LOCUS       40924_42484        7315 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pTac-VvSTS-At4CL1m, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7315)
  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 7315)
  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 7315)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7315)
  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..7315
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             8..1183
                     /note="Stilbene synthase 2 from Vitis vinifera. Accession#:
                     P51070"
     terminator      1435..1521
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      1613..1640
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        1660..1751
                     /label=AmpR promoter
     primer_bind     complement(2064..2080)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2325..2353
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    2361..2377
                     /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             2399..4081
                     /gene="4CL1"
                     /label=4CL1
                     /note="4-coumarate--CoA ligase 1 from Arabidopsis thaliana.
                     Accession#: Q42524"
     terminator      complement(4336..4379)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      4511..4538
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     CDS             4723..5535
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      6067..6612
                     /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."
     regulatory      7249..7277
                     /label=Tac promoter
                     /note="Tac promoter"
                     /regulatory_class="promoter"
     promoter        7249..7277
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    7285..7301
                     /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 gcaagtgtcg aggaattccg caatgctcag cgcgcaaagg gtcccgccac
       61 gatcctggcc attggaacag caacgcccga tcattgcgtg taccaatcgg actacgcgga
      121 tttttacttc cgcgtgacaa agtcagaaca catgactgca cttaaaaaga aattcaaccg
      181 tatttgcgat aagtccatga tcaagaaacg ctacatccac cttacagaag aaatgttaga
      241 ggaacatcct aacattggag cctatatggc accttcatta aacattcgcc aggaaattat
      301 taccgcggag gtccccaaac ttgggaagga ggccgcttta aaagctctta aggaatgggg
      361 tcagcctaag tctaaaatca cgcatttggt cttctgcact acatctgggg tggagatgcc
      421 cggagcagat tacaaattag cgaacttgtt gggacttgaa ccctccgtcc gtcgcgtcat
      481 gctgtaccat cagggctgct atgccggagg gactgtgctg cgcactgcta aagacttagc
      541 agaaaacaat gctggggctc gtgtcctggt tgtttgttcc gagatcaccg ttgtgacttt
      601 ccgcgggcca agtgaggatg ctcttgattc cttagtcggg caagctcttt tcggcgatgg
      661 ctcagcagcg gttattgtcg gctcagatcc agacattagc atcgagcgtc cactgtttca
      721 actggtgtca gcggctcaaa cctttatccc gaacagcgca ggagccattg caggcaactt
      781 acgcgaagtt ggattaacgt tccatctgtg gccgaatgtg cccaccctga ttagcgaaaa
      841 tattgaaaag tgcttaactc aggcatttga tccgttgggc atcagcgact ggaattcctt
      901 gttttggatc gcccatccag gggggcctgc gattttggac gccgtagaag ctaaactgaa
      961 tcttgacaag aaaaaattgg aggcaacacg tcatgtgctg tccgagtatg gcaacatgtc
     1021 cagtgcatgc gtgcttttca tcttggacga gatgcgtaag aaaagtttaa aaggggaacg
     1081 tgctacgacc ggggagggtt tagactgggg agtgttgttt ggttttggtc ccgggctgac
     1141 tatcgaaaca gtggtacttc attccattcc gatggttacc aattaatggt acccggggat
     1201 cctctagagt cgacctgcag gcatgcaagc ttggctgttt tggcggatga gagaagattt
     1261 tcagcctgat acagattaaa tcagaacgca gaagcggtct gataaaacag aatttgcctg
     1321 gcggcagtag cgcggtggtc ccacctgacc ccatgccgaa ctcagaagtg aaacgccgta
     1381 gcgccgatgg tagtgtgggg tctccccatg cgagagtagg gaactgccag gcatcaaata
     1441 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac
     1501 gctctcctga gtaggacaaa tccgccggga gcggatttga acgttgcgaa gcaacggccc
     1561 ggagggtggc gggcaggacg cccgccataa actgccaggc atcaaattaa gcagaaggcc
     1621 atcctgacgg atggcctttt tgcgtttcta caaactcttt ttgtttattt ttctaaatac
     1681 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     1741 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     1801 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     1861 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     1921 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     1981 cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     2041 agaatgactt ggttgagtaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     2101 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag gactgcacgg
     2161 tgcaccaatg cttctggcgt caggcagcca tcggaagctg tggtatggct gtgcaggtcg
     2221 taaatcactg cataattcgt gtcgctcaag gcgcactccc gttctggata atgttttttg
     2281 cgccgacatc ataacggttc tggcaaatat tctgaaatga gctgttgaca attaatcatc
     2341 ggctcgtata atgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagacatat
     2401 ggcgccacaa gaacaagcag tttctcaggt gatggagaaa cagagcaaca acaacaacag
     2461 tgacgtcatt ttccgatcaa agttaccgga tatttacatc ccgaaccacc tatctctcca
     2521 cgactacatc ttccaaaaca tctccgaatt cgccactaag ccttgcctaa tcaacggacc
     2581 aaccggccac gtgtacactt actccgacgt ccacgtcatc tcccgccaaa tcgccgccaa
     2641 ttttcacaaa ctcggcgtta accaaaacga cgtcgtcatg ctcctcctcc caaactgtcc
     2701 cgaattcgtc ctctctttcc tcgccgcctc cttccgcggc gcaaccgcca ccgccgcaaa
     2761 ccctttcttc actccggcgg agatagctaa acaagccaaa gcctccaaca ccaaactcat
