BUN21/pML300 (recipient strain)
(Bacterial strain #11675)

• Depositing Lab: Stephen Elledge
• Publication: Stegmeier et al Proc Natl Acad Sci U S A. 2005 Sep 13. 102(37):13212-7.

Backbone

• Vector backbone: F'
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Spectinomycin, 50 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): See comments.
• Growth instructions: Grow at 30'C. This plasmid contains a temperature sensitive mutant derivative of the pSC101 origin and will not replicate when bacteria are grown at 42'C.
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: Red-alpha, Red-beta, Gam
• Species: Bacteriophage lambda

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ sequencing primer: na

Resource Information

• Supplemental Documents:
  • Elledge-pPRIME

Terms and Licenses

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

Depositor Comments

pML300 carries the genes encoding the exo, beta, and gam recombination functions of bacteriophage lambda, as well as RecA, to render an E. coli strain recombination proficient. The lambda Red-alpha, Red-beta, and Gam proteins are under the rhaB promoter, induced by incubation with the sugar rhamnose. The bacterial strain is BW28705I (BUN21)/pML300 (recipient strain): lacIq rrnB3 delta-lacZ4787 hsdR514 delta-(araBAD)567 delta-(rhaBAD)568 galU95 delta-endA9::FRT delta-recA635::FRT umuC::ParaBAD-I-SceI-FRT + pML300(PrhaB-gam-beta-exo Ts(ori). Grow at 30'C in LB/Sp 50 ug/ml (spectinomycin resistant).