LOCUS       40924_6077        4674 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pBBR1TaC, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4674)
  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 4674)
  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  JOURNAL   Unpublished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 4674)
  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 4674)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 4674)
  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            "Unpublished"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..4674
                     /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)."
     misc_feature    72..127
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     terminator      330..416
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      508..535
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     protein_bind    complement(556..577)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(691..1323)
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQF"
     promoter        complement(1324..1426)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             complement(1576..2595)
                     /codon_start=1
                     /gene="mob"
                     /product="Mob"
                     /function="required for plasmid mobilization"
                     /label=mob
                     /protein_id="AYD60182.1"
                     /translation="MAAYAIMRCKKLAKMGNVAASLKHAYRERETPNADASRTPENEHW
                     AASSTDEAMGRLRELLPEKRRKDAVLAVEYVMTASPEWWKSASQEQQAAFFEKAHKWLA
                     DKYGADRIVTASIHRDETSPHMTAFVVPLTQDGRLSAKEFIGNKAQMTRDQTTFAAAVA
                     DLGLQRGIEGSKARHTRIQAFYEALERPPVGHVTISPQAVEPRAYAPQGLAEKLGISKR
                     VETPEAVADRLTKAVRQGYEPALQAAAGAREMRKKADQAQETARDLRERLKPVLDALGP
                     LNRDMQAKAAAIIKAVGEKLLTEQREVQRQKQAQRQQERGRAHFPEKCHLTSKKPLLS"
     gene            complement(1576..2595)
                     /gene="mob"
                     /label=mob
     rep_origin      2819..3590
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             3591..4250
                     /codon_start=1
                     /label=pBBR1 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica"
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
ORIGIN
        1 gttgacaatt aatcatcggc tcgtataatg tgtggaattg tgagcggata acaatttcac
       61 acaggaaaca ggatccgagc tcggtacccg gggatcctct agagtcgacc tgcaggcatg
      121 caagcttggc tgttttggcg gatgagagaa gattttcagc ctgatacaga ttaaatcaga
      181 acgcagaagc ggtctgataa aacagaattt gcctggcggc agtagcgcgg tggtcccacc
      241 tgaccccatg ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg tggggtctcc
      301 ccatgcgaga gtagggaact gccaggcatc aaataaaacg aaaggctcag tcgaaagact
      361 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgt
      421 cgggagcgga tttgaacgtt gcgaagcaac ggcccggagg gtggcgggca ggacgcccgc
      481 cataaactgc caggcatcaa attaagcaga aggccatcct gacggatggc cttttactag
      541 tgcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
      601 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
      661 aggcggtttg cgtattgggc gcatgcataa aaactgttgt aattcattaa gcattctgcc
      721 gacatggaag ccatcacaaa cggcatgatg aacctgaatc gccagcggca tcagcacctt
      781 gtcgccttgc gtataatatt tgcccatggt gaaaacgggg gcgaagaagt tgtccatatt
      841 ggccacgttt aaatcaaaac tggtgaaact cacccaggga ttggctgaga cgaaaaacat
      901 attctcaata aaccctttag ggaaataggc caggttttca ccgtaacacg ccacatcttg
      961 cgaatatatg tgtagaaact gccggaaatc gtcgtggtat tcactccaga gcgatgaaaa
     1021 cgtttcagtt tgctcatgga aaacggtgta acaagggtga acactatccc atatcaccag
     1081 ctcaccgtct ttcattgcca tacggaattc cggatgagca ttcatcaggc gggcaagaat
     1141 gtgaataaag gccggataaa acttgtgctt atttttcttt acggtcttta aaaaggccgt
     1201 aatatccagc tgaacggtct ggttataggt acattgagca actgactgaa atgcctcaaa
     1261 atgttcttta cgatgccatt gggatatatc aacggtggta tatccagtga tttttttctc
     1321 cattttagct tccttagctc ctgaaaatct cgataactca aaaaatacgc ccggtagtga
