pRB2949
(Plasmid #8672)

• Depositing Lab: David Botstein
• Publication: Anders et al Mol Biol Cell 2001 Dec;12(12):3973-86.

Backbone

• Vector backbone: pTS210
• Vector type: Yeast Expression

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: TUB1-828 under GAL1 promoter
• Alt name: TUB1-828
• Alt name: TUB1
• Species: S. cerevisiae (budding yeast)
• Mutation: D252A and E255A; dominant lethal
• Entrez Gene: TUB1 (a.k.a. YML085C)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: See comment (unknown if destroyed)
• 3′ cloning site: See comment (unknown if destroyed)
• 5′ sequencing primer: GAL1

Terms and Licenses

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

Depositor Comments

TUB1-828 was from Richards et al 2000 Mol. Biol. Cell 11, 1887-1903. It was PCRed, and the PCR products were digested with BamHI and SpeI and ligated to the BamHI and XbaI sites of pTS210. The vector plasmid pTS210, a YCp50-based construct, contained the GAL1/GAL10 promoter, a short polylinker (BamHI, HindIII, XbaI), and the ACT1 transcriptional terminator.

Please note that this construct is missing the first 8 amino acids of the TUB1 ORF. This deletion has no effect on the plasmid function as the 2 point mutations make this a dominant lethal allele.