pLA230
(Plasmid #11724)

• Depositing Lab: Lawrence Loeb
• Publication: Shinkai et al J Biol Chem. 2001 Dec 14. 276(50):46759-64.

Backbone

• Vector backbone: modified pGPS3
• Backbone manufacturer: New England Biolabs
• Backbone size w/o insert (bp): 2642
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: b-lactamase
• Alt name: beta-lactamase
• Alt name: ampicillin resistance
• Alt name: carbenicillin resistance
• Species: E. coli
• Insert Size (bp): 979
• Mutation: GAA for Glu26 changed to TAA (premature stop codon introduced in beta-lactamase gene).

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: AflIII (not destroyed)
• 3′ cloning site: KpnI (not destroyed)
• 5′ sequencing primer: n/a

Resource Information

• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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

Depositor Comments

Positive control for mutagenesis experiments: reversion of the stop codon results in ampicillin resistance.

The pGPS3 has a deletion from BglI to BglII that destroys the ampicillin resistance gene. The ampicillin resistance gene containing the stop codon was then cloned in between AflIII and KpnI.

Please also see pGFPuv from BD Biosciences as another control for forward mutation assays. Catalog # 6079-1 http://www.clontech.com/US/Products/Fluorescent_Proteins_and_Reporters/Fluorescent_Proteins_by_Name/GFP_GFPuv_Fluorescent_Proteins?sitex=10020:22372:US .