LOCUS       40924_42489        7311 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pTac-VvSTS-At4CL3, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7311)
  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 7311)
  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 7311)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7311)
  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..7311
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             8..1690
                     /gene="4CL3"
                     /label=4CL3
                     /note="4-coumarate--CoA ligase 3 from Arabidopsis thaliana.
                     Accession#: Q9S777"
     terminator      1942..2028
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      2120..2147
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        2167..2258
                     /label=AmpR promoter
     primer_bind     complement(2571..2587)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2830..3642
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      4174..4719
                     /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        5154..5182
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    5190..5206
                     /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             5228..6403
                     /note="Stilbene synthase 2 from Vitis vinifera. Accession#:
                     P51070"
     terminator      complement(6658..6701)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      6833..6860
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     regulatory      7245..7273
                     /label=Tac promoter
                     /note="Tac promoter"
                     /regulatory_class="promoter"
     promoter        7245..7273
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    7281..7297
                     /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 atcactgcag ctctacacga acctcagatt cacaaaccaa ccgatacatc
       61 cgtcgtctcc gatgatgtgc ttcctcattc tcctcctacg cctcgcattt tccggtcaaa
      121 gcttccggac attgacatcc ctaaccacct ccctctccac acttactgct tcgaaaaact
      181 ctcatctgtt tccgacaagc cttgtctcat cgttggctcc accgggaaaa gctacaccta
      241 cggcgaaaca cacctaatct gtcggagagt cgcttccggg ttatacaagt taggtatcag
      301 aaaaggtgac gtcatcatga tccttctcca aaactcagcc gagttcgttt tctccttcat
      361 gggtgcttcc atgatcggag ccgtctcaac caccgcaaac cctttctaca cttctcaaga
      421 gctttataaa cagcttaaat cttccggagc caaactcata atcactcact ctcaatacgt
      481 cgataaactt aaaaacctcg gagaaaacct cacgctgatc actaccgatg aaccaacacc
      541 ggagaattgt ctaccattct cgacactcat caccgacgac gaaacaaacc catttcaaga
      601 aaccgtcgat atcggcggtg acgatgcggc ggcgttacca ttctcatcgg gaacaacagg
      661 gttaccaaaa ggagttgttt tgacacacaa gagcttaatc acaagcgttg cacaacaagt
      721 agatggagat aacccaaatc tttacctcaa atcaaacgac gtcatactct gcgttttacc
      781 acttttccat atctactctc tcaatagcgt cctcctcaat tcactccgat ccggcgccac
      841 ggttctttta atgcataagt ttgagatcgg tgcgttattg gatcttattc aaagacatag
      901 agtgacaata gcggcgcttg taccgccgct agtgatcgct ctagctaaaa acccgacggt
      961 taattcttat gatctctcct ccgttagatt cgttctttcc ggtgcagctc ctttaggtaa
     1021 agaacttcaa gatagtcttc gtcgccgtct ccctcaagcc atccttggcc agggatatgg
     1081 tatgacggag gcaggaccag tgttgtcaat gagccttggg ttcgctaaag aaccgatacc
     1141 gacaaaatca ggttcatgtg ggactgtggt ccgaaacgca gagcttaaag tggttcacct
     1201 tgagacacgt ctctctcttg gatacaacca acctggtgag atttgtatcc gcggtcaaca
     1261 gatcatgaaa gagtacttga acgatccaga agccacttca gcaacaatag acgaagaagg
     1321 ttggcttcac acaggagaca ttggatatgt tgatgaagat gatgagattt tcattgttga
