LOCUS       40924_42519        6881 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pTacCDF-VvSTS-At4CL1m, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6881)
  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 6881)
  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 6881)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6881)
  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..6881
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        195..223
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    231..247
                     /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             269..1444
                     /note="Stilbene synthase 2 from Vitis vinifera. Accession#:
                     P51070"
     regulatory      1656..1684
                     /label=Tac promoter
                     /note="Tac promoter"
                     /regulatory_class="promoter"
     promoter        1656..1684
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    1692..1708
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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             1730..3412
                     /gene="4CL1"
                     /label=4CL1
                     /note="4-coumarate--CoA ligase 1 from Arabidopsis thaliana.
                     Accession#: Q42524"
     regulatory      3459..3604
                     /label=rrnBT1T2
                     /note="rrnBT1T2"
                     /regulatory_class="terminator"
     CDS             complement(3800..4588)
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
     promoter        complement(4589..4680)
                     /label=AmpR promoter
     rep_origin      complement(4728..5466)
                     /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(5642..5663)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(5679..6758)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(6759..6836)
                     /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 ggcgttgaca attaatcatc ggctcgtata atgtgtggaa ttgtgagcgg
      241 ataacaattt cacacaggaa acagacatat ggcaagtgtc gaggaattcc gcaatgctca
      301 gcgcgcaaag ggtcccgcca cgatcctggc cattggaaca gcaacgcccg atcattgcgt
      361 gtaccaatcg gactacgcgg atttttactt ccgcgtgaca aagtcagaac acatgactgc
      421 acttaaaaag aaattcaacc gtatttgcga taagtccatg atcaagaaac gctacatcca
      481 ccttacagaa gaaatgttag aggaacatcc taacattgga gcctatatgg caccttcatt
      541 aaacattcgc caggaaatta ttaccgcgga ggtccccaaa cttgggaagg aggccgcttt
      601 aaaagctctt aaggaatggg gtcagcctaa gtctaaaatc acgcatttgg tcttctgcac
      661 tacatctggg gtggagatgc ccggagcaga ttacaaatta gcgaacttgt tgggacttga
      721 accctccgtc cgtcgcgtca tgctgtacca tcagggctgc tatgccggag ggactgtgct
      781 gcgcactgct aaagacttag cagaaaacaa tgctggggct cgtgtcctgg ttgtttgttc
      841 cgagatcacc gttgtgactt tccgcgggcc aagtgaggat gctcttgatt ccttagtcgg
      901 gcaagctctt ttcggcgatg gctcagcagc ggttattgtc ggctcagatc cagacattag
      961 catcgagcgt ccactgtttc aactggtgtc agcggctcaa acctttatcc cgaacagcgc
     1021 aggagccatt gcaggcaact tacgcgaagt tggattaacg ttccatctgt ggccgaatgt
     1081 gcccaccctg attagcgaaa atattgaaaa gtgcttaact caggcatttg atccgttggg
     1141 catcagcgac tggaattcct tgttttggat cgcccatcca ggggggcctg cgattttgga
     1201 cgccgtagaa gctaaactga atcttgacaa gaaaaaattg gaggcaacac gtcatgtgct
     1261 gtccgagtat ggcaacatgt ccagtgcatg cgtgcttttc atcttggacg agatgcgtaa
     1321 gaaaagttta aaaggggaac gtgctacgac cggggagggt ttagactggg gagtgttgtt
