BW25141/pKD3      Source of Strain:
B.L. WannerOther Designations:
pKD3 Plasmid Markers/Mutations: oriR6Kγ
, bla Plasmid Comment:
template plasmid for gene disruption. The resistance gene is flanked by FRT sites. 2804 basepairs. GenBk Seq.AY48742 Chromosomal Markers: Δ(araD-araB)567
, hsdR514Strain Comments:
- This strain should be grown on rich medium with ampicillin (100 ug/ml), chloramphenicol (25 ug/ml), or both, at 37 C. pKD3 sequence: GenBank AY048742.
- 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.
- ΔuidA3::pir+-- Allows the replication of plasmids with an R6Kγ origin of replication
- recA1-- : Missense mutation, altered isoelectric point. Sequenced: G to A for Nuc. 720 (Gly 160 Asp).
- 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
- endA9(del-ins)::FRT was formerly called endABT333
- ΔlacZ4787(::rrnB-3) was formerly called Δ lacZWJ16
- ΔlacZ4787(::rrnB-3) was formerly called ::rrnB-4
- ΔuidA3::pir+ was formerly called uidA(Δ MluI)::pir+
- ::rrnB-3 = 4 copies of rrnB inserted
- del-ins = deletion-insertion
- ApR = ampicillin resistant
- Ter = terminator
- 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.