Description

This DNA is a pUC type vector possessing duplicated amber mutants on Kanamycin resistant gene. When transformed with this DNA, transformants of supE strains like JM109 can grow on plate containing Kanamycin, but transformants of sup ⁰ strains like MV1184 can not grow. This DNA is available to site-directed mutagenesis based on ODA method using this property.
Moreover, this DNA is available to cloning of a target gene and its expression using lac promoter when used on supE hosts like JM109, like as multi-cloning site as well as pUC18 /19 on lac Z gene, also Nde I restriction site (CATATG) at the position of initiation codon (ATG). It is possible to increase initiation efficiency of translation by using this Nde I site.

Storage

-20°C

Concentration

250-1,000 µg/ml

Chain length

2,204 bp

Purity

• Contains over 70% double-strand covalently closed circular form I (RF I).
• It is confirmed that the multi-cloning site is well-maintained upon sequence analysis by dideoxy method.
• It is confirmed that the vector contains a single cleavage site for EcoR I, Sac I, Kpn I, Sma I, BamH I, Xba I, Sal I, Pst I, Sph I and Hind III upon restriction analysis by the enzymes.

GenBank

	Entry Name	Accession No.
pKF18k	SYNAPG3G	D63846
pKF19k	SYNAPG3H	D63847

The sequence of pKF18k/19k registered at GenBank is 2,203 bp, lacks C before Nde I site. This product is 2,204 bp including C before Nde I site.

Intended usage

• Site-directed mutagenesis based on ODA method
• Cloning of a target gene and its expression using lac promoter
• DNA sequencing by using M13 primers
• Gene expression utilizing lac promoter

Form

10 mM Tris-HCl (pH8.0), 1 mM EDTA

Plasmid vector map of pKF18k-2/19k-2


The sequence of multi-cloning site of pKF18k-2/19k-2 is the same as pUC18/19.