctccagtga ctcccggtgt ggctgtcgat ctgagccata tccctactgc tgtgaaaatc
      241 aaaggttttt ctggtgaaga tgcgactccg gcgctggaag gcgcagatgt cgttcttatc
      301 tctgcaggtg tagcgcgtaa accgggtatg gatcgttccg acctgtttaa cgttaacgcc
      361 ggcatcgtga aaaacctggt acagcaagtt gcgaaaacct gcccgaaagc gtgcattggt
      421 attatcacta acccggttaa caccacagtt gcgattgctg ctgaagtgct gaaaaaagcc
      481 ggtgtttatg acaaaaacaa actgttcggc gttaccacgc tggatatcat tcgttccaac
      541 acctttgttg cggaactgaa aggcaaacag ccaggcgaag ttgaagtgcc ggttattggc
      601 ggtcactctg gtgttaccat tctgccgctg ctgtcacagg ttcctggcgt tagttttacc
      661 gagcaggaag tggctgatct gaccaaacgt atccagaacg cgggtactga ggtggttgaa
      721 gcgaaagccg gtggcgggtc tgcaaccctg tctatgggcc aggcagctgc acgttttggt
      781 ctgtctctgg ttcgtgcact gcagggcgaa caaggcgttg tcgaatgtgc ctacgttgaa
      841 ggcgacggtc agtacgcccg tttcttctct caaccgctgc tgctgggtaa aaacggcgtg
      901 gaagagcgta aatctatcgg taccctgagc gcatttgaac agaacgcgct ggaaggtatg
      961 ctggatacgc tcaagaaaga tatcgccctg ggcgaagagt tcgttaataa gtaatggatc
     1021 cgagctcggt acccggggat cctctagagt cgacctgcag gcatgcaagc ttggctgttt
     1081 tggcggatga gagaagattt tcagcctgat acagattaaa tcagaacgca gaagcggtct
     1141 gataaaacag aatttgcctg gcggcagtag cgcggtggtc ccacctgacc ccatgccgaa
     1201 ctcagaagtg aaacgccgta gcgccgatgg tagtgtgggg tctccccatg cgagagtagg
     1261 gaactgccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta
     1321 tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgtcggga gcggatttga
     1381 acgttgcgaa gcaacggccc ggagggtggc gggcaggacg cccgccataa actgccaggc
     1441 atcaaattaa gcagaaggcc atcctgacgg atggcctttt actagtgcct ggggtgccta
     1501 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     1561 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     1621 tgggcgcatg cataaaaact gttgtaattc attaagcatt ctgccgacat ggaagccatc
     1681 acaaacggca tgatgaacct gaatcgccag cggcatcagc accttgtcgc cttgcgtata
     1741 atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc atattggcca cgtttaaatc
     1801 aaaactggtg aaactcaccc agggattggc tgagacgaaa aacatattct caataaaccc
     1861 tttagggaaa taggccaggt tttcaccgta acacgccaca tcttgcgaat atatgtgtag
     1921 aaactgccgg aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt cagtttgctc
     1981 atggaaaacg gtgtaacaag ggtgaacact atcccatatc accagctcac cgtctttcat
     2041 tgccatacgg aattccggat gagcattcat caggcgggca agaatgtgaa taaaggccgg
     2101 ataaaacttg tgcttatttt tctttacggt ctttaaaaag gccgtaatat ccagctgaac
     2161 ggtctggtta taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg
     2221 ccattgggat atatcaacgg tggtatatcc agtgattttt ttctccattt tagcttcctt
     2281 agctcctgaa aatctcgata actcaaaaaa tacgcccggt agtgatctta tttcattatg
     2341 gtgaaagttg gaacctctta cgtgccgatc aacgtctcat tttcgccaaa agttggccca
     2401 gggcttcccg gtatcaacag ggacaccagg atttatttat tctgcgaagt gatcttccgt
     2461 cacaggtatt tattcgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     2521 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcgccc
     2581 gcgttcctgc tggcgctggg cctgtttctg gcgctggact tcccgctgtt ccgtcagcag
     2641 cttttcgccc acggccttga tgatcgcggc ggccttggcc tgcatatccc gattcaacgg
     2701 ccccagggcg tccagaacgg gcttcaggcg ctcccgaagg tctcgggccg tctcttgggc
     2761 ttgatcggcc ttcttgcgca tctcacgcgc tcctgcggcg gcctgtaggg caggctcata
     2821 cccctgccga accgcttttg tcagccggtc ggccacggct tccggcgtct caacgcgctt
