LOCUS       40924_42459        9940 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pTac-Pg6MSAS-sfp, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9940)
  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 9940)
  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 9940)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9940)
  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..9940
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             9..5330
                     /note="6-methylsalicylic acid synthase from Penicillium
                     patulum. Accession#: P22367"
     promoter        5451..5479
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    5487..5503
                     /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             5526..6197
                     /gene="sfp"
                     /label=sfp
                     /note="4'-phosphopantetheinyl transferase Sfp from Bacillus
                     subtilis (strain 168). Accession#: P39135"
     misc_feature    6201..6257
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     terminator      6460..6546
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      6638..6665
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        6685..6776
                     /label=AmpR promoter
     primer_bind     complement(7089..7105)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             7348..8160
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      8692..9237
                     /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      9874..9902
                     /label=Tac promoter
                     /note="Tac promoter"
                     /regulatory_class="promoter"
     promoter        9874..9902
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    9910..9926
                     /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 cagaattcat gcattccgct gcaacttcta catacccctc tgggaaaaca tccccagcac
       61 cagtcggaac ccctgggact gagtacagtg aatatgaatt ctccaacgat gtggcggtag
      121 tgggaatggc ctgccgcgta gcgggaggta accacaaccc tgaacttctc tggcaatcgc
      181 tcctcagcca gaagagtgca atgggtgaga ttccacccat gcggtgggag ccttactatc
      241 gtcgggatgc acgaaatgag aaattcctca agaacacaac gtcgcgtggc tatttcctag
      301 atcgcttgga ggactttgac tgtcagttct ttggcatctc ccctaaggag gccgagcaga
      361 tggaccccca acagcgagtc tctctggaag tggcatcgga agccctggaa gatgccggta
      421 ttcctgccaa gagtctgtct gggagtgaca cagcagtctt ctggggtgtt aactcagacg
      481 actactccaa gctagtgcta gaggatctgc caaatgtcga ggcgtggatg ggcattggca
      541 ctgcatactg tggcgtgccc aatcgcatct catatcacct caacttgatg ggtcctagca
      601 ccgccgtgga tgcggcgtgt gcttcgtctc tcgtcgccat acatcatgga gttcaggcca
      661 tccgactggg agaatcaaag gtggctattg tcggaggtgt caatgccctc tgcggcccag
      721 gactgacccg cgtcctcgac aaagccggcg ccatctcgtc tgacggatct tgcaagtcct
      781 tcgatgatga cgcccatggg tacgctagag gtgaaggtgc cggcgccctc gtcctcaaaa
      841 gtctgcaccg tgcattgctc gaccacgata atgtcctggc agtcattaag ggcagtgcgg
      901 tatgccagga tggcaagacc aacggcatca tggcccccaa ttccgtggcg caacagctag
      961 cagcaaacaa tgctctctct gctgcaaata tcgaccctca caccgtgcgc tatgtggaag
     1021 cacatgccac atccacgccc ttgggcgacc caactgaaat atccgccatt gcaagtgtct
     1081 acggcgccga ccgccccgcg gatgatccat gctacatcgg gtccatcaag cccaatattg
     1141 gtcacctgga ggctggtgcg ggagtcatgg gcttcatcaa agcagttctg gccatccaaa
     1201 agggcgtcct gccaccgcag gccaacttga caaagctcaa tagccggatc gactggaaga
     1261 cagccggtgt taaggtcgtt caagaggcca ccccgtggcc cgaatcagac cccatccgta
