LOCUS       40924_43893        5092 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pTrc-mdh, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5092)
  AUTHORS   Kim B, Binkley R, Kim HU, Lee SY.
  TITLE     Metabolic engineering of Escherichia coli for the enhanced 
            production of l-tyrosine
  JOURNAL   Biotechnol. Bioeng. (2018) In press
  PUBMED    30019750
REFERENCE   2  (bases 1 to 5092)
  AUTHORS   Yang D, Kim WJ, Yoo SM, Choi JH, Ha SH, Lee MH, Lee SY.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-JUN-2018) Dept. Chemical and Biomolecular Engineering,
            Korea Advanced Institute of Science and Technology, Daehak-ro 291, 
            Daejeon 34141, South Korea
REFERENCE   3  (bases 1 to 5092)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5092)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Biotechnol.
            Bioeng. (2018) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (15-JUN-2018) Dept. Chemical and Biomolecular Engineering, Korea 
            Advanced Institute of Science and Technology, Daehak-ro 291, Daejeon
            34141, South Korea"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5092
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        193..222
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    230..246
                     /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             267..1202
                     /gene="mdh"
                     /label=mdh
                     /note="Malate dehydrogenase from Escherichia coli O6:H1
                     (strain CFT073 / ATCC 700928 / UPEC). Accession#: P61890"
     terminator      1445..1531
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      1623..1650
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        1670..1761
                     /label=AmpR promoter
     CDS             1762..2619
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      2793..3381
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(3567..3707)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     promoter        3893..3970
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             3971..5050
                     /label=lacI
                     /note="lac repressor"
ORIGIN
        1 gtttgacagc ttatcatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc
       61 ggaagctgtg gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc
      121 gcactcccgt tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc
      181 tgaaatgagc tgttgacaat taatcatccg gctcgtataa tgtgtggaat tgtgagcgga
      241 taacaatttc acacaggaaa cagaccatga aagtcgcagt cctcggcgct gctggcggta
      301 ttggccaggc gcttgcacta ctgttaaaaa cccaactgcc ttcaggttca gaactctctc
      361 tgtatgatat cgctccagtg actcccggtg tggctgtcga tctgagccat atccctactg
      421 ctgtgaaaat caaaggtttt tctggtgaag atgcgactcc ggcgctggaa ggcgcagatg
      481 tcgttcttat ctctgcaggt gtagcgcgta aaccgggtat ggatcgttcc gacctgttta
      541 acgttaacgc cggcatcgtg aaaaacctgg tacagcaagt tgcgaaaacc tgcccgaaag
      601 cgtgcattgg tattatcact aacccggtta acaccacagt tgcgattgct gctgaagtgc
      661 tgaaaaaagc cggtgtttat gacaaaaaca aactgttcgg cgttaccacg ctggatatca
      721 ttcgttccaa cacctttgtt gcggaactga aaggcaaaca gccaggcgaa gttgaagtgc
      781 cggttattgg cggtcactct ggtgttacca ttctgccgct gctgtcacag gttcctggcg
      841 ttagttttac cgagcaggaa gtggctgatc tgaccaaacg tatccagaac gcgggtactg
      901 aggtggttga agcgaaagcc ggtggcgggt ctgcaaccct gtctatgggc caggcagctg
      961 cacgttttgg tctgtctctg gttcgtgcac tgcagggcga acaaggcgtt gtcgaatgtg
     1021 cctacgttga aggcgacggt cagtacgccc gtttcttctc tcaaccgctg ctgctgggta
     1081 aaaacggcgt ggaagagcgt aaatctatcg gtaccctgag cgcatttgaa cagaacgcgc
     1141 tggaaggtat gctggatacg ctcaagaaag atatcgccct gggcgaagag ttcgttaata
     1201 agtaatggat cctctagagt cgacctgcag gcatgcaagc ttggctgttt tggcggatga
     1261 gagaagattt tcagcctgat acagattaaa tcagaacgca gaagcggtct gataaaacag
     1321 aatttgcctg gcggcagtag cgcggtggtc ccacctgacc ccatgccgaa ctcagaagtg
     1381 aaacgccgta gcgccgatgg tagtgtgggg tctccccatg cgagagtagg gaactgccag
     1441 gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt
     1501 gtcggtgaac gctctcctga gtaggacaaa tccgccggga gcggatttga acgttgcgaa
     1561 gcaacggccc ggagggtggc gggcaggacg cccgccataa actgccaggc atcaaattaa
     1621 gcagaaggcc atcctgacgg atggcctttt tgcgtttcta caaactcttt ttgtttattt
     1681 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     1741 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     1801 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     1861 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     1921 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     1981 ctatgtggcg cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata
     2041 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     2101 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     2161 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     2221 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     2281 gacgagcgtg acaccacgat gcctacagca atggcaacaa cgttgcgcaa actattaact
     2341 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     2401 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     2461 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     2521 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     2581 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     2641 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     2701 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     2761 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     2821 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     2881 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     2941 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     3001 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     3061 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     3121 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     3181 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     3241 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     3301 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     3361 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     3421 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     3481 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     3541 tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc
     3601 ggtatttcac accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     3661 agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc
     3721 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     3781 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     3841 cgaggcagca gatcaattcg cgcgcgaagg cgaagcggca tgcatttacg ttgacaccat
     3901 cgaatggtgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga gtcaattcag
     3961 ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg gtgtctctta
     4021 tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa
     4081 agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac aacaactggc
     4141 gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc acgcgccgtc
     4201 gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc
     4261 gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca
     4321 acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca ttgctgtgga
     4381 agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga cacccatcaa
     4441 cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc tggtcgcatt
     4501 gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg
     4561 tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag cggaacggga
     4621 aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga atgagggcat
     4681 cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat
     4741 taccgagtcc gggctgcgcg ttggtgcgga tatctcggta gtgggatacg acgataccga
     4801 agacagctca tgttatatcc cgccgtcaac caccatcaaa caggattttc gcctgctggg
     4861 gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga agggcaatca
     4921 gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc
     4981 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     5041 aagcgggcag tgagcgcaac gcaattaatg tgagttagcg cgaattgatc tg
//
