LOCUS       40924_42474        5305 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pTac-Sc4CLm, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5305)
  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 5305)
  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 5305)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5305)
  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..5305
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             8..1576
                     /codon_start=1
                     /gene="Sc4CLm"
                     /product="4-coumarate:CoA ligase mutant"
                     /label=Sc4CLm
                     /note="A294G; derived from Streptomyces coelicolor"
                     /protein_id="AXN70034.1"
                     /translation="MFRSEYADVPPVDLPIHDAVLGGAAAFGSTPALIDGTDGTTLTYE
                     QVDRFHRRVAAALAETGVRKGDVLALHSPNTVAFPLAFYAATRAGASVTTVHPLATAEE
                     FAKQLKDSAARWIVTVSPLLSTARRAAELAGGVQEILVCDSAPGHRSLVDMLASTAPEP
                     SVAIDPAEDVAALPYSSGTTGTPKGVMLTHRQIATNLAQLEPSMPSAPGDRVLAVLPFF
                     HIYGLTALMNAPLRLGATVVVLPRFDLEQFLAAIQNHRITSLYVAPPIVLALAKHPLVA
                     DYDLSSLRYIVSGAAPLDARLAAACSQRLGLPPVGQAYGMTELSPGTHVVPLDAMADAP
                     PGTVGRLIAGTEMRIVSLTDPGTDLPAGESGEILIRGPQIMKGYLGRPDATAAMIDEEG
                     WLHTGDVGHVDADGWLFVVDRVKELIKYKGFQVAPAELEAHLLTHPGVADAAVVGAYDD
                     DGNEVPHAFVVRQPAAPGLAESEIMMYVAERVAPYKRVRRVTFVDAVPRAASGKILRRQ
                     LREPR"
     gene            8..1576
                     /gene="Sc4CLm"
                     /label=Sc4CLm
     terminator      1825..1911
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      2003..2030
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        2050..2141
                     /label=AmpR promoter
     primer_bind     complement(2454..2470)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2713..3525
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      4057..4602
                     /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        5239..5267
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    5275..5291
                     /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 ttccgcagcg agtacgcaga cgtcccgccc gtcgacctgc ccatccacga
       61 cgccgtgctc ggcggggccg ccgccttcgg gagcaccccg gcgctgatcg acggcaccga
      121 cggcaccacc ctcacctacg agcaggtgga ccggttccac cggcgcgtcg ccgccgccct
      181 cgccgagacc ggcgtgcgca agggcgacgt cctcgccctg cacagcccca acaccgtcgc
      241 cttccccctg gccttctacg ccgccacccg cgcgggcgcc tccgtcacca cggtgcatcc
      301 gctcgcgacg gcggaggagt tcgccaagca gctgaaggac agcgcggccc gctggatcgt
      361 caccgtctca ccgctcctgt ccaccgcccg ccgggccgcc gaactcgcgg gcggcgtcca
      421 ggagatcctg gtctgcgaca gcgcgcccgg tcaccgctcc ctcgtcgaca tgctggcctc
      481 gaccgcgccc gaaccgtccg tcgccatcga cccggccgag gacgtcgccg ccctgccgta
      541 ctcctcgggc accaccggca cccccaaggg cgtcatgctc acacaccggc agatcgccac
      601 caacctcgcc cagctcgaac cgtcgatgcc gtccgcgccc ggcgaccgcg tcctcgccgt
      661 gctgccgttc ttccacatct acggcctgac cgccctgatg aacgccccgc tccggctcgg
      721 cgccaccgtc gtggtcctgc cccgcttcga cctggagcag ttcctcgccg ccatccagaa
      781 ccaccgcatc accagcctgt acgtcgcccc gccgatcgtc ctggccctcg ccaaacaccc
      841 cctggtcgcc gactacgacc tctcctcgct gaggtacatc gtcagcggcg ccgccccgct
      901 cgacgcgcgt ctcgccgccg cctgctcgca gcggctcggc ctgccgcccg tcggccaggc
      961 ctacggcatg accgaactgt ccccgggcac ccacgtcgtc cccctggacg cgatggccga
     1021 cgcgccgccc ggcaccgtcg gcaggctcat cgcgggcacc gagatgcgca tcgtctccct
     1081 caccgacccg ggcacggacc tccccgccgg agagtccggg gagatcctca tccgcggccc
     1141 ccagatcatg aagggctacc tgggccgccc cgacgccacc gccgccatga tcgacgagga
     1201 gggctggctg cacaccgggg acgtcggaca cgtcgacgcc gacggctggc tgttcgtcgt
     1261 cgaccgcgtc aaggaactga tcaagtacaa gggcttccag gtggcccccg ccgaactgga
     1321 ggcccacctg ctcacccacc ccggcgtcgc cgacgcggcc gtcgtcggcg cctacgacga
     1381 cgacggcaac gaggtaccgc acgccttcgt cgtccgccag ccggccgcac ccggcctcgc
     1441 ggagagcgag atcatgatgt acgtcgccga acgcgtcgcc ccctacaaac gcgtccgccg
     1501 ggtcaccttc gtcgacgccg tcccccgcgc cgcctccggc aagatcctcc gccgacagct
     1561 cagggagccg cgataatggt acccggggat cctctagagt cgacctgcag gcatgcaagc
     1621 ttggctgttt tggcggatga gagaagattt tcagcctgat acagattaaa tcagaacgca