     1321 gagcaggtgt ctgctcgtat gggtacggag gtactgtttc tcacgcggtg atcgaagaat
     1381 tcagccctat cctgcagcca gacccactag gtaatggagc agtcagcgga ccaggccttc
     1441 ttttattgtc cggaccccag gagaaacgtt tagccttgca ggccaaaaca ttgcgcgact
     1501 ggatgactgc cgaaggaaag gatcacaatt tgagtgatat tctcactact ctagctactc
     1561 gacgagatca ccacgactac cgagcggcgc ttgtagttga tgactaccgt gatgctgagc
     1621 aagtcctgca atcactggcc aatggggtcg accatacatt cacgacccag agccgtgtac
     1681 tgggctcaga catcagcaag gacgttgtct gggtgttttc tgggcacggt gcgcagtggc
     1741 cggacatggg caagcaactt attcacaacc cggtcttctt cgcagccatc cagccactcg
     1801 atgagttgat ccaagccgag attgggctgt cccctattga gctactccgg acgggtgact
     1861 ttgagtcgtc tgatcgggta cagatcctta cttatgtcat gcaaattggt ctcagcgccc
     1921 tgcttcaaag caatggcatc actccacagg ccgtgattgg acactccgtg ggcgaaatcg
     1981 cggccagtgt tgtggctgga gcactatccc cagcagaggg tgcgcttatt gtcacccgca
     2041 gagcgttgct gtaccgccag gttatgggca agggcggcat gatccttgtc aacctcccta
     2101 gtgccgagac tgaagagatc ttaggttccc gatcagacct ggttgtggcc attgactcgt
     2161 cgccatcttc ctgtgtggtg gctggagata aggagcttgt ggccgaaaca gcagaggctc
     2221 tcaaggcacg aggcgtgaag acattcactg tgaagagtga tattgccttc cacagtccaa
     2281 ccttgaatgg gctggtcgat cctcttcggg atgtgctcgc agagactctg tcccctgtga
     2341 gtcccaacgt caagttgtat tccaccgcac tggcagaccc ccgcggccaa gacctgcgcg
     2401 acgtggaata ctgggccggc aacatggtca accgcgtgcg tctcacttcg gctgtcaagg
     2461 ccgctgttga ggacggatat cgtctcttcc ttgaagtctc aacccaccca gtggtgtctc
     2521 actctatcaa cgagacacta atggatgcag gcatggaaga ctttgcggtc atcccaactc
     2581 tccttcggaa aaagccaact gagaagcata tcctacacag cattgcacag cttcactgcc
     2641 gcggtgcgga agtgaactgg gcggcacaga tgcccggacg ctgggccact ggagttccca
     2701 ccacgacctg gatgcataaa cccatctggc gcaagataga gactgcgccg cttcacactg
     2761 gtctcacgca tgatgtcgag aagcacacac ttctgggaca gcgtatccca gtgcccggca
     2821 ccgacaccta tgtctacaca acccggctgg ataacgacac aaagcccttc ccgggcagcc
     2881 accctctgca cggcaccgag atcgttccgg ctgcaggttt gatcaatact ttcttgaagg
     2941 gcacaggcgg ccagatgctg cagaacgttg ttctccgcgt tcccgttgct attaacgcac
     3001 cacgatccgt ccaggtcgta gtgcagcagg atcaagtcaa ggttgtgtct cgactgatcc
     3061 cgagtgagcc ctcgcagttg gatgacgatg cctcctgggt cactcacacc actgcatact
     3121 gggatcggaa ggttgctggt tccgaagatc gcatcgactt cgctgccgtc aaatcgcgac
     3181 tggtaaccaa actagcagac aacttttcga ttgactacct ggacaaggtg ggtgtatctg
     3241 ccatgggatt cccatgggcg gtaacagagc actaccgcaa tgacaaggag atgctggcac
     3301 gtgttgatgt caatcctgcc atctcgggag atgcccccct gccctgggat tcttcatcct
     3361 gggccccagt tctcgacgct gccacctccg tcggttcaac catcttcccg acacctgcac
     3421 ttcggatgcc cgcccagatt gagcgggtag aggtcttcac ctcgcaggat ccgcccaaga
     3481 tcagctggct ctacgtccag gaggcctccg attcggtgcc cacctctcac gttagcgtgg
     3541 tgagcgaggc gggtgaggtc cttgccaaat tcacagccat gcggttctcc gagattgaag
     3601 gcactcctgg cgtcagtggc agcatggaga gtctcgtgca ccagatcgct tggccaccag
     3661 ccaccccggc cgaagagccc ttgtccatcg agacagtgat cctggtctcg cccgatgcta
     3721 ccaccagggc cctgtatgcc gcaagtctgc cgacccgggt taactccttc cagttctcct
     3781 ctacacagga gttcttcagc aacgcttcat cccttcccct ggagaaggga accgtggtga
     3841 cctacatccc cggtgaggtc gcctcgctgg ccgaggttcc cgccgcatcc gagtcattca
     3901 cttggaactt gctcgaacta attaaattca ccgtcaatgg ctcactgcct atcaaagtct
     3961 tcactcttac cgcgaacatc ggcgagggtc aaacacctac cgctctagcc cagtccccat
     4021 tgtacggcct ggcacgggtc atcgccagtg agcaccctga tctaggaacc ctcatcgacg
     4081 ttgaagagcc cgtcatcccc ctgtcgacga tgcgatacat ccagggggca gacatcatcc
