LOCUS       40924_42494        7338 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pTac-VvSTS-At4CL4, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7338)
  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 7338)
  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 7338)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7338)
  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..7338
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             8..1717
                     /gene="4CL4"
                     /label=4CL4
                     /note="4-coumarate--CoA ligase 4 from Arabidopsis thaliana.
                     Accession#: Q9LU36"
     terminator      1969..2055
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      2147..2174
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        2194..2285
                     /label=AmpR promoter
     primer_bind     complement(2598..2614)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2857..3669
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      4201..4746
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."
     promoter        5181..5209
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    5217..5233
                     /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             5255..6430
                     /note="Stilbene synthase 2 from Vitis vinifera. Accession#:
                     P51070"
     terminator      complement(6685..6728)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      6860..6887
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     regulatory      7272..7300
                     /label=Tac promoter
                     /note="Tac promoter"
                     /regulatory_class="promoter"
     promoter        7272..7300
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    7308..7324
                     /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)."
ORIGIN
        1 cagacatatg gtgctccaac aacaaacgca ttttctcaca aagaagatag atcaagaaga
       61 cgaagaagaa gaaccttctc atgatttcat tttccgatct aagcttcctg atatcttcat
      121 ccctaaccac ctccctctca ccgattacgt cttccaaagg ttctccggcg atggcgacgg
      181 agattcctca acaacctgta tcatcgacgg tgccactggc cgtattttga cctacgccga
      241 tgtacaaacc aacatgcgga ggatagctgc cggaatccac cggctcggga tccgccacgg
      301 agacgtcgta atgcttctcc tcccgaactc gccggagttt gctctgtctt ttcttgccgt
      361 tgcttacctt ggagccgttt ccactaccgc gaatccgttc tacactcagc cggagatcgc
      421 taaacaggct aaagcctccg ccgcgaagat gatcatcacg aagaaatgtt tagtcgataa
      481 actaacaaac ctcaaaaacg acggcgtttt gatcgtttgc ctagacgatg acggtgacaa
      541 cggcgttgtt tcgtcgagtg atgatggttg cgtgagtttc acggaactga ctcaagcgga
      601 cgaaacagaa cttctaaaac ctaaaatatc gccggaagac acggtggcga tgccgtactc
      661 ttccggaaca acgggacttc caaagggagt gatgattact cacaagggtt tagttacaag
      721 catcgctcag aaagtcgacg gagaaaaccc taaccttaac ttcacggcca atgacgtcat
      781 cctctgtttt ctcccaatgt ttcacattta cgctctcgac gcgttgatgc tttcggcgat
      841 gagaaccggt gcagcgttac tgatcgtgcc gaggttcgag ttgaatctgg tgatggagct
      901 gattcaaagg tacaaagtca cggttgtgcc ggtggctcct ccggtggttt tagcattcat
      961 taaatcccct gagacggaga gatacgactt gagctcggtg agaattatgc tatcaggtgc
     1021 agctacacta aagaaggagc ttgaagacgc cgtgcgactc aagtttccta atgccatttt
     1081 tggtcagggt tatggaatga ccgagtccgg aacggtagct aagtcattag cgtttgcaaa
     1141 gaacccattt aaaaccaagt ccggtgcatg tgggactgtg atcagaaacg ccgagatgaa
     1201 agtggtcgat accgaaaccg gaatctcctt gccgcgcaac aaatctggcg agatctgcgt
     1261 ccgaggtcat cagctcatga aaggttattt gaatgatccg gaagctactg cacgaaccat
     1321 agacaaagac gggtggttac atacaggaga tattgggttt gtggatgatg atgatgagat
