pWKS130 vector (V002133)

LOCUS       40924_46568        6811 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pWKS130, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6811)
  AUTHORS   Wang RF, Kushner SR.
  TITLE     Construction of versatile low-copy-number vectors for cloning, 
            sequencing and gene expression in Escherichia coli
  JOURNAL   Gene 100, 195-199 (1991)
  PUBMED    2055470
REFERENCE   2  (bases 1 to 6811)
  AUTHORS   Obrist MW, Miller VL.
  TITLE     Low copy expression vectors for use in Yersinia sp. and related 
            organisms
  JOURNAL   Plasmid 68 (1), 33-42 (2012)
  PUBMED    22445322
REFERENCE   3  (bases 1 to 6811)
  AUTHORS   Obrist MW, Miller VL.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-DEC-2011) Microbiology and Immunology, University of 
            North Carolina at Chapel Hill, 116 Manning Drive, 505 Mary Ellen 
            Jones, CB# 7290, Chapel Hill, NC 27599, USA
REFERENCE   4  (bases 1 to 6811)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 6811)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Gene 100, 
            195-199 (1991)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Plasmid"; 
            date: "2012"; volume: "68"; issue: "1"; pages: "33-42"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (14-DEC-2011) Microbiology and Immunology, University of North 
            Carolina at Chapel Hill, 116 Manning Drive, 505 Mary Ellen Jones, 
            CB# 7290, Chapel Hill, NC 27599, USA"
COMMENT     SGRef: number: 4; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Assembly Method :: DNASTAR Lasergene SeqMan Pro v. 7.2.1 (1) Intel 
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6811
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    240..261
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        276..306
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    314..330
                     /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)."
     primer_bind     338..354
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        375..393
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    complement(406..513)
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     promoter        complement(522..540)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(550..566)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      708..1136
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(2454..3266)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(4606..5553)
                     /codon_start=1
                     /label=Rep101
                     /note="RepA protein needed for replication with the pSC101 
                     origin"
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      complement(5601..5823)
                     /direction=LEFT
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"