LOCUS       40924_44564        7684 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pTY13, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7684)
  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 7684)
  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 7684)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7684)
  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..7684
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        70..99
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    107..123
                     /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             143..1021
                     /gene="tyrC"
                     /label=tyrC
                     /note="Cyclohexadienyl dehydrogenase from Zymomonas mobilis
                     subsp. mobilis (strain ATCC 31821 / ZM4 / CP4). Accession#:
                     Q04983"
     terminator      1277..1363
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      1455..1482
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        1547..1575
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    1583..1599
                     /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             1622..2671
                     /gene="aroG"
                     /label=aroG
                     /note="Phospho-2-dehydro-3-deoxyheptonate aldolase, 
                     Phe-sensitive from Escherichia coli (strain K12). 
                     Accession#: P0AB91"
     CDS             2712..3233
                     /gene="aroL"
                     /label=aroL
                     /note="Shikimate kinase 2 from Escherichia coli (strain
                     55989 / EAEC). Accession#: B7L543"
     regulatory      3337..3662
                     /label=rrnBT1T2
                     /note="rrnBT1T2"
                     /regulatory_class="terminator"
     terminator      3457..3543
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      3635..3662
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(3978..4021)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      4153..4180
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        4200..4291
                     /label=AmpR promoter
     primer_bind     complement(4604..4620)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             4863..5675
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      6437..6982
                     /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      7618..7646
                     /label=Tac promoter
                     /note="Tac promoter"
                     /regulatory_class="promoter"
     promoter        7618..7646
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    7654..7670
                     /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 cagaattcca ttactcgcat ccattctcag gctgtctcgt ctcgtctcga tatattctga
       61 aatgagctgt tgacaattaa tcatccggct cgtataatgt gtggaattgt gagcggataa
      121 caatttcaca caggaaacag acatgaccgt ctttaagcat attgccatta tcggattagg
      181 actgatcggt tcctctgcgg cacgggcaac aaaggcctat tgtcctgatg taacggtcag
      241 tctctatgac aaaagcgaat ttgtccgcga cagagctaga gcgctcaatc tcggcgacaa
      301 tgtcaccgat gatattcaag atgcggttcg tgaggctgat ctggtgctgc tatgcgtgcc
      361 agtcagggca atgggtatcg tcgcggcagc gatggcaccg gcactgaaaa aagacgttat
      421 tatctgcgat acaggttcgg taaaagtcag cgttataaaa acgctgcaag acaatttacc
      481 caatcacatt attgttccca gccatccttt ggctgggact gaaaataacg gacccgatgc
      541 cggttttgct gaattattcc aagatcatcc tgttattttg acccccgatg cccatacacc
      601 agcacaggct atcgcctata tcgccgatta ttgggaagaa attggtgggc gtatcaatct
      661 gatgtcggcg gaacatcacg atcacgtttt ggcgcttacc agccatttgc ctcatgtcat
      721 tgcataccaa cttatcggga tggtatcggg ttatgagaaa aaaagccgga cacccattat
      781 gcgttattcg gcaggcagct ttcgggatgc gacgcgggta gcggcttcgg aaccgcgtct
      841 ctggcaagat attatgctgg aaaatgcgcc cgccctttta ccagtgctgg atcattttat
      901 cgcagatctc aaaaaattgc ggacagccat tgcttcgcaa gatggggatt atcttcttga
      961 gcatttcaaa gaatcgcaga aagcgcgttt agccttaaaa acagaccacg atattcaccc
