LOCUS       40924_43938        7359 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pTrcCDF-At4CL1m-AtCHI-PhCHS, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7359)
  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 7359)
  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 7359)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7359)
  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..7359
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        175..204
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    212..228
                     /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             249..1931
                     /gene="4CL1"
                     /label=4CL1
                     /note="4-coumarate--CoA ligase 1 from Arabidopsis thaliana.
                     Accession#: Q42524"
     CDS             1960..2697
                     /gene="CHI1"
                     /label=CHI1
                     /note="Chalcone--flavanone isomerase 1 from Arabidopsis 
                     thaliana. Accession#: P41088"
     CDS             2760..3926
                     /gene="CHSA"
                     /label=CHSA
                     /note="Chalcone synthase A from Petunia hybrida.
                     Accession#: P08894"
     regulatory      3957..4104
                     /label=rrnBT1T2
                     /note="rrnBT1T2"
                     /regulatory_class="terminator"
     CDS             complement(4278..5066)
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
     promoter        complement(5067..5158)
                     /label=AmpR promoter
     rep_origin      complement(5206..5944)
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="Plasmids containing the CloDF13 (CDF) origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     protein_bind    complement(6120..6141)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(6157..7236)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(7237..7314)
                     /label=lacI promoter
ORIGIN
        1 gactgcacgg tgcaccaatg cttctggcgt caggcagcca tcggaagctg tggtatggct
       61 gtgcaggtcg taaatcactg cataattcgt gtcgctcaag gcgcactccc gttctggata
      121 atgttttttg cgccgacatc ataacggttc tggcaaatat tctgaaatga gctgttgaca
      181 attaatcatc cggctcgtat aatgtgtgga attgtgagcg gataacaatt tcacacagga
      241 aacagaccat ggcgccacaa gaacaagcag tttctcaggt gatggagaaa cagagcaaca
      301 acaacaacag tgacgtcatt ttccgatcaa agttaccgga tatttacatc ccgaaccacc
      361 tatctctcca cgactacatc ttccaaaaca tctccgaatt cgccactaag ccttgcctaa
      421 tcaacggacc aaccggccac gtgtacactt actccgacgt ccacgtcatc tcccgccaaa
      481 tcgccgccaa ttttcacaaa ctcggcgtta accaaaacga cgtcgtcatg ctcctcctcc
      541 caaactgtcc cgaattcgtc ctctctttcc tcgccgcctc cttccgcggc gcaaccgcca
      601 ccgccgcaaa ccctttcttc actccggcgg agatagctaa acaagccaaa gcctccaaca
      661 ccaaactcat aatcaccgaa gctcgttacg tcgacaaaat caaaccactt caaaacgacg
      721 acggagtagt catcgtctgc atcgacgaca acgaatccgt gccaatccct gaaggctgcc
      781 tccgcttcac cgagttgact cagtcgacaa ccgaggcatc agaagtcatc gactcggtgg
      841 agatttcacc ggacgacgtg gtggcactac cttactcctc tggcacgacg ggattaccaa
      901 aaggagtgat gctgactcac aagggactag tcacgagcgt tgctcagcaa gtcgacggcg
      961 agaacccgaa tctttatttc cacagcgatg acgtcctact ctgtgttttg cccatgtttc
     1021 atatctacgc tttgaactcg atcatgttgt gtggtcttag agttggtgcg gcgattctga
     1081 taatgccgaa gtttgagatc aatctgctat tggagctgat ccagaggtgt aaagtgacgg
     1141 tggctccgat ggttccgccg attgtgttgg ccattgcgaa gtcttcggag acggagaagt
     1201 atgatttgag ctcgataaga gtggtgaaat ctggtgctgc tcctcttggt aaagaacttg
     1261 aagatgccgt taatgccaag tttcctaatg ccaaactcgg tcagggatac ggaatgacgg
