CGSC Strain#: 7669
Strain Designation: MG1655/pKD46      Source of Strain: B.L. Wanner
Sex: F- Episome/Plasmid: pKD46
   Plasmid Markers/Mutations: repA101(ts), araBp-gam-bet-exo, oriR101, bla(ApR), araC , bla , [tL3]
   Plasmid Comment: Lambda Red recombinase expression plasmid -- includes 2154 nucleotides (31088-33241) of phage Lambda, which includes the terminator downstream frp, exo, tL3 and a small ORF.GenBk-AY048746
Chromosomal Markers: λ^-, rph-1
Strain Comments:

• pKD46 is more efficient than pKD20 for making gene disruptions. pKD46 sequence: GenBank AY048746
• The pKD46 plasmid seems to be less temperature sensitive when in this host. We are currently investigating this phenomenon.
• This strain should be grown on rich media with ampicillin (100 ug/ml) at 30 C.
• rph-1-- is a 1 bp deletion that results in frameshift over last 15 codons and has polar effect on pyrE leading to suboptimal pyrimidine levels on minimal medium.(Jensen 1993 JBact.175:3401)
• ApR = ampicillin resistant
• ts = Temperature Sensitive

References:

• Datsenko, KA, BL Wanner 2000. One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products. Proc. Natl. Acad. Sci. U.S.A. 97(12):6640-5.