pKF 18k-2 vector (V012455)

LOCUS       pKF_18k-2.        2204 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Bacterial cloning and site-directed mutagenesis vector with a 
            kanamycin resistance gene containing amber stop codons. The MCS is 
            reversed in pKF 19k-2.
ACCESSION   .
VERSION     .
KEYWORDS    pKF 18k-2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2204)
  AUTHORS   Hashimoto-Gotoh T, Yasojima K, Tsujimura A.
  TITLE     Plasmids with a kanamycin-resistance gene for site-directed 
            mutagenesis using the oligodeoxyribonucleotide-directed dual amber 
            method.
  JOURNAL   Gene 1995;167:333-4.
  PUBMED    8566803
REFERENCE   2  (bases 1 to 2204)
  AUTHORS   TaKaRa
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 2204)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Gene"; 
            date: "1995"; volume: "167"; pages: "333-4"
COMMENT     SGRef: number: 2; type: "Journal Article"
COMMENT     An NdeI site overlaps the lacZ-alpha start codon.
FEATURES             Location/Qualifiers
     source          1..2204
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    18..74
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(75..91)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(310..1125)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=aph(3')-Ia
                     /note="KanR (with amber stop codons)"
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSKPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLA*A*SRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      1450..2038
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2095..2116
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2131..2161
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2169..2185
                     /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)."