CGSC Strain#: 14236
Strain Designation: BW25113/pKD46/pB15      Source of Strain: J.E. Wertz
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: Δ(araD-araB)567, ΔlacZ4787(::rrnB-3), λ^-, rph-1, Δ(rhaD-rhaB)568, hsdR514
Strain 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 rhaBAD_LD78 and Δ araBAD_AH33
• This strain also contains the conjugative pB15 plasmid from PET297
• 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 araBAD_AH33
• Δ(rhaD-rhaB)568 was formerly called Δ rhaBAD_LD78
• ΔlacZ4787(::rrnB-3) was formerly called ::rrnB-4
• ΔlacZ4787(::rrnB-3) was formerly called Δ lacZ_WJ16
• ::rrnB-3 = 4 copies of rrnB inserted
• 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.
• Turner, PE, ES Williams, C Okeke, VS Cooper, S Duffy, JE Wertz 2014. Antibiotic resistance correlates with transmission in plasmid evolution. Evolution 68(12):3368-80.