     1021 ttaaagctcg gtacccgggg atcctctaga gtcgacctgc aggcatgcaa gcttggctgt
     1081 tttggcggat gagagaagat tttcagcctg atacagatta aatcagaacg cagaagcggt
     1141 ctgataaaac agaatttgcc tggcggcagt agcgcggtgg tcccacctga ccccatgccg
     1201 aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta
     1261 gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
     1321 tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt
     1381 gaacgttgcg aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag
     1441 gcatcaaatt aagcagaagg ccatcctgac ggatggcctt ttatcgctgg gagttcgtag
     1501 acggaaacaa acgcagaatc caagctgcat attctgaaat gagctgttga caattaatca
     1561 tcggctcgta taatgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagaatt
     1621 catgaattat cagaacgacg atttacgcat caaagaaatc aaagagttac ttcctcctgt
     1681 cgcattgctg gaaaaattcc ccgctactga aaatgccgcg aatacggttg cccatgcccg
     1741 aaaagcgatc cataagatcc tgaaaggtaa tgatgatcgc ctgttggttg tgattggccc
     1801 atgctcaatt catgatcctg tcgcggcaaa agagtatgcc actcgcttgc tggcgctgcg
     1861 tgaagagctg aaagatgagc tggaaatcgt aatgcgcgtc tattttgaaa agccgcgtac
     1921 cacggtgggc tggaaagggc tgattaacga tccgcatatg gataatagct tccagatcaa
     1981 cgacggtctg cgtatagccc gtaaattgct gcttgatatt aacgacagcg gtctgccagc
     2041 ggcaggtgag tttctcaata tgatcacccc acaatatctc gctgacctga tgagctgggg
     2101 cgcaattggc gcacgtacca ccgaatcgca ggtgcaccgc gaactggcat cagggctttc
     2161 ttgtccggtc ggcttcaaaa atggcaccga cggtacgatt aaagtggcta tcgatgccat
     2221 taatgccgcc ggtgcgccgc actgcttcct gtccgtaacg aaatgggggc attcggcgat
     2281 tgtgaatacc agcggtaacg gcgattgcca tatcattctg cgcggcggta aagagcctaa
     2341 ctacagcgcg aagcacgttg ctgaagtgaa agaagggctg aacaaagcag gcctgccagc
     2401 acaggtgatg atcgatttca gccatgctaa ctcgtccaaa caattcaaaa agcagatgga
     2461 tgtttgtgct gacgtttgcc agcagattgc cggtggcgaa aaggccatta ttggcgtgat
     2521 ggtggaaagc catctggtgg aaggcaatca gagcctcgag agcggggagc cgctggccta
     2581 cggtaagagc atcaccgatg cctgcatcgg ctgggaagat accgatgctc tgttacgtca
     2641 actggcgaat gcagtaaaag cgcgtcgcgg gtaaccaggc atcaaataaa acgactagaa
     2701 ggaggttatt catgacacaa cctctttttc tgatcgggcc tcggggctgt ggtaaaacaa
     2761 cggtcggaat ggcccttgcc gattcgctta accgtcggtt tgtcgatacc gatcagtggt
     2821 tgcaatcaca gctcaatatg acggtcgcgg agatcgtcga aagggaagag tgggcgggat
     2881 ttcgcgccag agaaacggcg gcgctggaag cggtaactgc gccatccacc gttatcgcta
     2941 caggcggcgg cattattctg acggaattta atcgtcactt catgcaaaat aacgggatcg
     3001 tggtttattt gtgtgcgcca gtatcagtcc tggttaaccg actgcaagct gcaccggaag
     3061 aagatttacg gccaacctta acgggaaaac cgctgagcga agaagttcag gaagtgctgg
     3121 aagaacgcga tgcgctatat cgcgaagttg cgcatattat catcgacgca acaaacgaac
     3181 ccagccaggt gatttctgaa attcgcagcg ccctggcaca gacgatcaat tgttgactgc
     3241 aggcatgcaa gcttggctgt tttggcggat gagagaagat tttcagcctg atacagatta
     3301 aatcagaacg cagaagcggt ctgataaaac agaatttgcc tggcggcagt agcgcggtgg
     3361 tcccacctga ccccatgccg aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg
     3421 ggtctcccca tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg
     3481 aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca
     3541 aatccgccgg gagcggattt gaacgttgcg aagcaacggc ccggagggtg gcgggcagga
     3601 cgcccgccat aaactgccag gcatcaaatt aagcagaagg ccatcctgac ggatggcctt
     3661 ttgcagcact gaaggtcctc aatcgcactg gaaacatcaa ggtcgtacgt agtaccagga
     3721 tacatagatt accacaactc cgagcccttc caccctgcag gcatgcaagc ttggctgttt
     3781 tggcggatga gagaagattt tcagcctgat acagattaaa tcagaacgca gaagcggtct
     3841 gataaaacag aatttgcctg gcggcagtag cgcggtggtc ccacctgacc ccatgccgaa
     3901 ctcagaagtg aaacgccgta gcgccgatgg tagtgtgggg tctccccatg cgagagtagg
     3961 gaactgccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta
     4021 tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccggga gcggatttga
     4081 acgttgcgaa gcaacggccc ggagggtggc gggcaggacg cccgccataa actgccaggc
     4141 atcaaattaa gcagaaggcc atcctgacgg atggcctttt tgcgtttcta caaactcttt
