pXP420 v2 (repaired Amp promoter)
(Plasmid #86920)

• 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.
• Depositing Lab: Gary Yellen
• Publication: pXP420 repaired Amp promoter (unpublished)

Backbone

• Vector backbone: pUC18
• Backbone manufacturer: Sandmeyer lab (Addgene plasmid 26844)
• Modifications to backbone: Ampicillin 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 type: Yeast Expression, Cre/Lox
• Promoter: TEF1
• Selectable markers: HIS3

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ sequencing primer: TEF_seq (5'-ggtcttcaatttctcaagtttc-3')
• 3′ sequencing primer: CYC1 (5'-GCGTGAATGTAAGCGTGAC-3')

Resource Information

• Supplemental Documents:
  • pXP420-Sandmeyer-RepairedAmpPro.gb

Terms and Licenses

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

Depositor Comments

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.