LOCUS       40924_43863        7827 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pTrc-At4CL1m-AtCHI-PhCHS, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7827)
  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 7827)
  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 7827)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7827)
  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..7827
                     /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..1949
                     /gene="4CL1"
                     /label=4CL1
                     /note="4-coumarate--CoA ligase 1 from Arabidopsis thaliana.
                     Accession#: Q42524"
     CDS             1978..2715
                     /gene="CHI1"
                     /label=CHI1
                     /note="Chalcone--flavanone isomerase 1 from Arabidopsis 
                     thaliana. Accession#: P41088"
     CDS             2778..3944
                     /gene="CHSA"
                     /label=CHSA
                     /note="Chalcone synthase A from Petunia hybrida.
                     Accession#: P08894"
     terminator      4180..4266
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      4358..4385
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        4405..4496
                     /label=AmpR promoter
     CDS             4497..5354
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      5528..6116
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(6302..6442)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     promoter        6628..6705
                     /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             6706..7785
                     /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 cagaccatgg cgccacaaga acaagcagtt tctcaggtga
      301 tggagaaaca gagcaacaac aacaacagtg acgtcatttt ccgatcaaag ttaccggata
      361 tttacatccc gaaccaccta tctctccacg actacatctt ccaaaacatc tccgaattcg
      421 ccactaagcc ttgcctaatc aacggaccaa ccggccacgt gtacacttac tccgacgtcc
      481 acgtcatctc ccgccaaatc gccgccaatt ttcacaaact cggcgttaac caaaacgacg
      541 tcgtcatgct cctcctccca aactgtcccg aattcgtcct ctctttcctc gccgcctcct
      601 tccgcggcgc aaccgccacc gccgcaaacc ctttcttcac tccggcggag atagctaaac
      661 aagccaaagc ctccaacacc aaactcataa tcaccgaagc tcgttacgtc gacaaaatca
      721 aaccacttca aaacgacgac ggagtagtca tcgtctgcat cgacgacaac gaatccgtgc
      781 caatccctga aggctgcctc cgcttcaccg agttgactca gtcgacaacc gaggcatcag
      841 aagtcatcga ctcggtggag atttcaccgg acgacgtggt ggcactacct tactcctctg
      901 gcacgacggg attaccaaaa ggagtgatgc tgactcacaa gggactagtc acgagcgttg
      961 ctcagcaagt cgacggcgag aacccgaatc tttatttcca cagcgatgac gtcctactct
     1021 gtgttttgcc catgtttcat atctacgctt tgaactcgat catgttgtgt ggtcttagag
     1081 ttggtgcggc gattctgata atgccgaagt ttgagatcaa tctgctattg gagctgatcc
     1141 agaggtgtaa agtgacggtg gctccgatgg ttccgccgat tgtgttggcc attgcgaagt
     1201 cttcggagac ggagaagtat gatttgagct cgataagagt ggtgaaatct ggtgctgctc
     1261 ctcttggtaa agaacttgaa gatgccgtta atgccaagtt tcctaatgcc aaactcggtc
     1321 agggatacgg aatgacggaa gcaggtccag tgctagcaat gtcgttaggt tttgcaaagg
     1381 aaccttttcc ggttaagtca ggagcttgtg gtactgttgt aagaaatgct gagatgaaaa
     1441 tagttgatcc agacaccgga gattctcttt cgaggaaaca acccggtgag atttgtattc
     1501 gtggtcacca gatcatgaaa ggttacctca acaatccggc agctacagca gagaccattg
     1561 ataaagacgg ttggcttcat actggagata ttggattgat cgatgacgat gacgagcttt
     1621 tcatcgttga tcgattgaaa gaacttgtca agtataaagg ttttcaggta gctccggctg
     1681 agctagaggc tttgctcatc ggtcatcctg acattactga tgttgctgtt gtagcaatga
     1741 aagaagaagc agctggtgaa gttcctgttg catttgtggt gaaatcgaag gattcggagt
     1801 tatcagaaga tgatgtgaag caattcgtgt cgaaacaggt tgtgttttac aagagaatca
     1861 acaaagtgtt cttcactgaa tccattccta aagctccatc agggaagata ttgaggaaag
     1921 atctgagggc aaaactagca aatggattgt aattaacttt aagaaggaga tatacatatg
     1981 tcttcatcca acgcctgcgc ctctccgtca ccgttccccg ccgtcacgaa gcttcatgta
     2041 gactccgtca cgtttgtacc gtccgtcaag tcaccggcct cctccaatcc attattcctc
     2101 ggcggcgccg gtgtccgagg ccttgatatc caaggtaaat tcgtgatctt caccgtcatt
     2161 ggagtatacc tagagggtaa cgccgttcct tctctatctg tcaagtggaa gggaaaaact
     2221 acggaggagc taacagaatc tatcccgttc ttccgtgaaa tagtcaccgg tgcgtttgag
     2281 aagtttatca aggtgacaat gaaactgccg ttaacgggac aacaatattc ggagaaagtg
     2341 acggagaatt gtgtggctat atggaaacaa ttagggcttt atacggactg tgaagctaaa
     2401 gctgtggaga agttcttgga gatcttcaag gaagaaacat tccctcccgg ttcatcgatc
     2461 ctcttcgctc tctcccctac cggctctctt acggttgcgt tttcgaaaga tgatagtatc
     2521 cctgaaaccg ggatcgctgt gatcgagaac aaattgttgg cggaggcggt tctggaatct
     2581 atcatcggga agaacggtgt gtcacctggc actaggttaa gtgttgcaga aagattatct
     2641 cagctaatga tgaagaacaa ggacgaaaag gaagttagtg atcactctgt tgaggaaaaa
     2701 ctagccaaag agaactgagg atccataaca attccccatc ttagtatatt agttaagtat
     2761 aagaaggaga tatacatatg gttacggtgg aagaataccg caaagctcaa cgcgctgaag
     2821 gcccggcgac ggtgatggcg attggcacgg caaccccgac gaactgtgtt gatcagagca
     2881 cctatccgga ctattacttt cgtatcacca actctgaaca taaaacggat ctgaaagaaa
     2941 aattcaaacg tatgtgcgaa aaaagcatga tcaaaaaacg ctatatgcac ctgaccgaag
     3001 aaattctgaa agaaaatccg agcatgtgtg aatacatggc accgtctctg gatgctcgcc
     3061 aggacattgt ggttgtcgaa gtgccgaaac tgggtaaaga agcggcccag aaagcgatca
     3121 aagaatgggg ccaaccgaaa tcaaaaatta cccatctggt cttttgcacc acgtcgggtg
     3181 tggatatgcc gggttgtgac tatcaactga cgaaactgct gggtctgcgt ccgagcgtga
