YIp211-P30-His-pol30(K127/164R)-3'
(Plasmid #99549)

• Purpose: Yeast integrative vector for expression of His-tagged POL30 (PCNA) mutant K127/164R; URA3 marker
• Depositing Lab: Helle Ulrich
• Publication: Davies et al Methods Mol Biol. 2012;920:543-67. doi: 10.1007/978-1-61779-998-3_36.

Backbone

• Vector backbone: YIplac211
• Backbone manufacturer: Gietz & Sugino (1988) Gene 74:527-34
• Backbone size w/o insert (bp): 3797
• Vector type: Yeast Expression
• Selectable markers: URA3

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: POL30
• Alt name: PCNA
• Species: S. cerevisiae (budding yeast)
• Insert Size (bp): 2502
• Mutation: K127R, K164R
• Entrez Gene: POL30 (a.k.a. YBR088C)
• Promoter: POL30
• Tag / Fusion Protein:
  • 6xHis (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: EcoRI (unknown if destroyed)
• 3′ cloning site: SphI (unknown if destroyed)
• 5′ sequencing primer: M13fwd
• 3′ sequencing primer: M13rev

Terms and Licenses

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

Depositor Comments

The ORF is followed by 1129 bp of the POL30 3' region; digest with StuI for integration into URA3 or with PmlI for integration upstream of endogenous POL30 (for exchange of alleles by 5FOA selection.) Please see secondary publication PMID: 19107412 for more information about this plasmid.