pET-Sgr_rppA_whiE vector (V006500)

Basic Vector Information

Vector Name:
pET-Sgr_rppA_whiE
Antibiotic Resistance:
Kanamycin
Length:
6887 bp
Type:
Cloning vector
Replication origin:
ori
Source/Author:
Kim B, Binkley R, Kim HU, Lee SY.

pET-Sgr_rppA_whiE vector Map

pET-Sgr_rppA_whiE6887 bp3006009001200150018002100240027003000330036003900420045004800510054005700600063006600f1 oriKanRoribomropCAP binding sitelacIlacI promoterT7 promoterlac operatorRBS1,3,6,8-tetrahydroxynaphthalene synthaseRBSwhiE_ORFII6xHisT7 terminator

pET-Sgr_rppA_whiE vector Sequence

LOCUS       V006500                 6887 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V006500
VERSION     V006500
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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6887)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Biotechnol.
            Bioeng. (2018) In press"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (14-JUN-2018) Dept. Chemical and Biomolecular Engineering, Korea
            Advanced Institute of Science and Technology, Daehak-ro 291, Daejeon
            34141, South Korea"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6887
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      12..467
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(563..1375)
                     /label="KanR"
                     /note="aminoglycoside phosphotransferase"
     rep_origin      1497..2085
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     misc_feature    complement(2271..2413)
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     CDS             complement(2518..2706)
                     /label="rop"
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     protein_bind    complement(3481..3502)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(3518..4597)
                     /label="lacI"
                     /note="lac repressor"
     promoter        complement(4598..4675)
                     /label="lacI promoter"
     promoter        4988..5006
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5007..5031
                     /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)."
     RBS             5046..5068
                     /label="RBS"
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5076..6191
                     /gene="rppA"
                     /label="1,3,6,8-tetrahydroxynaphthalene synthase"
                     /note="1,3,6,8-tetrahydroxynaphthalene synthase from
                     Streptomyces griseus. Accession#: Q54240"
     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
                     /label="6xHis"
                     /note="6xHis affinity tag"
     terminator      6815..6862
                     /label="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"

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