LOCUS       40924_43868        5800 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pTrc-fumA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5800)
  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 5800)
  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 5800)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5800)
  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..5800
                     /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..1910
                     /gene="fumA"
                     /label=fumA
                     /note="Fumarate hydratase class I, aerobic from Escherichia
                     coli (strain K12). Accession#: P0AC33"
     terminator      2153..2239
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      2331..2358
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        2378..2469
                     /label=AmpR promoter
     CDS             2470..3327
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      3501..4089
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(4275..4415)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     promoter        4601..4678
                     /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             4679..5758
                     /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 cagaccatgt caaacaaacc ctttcattat caggctcctt
      301 ttccactcaa aaaagatgat actgagtatt acctgctaac cagcgaacac gttagcgtat
      361 ctgaatttga agggcaggag attttgaaag tcgcacccga agcgttaact ctattggcgc
      421 gccaggcgtt tcatgatgcg tcgttcatgc tgcgtccggc gcaccaacaa caggtggccg
      481 acattctgcg tgacccggag gccagcgaaa atgataaata tgtggcgctg caattcctgc
      541 gtaactccga catcgcggcg aaaggcgttc tgccaacctg tcaggatacc ggcaccgcga
      601 ttattgttgg taaaaaaggg cagcgtgtat ggaccggtgg tggtgatgaa gcggcgctgg
      661 cgcgcggtgt ctataacact tatatcgaag ataatctgcg ctactcgcaa aacgcgccgc
      721 tggatatgta taaagaagtg aataccggca ccaatctgcc agcgcagatc gatctttatg
      781 ccgttgatgg cgacgaatac aaattcctct gtatcgccaa aggtggtggt tcggcaaaca
      841 agacgtatct ctatcaggaa accaaagcgt tactgacgcc ggggaaactg aaaaattacc
      901 tggttgagaa gatgcgcacg ctgggtacgg cggcctgtcc tccgtatcat attgcgttcg
      961 ttattggtgg aacttctgca gaaacgaacc ttaaaacggt gaaactggct tccgcgaaat
     1021 actatgatga actgccaacg gaagggaatg agcacggtca ggcgttccgc gatgtggaac
     1081 tggaaaaaga attgctgatc gaagcgcaaa atcttggtct gggtgcgcag tttggtggta
     1141 aatacttcgc tcacgacatc cgcgtgattc gcctgccacg tcacggcgca tcctgcccgg
     1201 tcggtatggg cgtctcctgc tctgctgacc gtaatatcaa agcgaagatc aaccgtcagg
     1261 ggatctggat cgaaaaactg gaacataatc caggcaaata tatcccggaa gagctgcgca
     1321 aagcgggaga aggcgaagcg gtgcgcgttg accttaaccg tccgatgaaa gagatcctcg
     1381 cacagttgtc gcagtatccc gtttctacac gcttatcgct taacggcacg attatcgtcg
     1441 gtcgtgatat tgctcacgcc aaactgaaag agcggatgga taacggtgaa gggctgccgc
     1501 agtacatcaa agatcatccg atttactacg cgggtccggc caaaacgccg gaaggttatg
     1561 cctccggttc tcttggccca acgaccgccg gacggatgga ttcttatgtc gatcaactgc
     1621 aagcgcaggg cggaagtatg atcatgctgg cgaaaggcaa ccgcagccag caggtgacgg
     1681 atgcctgtaa aaaacacggc ggcttctacc ttggcagtat cggtggtccg gccgctgtat
     1741 tggcgcaggg aagtattaag agcctggaat gtgttgaata tccggaactg ggaatggaag
     1801 ccatctggaa aattgaagtg gaagatttcc cggcgtttat ccttgtggat gataaaggaa
     1861 atgacttctt ccagcagata caactcacac aatgcacccg ctgtgtgaaa taatggatcc
     1921 tctagagtcg acctgcaggc atgcaagctt ggctgttttg gcggatgaga gaagattttc
     1981 agcctgatac agattaaatc agaacgcaga agcggtctga taaaacagaa tttgcctggc
     2041 ggcagtagcg cggtggtccc acctgacccc atgccgaact cagaagtgaa acgccgtagc
     2101 gccgatggta gtgtggggtc tccccatgcg agagtaggga actgccaggc atcaaataaa
     2161 acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc
     2221 tctcctgagt aggacaaatc cgccgggagc ggatttgaac gttgcgaagc aacggcccgg
     2281 agggtggcgg gcaggacgcc cgccataaac tgccaggcat caaattaagc agaaggccat
     2341 cctgacggat ggcctttttg cgtttctaca aactcttttt gtttattttt ctaaatacat
     2401 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     2461 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     2521 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     2581 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     2641 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     2701 gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     2761 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     2821 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     2881 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     2941 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     3001 accacgatgc ctacagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     3061 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     3121 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     3181 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     3241 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     3301 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     3361 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     3421 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     3481 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     3541 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     3601 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     3661 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     3721 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     3781 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     3841 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     3901 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     3961 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     4021 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     4081 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     4141 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     4201 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     4261 gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     4321 cgcatatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca
     4381 ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg
     4441 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     4501 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagcaga
     4561 tcaattcgcg cgcgaaggcg aagcggcatg catttacgtt gacaccatcg aatggtgcaa
     4621 aacctttcgc ggtatggcat gatagcgccc ggaagagagt caattcaggg tggtgaatgt
     4681 gaaaccagta acgttatacg atgtcgcaga gtatgccggt gtctcttatc agaccgtttc
     4741 ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg cgggaaaaag tggaagcggc
     4801 gatggcggag ctgaattaca ttcccaaccg cgtggcacaa caactggcgg gcaaacagtc
     4861 gttgctgatt ggcgttgcca cctccagtct ggccctgcac gcgccgtcgc aaattgtcgc
     4921 ggcgattaaa tctcgcgccg atcaactggg tgccagcgtg gtggtgtcga tggtagaacg
     4981 aagcggcgtc gaagcctgta aagcggcggt gcacaatctt ctcgcgcaac gcgtcagtgg
     5041 gctgatcatt aactatccgc tggatgacca ggatgccatt gctgtggaag ctgcctgcac
     5101 taatgttccg gcgttatttc ttgatgtctc tgaccagaca cccatcaaca gtattatttt
     5161 ctcccatgaa gacggtacgc gactgggcgt ggagcatctg gtcgcattgg gtcaccagca
     5221 aatcgcgctg ttagcgggcc cattaagttc tgtctcggcg cgtctgcgtc tggctggctg
     5281 gcataaatat ctcactcgca atcaaattca gccgatagcg gaacgggaag gcgactggag
     5341 tgccatgtcc ggttttcaac aaaccatgca aatgctgaat gagggcatcg ttcccactgc
     5401 gatgctggtt gccaacgatc agatggcgct gggcgcaatg cgcgccatta ccgagtccgg
     5461 gctgcgcgtt ggtgcggata tctcggtagt gggatacgac gataccgaag acagctcatg
     5521 ttatatcccg ccgtcaacca ccatcaaaca ggattttcgc ctgctggggc aaaccagcgt
     5581 ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag ggcaatcagc tgttgcccgt
     5641 ctcactggtg aaaagaaaaa ccaccctggc gcccaatacg caaaccgcct ctccccgcgc
     5701 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     5761 agcgcaacgc aattaatgtg agttagcgcg aattgatctg
//