     2821 aatcaccgaa gctcgttacg tcgacaaaat caaaccactt caaaacgacg acggagtagt
     2881 catcgtctgc atcgacgaca acgaatccgt gccaatccct gaaggctgcc tccgcttcac
     2941 cgagttgact cagtcgacaa ccgaggcatc agaagtcatc gactcggtgg agatttcacc
     3001 ggacgacgtg gtggcactac cttactcctc tggcacgacg ggattaccaa aaggagtgat
     3061 gctgactcac aagggactag tcacgagcgt tgctcagcaa gtcgacggcg agaacccgaa
     3121 tctttatttc cacagcgatg acgtcctact ctgtgttttg cccatgtttc atatctacgc
     3181 tttgaactcg atcatgttgt gtggtcttag agttggtgcg gcgattctga taatgccgaa
     3241 gtttgagatc aatctgctat tggagctgat ccagaggtgt aaagtgacgg tggctccgat
     3301 ggttccgccg attgtgttgg ccattgcgaa gtcttcggag acggagaagt atgatttgag
     3361 ctcgataaga gtggtgaaat ctggtgctgc tcctcttggt aaagaacttg aagatgccgt
     3421 taatgccaag tttcctaatg ccaaactcgg tcagggatac ggaatgacgg aagcaggtcc
     3481 agtgctagca atgtcgttag gttttgcaaa ggaacctttt ccggttaagt caggagcttg
     3541 tggtactgtt gtaagaaatg ctgagatgaa aatagttgat ccagacaccg gagattctct
     3601 ttcgaggaaa caacccggtg agatttgtat tcgtggtcac cagatcatga aaggttacct
     3661 caacaatccg gcagctacag cagagaccat tgataaagac ggttggcttc atactggaga
     3721 tattggattg atcgatgacg atgacgagct tttcatcgtt gatcgattga aagaacttgt
     3781 caagtataaa ggttttcagg tagctccggc tgagctagag gctttgctca tcggtcatcc
     3841 tgacattact gatgttgctg ttgtcgcaat gaaagaagaa gcagctggtg aagttcctgt
     3901 tgcatttgtg gtgaaatcga aggattcgga gttatcagaa gatgatgtga agcaattcgt
     3961 gtcgaaacag gttgtgtttt acaagagaat caacaaagtg ttcttcactg aatccattcc
     4021 taaagctcca tcagggaaga tattgaggaa agatctgagg gcaaaactag caaatggatt
     4081 gtaatggtac ccggggatcc tctagagtcg acctgcaggc atgcaagctt ggctgttttg
     4141 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga
     4201 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact
     4261 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga
     4321 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc
     4381 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac
     4441 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat
     4501 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca ctggcgtaat
     4561 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gggggggggg ggaaagccac
     4621 gttgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc atcatgaaca
     4681 ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca tattcaacgg
     4741 gaaacgtctt gctcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat
     4801 aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag
     4861 cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca
     4921 gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat
     4981 tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatccccgg gaaaacagca
     5041 ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg
     5101 ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta
     5161 tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt
     5221 gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca taagcttttg
     5281 ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt
     5341 gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac
     5401 caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg
     5461 ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg
     5521 ctcgatgagt ttttctaagc attggtaacc tggtatgagt cagcaacacc ttcttcacga
     5581 ggcagacctc agcgcccccc ccccccgcaa actattaact ggcgaactac ttactctagc
     5641 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     5701 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     5761 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     5821 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     5881 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     5941 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     6001 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagacccct taataagatg
     6061 atcttcttga gatcgttttg gtctgcgcgt aatctcttgc tctgaaaacg aaaaaaccgc
     6121 cttgcagggc ggtttttcga aggttctctg agctaccaac tctttgaacc gaggtaactg
     6181 gcttggagga gcgcagtcac caaaacttgt cctttcagtt tagccttaac cggcgcatga
     6241 cttcaagact aactcctcta aatcaattac cagtggctgc tgccagtggt gcttttgcat
     6301 gtctttccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcggactgaa
     6361 cggggggttc gtgcatacag tccagcttgg agcgaactgc ctacccggaa ctgagtgtca
     6421 ggcgtggaat gagacaaacg cggccataac agcggaatga caccggtaaa ccgaaaggca
     6481 ggaacaggag agcgcacgag ggagccgcca gggggaaacg cctggtatct ttatagtcct
     6541 gtcgggtttc gccaccactg atttgagcgt cagatttcgt gatgcttgtc aggggggcgg
     6601 agcctatgga aaaacggctt tgccgcggcc ctctcacttc cctgttaagt atcttcctgg
     6661 catcttccag gaaatctccg ccccgttcgt aagccatttc cgctcgccgc agtcgaacga
     6721 ccgagcgtag cgagtcagtg agcgaggaag cggaatatat cctgtatcac atattctgct
     6781 gacgcaccgg tgcagccttt tttctcctgc cacatgaagc acttcactga caccctcatc
     6841 agtgccaaca tagtaagcca gtatacactc cgctagcaag gagatggcgc ccaacagtcc
     6901 cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg
     6961 gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt
     7021 ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atccggagct tatcgactgc
     7081 acggtgcacc aatgcttctg gcgtcaggca gccatcggaa gctgtggtat ggctgtgcag
     7141 gtcgtaaatc actgcataat tcgtgtcgct caaggcgcac tcccgttctg gataatgttt
     7201 tttgcgccga catcataacg gttctggcaa atattctgaa atgagctgtt gacaattaat
     7261 catcggctcg tataatgtgt ggaattgtga gcggataaca atttcacaca ggaaa
//
