|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||11724||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
Vector backbonemodified pGPS3
Backbone manufacturerNew England Biolabs
- Backbone size w/o insert (bp) 2642
Vector typeBacterial Expression
Growth in Bacteria
Copy numberHigh Copy
Alt nameampicillin resistance
Alt namecarbenicillin resistance
Insert Size (bp)979
MutationGAA for Glu26 changed to TAA (premature stop codon introduced in beta-lactamase gene).
- Cloning method Restriction Enzyme
- 5′ cloning site AflIII (not destroyed)
- 3′ cloning site KpnI (not destroyed)
- 5′ sequencing primer n/a (Common Sequencing Primers)
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 .
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:pLA230 was a gift from Lawrence Loeb (Addgene plasmid # 11724 ; http://n2t.net/addgene:11724 ; RRID:Addgene_11724)
For your References section:In vivo mutagenesis by Escherichia coli DNA polymerase I. Ile(709) in motif A functions in base selection. Shinkai A, Loeb LA. J Biol Chem. 2001 Dec 14. 276(50):46759-64. 10.1074/jbc.M104780200 PubMed 11602576