     2881 tgagattccc agcttttcgg ccaatccctg cggtgcatag gcgcgtggct cgaccgcttg
     2941 cgggctgatg gtgacgtggc ccactggtgg ccgctccagg gcctcgtaga acgcctgaat
     3001 gcgcgtgtga cgtgccttgc tgccctcgat gccccgttgc agccctagat cggccacagc
     3061 ggccgcaaac gtggtctggt cgcgggtcat ctgcgctttg ttgccgatga actccttggc
     3121 cgacagcctg ccgtcctgcg tcagcggcac cacgaacgcg gtcatgtgcg ggctggtttc
     3181 gtcacggtgg atgctggccg tcacgatgcg atccgccccg tacttgtccg ccagccactt
     3241 gtgcgccttc tcgaagaacg ccgcctgctg ttcttggctg gccgacttcc accattccgg
     3301 gctggccgtc atgacgtact cgaccgccaa cacagcgtcc ttgcgccgct tctctggcag
     3361 caactcgcgc agtcggccca tcgcttcatc ggtgctgctg gccgcccagt gctcgttctc
     3421 tggcgtcctg ctggcgtcag cgttgggcgt ctcgcgctcg cggtaggcgt gcttgagact
     3481 ggccgccacg ttgcccattt tcgccagctt cttgcatcgc atgatcgcgt atgccgccat
     3541 gcctgcccct cccttttggt gtccaaccgg ctcgacgggg gcagcgcaag gcggtgcctc
     3601 cggcgggcca ctcaatgctt gagtatactc actagacttt gcttcgcaaa gtcgtgaccg
     3661 cctacggcgg ctgcggcgcc ctacgggctt gctctccggg cttcgccctg cgcggtcgct
     3721 gcgctccctt gccagcccgt ggatatgtgg acgatggccg cgagcggcca ccggctggct
     3781 cgcttcgctc ggcccgtgga caaccctgct ggacaagctg atggacaggc tgcgcctgcc
     3841 cacgagcttg accacaggga ttgcccaccg gctacccagc cttcgaccac atacccaccg
     3901 gctccaactg cgcggcctgc ggccttgccc catcaatttt tttaattttc tctggggaaa
     3961 agcctccggc ctgcggcctg cgcgcttcgc ttgccggttg gacaccaagt ggaaggcggg
     4021 tcaaggctcg cgcagcgacc gcgcagcggc ttggccttga cgcgcctgga acgacccaag
     4081 cctatgcgag tgggggcagt cgaagggcga agcccgcccg cctgcccccc gagcctcacg
     4141 gcggcgagtg cgggggttcc aagggggcag cgccaccttg ggcaaggccg aaggccgcgc
     4201 agtcgatcaa caagccccgg aggggccact ttttgccgga gggggagccg cgccgaaggc
     4261 gtgggggaac cccgcagggg tgcccttctt tgggcaccaa agaactagat atagggcgaa
     4321 atgcgaaaga cttaaaaatc aacaacttaa aaaagggggg tacgcaacag ctcattgcgg
     4381 caccccccgc aatagctcat tgcgtaggtt aaagaaaatc tgtaattgac tgccactttt
     4441 acgcaacgca taattgttgt cgcgctgccg aaaagttgca gctgattgcg catggtgccg
     4501 caaccgtgcg gcacccctac cgcatggaga taagcatggc cacgcagtcc agagaaatcg
     4561 gcattcaagc caagaacaag cccggtcact gggtgcaaac ggaacgcaaa gcgcatgagg
     4621 cgtgggccgg gcttattgcg aggaaaccca cggcggcaat gctgctgcat cacctcgtgg
     4681 cgcagatggg ccaccagaac gccgtggtgg tcagccagaa gacactttcc aagctcatcg
     4741 gacgttcttt gcggacggtc caatacgcag tcaaggactt ggtggccgag cgctggatct
     4801 ccgtcgtgaa gctcaacggc cccggcaccg tgtcggccta cgtggtcaat gaccgcgtgg
     4861 cgtggggcca gccccgcgac cagttgcgcc tgtcggtgtt cagtgccgcc gtggtggttg
     4921 atcacgacga ccaggacgaa tcgctgttgg ggcatggcga cctgcgccgc atcccgaccc
     4981 tgtatccggg cgagcagcaa ctaccgaccg gccccggcga ggagccgccc agccagcccg
     5041 gcattccggg catggaacca gacctgccag ccttgaccga aacggaggaa tgggaacggc
     5101 gcgggcagca gcgcctgccg atgcccgatg agccgtgttt tctggacgat ggcgagccgt
     5161 tggagccgcc gacacgggtc acgctgccgc gccggtagca cttgggttgc gcagcaaccc
     5221 gtaagtgcgc tgttccagac tatcggctgt agccgcctcg ccgccctata ccttgtctgc
     5281 ctccccgcgt tgcgtcgcgg tgcatggagc cgggccacct cgacctgaat ggaagccggc
     5341 ggcacctcgc taacggattc accgttttta tcaggctctg ggaggcagaa taaatgatca
     5401 tatcgtcaat tattacctcc acggggagag cctgagcaaa ctggcctcag gcatttgaga
     5461 agcacacggt cacactgctt ccggtagtca ataaaccggt aaaccagcaa tagacataag
     5521 cggctattta acgaccctgc cctgaaccga cgaccgggtc gaatttgctt tcgaatttct
     5581 gccattcatc cgcttattat cacttattca ggcgtagca
//
