LOCUS       Exported                6887 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pET-Sgr_rppA_whiE, complete sequence.
ACCESSION   MH488907
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6887)
  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 6887)
  AUTHORS   Yang D, Kim WJ, Yoo SM, Choi JH, Ha SH, Lee MH, Lee SY.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-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 6887)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6887
                     /organism="Cloning vector pET-Sgr_rppA_whiE"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
                     /db_xref="taxon:2301146"
     rep_origin      12..467
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(560..1375)
                     /codon_start=1
                     /product="kanamycin resistance protein"
                     /label=kanamycin resistance protein
                     /protein_id="AXN69954.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      1497..2085
                     /label=pBR322
                     /note="pBR322"
     misc_feature    2271..2413
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(2515..2706)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3515..4597)
                     /codon_start=1
                     /gene="lacIq"
                     /product="lac repressor protein"
                     /label=lacIq
                     /protein_id="AXN69955.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     gene            complement(3515..4597)
                     /gene="lacIq"
                     /label=lacIq
     promoter        complement(4598..4675)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        4988..5006
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      4988..5004
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     protein_bind    5007..5031
                     /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)."
     RBS             5046..5068
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5076..6194
                     /codon_start=1
                     /gene="rppA"
                     /product="1,3,6,8-tetrahydroxynaphthalene synthase"
                     /label=rppA
                     /note="derived from Streptomyces griseus"
                     /protein_id="AXN69956.1"
                     /translation="MATLCRPAIAVPEHVITMQQTLDLARETHAGHPQRDLVLRLIQNT
                     GVQTRHLVQPIEKTLAHPGFEVRNQVYEAEAKTRVPEVVRRALANAETEPSEIDLIVYV
                     SCTGFMMPSLTAWIINSMGFRPETRQLPIAQLGCAAGGAAINRAHDFCVAYPDSNVLIV
                     SCEFCSLCYQPTDIGVGSLLSNGLFGDALSAAVVRGQGGTGMRLERNGSHLVPDTEDWI
                     SYAVRDTGFHFQLDKRVPGTMEMLAPVLLDLVDLHGWSVPNMDFFIVHAGGPRILDDLC
                     HFLDLPPEMFRYSRATLTERGNIASSVVFDALARLFDDGGAAESAQGLIAGFGPGITAE
                     VAVGSWAKEGLGADVGRDLDELELTAGVALSG"
     gene            5076..6194
                     /gene="rppA"
                     /label=rppA
     RBS             6208..6230
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             6235..6708
                     /codon_start=1
                     /gene="whiE_ORFII"
                     /product="monooxygenase"
                     /label=whiE_ORFII
                     /note="derived from Streptomyces coelicolor"
                     /protein_id="AXN69957.1"
                     /translation="MTDQQVRIVSLGDIAPNRRRGGDLRALLTPTTAGSTSGFMGVAIV
                     QPGDRIAEHYHPYSEEFVYVTEGALEVDLDGVPHSLGTGQGLLIPQDMRHRFRNAGDVE
                     ARLVFHLGPLAPRPELGHVDTEDAEGNALPCDGAHAGHAAGHERPAPAAEVVS"
     gene            6235..6708
                     /gene="whiE_ORFII"
                     /label=whiE_ORFII
     CDS             6731..6748
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      6815..6862
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"