     4201 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     4261 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     4321 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     4381 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     4441 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     4501 taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag agcaactcgg
     4561 tcgccgcata cactattctc agaatgactt ggttgagtaa ttcactggcc gtcgttttac
     4621 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     4681 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     4741 gggggggggg ggaaagccac gttgtgtctc aaaatctctg atgttacatt gcacaagata
     4801 aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg
     4861 ttatgagcca tattcaacgg gaaacgtctt gctcgaggcc gcgattaaat tccaacatgg
     4921 atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa
     4981 tctatcgatt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta
     5041 gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc
     5101 ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg
     5161 cgatccccgg gaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata
     5221 ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc
     5281 cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg aataacggtt
     5341 tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga
     5401 aagaaatgca taagcttttg ccattctcac cggattcagt cgtcactcat ggtgatttct
     5461 cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag
     5521 tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt
     5581 ctccttcatt acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata
     5641 aattgcagtt tcatttgatg ctcgatgagt ttttctaatc agaattggtt aattggttgt
     5701 aacactggca gagcattacg ctgacttgac gggacggcgg ctttgttgaa taaatcgaac
     5761 ttttgctgag ttgaaggatc agatcacgca tcttcccgac aacgcagacc gttccgtggc
     5821 aaagcaaaag ttcaaaatca ccaactggtc cacctacaac aaagctctca tcaaccgtgg
     5881 ctccctcact ttctggctgg atgatggggc gattcaggcc tggtatgagt cagcaacacc
     5941 ttcttcacga ggcagacctc agcgcccccc ccccccgcaa actattaact ggcgaactac
     6001 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     6061 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     6121 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     6181 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     6241 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     6301 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     6361 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagacccct
     6421 taataagatg atcttcttga gatcgttttg gtctgcgcgt aatctcttgc tctgaaaacg
     6481 aaaaaaccgc cttgcagggc ggtttttcga aggttctctg agctaccaac tctttgaacc
     6541 gaggtaactg gcttggagga gcgcagtcac caaaacttgt cctttcagtt tagccttaac
     6601 cggcgcatga cttcaagact aactcctcta aatcaattac cagtggctgc tgccagtggt
     6661 gcttttgcat gtctttccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     6721 tcggactgaa cggggggttc gtgcatacag tccagcttgg agcgaactgc ctacccggaa
     6781 ctgagtgtca ggcgtggaat gagacaaacg cggccataac agcggaatga caccggtaaa
     6841 ccgaaaggca ggaacaggag agcgcacgag ggagccgcca gggggaaacg cctggtatct
     6901 ttatagtcct gtcgggtttc gccaccactg atttgagcgt cagatttcgt gatgcttgtc
     6961 aggggggcgg agcctatgga aaaacggctt tgccgcggcc ctctcacttc cctgttaagt
     7021 atcttcctgg catcttccag gaaatctccg ccccgttcgt aagccatttc cgctcgccgc
     7081 agtcgaacga ccgagcgtag cgagtcagtg agcgaggaag cggaatatat cctgtatcac
     7141 atattctgct gacgcaccgg tgcagccttt tttctcctgc cacatgaagc acttcactga
     7201 caccctcatc agtgccaaca tagtaagcca gtatacactc cgctagcaag gagatggcgc
     7261 ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga
     7321 gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa
     7381 ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atccgggctt
     7441 atcgactgca cggtgcacca atgcttctgg cgtcaggcag ccatcggaag ctgtggtatg
     7501 gctgtgcagg tcgtaaatca ctgcataatt cgtgtcgctc aaggcgcact cccgttctgg
     7561 ataatgtttt ttgcgccgac atcataacgg ttctggcaaa tattctgaaa tgagctgttg
     7621 acaattaatc atcggctcgt ataatgtgtg gaattgtgag cggataacaa tttcacacag
     7681 gaaa
//