     1381 cttcattgtt gatcggttga aggaactgat caaattcaaa ggctaccaag tggctccagc
     1441 tgagcttgaa gcattgctta tttctcatcc ttctatcgat gatgccgcag ttgttgcaat
     1501 gaaggatgaa gtagctgatg aggttccagt agcatttgta gcaagatctc aagggtctca
     1561 gttaactgaa gatgatgtca agagttacgt aaacaaacag gtggttcact acaagcggat
     1621 taagatggtg tttttcatag aagttatacc aaaagcagtt tcgggcaaga ttctgaggaa
     1681 ggatctgcga gctaaattgg aaaccatgtg ctctaaataa tggtacccgg ggatcctcta
     1741 gagtcgacct gcaggcatgc aagcttggct gttttggcgg atgagagaag attttcagcc
     1801 tgatacagat taaatcagaa cgcagaagcg gtctgataaa acagaatttg cctggcggca
     1861 gtagcgcggt ggtcccacct gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg
     1921 atggtagtgt ggggtctccc catgcgagag tagggaactg ccaggcatca aataaaacga
     1981 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc
     2041 ctgagtagga caaatccgcc gggagcggat ttgaacgttg cgaagcaacg gcccggaggg
     2101 tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg
     2161 acggatggcc tttttgcgtt tctacaaact ctttttgttt atttttctaa atacattcaa
     2221 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     2281 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     2341 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     2401 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     2461 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     2521 tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     2581 acttggttga gtaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     2641 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     2701 gaggcccgca ccgatcgccc ttcccaacag ttgcgggggg ggggggaaag ccacgttgtg
     2761 tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg aacaataaaa
     2821 ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca acgggaaacg
     2881 tcttgctcga ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg
     2941 gctcgcgata atgtcgggca atcaggtgcg acaatctatc gattgtatgg gaagcccgat
     3001 gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag
     3061 atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc
     3121 cgtactcctg atgatgcatg gttactcacc actgcgatcc ccgggaaaac agcattccag
     3181 gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg
     3241 cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt
     3301 ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac
     3361 gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataagct tttgccattc
     3421 tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag
     3481 gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat
     3541 cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt
     3601 caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat
     3661 gagtttttct aagcattggt aacctggtat gagtcagcaa caccttcttc acgaggcaga
     3721 cctcagcgcc cccccccccc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     3781 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     3841 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     3901 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     3961 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     4021 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     4081 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     4141 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccttaataa gatgatcttc
     4201 ttgagatcgt tttggtctgc gcgtaatctc ttgctctgaa aacgaaaaaa ccgccttgca
     4261 gggcggtttt tcgaaggttc tctgagctac caactctttg aaccgaggta actggcttgg
     4321 aggagcgcag tcaccaaaac ttgtcctttc agtttagcct taaccggcgc atgacttcaa
     4381 gactaactcc tctaaatcaa ttaccagtgg ctgctgccag tggtgctttt gcatgtcttt
