LOCUS       40924_42429        6610 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pTac-At4CL1m-fus-VvSTS, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6610)
  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 6610)
  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 6610)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6610)
  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..6610
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             8..1690
                     /gene="4CL1"
                     /label=4CL1
                     /note="4-coumarate--CoA ligase 1 from Arabidopsis thaliana.
                     Accession#: Q42524"
     misc_feature    1691..1702
                     /label=similar to glycine-serine linker
                     /note="similar to glycine-serine linker"
     CDS             1703..2878
                     /note="Stilbene synthase 2 from Vitis vinifera. Accession#:
                     P51070"
     terminator      3130..3216
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      3308..3335
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        3355..3446
                     /label=AmpR promoter
     primer_bind     complement(3759..3775)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             4018..4830
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      5362..5907
                     /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        6544..6572
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    6580..6596
                     /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)."
ORIGIN
        1 cagacatatg gcgccacaag aacaagcagt ttctcaggtg atggagaaac agagcaacaa
       61 caacaacagt gacgtcattt tccgatcaaa gttaccggat atttacatcc cgaaccacct
      121 atctctccac gactacatct tccaaaacat ctccgaattc gccactaagc cttgcctaat
      181 caacggacca accggccacg tgtacactta ctccgacgtc cacgtcatct cccgccaaat
      241 cgccgccaat tttcacaaac tcggcgttaa ccaaaacgac gtcgtcatgc tcctcctccc
      301 aaactgtccc gaattcgtcc tctctttcct cgccgcctcc ttccgcggcg caaccgccac
      361 cgccgcaaac cctttcttca ctccggcgga gatagctaaa caagccaaag cctccaacac
      421 caaactcata atcaccgaag ctcgttacgt cgacaaaatc aaaccacttc aaaacgacga
      481 cggagtagtc atcgtctgca tcgacgacaa cgaatccgtg ccaatccctg aaggctgcct
      541 ccgcttcacc gagttgactc agtcgacaac cgaggcatca gaagtcatcg actcggtgga
      601 gatttcaccg gacgacgtgg tggcactacc ttactcctct ggcacgacgg gattaccaaa
      661 aggagtgatg ctgactcaca agggactagt cacgagcgtt gctcagcaag tcgacggcga
      721 gaacccgaat ctttatttcc acagcgatga cgtcctactc tgtgttttgc ccatgtttca
      781 tatctacgct ttgaactcga tcatgttgtg tggtcttaga gttggtgcgg cgattctgat
      841 aatgccgaag tttgagatca atctgctatt ggagctgatc cagaggtgta aagtgacggt
      901 ggctccgatg gttccgccga ttgtgttggc cattgcgaag tcttcggaga cggagaagta
      961 tgatttgagc tcgataagag tggtgaaatc tggtgctgct cctcttggta aagaacttga
     1021 agatgccgtt aatgccaagt ttcctaatgc caaactcggt cagggatacg gaatgacgga
     1081 agcaggtcca gtgctagcaa tgtcgttagg ttttgcaaag gaaccttttc cggttaagtc
     1141 aggagcttgt ggtactgttg taagaaatgc tgagatgaaa atagttgatc cagacaccgg
     1201 agattctctt tcgaggaaac aacccggtga gatttgtatt cgtggtcacc agatcatgaa
     1261 aggttacctc aacaatccgg cagctacagc agagaccatt gataaagacg gttggcttca
     1321 tactggagat attggattga tcgatgacga tgacgagctt ttcatcgttg atcgattgaa
     1381 agaacttgtc aagtataaag gttttcaggt agctccggct gagctagagg ctttgctcat
     1441 cggtcatcct gacattactg atgttgctgt tgtcgcaatg aaagaagaag cagctggtga
     1501 agttcctgtt gcatttgtgg tgaaatcgaa ggattcggag ttatcagaag atgatgtgaa
     1561 gcaattcgtg tcgaaacagg ttgtgtttta caagagaatc aacaaagtgt tcttcactga
     1621 atccattcct aaagctccat cagggaagat attgaggaaa gatctgaggg caaaactagc
     1681 aaatggattg ggtggtggta gtatggcaag tgtcgaggaa ttccgcaatg ctcagcgcgc
     1741 aaagggtccc gccacgatcc tggccattgg aacagcaacg cccgatcatt gcgtgtacca
     1801 atcggactac gcggattttt acttccgcgt gacaaagtca gaacacatga ctgcacttaa
     1861 aaagaaattc aaccgtattt gcgataagtc catgatcaag aaacgctaca tccaccttac
     1921 agaagaaatg ttagaggaac atcctaacat tggagcctat atggcacctt cattaaacat
     1981 tcgccaggaa attattaccg cggaggtccc caaacttggg aaggaggccg ctttaaaagc
     2041 tcttaaggaa tggggtcagc ctaagtctaa aatcacgcat ttggtcttct gcactacatc
     2101 tggggtggag atgcccggag cagattacaa attagcgaac ttgttgggac ttgaaccctc
     2161 cgtccgtcgc gtcatgctgt accatcaggg ctgctatgcc ggagggactg tgctgcgcac
     2221 tgctaaagac ttagcagaaa acaatgctgg ggctcgtgtc ctggttgttt gttccgagat
     2281 caccgttgtg actttccgcg ggccaagtga ggatgctctt gattccttag tcgggcaagc
     2341 tcttttcggc gatggctcag cagcggttat tgtcggctca gatccagaca ttagcatcga
     2401 gcgtccactg tttcaactgg tgtcagcggc tcaaaccttt atcccgaaca gcgcaggagc
     2461 cattgcaggc aacttacgcg aagttggatt aacgttccat ctgtggccga atgtgcccac
