LOCUS       Exported                4828 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pTac-His-Sgr_rppA, complete sequence.
ACCESSION   MH488951
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4828)
  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 4828)
  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 4828)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4828
                     /organism="Cloning vector pTac-His-Sgr_rppA"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
                     /db_xref="taxon:2301157"
     regulatory      175..203
                     /regulatory_class="promoter"
                     /note="Tac promoter"
     promoter        175..203
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    211..227
                     /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             246..1382
                     /codon_start=1
                     /gene="rppA"
                     /product="1,3,6,8-tetrahydroxynaphthalene synthase"
                     /label=rppA
                     /note="N-terminal 6XHis-tagged; derived from Streptomyces 
                     griseus"
                     /protein_id="AXN70078.1"
                     /translation="MHHHHHHATLCRPAIAVPEHVITMQQTLDLARETHAGHPQRDLVL
                     RLIQNTGVQTRHLVQPIEKTLAHPGFEVRNQVYEAEAKTRVPEVVRRALANAETEPSEI
                     DLIVYVSCTGFMMPSLTAWIINSMGFRPETRQLPIAQLGCAAGGAAINRAHDFCVAYPD
                     SNVLIVSCEFCSLCYQPTDIGVGSLLSNGLFGDALSAAVVRGQGGTGMRLERNGSHLVP
                     DTEDWISYAVRDTGFHFQLDKRVPGTMEMLAPVLLDLVDLHGWSVPNMDFFIVHAGGPR
                     ILDDLCHFLDLPPEMFRYSRATLTERGNIASSVVFDALARLFDDGGAAESAQGLIAGFG
                     PGITAEVAVGSWAKEGLGADVGRDLDELELTAGVALSG"
     gene            246..1382
                     /gene="rppA"
                     /label=rppA
     CDS             249..266
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     regulatory      1426..1573
                     /regulatory_class="terminator"
                     /note="rrnBT1T2"
     CDS             complement(1744..2535)
                     /codon_start=1
                     /product="spectinomycin resistance protein"
                     /label=spectinomycin resistance protein
                     /protein_id="AXN70079.1"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(2536..2627)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2675..3413)
                     /direction=LEFT
                     /label=CDF
                     /note="CDF"
     CDS             complement(3623..4705)
                     /codon_start=1
                     /gene="lacIq"
                     /product="lac repressor protein"
                     /label=lacIq
                     /protein_id="AXN70080.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     gene            complement(3623..4705)
                     /gene="lacIq"
                     /label=lacIq
     promoter        complement(4706..4783)
                     /gene="lacI"
                     /label=lacI promoter