     3241 aacgcctgat gatgtaccag caaggctgct tcgcaggcgg taccgttctg cgtctggcga
     3301 aagatctggc cgaaaacaat aaaggtgcgc gtgttctggt ggtgtgtagt gaaatcaccg
     3361 ctgttacgtt tcgtggtccg aacgatacgc acctggactc cctggttggc caggccctgt
     3421 tcggtgatgg tgcaggtgcc attatcattg gtagcgaccc gattccgggc gttgaacgtc
     3481 cgctgtttga actggtcagc gcagctcaaa ccctgctgcc ggatagccac ggcgcaattg
     3541 acggtcacct gcgtgaagtc ggtctgacgt tccatctgct gaaagatgtg ccgggcctga
     3601 tctcaaaaaa cattgaaaaa agcctggaag aagcgtttcg cccgctgagt atctccgatt
     3661 ggaacagcct gttctggatt gcacatccgg gcggcccggc aatcctggac caggtcgaaa
     3721 ttaaactggg tctgaaaccg gaaaaactga aagcgacccg taatgttctg tcaaactacg
     3781 gcaatatgag ctctgcctgc gtcctgttta ttctggatga aatgcgcaaa gcatcggcta
     3841 aagaaggtct gggcaccacg ggtgaaggcc tggaatgggg cgtgctgttc ggctttggtc
     3901 cgggtctgac ggtggaaacg gtggttctgc atagtgtggc tacctaatct agagtcgacc
     3961 tgcaggcatg caagcttggc tgttttggcg gatgagagaa gattttcagc ctgatacaga
     4021 ttaaatcaga acgcagaagc ggtctgataa aacagaattt gcctggcggc agtagcgcgg
     4081 tggtcccacc tgaccccatg ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg
     4141 tggggtctcc ccatgcgaga gtagggaact gccaggcatc aaataaaacg aaaggctcag
     4201 tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg
     4261 acaaatccgc cgggagcgga tttgaacgtt gcgaagcaac ggcccggagg gtggcgggca
     4321 ggacgcccgc cataaactgc caggcatcaa attaagcaga aggccatcct gacggatggc
     4381 ctttttgcgt ttctacaaac tctttttgtt tatttttcta aatacattca aatatgtatc
     4441 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     4501 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     4561 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     4621 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     4681 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg
     4741 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     4801 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     4861 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     4921 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     4981 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgccta
     5041 cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     5101 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     5161 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     5221 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     5281 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     5341 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     5401 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     5461 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     5521 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     5581 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     5641 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     5701 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     5761 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     5821 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     5881 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     5941 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     6001 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     6061 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     6121 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     6181 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     6241 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     6301 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca
     6361 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct
     6421 acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg
     6481 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     6541 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagcagatca attcgcgcgc
     6601 gaaggcgaag cggcatgcat ttacgttgac accatcgaat ggtgcaaaac ctttcgcggt
     6661 atggcatgat agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa accagtaacg
     6721 ttatacgatg tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg cgtggtgaac
     6781 caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat ggcggagctg
     6841 aattacattc ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt gctgattggc
     6901 gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc gattaaatct
     6961 cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag cggcgtcgaa
     7021 gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg tcagtgggct gatcattaac
     7081 tatccgctgg atgaccagga tgccattgct gtggaagctg cctgcactaa tgttccggcg
     7141 ttatttcttg atgtctctga ccagacaccc atcaacagta ttattttctc ccatgaagac
     7201 ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc accagcaaat cgcgctgtta
     7261 gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca taaatatctc
     7321 actcgcaatc aaattcagcc gatagcggaa cgggaaggcg actggagtgc catgtccggt
     7381 tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat gctggttgcc
     7441 aacgatcaga tggcgctggg cgcaatgcgc gccattaccg agtccgggct gcgcgttggt
     7501 gcggatatct cggtagtggg atacgacgat accgaagaca gctcatgtta tatcccgccg
     7561 tcaaccacca tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga ccgcttgctg
     7621 caactctctc agggccaggc ggtgaagggc aatcagctgt tgcccgtctc actggtgaaa
     7681 agaaaaacca ccctggcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     7741 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     7801 taatgtgagt tagcgcgaat tgatctg
//
