pEVA1
(Plasmid #58953)

• Purpose: Contains a synthetic DNA sequence, VAR1u, that specifies the Var1 protein. The Cox4 targeting signal is fused to the N-terminus to direct import of Var1. Complements var1 mutations in mtDNA.
• Depositing Labs: Thomas Fox, Thomas Mason
• Publication: Sanchirico et al EMBO J. 1998 Oct 1;17(19):5796-804.

Backbone

• Vector backbone: Yep13
• Modifications to backbone: Yep13 (2-micron yeast origin, LEU2) modified by BENDER, A., and J. R. PRINGLE, 1991 Mol. Cell. Biol. 11: 1295-1305, to contain ADE3. Received from them as pTSV30a.
• Vector type: Yeast Expression ; Yeast Ecoli shuttle vector
• Selectable markers: LEU2 ; ADE3

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: VAR1u, VAR1 recoded for expression from the nucleus
• Alt name: preCox4-Var1 fusion for targeting to mitochondria
• Species: S. cerevisiae (budding yeast)
• Insert Size (bp): 3100
• Mutation: Synthetic gene in standard genetic code
• GenBank ID: AJ010480.1
• Promoter: ADH1
• Tag / Fusion Protein:
  • preCox4 mitochondrial matrix targeting signal (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: SalI (not destroyed)
• 3′ cloning site: SalI (not destroyed)
• 5′ sequencing primer: none
• 3′ sequencing primer: none

Resource Information

• A portion of this plasmid was derived from a plasmid made by: From collaborators Marie Sanchirico and Thomas L. Mason, then at University of Massachusetts Amherst.

Terms and Licenses

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