pRS426-GFP-COF1
(Plasmid #31844)

• Depositing Lab: David Drubin
• Publication: Okreglak et al J Cell Biol. 2007 Sep 24;178(7):1251-64. Epub 2007 Sep 17.

Backbone

• Vector backbone: pRS426
• Backbone manufacturer: ATCC
• Backbone size w/o insert (bp): 5726
• Vector type: Yeast Expression

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: Cofilin
• Alt name: Cof1p
• Alt name: COF1
• Species: S. cerevisiae (budding yeast)
• Entrez Gene: COF1 (a.k.a. YLL050C)
• Promoter: T7
• Tag / Fusion Protein:
  • GFP (S65T) (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: None (unknown if destroyed)
• 3′ cloning site: None (unknown if destroyed)
• 5′ sequencing primer: T7

Terms and Licenses

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

Depositor Comments

The GFP-cofilin construct has S65T-GFP and a 12–amino acid linker (GHGTGSTGSGSS) inserted in between amino acids N74 and G75 of cofilin. Note that the GFP is NOT N-terminal.

A missense mutation was detected by the lab in sequencing: converting the third glycine in the 5′ GFP linker into an arginine. This was the construct used in the paper.