     1381 tcgacttaaa gaagtcatca agttcaaagg ctttcaggtc cctccagccg agctagagag
     1441 tttgctaatc aatcaccatt caattgccga tgcagctgtt gttccgcaaa atgatgaagt
     1501 cgctggggaa gttccggtgg ctttcgtggt gcgatcaaat ggaaatgata tcactgaaga
     1561 agatgttaaa gaatatgttg ccaaacaggt ggtgttctat aagagattgc acaaggtctt
     1621 ctttgttgct tccattccta aatctccttc gggaaaaata ctaagaaagg acctcaaagc
     1681 taagctttgt taatggtacc cggggatcct ctagagtcga cctgcaggca tgcaagcttg
     1741 gctgttttgg cggatgagag aagattttca gcctgataca gattaaatca gaacgcagaa
     1801 gcggtctgat aaaacagaat ttgcctggcg gcagtagcgc ggtggtccca cctgacccca
     1861 tgccgaactc agaagtgaaa cgccgtagcg ccgatggtag tgtggggtct ccccatgcga
     1921 gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt
     1981 cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg
     2041 gatttgaacg ttgcgaagca acggcccgga gggtggcggg caggacgccc gccataaact
     2101 gccaggcatc aaattaagca gaaggccatc ctgacggatg gcctttttgc gtttctacaa
     2161 actctttttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     2221 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     2281 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     2341 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     2401 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     2461 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc
     2521 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtaattc actggccgtc
     2581 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     2641 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     2701 cagttgcggg ggggggggga aagccacgtt gtgtctcaaa atctctgatg ttacattgca
     2761 caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca
     2821 aggggtgtta tgagccatat tcaacgggaa acgtcttgct cgaggccgcg attaaattcc
     2881 aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt
     2941 gcgacaatct atcgattgta tgggaagccc gatgcgccag agttgtttct gaaacatggc
     3001 aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa
     3061 tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc
     3121 accactgcga tccccgggaa aacagcattc caggtattag aagaatatcc tgattcaggt
     3181 gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt
     3241 aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat
     3301 aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa
     3361 gtctggaaag aaatgcataa gcttttgcca ttctcaccgg attcagtcgt cactcatggt
     3421 gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt
     3481 ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt
     3541 gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat
     3601 atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaagcatt ggtaacctgg
     3661 tatgagtcag caacaccttc ttcacgaggc agacctcagc gccccccccc cccgcaaact
     3721 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     3781 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     3841 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     3901 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     3961 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     4021 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     4081 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     4141 ctgagcgtca gaccccttaa taagatgatc ttcttgagat cgttttggtc tgcgcgtaat
     4201 ctcttgctct gaaaacgaaa aaaccgcctt gcagggcggt ttttcgaagg ttctctgagc
     4261 taccaactct ttgaaccgag gtaactggct tggaggagcg cagtcaccaa aacttgtcct
     4321 ttcagtttag ccttaaccgg cgcatgactt caagactaac tcctctaaat caattaccag
     4381 tggctgctgc cagtggtgct tttgcatgtc tttccgggtt ggactcaaga cgatagttac
