BW25113/pKD46      Source of Strain:
B.L. WannerOther Designations:
pKD46 Plasmid Markers/Mutations: repA101
, [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: Δ(araD-araB)567
, hsdR514Strain Comments:
- pKD46 is more efficient than pKD20 for making gene disruptions. pKD46 sequence: GenBank AY048746. See also pKD78(CmR) and pKD119(TetR).
- The ara and rha deletions were published as Δ rhaBADLD78 and Δ araBADAH33
- This strain should be grown on rich media with ampicillin (100 ug/ml at 30 C).
- Δ(araD-araB)567-- This deletion extends from ~25 bp upstream of the araB start codon to ~8 bp into the beginning of the araD gene
- ΔlacZ4787(::rrnB-3)-- : 4 tandem copies of the rrnB transcriptional terminator inserted by gene replacement into the region extending from near the SacII site near the N-terminus of lacZ through the promoter.
- 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)
- Δ(araD-araB)567 was formerly called Δ araBADAH33
- Δ(rhaD-rhaB)568 was formerly called Δ rhaBADLD78
- ΔlacZ4787(::rrnB-3) was formerly called Δ lacZWJ16
- ΔlacZ4787(::rrnB-3) was formerly called ::rrnB-4
- ::rrnB-3 = 4 copies of rrnB inserted
- ApR = ampicillin resistant
- ts = Temperature Sensitive
- 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.