BW25113/pKD20      Source of Strain:
pKD20 Plasmid Markers/Mutations: repA101
, 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: Δ(araD-araB)567
, hsdR514Strain Comments:
- pKD20 sequence: GenBank AY048745. Send pKD46 instead. It is more efficient.
- pKD46 is more efficient than pKD20 for making gene disruptions. See BW25113/pKD46.
- This strain should be grown on rich medium with ampicillin (100ug/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.