     4441 cggataaggc gcagcggtcg gactgaacgg ggggttcgtg catacagtcc agcttggagc
     4501 gaactgccta cccggaactg agtgtcaggc gtggaatgag acaaacgcgg ccataacagc
     4561 ggaatgacac cggtaaaccg aaaggcagga acaggagagc gcacgaggga gccgccaggg
     4621 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc accactgatt tgagcgtcag
     4681 atttcgtgat gcttgtcagg ggggcggagc ctatggaaaa acggctttgc cgcggccctc
     4741 tcacttccct gttaagtatc ttcctggcat cttccaggaa atctccgccc cgttcgtaag
     4801 ccatttccgc tcgccgcagt cgaacgaccg agcgtagcga gtcagtgagc gaggaagcgg
     4861 aatatatcct gtatcacata ttctgctgac gcaccggtgc agcctttttt ctcctgccac
     4921 atgaagcact tcactgacac cctcatcagt gccaacatag taagccagta tacactccgg
     4981 actgcacggt gcaccaatgc ttctggcgtc aggcagccat cggaagctgt ggtatggctg
     5041 tgcaggtcgt aaatcactgc ataattcgtg tcgctcaagg cgcactcccg ttctggataa
     5101 tgttttttgc gccgacatca taacggttct ggcaaatatt ctgaaatgag ctgttgacaa
     5161 ttaatcatcg gctcgtataa tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     5221 cagacatatg gcaagtgtcg aggaattccg caatgctcag cgcgcaaagg gtcccgccac
     5281 gatcctggcc attggaacag caacgcccga tcattgcgtg taccaatcgg actacgcgga
     5341 tttttacttc cgcgtgacaa agtcagaaca catgactgca cttaaaaaga aattcaaccg
     5401 tatttgcgat aagtccatga tcaagaaacg ctacatccac cttacagaag aaatgttaga
     5461 ggaacatcct aacattggag cctatatggc accttcatta aacattcgcc aggaaattat
     5521 taccgcggag gtccccaaac ttgggaagga ggccgcttta aaagctctta aggaatgggg
     5581 tcagcctaag tctaaaatca cgcatttggt cttctgcact acatctgggg tggagatgcc
     5641 cggagcagat tacaaattag cgaacttgtt gggacttgaa ccctccgtcc gtcgcgtcat
     5701 gctgtaccat cagggctgct atgccggagg gactgtgctg cgcactgcta aagacttagc
     5761 agaaaacaat gctggggctc gtgtcctggt tgtttgttcc gagatcaccg ttgtgacttt
     5821 ccgcgggcca agtgaggatg ctcttgattc cttagtcggg caagctcttt tcggcgatgg
     5881 ctcagcagcg gttattgtcg gctcagatcc agacattagc atcgagcgtc cactgtttca
     5941 actggtgtca gcggctcaaa cctttatccc gaacagcgca ggagccattg caggcaactt
     6001 acgcgaagtt ggattaacgt tccatctgtg gccgaatgtg cccaccctga ttagcgaaaa
     6061 tattgaaaag tgcttaactc aggcatttga tccgttgggc atcagcgact ggaattcctt
     6121 gttttggatc gcccatccag gggggcctgc gattttggac gccgtagaag ctaaactgaa
     6181 tcttgacaag aaaaaattgg aggcaacacg tcatgtgctg tccgagtatg gcaacatgtc
     6241 cagtgcatgc gtgcttttca tcttggacga gatgcgtaag aaaagtttaa aaggggaacg
     6301 tgctacgacc ggggagggtt tagactgggg agtgttgttt ggttttggtc ccgggctgac
     6361 tatcgaaaca gtggtacttc attccattcc gatggttacc aattaatggt acccggggat
     6421 cctctagagt cgacctgcag gcatgcaagc ttggctgttt tggcggatga gagaagattt
     6481 tcagcctgat acagattaaa tcagaacgca gaagcggtct gataaaacag aatttgcctg
     6541 gcggcagtag cgcggtggtc ccacctgacc ccatgccgaa ctcagaagtg aaacgccgta
     6601 gcgccgatgg tagtgtgggg tctccccatg cgagagtagg gaactgccag gcatcaaata
     6661 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac
     6721 gctctcctga gtaggacaaa tccgccggga gcggatttga acgttgcgaa gcaacggccc
     6781 ggagggtggc gggcaggacg cccgccataa actgccaggc atcaaattaa gcagaaggcc
     6841 atcctgacgg atggcctttt tgcgtttcta cacaaggaga tggcgcccaa cagtcccccg
     6901 gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc gaagtggcga
     6961 gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg
     7021 ccggtgatgc cggccacgat gcgtccggcg tagaggatcc ggagcttatc gactgcacgg
     7081 tgcaccaatg cttctggcgt caggcagcca tcggaagctg tggtatggct gtgcaggtcg
     7141 taaatcactg cataattcgt gtcgctcaag gcgcactccc gttctggata atgttttttg
     7201 cgccgacatc ataacggttc tggcaaatat tctgaaatga gctgttgaca attaatcatc
     7261 ggctcgtata atgtgtggaa ttgtgagcgg ataacaattt cacacaggaa a
//