     1381 tcttatttca ttatggtgaa agttggaacc tcttacgtgc cgatcaacgt ctcattttcg
     1441 ccaaaagttg gcccagggct tcccggtatc aacagggaca ccaggattta tttattctgc
     1501 gaagtgatct tccgtcacag gtatttattc gaagacgaaa gggcctcgtg atacgcctat
     1561 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     1621 gaaatgtgcg cgcccgcgtt cctgctggcg ctgggcctgt ttctggcgct ggacttcccg
     1681 ctgttccgtc agcagctttt cgcccacggc cttgatgatc gcggcggcct tggcctgcat
     1741 atcccgattc aacggcccca gggcgtccag aacgggcttc aggcgctccc gaaggtctcg
     1801 ggccgtctct tgggcttgat cggccttctt gcgcatctca cgcgctcctg cggcggcctg
     1861 tagggcaggc tcatacccct gccgaaccgc ttttgtcagc cggtcggcca cggcttccgg
     1921 cgtctcaacg cgctttgaga ttcccagctt ttcggccaat ccctgcggtg cataggcgcg
     1981 tggctcgacc gcttgcgggc tgatggtgac gtggcccact ggtggccgct ccagggcctc
     2041 gtagaacgcc tgaatgcgcg tgtgacgtgc cttgctgccc tcgatgcccc gttgcagccc
     2101 tagatcggcc acagcggccg caaacgtggt ctggtcgcgg gtcatctgcg ctttgttgcc
     2161 gatgaactcc ttggccgaca gcctgccgtc ctgcgtcagc ggcaccacga acgcggtcat
     2221 gtgcgggctg gtttcgtcac ggtggatgct ggccgtcacg atgcgatccg ccccgtactt
     2281 gtccgccagc cacttgtgcg ccttctcgaa gaacgccgcc tgctgttctt ggctggccga
     2341 cttccaccat tccgggctgg ccgtcatgac gtactcgacc gccaacacag cgtccttgcg
     2401 ccgcttctct ggcagcaact cgcgcagtcg gcccatcgct tcatcggtgc tgctggccgc
     2461 ccagtgctcg ttctctggcg tcctgctggc gtcagcgttg ggcgtctcgc gctcgcggta
     2521 ggcgtgcttg agactggccg ccacgttgcc cattttcgcc agcttcttgc atcgcatgat
     2581 cgcgtatgcc gccatgcctg cccctccctt ttggtgtcca accggctcga cgggggcagc
     2641 gcaaggcggt gcctccggcg ggccactcaa tgcttgagta tactcactag actttgcttc
     2701 gcaaagtcgt gaccgcctac ggcggctgcg gcgccctacg ggcttgctct ccgggcttcg
     2761 ccctgcgcgg tcgctgcgct cccttgccag cccgtggata tgtggacgat ggccgcgagc
     2821 ggccaccggc tggctcgctt cgctcggccc gtggacaacc ctgctggaca agctgatgga
     2881 caggctgcgc ctgcccacga gcttgaccac agggattgcc caccggctac ccagccttcg
     2941 accacatacc caccggctcc aactgcgcgg cctgcggcct tgccccatca atttttttaa
     3001 ttttctctgg ggaaaagcct ccggcctgcg gcctgcgcgc ttcgcttgcc ggttggacac
     3061 caagtggaag gcgggtcaag gctcgcgcag cgaccgcgca gcggcttggc cttgacgcgc
     3121 ctggaacgac ccaagcctat gcgagtgggg gcagtcgaag ggcgaagccc gcccgcctgc
     3181 cccccgagcc tcacggcggc gagtgcgggg gttccaaggg ggcagcgcca ccttgggcaa
     3241 ggccgaaggc cgcgcagtcg atcaacaagc cccggagggg ccactttttg ccggaggggg
     3301 agccgcgccg aaggcgtggg ggaaccccgc aggggtgccc ttctttgggc accaaagaac
     3361 tagatatagg gcgaaatgcg aaagacttaa aaatcaacaa cttaaaaaag gggggtacgc
     3421 aacagctcat tgcggcaccc cccgcaatag ctcattgcgt aggttaaaga aaatctgtaa
     3481 ttgactgcca cttttacgca acgcataatt gttgtcgcgc tgccgaaaag ttgcagctga
     3541 ttgcgcatgg tgccgcaacc gtgcggcacc cctaccgcat ggagataagc atggccacgc
     3601 agtccagaga aatcggcatt caagccaaga acaagcccgg tcactgggtg caaacggaac
     3661 gcaaagcgca tgaggcgtgg gccgggctta ttgcgaggaa acccacggcg gcaatgctgc
     3721 tgcatcacct cgtggcgcag atgggccacc agaacgccgt ggtggtcagc cagaagacac
     3781 tttccaagct catcggacgt tctttgcgga cggtccaata cgcagtcaag gacttggtgg
     3841 ccgagcgctg gatctccgtc gtgaagctca acggccccgg caccgtgtcg gcctacgtgg
     3901 tcaatgaccg cgtggcgtgg ggccagcccc gcgaccagtt gcgcctgtcg gtgttcagtg
     3961 ccgccgtggt ggttgatcac gacgaccagg acgaatcgct gttggggcat ggcgacctgc
     4021 gccgcatccc gaccctgtat ccgggcgagc agcaactacc gaccggcccc ggcgaggagc
     4081 cgcccagcca gcccggcatt ccgggcatgg aaccagacct gccagccttg accgaaacgg
     4141 aggaatggga acggcgcggg cagcagcgcc tgccgatgcc cgatgagccg tgttttctgg
     4201 acgatggcga gccgttggag ccgccgacac gggtcacgct gccgcgccgg tagcacttgg
     4261 gttgcgcagc aacccgtaag tgcgctgttc cagactatcg gctgtagccg cctcgccgcc
     4321 ctataccttg tctgcctccc cgcgttgcgt cgcggtgcat ggagccgggc cacctcgacc
     4381 tgaatggaag ccggcggcac ctcgctaacg gattcaccgt ttttatcagg ctctgggagg
     4441 cagaataaat gatcatatcg tcaattatta cctccacggg gagagcctga gcaaactggc
     4501 ctcaggcatt tgagaagcac acggtcacac tgcttccggt agtcaataaa ccggtaaacc
     4561 agcaatagac ataagcggct atttaacgac cctgccctga accgacgacc gggtcgaatt
     4621 tgctttcgaa tttctgccat tcatccgctt attatcactt attcaggcgt agca
//