     1381 tggttttggt cccgggctga ctatcgaaac agtggtactt cattccattc cgatggttac
     1441 caattaatgg tacccgggga tcctctagag tcgacctgca ggactgcacg gtgcaccaat
     1501 gcttctggcg tcaggcagcc atcggaagct gtggtatggc tgtgcaggtc gtaaatcact
     1561 gcataattcg tgtcgctcaa ggcgcactcc cgttctggat aatgtttttt gcgccgacat
     1621 cataacggtt ctggcaaata ttctgaaatg agctgttgac aattaatcat cggctcgtat
     1681 aatgtgtgga attgtgagcg gataacaatt tcacacagga aacagacata tggcgccaca
     1741 agaacaagca gtttctcagg tgatggagaa acagagcaac aacaacaaca gtgacgtcat
     1801 tttccgatca aagttaccgg atatttacat cccgaaccac ctatctctcc acgactacat
     1861 cttccaaaac atctccgaat tcgccactaa gccttgccta atcaacggac caaccggcca
     1921 cgtgtacact tactccgacg tccacgtcat ctcccgccaa atcgccgcca attttcacaa
     1981 actcggcgtt aaccaaaacg acgtcgtcat gctcctcctc ccaaactgtc ccgaattcgt
     2041 cctctctttc ctcgccgcct ccttccgcgg cgcaaccgcc accgccgcaa accctttctt
     2101 cactccggcg gagatagcta aacaagccaa agcctccaac accaaactca taatcaccga
     2161 agctcgttac gtcgacaaaa tcaaaccact tcaaaacgac gacggagtag tcatcgtctg
     2221 catcgacgac aacgaatccg tgccaatccc tgaaggctgc ctccgcttca ccgagttgac
     2281 tcagtcgaca accgaggcat cagaagtcat cgactcggtg gagatttcac cggacgacgt
     2341 ggtggcacta ccttactcct ctggcacgac gggattacca aaaggagtga tgctgactca
     2401 caagggacta gtcacgagcg ttgctcagca agtcgacggc gagaacccga atctttattt
     2461 ccacagcgat gacgtcctac tctgtgtttt gcccatgttt catatctacg ctttgaactc
     2521 gatcatgttg tgtggtctta gagttggtgc ggcgattctg ataatgccga agtttgagat
     2581 caatctgcta ttggagctga tccagaggtg taaagtgacg gtggctccga tggttccgcc
     2641 gattgtgttg gccattgcga agtcttcgga gacggagaag tatgatttga gctcgataag
     2701 agtggtgaaa tctggtgctg ctcctcttgg taaagaactt gaagatgccg ttaatgccaa
     2761 gtttcctaat gccaaactcg gtcagggata cggaatgacg gaagcaggtc cagtgctagc
     2821 aatgtcgtta ggttttgcaa aggaaccttt tccggttaag tcaggagctt gtggtactgt
     2881 tgtaagaaat gctgagatga aaatagttga tccagacacc ggagattctc tttcgaggaa
     2941 acaacccggt gagatttgta ttcgtggtca ccagatcatg aaaggttacc tcaacaatcc
     3001 ggcagctaca gcagagacca ttgataaaga cggttggctt catactggag atattggatt
     3061 gatcgatgac gatgacgagc ttttcatcgt tgatcgattg aaagaacttg tcaagtataa
     3121 aggttttcag gtagctccgg ctgagctaga ggctttgctc atcggtcatc ctgacattac
     3181 tgatgttgct gttgtcgcaa tgaaagaaga agcagctggt gaagttcctg ttgcatttgt
     3241 ggtgaaatcg aaggattcgg agttatcaga agatgatgtg aagcaattcg tgtcgaaaca
     3301 ggttgtgttt tacaagagaa tcaacaaagt gttcttcact gaatccattc ctaaagctcc
     3361 atcagggaag atattgagga aagatctgag ggcaaaacta gcaaatggat tgtaatggta
     3421 cccggggatc ctctagagtc gacctgcagg catgcaagct tggctgtttt ggcggatgag
     3481 agaagatttt cagcctgata cagattaaat cagaacgcag aagcggtctg ataaaacaga
     3541 atttgcctgg cggcagtagc gcggtggtcc cacctgaccc catgccgaac tcagaagtga
     3601 aacgccgaac tcagaagtga aacgccctga aacctcaggc atttgagaag cacacggtca
     3661 cactgcttcc ggtagtcaat aaaccggtaa accagcaata gacataagcg gctatttaac
     3721 gaccctgccc tgaaccgacg accgggtcat cgtggccgga tcttgcggcc cctcggcttg
     3781 aacgaattgt tagacattat ttgccgacta ccttggtgat ctcgcctttc acgtagtgga
     3841 caaattcttc caactgatct gcgcgcgagg ccaagcgatc ttcttcttgt ccaagataag
     3901 cctgtctagc ttcaagtatg acgggctgat actgggccgg caggcgctcc attgcccagt
     3961 cggcagcgac atccttcggc gcgattttgc cggttactgc gctgtaccaa atgcgggaca
     4021 acgtaagcac tacatttcgc tcatcgccag cccagtcggg cggcgagttc catagcgtta
     4081 aggtttcatt tagcgcctca aatagatcct gttcaggaac cggatcaaag agttcctccg
