pET-AaPKS4 vector (V006510)

LOCUS       V006510                 6485 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V006510
VERSION     V006510
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6485)
  AUTHORS   Yang D, Kim WJ, Yoo SM, Choi JH, Ha SH, Lee MH, Lee SY.
  TITLE     Repurposing type III polyketide synthase as a malonyl-CoA biosensor
            for metabolic engineering in bacteria
  JOURNAL   Proc. Natl. Acad. Sci. U.S.A. (2018) In press
   PUBMED   30232266
REFERENCE   2  (bases 1 to 6485)
  AUTHORS   Yang D, Kim WJ, Yoo SM, Choi JH, Ha SH, Lee MH, Lee SY.
  TITLE     Novel Enzyme-coupled Malonyl-CoA Biosensor based on Type III
            Polyketide Synthase and Use of the same (KR 10-2018-0066323)
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 6485)
  AUTHORS   Yang D, Kim WJ, Yoo SM, Choi JH, Ha SH, Lee MH, Lee SY.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-JUL-2018) Dept. Chemical and Biomolecular Engineering,
            Korea Advanced Institute of Science and Technology, Daehak-ro 291,
            Daejeon 34141, South Korea
REFERENCE   4  (bases 1 to 6485)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 6485)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc. Natl.
            Acad. Sci. U.S.A. (2018) In press"
            SGRef: number: 2; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 3; type: "Journal Article"; journalName: "Submitted
            (19-JUL-2018) Dept. Chemical and Biomolecular Engineering, Korea
            Advanced Institute of Science and Technology, Daehak-ro 291, Daejeon
            34141, South Korea"
            SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6485
                     /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..6284
                     /gene="PKS4"
                     /label="Octaketide synthase 2"
                     /note="Octaketide synthase 2 from Aloe arborescens.
                     Accession#: C4NF90"
     CDS             6329..6346
                     /label="6xHis"
                     /note="6xHis affinity tag"
     terminator      6413..6460
                     /label="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"