CGSC Strain#: 7668
Strain Designation: MG1655/pKD20      Source of Strain: B.L. Wanner
Sex: F- Episome/Plasmid: pKD20
   Plasmid Markers/Mutations: repA101(ts), araBp-gam-bet-exo, oriR101, bla(ApR), araC , bla
   Plasmid Comment: Lambda Red recombinase expression plasmid. This plasmid includes 2154 nucleotides (31088-33241) of phage Lambda. 6078 basepairs. LESS EFFICIENT than pKD46. Send pKD46. Genbank AY048745, pKD20.
Chromosomal Markers: λ^-, rph-1
Strain Comments:

• Grow this strain at 30C in presence of ampicillin (100ug/ml for rich media).
• pKD20 sequence: GenBank AY048745
• 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.