pXP420 v2 (repaired Amp promoter)
Purpose(Empty Backbone) Shuttle vector to facilitate gene expression for metabolic engineering in S. cerevisiae. Contains TEF1 promoter and CYC1 terminator for insertion of gene of interest. Contains HIS3 selection marker.
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||86920||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
Backbone manufacturerSandmeyer lab (Addgene plasmid 26844)
Modifications to backboneAmpicillin promoter was corrected to repair a split in the promoter that occurred during the original construction of pX420 using the Ssp1 site for cloning, as described in Fang et al., Yeast. 2011 Feb;28(2):123-36.
Vector typeYeast Expression, Cre/Lox
- Promoter TEF1
Growth in Bacteria
Copy numberHigh Copy
- Cloning method Restriction Enzyme
- 5′ sequencing primer TEF_seq (5'-ggtcttcaatttctcaagtttc-3')
- 3′ sequencing primer CYC1 (5'-GCGTGAATGTAAGCGTGAC-3') (Common Sequencing Primers)
Please note that the corrected Ampicillin promoter may improve the bacterial resistance of the plasmid to ampicillin compared to the original pXP420 (Addgene plasmid# 26844).
The HIS3 selection marker in this plasmid contains a V132A mutation in imidazoleglycerol-phosphate dehydratase. Yeast transformants grow normally on SC-HIS plates, so this mutation does not inhibit gene function.
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:pXP420 v2 (repaired Amp promoter) was a gift from Gary Yellen (Addgene plasmid # 86920 ; http://n2t.net/addgene:86920 ; RRID:Addgene_86920)