     4141 ccgctggacc taccaaggca acgctatgtt ctcttgcttt tgtcagcaag atagccagat
     4201 caatgtcgat cgtggctggc tcgaagatac ctgcaagaat gtcattgcgc tgccattctc
     4261 caaattgcag ttcgcgctta gctggataac gccacggaat gatgtcgtcg tgcacaacaa
     4321 tggtgacttc tacagcgcgg agaatctcgc tctctccagg ggaagccgaa gtttccaaaa
     4381 ggtcgttgat caaagctcgc cgcgttgttt catcaagcct tacggtcacc gtaaccagca
     4441 aatcaatatc actgtgtggc ttcaggccgc catccactgc ggagccgtac aaatgtacgg
     4501 ccagcaacgt cggttcgaga tggcgctcga tgacgccaac tacctctgat agttgagtcg
     4561 atacttcggc gatcaccgct tccctcatac tcttcctttt tcaatattat tgaagcattt
     4621 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     4681 tagctagctc actcggtcgc tacgctccgg gcgtgagact gcggcgggcg ctgcggacac
     4741 atacaaagtt acccacagat tccgtggata agcaggggac taacatgtga ggcaaaacag
     4801 cagggccgcg ccggtggcgt ttttccatag gctccgccct cctgccagag ttcacataaa
     4861 cagacgcttt tccggtgcat ctgtgggagc cgtgaggctc aaccatgaat ctgacagtac
     4921 gggcgaaacc cgacaggact taaagatccc caccgtttcc ggcgggtcgc tccctcttgc
     4981 gctctcctgt tccgaccctg ccgtttaccg gatacctgtt ccgcctttct cccttacggg
     5041 aagtgtggcg ctttctcata gctcacacac tggtatctcg gctcggtgta ggtcgttcgc
     5101 tccaagctgg gctgtaagca agaactcccc gttcagcccg actgctgcgc cttatccggt
     5161 aactgttcac ttgagtccaa cccggaaaag cacggtaaaa cgccactggc agcagccatt
     5221 ggtaactggg agttcgcaga ggatttgttt agctaaacac gcggttgctc ttgaagtgtg
     5281 cgccaaagtc cggctacact ggaaggacag atttggttgc tgtgctctgc gaaagccagt
     5341 taccacggtt aagcagttcc ccaactgact taaccttcga tcaaaccacc tccccaggtg
     5401 gttttttcgt ttacagggca aaagattacg cgcagaaaaa aaggatctca agaagatcct
     5461 ttgatctttt ctactgaacc gctctagatt tcagtgcaat ttatctcttc aaatgtagca
     5521 cctgaagtca gccccatacg atataagttg taattctcat gttagtcatg ccccgcgccc
     5581 accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgagat cccggtgcct
     5641 aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     5701 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     5761 ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg attgcccttc
     5821 accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc cagcaggcga
     5881 aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc ggtatcgtcg
     5941 tatcccacta ccgagatgtc cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt
     6001 gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat gccctcattc
     6061 agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc ccgttccgct
     6121 atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg cagacgcgcc
     6181 gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga
     6241 tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt gatgggtgtc
     6301 tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc cacagcaatg
     6361 gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg cgcgagaaga
     6421 ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg
     6481 ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga cggcgcgtgc
     6541 agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc cagttgttgt
     6601 gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt ttcccgcgtt
     6661 ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata agagacaccg
     6721 gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct gaattgactc
     6781 tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat ggtgtccggg
     6841 atctcgacgc tctcccttat gcgactcctg cattaggaaa t
//