     4141 gaatcaacga cggaatcgct cgcacctccc ggttccgcag cctgccgcgc aacaagctgc
     4201 tccctgccag cgaaggtcct cgcctgcttc cccgccccga gggcacatac ctgatcaccg
     4261 gtggtcttgg cgtcctcggt ctcgaggtcg ctgacttcct ggtcgagaag ggtgccagac
     4321 gtctgctgct catctcgcga cgcgcccttc cccctcgccg aacctgggac caggttagcg
     4381 aggatctcca gccgaccatc gccaagatcc gcctcctgga gagtcgcggc gcctcggtcc
     4441 atgtcctgcc cctggacatc accaaaccgg atgcggtcga gcagcttacc acggcgttgg
     4501 accgactttc cctaccttcg gtccagggtg tcgttcacgc tgccggtgtg ctggacaacg
     4561 agctagtgat gcagaccact cgcgacgcct tcaaccgggt tctcgcaccc aagattgcgg
     4621 gcgcgctagc cctgcatgaa gtcttccccc ctaagagcgt cgacttcttc gtcatgttct
     4681 cctcttgtgg aaacctcgtc gggttcactg gtcaagcttc ctacggcagt ggtaacgcct
     4741 tcttggatac actagccacc caccgtgcgc gcctaggtga cgcagccgtg tccttccagt
     4801 ggacatcgtg gcgtggcctt ggcatgggtg ccagcacaga ctttatcaac gctgaattgg
     4861 aatccaaagg aatcacggac gtgacgcgcg atgaggcctt tgctgcatgg cagcatctcg
     4921 ccaagtatga tatggaccac ggagttgttt tgcgaagccg tgcctttgag gacggggaac
     4981 cgattccagt gtctattctc aatgatatcg cggtgcgacg cgtgggcact gtgtctaaca
     5041 cctcgccggc ggctgccggt tccagcgacg cggtgcctac gtctggaccg gagcttaagg
     5101 cttatttgga cgagaagatc aggggctgtg tggccaaggt gttacagatg actgctgagg
     5161 atgtggattc aaaggcagct ctggccgatc tgggtgttga ctcagtcatg acagtcactc
     5221 tgcgtcgcca gctgcagcta acactgaaga ttgcggtgcc ccctacattg acgtggagtc
     5281 atccgactgt cagccatctg gcggtgtggt ttgctgagaa gcttgccaaa taatgagctc
     5341 ggtacccggg gatcctctag agtcgacctg caggcatgcc actcccgttc tggataatgt
     5401 tttttgcgcc gacatcataa cggttctggc aaatattctg aaatgagctg ttgacaatta
     5461 atcatcggct cgtataatgt gtggaattgt gagcggataa caatttcaca caggaaacag
     5521 aattcatgaa gatttacgga atttatatgg accgcccgct ttcacaggaa gaaaatgaac
     5581 ggttcatgtc tttcatatca cctgaaaaac gggagaaatg ccggagattt tatcataaag
     5641 aagatgctca ccgcaccctg ctgggagatg tgctcgttcg ctcagtcata agcaggcagt
     5701 atcagttgga caaatccgat atccgcttta gcacgcagga atacgggaag ccgtgcatcc
     5761 ctgatcttcc cgacgctcat ttcaacattt ctcactccgg acgctgggtc atttgcgcgt
     5821 ttgattcaca gccgatcggc atagatatcg aaaaaacgaa accgatcagc cttgagatcg
     5881 ccaagcgctt cttttcaaaa acagagtaca gcgacctttt agcaaaagac aaggacgagc
     5941 agacagacta tttttatcat ctatggtcaa tgaaagaaag ctttatcaaa caggaaggca
     6001 aaggcttatc gcttccgctt gattcctttt cagtgcgcct gcaccaggac ggacaagtat
     6061 ccattgagct tccggacagc cattccccat gctatatcaa aacgtatgag gtcgatcccg
     6121 gctacaaaat ggctgtatgc gccgtacacc ctgatttccc cgaggatatc acaatggtct
     6181 cgtacgaaga gcttttataa gaattcgagc tcggtacccg gggatcctct agagtcgacc
     6241 tgcaggcatg caagcttggc tgttttggcg gatgagagaa gattttcagc ctgatacaga
     6301 ttaaatcaga acgcagaagc ggtctgataa aacagaattt gcctggcggc agtagcgcgg
     6361 tggtcccacc tgaccccatg ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg
     6421 tggggtctcc ccatgcgaga gtagggaact gccaggcatc aaataaaacg aaaggctcag
     6481 tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg
     6541 acaaatccgc cgggagcgga tttgaacgtt gcgaagcaac ggcccggagg gtggcgggca
     6601 ggacgcccgc cataaactgc caggcatcaa attaagcaga aggccatcct gacggatggc
     6661 ctttttgcgt ttctacaaac tctttttgtt tatttttcta aatacattca aatatgtatc
     6721 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     6781 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     6841 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     6901 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     6961 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg
