EC1000
(Bacterial strain #71852)

• Purpose: RepA+ MC1000, KmR , carrying a single copy of the pWV01 repA gene in the glgB gene; host for pOR128-based plasmids
• Depositing Labs: Todd Klaenhammer, Jan Kok
• Publication: Leenhouts et al Mol Gen Genet. 1996 Nov 27;253(1-2):217-24.

Backbone

• Vector backbone: NA

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): This is a strain
• Growth instructions: Grow in 40 micrograms/mL kanamycin to maintain and provide chromosomal repA in trans for replication of pORI based plasmids
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: Carries a single chromosomal copy of the pWV01 repA gene in the glgB gene

Cloning Information

• Cloning method: Unknown
• 5′ sequencing primer: NA

Resource Information

• Supplemental Documents:
  • Verification gel for strain EC1000
• A portion of this plasmid was derived from a plasmid made by: Jan Kok

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

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
Strain Comments:
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.