PurposeRepA+ MC1000, KmR , carrying a single copy of the pWV01 repA gene in the glgB gene; host for pOR128-based plasmids
Full plasmid sequence is not available for this item.
|Item||Catalog #||Description||Quantity||Price (USD)|
|Bacterial Strain||71852||Bacteria in agar stab||1||$75|
This material is available to academics and nonprofits only.
Growth in Bacteria
Growth Strain(s)This is a strain
Growth instructionsGrow in 40 micrograms/mL kanamycin to maintain and provide chromosomal repA in trans for replication of pORI based plasmids
Gene/Insert nameCarries a single chromosomal copy of the pWV01 repA gene in the glgB gene
- Cloning method Unknown
- 5′ sequencing primer NA (Common Sequencing Primers)
The strain was made with a chromosomal insertion into MC1000 from M.J. Casadaban (Casadaban, MJ, SN Cohen 1980. Analysis of gene control signals by DNA fusion and cloning in Escherichia coli. J. Mol. Biol. 138(2):179-207.):
Sex: F- Chromosomal Markers: Δ(araA-leu)7697, [araD139]B/r, Δ(codB-lacI)3, galK16, galE15(GalS), λ-, e14-, relA1, rpsL150(strR), spoT1, mcrB1
The ara region in this strain was transduced in from E. coliB.
Δ(araA-leu)7697-- Shown by Durfee et al. to be a deletion of the first 295 codons of hepA to the first 82 codons of fruR.
Δ(araA-leu)7697-- Gerdes et al. show this deletion to cover from 63.4 kb to 89 kb, from polB through fruR.
[araD139]B/r-- The thr-leu region was transduced from an E. coli B/r strain (SB3118) in early steps of strain derivation; ara or in some strains ara-leu or thr-leu are from B.
Δ(codB-lacI)3-- is a deletion of the entire lac operon, from the strain Hfr 3000 X74 of Jacob. Refs. on extent of deletion: Sung et al.1987,J.Bact.169:2639; Gerdes et al.2001
Δ(codB-lacI)3-- Shown by Ferrandez et al. 1997 to extend to mhpD
Δ(codB-lacI)3-- Shown by Durfee et al. to extend from codon 292 of yahG through mhpE
galK16-- An IS2 insertion resulting in the truncation of the protein at codon 56
galE15(GalS)-- G to A transition resulting in an S122F amino acid substitution
galE15(GalS)-- is Brenner's gal-U38, isolated by UV mutagenesis from his strain CA1.
e14--- : Loss of the e14 element makes the strain McrA-.
rpsL150(strR)-- T to C transition mutation resulting in a K43R amino acid substitution.
spoT1-- In addition to the 6bp insertion, there is a C to T transition resulting in an H257Y substitution.
Δ(araA-leu)7697 was formerly called Δ(hepA-fruR)7697
Δ(codB-lacI)3 was formerly called Δ(lac)3
Δ(codB-lacI)3 was formerly called Δlac74
Δ(codB-lacI)3 was formerly called DE(lac)6
Δ(codB-lacI)3 was formerly called X74
Δ(codB-lacI)3 was formerly called Δ(yahG-mhpE)
GalS = UDPGal accum.toxic if GalKRP+
strR = streptomycin resist.
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
For your Materials & Methods section:EC1000 was a gift from Todd Klaenhammer & Jan Kok (Addgene plasmid # 71852)
For your References section:A general system for generating unlabelled gene replacements in bacterial chromosomes. Leenhouts K, Buist G, Bolhuis A, ten Berge A, Kiel J, Mierau I, Dabrowska M, Venema G, Kok J. Mol Gen Genet. 1996 Nov 27;253(1-2):217-24. PubMed 9003306