     1321 aagcaggtcc agtgctagca atgtcgttag gttttgcaaa ggaacctttt ccggttaagt
     1381 caggagcttg tggtactgtt gtaagaaatg ctgagatgaa aatagttgat ccagacaccg
     1441 gagattctct ttcgaggaaa caacccggtg agatttgtat tcgtggtcac cagatcatga
     1501 aaggttacct caacaatccg gcagctacag cagagaccat tgataaagac ggttggcttc
     1561 atactggaga tattggattg atcgatgacg atgacgagct tttcatcgtt gatcgattga
     1621 aagaacttgt caagtataaa ggttttcagg tagctccggc tgagctagag gctttgctca
     1681 tcggtcatcc tgacattact gatgttgctg ttgtagcaat gaaagaagaa gcagctggtg
     1741 aagttcctgt tgcatttgtg gtgaaatcga aggattcgga gttatcagaa gatgatgtga
     1801 agcaattcgt gtcgaaacag gttgtgtttt acaagagaat caacaaagtg ttcttcactg
     1861 aatccattcc taaagctcca tcagggaaga tattgaggaa agatctgagg gcaaaactag
     1921 caaatggatt gtaattaact ttaagaagga gatatacata tgtcttcatc caacgcctgc
     1981 gcctctccgt caccgttccc cgccgtcacg aagcttcatg tagactccgt cacgtttgta
     2041 ccgtccgtca agtcaccggc ctcctccaat ccattattcc tcggcggcgc cggtgtccga
     2101 ggccttgata tccaaggtaa attcgtgatc ttcaccgtca ttggagtata cctagagggt
     2161 aacgccgttc cttctctatc tgtcaagtgg aagggaaaaa ctacggagga gctaacagaa
     2221 tctatcccgt tcttccgtga aatagtcacc ggtgcgtttg agaagtttat caaggtgaca
     2281 atgaaactgc cgttaacggg acaacaatat tcggagaaag tgacggagaa ttgtgtggct
     2341 atatggaaac aattagggct ttatacggac tgtgaagcta aagctgtgga gaagttcttg
     2401 gagatcttca aggaagaaac attccctccc ggttcatcga tcctcttcgc tctctcccct
     2461 accggctctc ttacggttgc gttttcgaaa gatgatagta tccctgaaac cgggatcgct
     2521 gtgatcgaga acaaattgtt ggcggaggcg gttctggaat ctatcatcgg gaagaacggt
     2581 gtgtcacctg gcactaggtt aagtgttgca gaaagattat ctcagctaat gatgaagaac
     2641 aaggacgaaa aggaagttag tgatcactct gttgaggaaa aactagccaa agagaactga
     2701 ggatccataa caattcccca tcttagtata ttagttaagt ataagaagga gatatacata
     2761 tggttacggt ggaagaatac cgcaaagctc aacgcgctga aggcccggcg acggtgatgg
     2821 cgattggcac ggcaaccccg acgaactgtg ttgatcagag cacctatccg gactattact
     2881 ttcgtatcac caactctgaa cataaaacgg atctgaaaga aaaattcaaa cgtatgtgcg
     2941 aaaaaagcat gatcaaaaaa cgctatatgc acctgaccga agaaattctg aaagaaaatc
     3001 cgagcatgtg tgaatacatg gcaccgtctc tggatgctcg ccaggacatt gtggttgtcg
     3061 aagtgccgaa actgggtaaa gaagcggccc agaaagcgat caaagaatgg ggccaaccga
     3121 aatcaaaaat tacccatctg gtcttttgca ccacgtcggg tgtggatatg ccgggttgtg
     3181 actatcaact gacgaaactg ctgggtctgc gtccgagcgt gaaacgcctg atgatgtacc
     3241 agcaaggctg cttcgcaggc ggtaccgttc tgcgtctggc gaaagatctg gccgaaaaca
     3301 ataaaggtgc gcgtgttctg gtggtgtgta gtgaaatcac cgctgttacg tttcgtggtc
     3361 cgaacgatac gcacctggac tccctggttg gccaggccct gttcggtgat ggtgcaggtg
     3421 ccattatcat tggtagcgac ccgattccgg gcgttgaacg tccgctgttt gaactggtca
     3481 gcgcagctca aaccctgctg ccggatagcc acggcgcaat tgacggtcac ctgcgtgaag
     3541 tcggtctgac gttccatctg ctgaaagatg tgccgggcct gatctcaaaa aacattgaaa
     3601 aaagcctgga agaagcgttt cgcccgctga gtatctccga ttggaacagc ctgttctgga
     3661 ttgcacatcc gggcggcccg gcaatcctgg accaggtcga aattaaactg ggtctgaaac
     3721 cggaaaaact gaaagcgacc cgtaatgttc tgtcaaacta cggcaatatg agctctgcct
     3781 gcgtcctgtt tattctggat gaaatgcgca aagcatcggc taaagaaggt ctgggcacca
     3841 cgggtgaagg cctggaatgg ggcgtgctgt tcggctttgg tccgggtctg acggtggaaa
     3901 cggtggttct gcatagtgtg gctacctaat ctagagtcga cctgcaggca tgcaagcttg
     3961 gctgttttgg cggatgagag aagattttca gcctgataca gattaaatca gaacgcagaa
     4021 gcggtctgat aaaacagaat ttgcctggcg gcagtagcgc ggtggtccca cctgacccca
     4081 tgccgaactc agaagtgaaa cgccctgaaa cctcaggcat ttgagaagca cacggtcaca
     4141 ctgcttccgg tagtcaataa accggtaaac cagcaataga cataagcggc tatttaacga
     4201 ccctgccctg aaccgacgac cgggtcatcg tggccggatc ttgcggcccc tcggcttgaa
     4261 cgaattgtta gacattattt gccgactacc ttggtgatct cgcctttcac gtagtggaca
