CGSC Strain#: 14162
Strain Designation: BW25113/pWTZ594      Source of Strain:
Sex: F- Episome/Plasmid: pWTZ594
   Plasmid Markers/Mutations: bla(ApR), lacZp-GFPuv(gfp), ori[pMB1]
   Plasmid Comment: This plasmid was constructed by fusing the GFPuv gene behind the lacZ promoter in pBluescript KS+
Chromosomal Markers: Δ(araD-araB)567, ΔlacZ4787(::rrnB-3), λ^-, rph-1, Δ(rhaD-rhaB)568, hsdR514
Strain Comments:

• This is the parent strain for the Keio Collection of single gene knockouts that contains the GFPuv expressing pWTZ594 plasmid
• This strain was originally reported to be lacI^Q, but it was later shown to be lacI⁺ (Baba 2006 ref. Supplementary Table 1)
• 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
• gfp = green fluorescent protein

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.
• Baba, T., T. Ara, M. Hasegawa, Y. Takai, Y. Okumura, M. Baba, K.A. Datsenko, M. Tomita, B.L. Wanner, H. Mori 2006. Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants: the Keio collection. Mol Syst Biol 2:1-11