     1681 gaagcggtct gataaaacag aatttgcctg gcggcagtag cgcggtggtc ccacctgacc
     1741 ccatgccgaa ctcagaagtg aaacgccgta gcgccgatgg tagtgtgggg tctccccatg
     1801 cgagagtagg gaactgccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc
     1861 tttcgtttta tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccggga
     1921 gcggatttga acgttgcgaa gcaacggccc ggagggtggc gggcaggacg cccgccataa
     1981 actgccaggc atcaaattaa gcagaaggcc atcctgacgg atggcctttt tgcgtttcta
     2041 caaactcttt ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     2101 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     2161 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     2221 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     2281 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     2341 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag
     2401 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtaa ttcactggcc
     2461 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     2521 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     2581 caacagttgc gggggggggg ggaaagccac gttgtgtctc aaaatctctg atgttacatt
     2641 gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat
     2701 acaaggggtg ttatgagcca tattcaacgg gaaacgtctt gctcgaggcc gcgattaaat
     2761 tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca
     2821 ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat
     2881 ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg
     2941 gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta
     3001 ctcaccactg cgatccccgg gaaaacagca ttccaggtat tagaagaata tcctgattca
     3061 ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt
     3121 tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg
     3181 aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa
     3241 caagtctgga aagaaatgca taagcttttg ccattctcac cggattcagt cgtcactcat
     3301 ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat
     3361 gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc
     3421 ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat tgataatcct
     3481 gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaagc attggtaacc
     3541 tggtatgagt cagcaacacc ttcttcacga ggcagacctc agcgcccccc ccccccgcaa
     3601 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     3661 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     3721 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     3781 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     3841 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     3901 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     3961 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     4021 ccactgagcg tcagacccct taataagatg atcttcttga gatcgttttg gtctgcgcgt
     4081 aatctcttgc tctgaaaacg aaaaaaccgc cttgcagggc ggtttttcga aggttctctg
     4141 agctaccaac tctttgaacc gaggtaactg gcttggagga gcgcagtcac caaaacttgt
     4201 cctttcagtt tagccttaac cggcgcatga cttcaagact aactcctcta aatcaattac
     4261 cagtggctgc tgccagtggt gcttttgcat gtctttccgg gttggactca agacgatagt
     4321 taccggataa ggcgcagcgg tcggactgaa cggggggttc gtgcatacag tccagcttgg
     4381 agcgaactgc ctacccggaa ctgagtgtca ggcgtggaat gagacaaacg cggccataac
     4441 agcggaatga caccggtaaa ccgaaaggca ggaacaggag agcgcacgag ggagccgcca
     4501 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccaccactg atttgagcgt
     4561 cagatttcgt gatgcttgtc aggggggcgg agcctatgga aaaacggctt tgccgcggcc
     4621 ctctcacttc cctgttaagt atcttcctgg catcttccag gaaatctccg ccccgttcgt
     4681 aagccatttc cgctcgccgc agtcgaacga ccgagcgtag cgagtcagtg agcgaggaag
     4741 cggaatatat cctgtatcac atattctgct gacgcaccgg tgcagccttt tttctcctgc
     4801 cacatgaagc acttcactga caccctcatc agtgccaaca tagtaagcca gtatacactc
     4861 cgctagcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac
     4921 gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc
     4981 ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc
     5041 ggcgtagagg atccggagct tatcgactgc acggtgcacc aatgcttctg gcgtcaggca
     5101 gccatcggaa gctgtggtat ggctgtgcag gtcgtaaatc actgcataat tcgtgtcgct
     5161 caaggcgcac tcccgttctg gataatgttt tttgcgccga catcataacg gttctggcaa
     5221 atattctgaa atgagctgtt gacaattaat catcggctcg tataatgtgt ggaattgtga
     5281 gcggataaca atttcacaca ggaaa
//