     7021 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     7081 agtaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     7141 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     7201 accgatcgcc cttcccaaca gttgcggggg gggggggaaa gccacgttgt gtctcaaaat
     7261 ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa actgtctgct
     7321 tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac gtcttgctcg
     7381 aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat
     7441 aatgtcgggc aatcaggtgc gacaatctat cgattgtatg ggaagcccga tgcgccagag
     7501 ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga
     7561 ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct
     7621 gatgatgcat ggttactcac cactgcgatc cccgggaaaa cagcattcca ggtattagaa
     7681 gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg
     7741 cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag
     7801 gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat
     7861 ggctggcctg ttgaacaagt ctggaaagaa atgcataagc ttttgccatt ctcaccggat
     7921 tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta
     7981 ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc
     8041 ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat
     8101 ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc
     8161 taagcattgg taacctggta tgagtcagca acaccttctt cacgaggcag acctcagcgc
     8221 cccccccccc cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     8281 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     8341 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     8401 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     8461 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     8521 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     8581 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     8641 acgtgagttt tcgttccact gagcgtcaga ccccttaata agatgatctt cttgagatcg
     8701 ttttggtctg cgcgtaatct cttgctctga aaacgaaaaa accgccttgc agggcggttt
     8761 ttcgaaggtt ctctgagcta ccaactcttt gaaccgaggt aactggcttg gaggagcgca
     8821 gtcaccaaaa cttgtccttt cagtttagcc ttaaccggcg catgacttca agactaactc
     8881 ctctaaatca attaccagtg gctgctgcca gtggtgcttt tgcatgtctt tccgggttgg
     8941 actcaagacg atagttaccg gataaggcgc agcggtcgga ctgaacgggg ggttcgtgca
     9001 tacagtccag cttggagcga actgcctacc cggaactgag tgtcaggcgt ggaatgagac
     9061 aaacgcggcc ataacagcgg aatgacaccg gtaaaccgaa aggcaggaac aggagagcgc
     9121 acgagggagc cgccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     9181 cactgatttg agcgtcagat ttcgtgatgc ttgtcagggg ggcggagcct atggaaaaac
     9241 ggctttgccg cggccctctc acttccctgt taagtatctt cctggcatct tccaggaaat
     9301 ctccgccccg ttcgtaagcc atttccgctc gccgcagtcg aacgaccgag cgtagcgagt
     9361 cagtgagcga ggaagcggaa tatatcctgt atcacatatt ctgctgacgc accggtgcag
     9421 ccttttttct cctgccacat gaagcacttc actgacaccc tcatcagtgc caacatagta
     9481 agccagtata cactccgcta gcaaggagat ggcgcccaac agtcccccgg ccacggggcc
     9541 tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc
     9601 cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc
     9661 ggccacgatg cgtccggcgt agaggatccg gagcttatcg actgcacggt gcaccaatgc
     9721 ttctggcgtc aggcagccat cggaagctgt ggtatggctg tgcaggtcgt aaatcactgc
     9781 ataattcgtg tcgctcaagg cgcactcccg ttctggataa tgttttttgc gccgacatca
     9841 taacggttct ggcaaatatt ctgaaatgag ctgttgacaa ttaatcatcg gctcgtataa
     9901 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
//