     4321 aattcttcca actgatctgc gcgcgaggcc aagcgatctt cttcttgtcc aagataagcc
     4381 tgtctagctt caagtatgac gggctgatac tgggccggca ggcgctccat tgcccagtcg
     4441 gcagcgacat ccttcggcgc gattttgccg gttactgcgc tgtaccaaat gcgggacaac
     4501 gtaagcacta catttcgctc atcgccagcc cagtcgggcg gcgagttcca tagcgttaag
     4561 gtttcattta gcgcctcaaa tagatcctgt tcaggaaccg gatcaaagag ttcctccgcc
     4621 gctggaccta ccaaggcaac gctatgttct cttgcttttg tcagcaagat agccagatca
     4681 atgtcgatcg tggctggctc gaagatacct gcaagaatgt cattgcgctg ccattctcca
     4741 aattgcagtt cgcgcttagc tggataacgc cacggaatga tgtcgtcgtg cacaacaatg
     4801 gtgacttcta cagcgcggag aatctcgctc tctccagggg aagccgaagt ttccaaaagg
     4861 tcgttgatca aagctcgccg cgttgtttca tcaagcctta cggtcaccgt aaccagcaaa
     4921 tcaatatcac tgtgtggctt caggccgcca tccactgcgg agccgtacaa atgtacggcc
     4981 agcaacgtcg gttcgagatg gcgctcgatg acgccaacta cctctgatag ttgagtcgat
     5041 acttcggcga tcaccgcttc cctcatactc ttcctttttc aatattattg aagcatttat
     5101 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     5161 gctagctcac tcggtcgcta cgctccgggc gtgagactgc ggcgggcgct gcggacacat
     5221 acaaagttac ccacagattc cgtggataag caggggacta acatgtgagg caaaacagca
     5281 gggccgcgcc ggtggcgttt ttccataggc tccgccctcc tgccagagtt cacataaaca
     5341 gacgcttttc cggtgcatct gtgggagccg tgaggctcaa ccatgaatct gacagtacgg
     5401 gcgaaacccg acaggactta aagatcccca ccgtttccgg cgggtcgctc cctcttgcgc
     5461 tctcctgttc cgaccctgcc gtttaccgga tacctgttcc gcctttctcc cttacgggaa
     5521 gtgtggcgct ttctcatagc tcacacactg gtatctcggc tcggtgtagg tcgttcgctc
     5581 caagctgggc tgtaagcaag aactccccgt tcagcccgac tgctgcgcct tatccggtaa
     5641 ctgttcactt gagtccaacc cggaaaagca cggtaaaacg ccactggcag cagccattgg
     5701 taactgggag ttcgcagagg atttgtttag ctaaacacgc ggttgctctt gaagtgtgcg
     5761 ccaaagtccg gctacactgg aaggacagat ttggttgctg tgctctgcga aagccagtta
     5821 ccacggttaa gcagttcccc aactgactta accttcgatc aaaccacctc cccaggtggt
     5881 tttttcgttt acagggcaaa agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5941 gatcttttct actgaaccgc tctagatttc agtgcaattt atctcttcaa atgtagcacc
     6001 tgaagtcagc cccatacgat ataagttgta attctcatgt tagtcatgcc ccgcgcccac
     6061 cggaaggagc tgactgggtt gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa
     6121 tgagtgagct aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     6181 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     6241 gggcgccagg gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac
     6301 cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa
     6361 atcctgtttg atggtggtta acggcgggat ataacatgag ctgtcttcgg tatcgtcgta
     6421 tcccactacc gagatgtccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc
     6481 gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag
     6541 catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat
     6601 cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga
     6661 gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg
     6721 ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg
     6781 gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc
     6841 atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt
     6901 gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct
     6961 ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag
     7021 ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc
     7081 cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt
     7141 cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc
     7201 atactctgcg acatcgtata acgttactgg tttcacattc accaccctga attgactctc
     7261 ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat
     7321 ctcgacgctc tcccttatgc gactcctgca ttaggaaat
//