     4441 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcggac tgaacggggg
     4501 gttcgtgcat acagtccagc ttggagcgaa ctgcctaccc ggaactgagt gtcaggcgtg
     4561 gaatgagaca aacgcggcca taacagcgga atgacaccgg taaaccgaaa ggcaggaaca
     4621 ggagagcgca cgagggagcc gccaggggga aacgcctggt atctttatag tcctgtcggg
     4681 tttcgccacc actgatttga gcgtcagatt tcgtgatgct tgtcaggggg gcggagccta
     4741 tggaaaaacg gctttgccgc ggccctctca cttccctgtt aagtatcttc ctggcatctt
     4801 ccaggaaatc tccgccccgt tcgtaagcca tttccgctcg ccgcagtcga acgaccgagc
     4861 gtagcgagtc agtgagcgag gaagcggaat atatcctgta tcacatattc tgctgacgca
     4921 ccggtgcagc cttttttctc ctgccacatg aagcacttca ctgacaccct catcagtgcc
     4981 aacatagtaa gccagtatac actccggact gcacggtgca ccaatgcttc tggcgtcagg
     5041 cagccatcgg aagctgtggt atggctgtgc aggtcgtaaa tcactgcata attcgtgtcg
     5101 ctcaaggcgc actcccgttc tggataatgt tttttgcgcc gacatcataa cggttctggc
     5161 aaatattctg aaatgagctg ttgacaatta atcatcggct cgtataatgt gtggaattgt
     5221 gagcggataa caatttcaca caggaaacag acatatggca agtgtcgagg aattccgcaa
     5281 tgctcagcgc gcaaagggtc ccgccacgat cctggccatt ggaacagcaa cgcccgatca
     5341 ttgcgtgtac caatcggact acgcggattt ttacttccgc gtgacaaagt cagaacacat
     5401 gactgcactt aaaaagaaat tcaaccgtat ttgcgataag tccatgatca agaaacgcta
     5461 catccacctt acagaagaaa tgttagagga acatcctaac attggagcct atatggcacc
     5521 ttcattaaac attcgccagg aaattattac cgcggaggtc cccaaacttg ggaaggaggc
     5581 cgctttaaaa gctcttaagg aatggggtca gcctaagtct aaaatcacgc atttggtctt
     5641 ctgcactaca tctggggtgg agatgcccgg agcagattac aaattagcga acttgttggg
     5701 acttgaaccc tccgtccgtc gcgtcatgct gtaccatcag ggctgctatg ccggagggac
     5761 tgtgctgcgc actgctaaag acttagcaga aaacaatgct ggggctcgtg tcctggttgt
     5821 ttgttccgag atcaccgttg tgactttccg cgggccaagt gaggatgctc ttgattcctt
     5881 agtcgggcaa gctcttttcg gcgatggctc agcagcggtt attgtcggct cagatccaga
     5941 cattagcatc gagcgtccac tgtttcaact ggtgtcagcg gctcaaacct ttatcccgaa
     6001 cagcgcagga gccattgcag gcaacttacg cgaagttgga ttaacgttcc atctgtggcc
     6061 gaatgtgccc accctgatta gcgaaaatat tgaaaagtgc ttaactcagg catttgatcc
     6121 gttgggcatc agcgactgga attccttgtt ttggatcgcc catccagggg ggcctgcgat
     6181 tttggacgcc gtagaagcta aactgaatct tgacaagaaa aaattggagg caacacgtca
     6241 tgtgctgtcc gagtatggca acatgtccag tgcatgcgtg cttttcatct tggacgagat
     6301 gcgtaagaaa agtttaaaag gggaacgtgc tacgaccggg gagggtttag actggggagt
     6361 gttgtttggt tttggtcccg ggctgactat cgaaacagtg gtacttcatt ccattccgat
     6421 ggttaccaat taatggtacc cggggatcct ctagagtcga cctgcaggca tgcaagcttg
     6481 gctgttttgg cggatgagag aagattttca gcctgataca gattaaatca gaacgcagaa
     6541 gcggtctgat aaaacagaat ttgcctggcg gcagtagcgc ggtggtccca cctgacccca
     6601 tgccgaactc agaagtgaaa cgccgtagcg ccgatggtag tgtggggtct ccccatgcga
     6661 gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt
     6721 cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg
     6781 gatttgaacg ttgcgaagca acggcccgga gggtggcggg caggacgccc gccataaact
     6841 gccaggcatc aaattaagca gaaggccatc ctgacggatg gcctttttgc gtttctacac
     6901 aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa
     6961 caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata
     7021 taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag
     7081 aggatccgga gcttatcgac tgcacggtgc accaatgctt ctggcgtcag gcagccatcg
     7141 gaagctgtgg tatggctgtg caggtcgtaa atcactgcat aattcgtgtc gctcaaggcg
     7201 cactcccgtt ctggataatg ttttttgcgc cgacatcata acggttctgg caaatattct
     7261 gaaatgagct gttgacaatt aatcatcggc tcgtataatg tgtggaattg tgagcggata
     7321 acaatttcac acaggaaa
//