     2521 cctgattagc gaaaatattg aaaagtgctt aactcaggca tttgatccgt tgggcatcag
     2581 cgactggaat tccttgtttt ggatcgccca tccagggggg cctgcgattt tggacgccgt
     2641 agaagctaaa ctgaatcttg acaagaaaaa attggaggca acacgtcatg tgctgtccga
     2701 gtatggcaac atgtccagtg catgcgtgct tttcatcttg gacgagatgc gtaagaaaag
     2761 tttaaaaggg gaacgtgcta cgaccgggga gggtttagac tggggagtgt tgtttggttt
     2821 tggtcccggg ctgactatcg aaacagtggt acttcattcc attccgatgg ttaccaatta
     2881 atggtacccg gggatcctct agagtcgacc tgcaggcatg caagcttggc tgttttggcg
     2941 gatgagagaa gattttcagc ctgatacaga ttaaatcaga acgcagaagc ggtctgataa
     3001 aacagaattt gcctggcggc agtagcgcgg tggtcccacc tgaccccatg ccgaactcag
     3061 aagtgaaacg ccgtagcgcc gatggtagtg tggggtctcc ccatgcgaga gtagggaact
     3121 gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
     3181 tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgggagcgga tttgaacgtt
     3241 gcgaagcaac ggcccggagg gtggcgggca ggacgcccgc cataaactgc caggcatcaa
     3301 attaagcaga aggccatcct gacggatggc ctttttgcgt ttctacaaac tctttttgtt
     3361 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     3421 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     3481 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     3541 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     3601 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     3661 ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc
     3721 gcatacacta ttctcagaat gacttggttg agtaattcac tggccgtcgt tttacaacgt
     3781 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     3841 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcggggg
     3901 gggggggaaa gccacgttgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat
     3961 atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgttatg
     4021 agccatattc aacgggaaac gtcttgctcg aggccgcgat taaattccaa catggatgct
     4081 gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat
     4141 cgattgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt
     4201 gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt
     4261 ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc
     4321 cccgggaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt
     4381 gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt
     4441 aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt
     4501 gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa
     4561 atgcataagc ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt
     4621 gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga
     4681 atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct
     4741 tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg
     4801 cagtttcatt tgatgctcga tgagtttttc taagcattgg taacctggta tgagtcagca
     4861 acaccttctt cacgaggcag acctcagcgc cccccccccc cgcaaactat taactggcga
     4921 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     4981 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     5041 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     5101 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     5161 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     5221 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     5281 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     5341 ccccttaata agatgatctt cttgagatcg ttttggtctg cgcgtaatct cttgctctga
     5401 aaacgaaaaa accgccttgc agggcggttt ttcgaaggtt ctctgagcta ccaactcttt
     5461 gaaccgaggt aactggcttg gaggagcgca gtcaccaaaa cttgtccttt cagtttagcc
     5521 ttaaccggcg catgacttca agactaactc ctctaaatca attaccagtg gctgctgcca
     5581 gtggtgcttt tgcatgtctt tccgggttgg actcaagacg atagttaccg gataaggcgc
     5641 agcggtcgga ctgaacgggg ggttcgtgca tacagtccag cttggagcga actgcctacc
     5701 cggaactgag tgtcaggcgt ggaatgagac aaacgcggcc ataacagcgg aatgacaccg
     5761 gtaaaccgaa aggcaggaac aggagagcgc acgagggagc cgccaggggg aaacgcctgg
     5821 tatctttata gtcctgtcgg gtttcgccac cactgatttg agcgtcagat ttcgtgatgc
     5881 ttgtcagggg ggcggagcct atggaaaaac ggctttgccg cggccctctc acttccctgt
     5941 taagtatctt cctggcatct tccaggaaat ctccgccccg ttcgtaagcc atttccgctc
     6001 gccgcagtcg aacgaccgag cgtagcgagt cagtgagcga ggaagcggaa tatatcctgt
     6061 atcacatatt ctgctgacgc accggtgcag ccttttttct cctgccacat gaagcacttc
     6121 actgacaccc tcatcagtgc caacatagta agccagtata cactccgcta gcaaggagat
     6181 ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct
     6241 catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc
     6301 agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatccg
     6361 gagcttatcg actgcacggt gcaccaatgc ttctggcgtc aggcagccat cggaagctgt
     6421 ggtatggctg tgcaggtcgt aaatcactgc ataattcgtg tcgctcaagg cgcactcccg
     6481 ttctggataa tgttttttgc gccgacatca taacggttct ggcaaatatt ctgaaatgag
     6541 ctgttgacaa ttaatcatcg gctcgtataa tgtgtggaat tgtgagcgga taacaatttc
     6601 acacaggaaa
//
