pRS423-mitoT
(Plasmid #174525)

• Purpose: Expresses yeast variant of mitoT synthetic intermembrane tether bridging the inner and outer mitochondrial membranes
• Depositing Lab: Oleh Khalimonchuk
• Publication: Dietz et al Redox Biol. 2021 Oct;46:102125. doi: 10.1016/j.redox.2021.102125. Epub 2021 Sep 10.

Backbone

• Vector backbone: pRS423
• Backbone size w/o insert (bp): 6410
• Total vector size (bp): 7535
• Vector type: Yeast Expression, Synthetic Biology
• 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

Gene/Insert

• Gene/Insert name: Yeast mitochondrial intermembrane tether
• Alt name: mitoT
• Species: S. cerevisiae (budding yeast), Synthetic; E.coli
• Insert Size (bp): 1125
• Promoter: MET25
• Tag / Fusion Protein:
  • eGFP (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (not destroyed)
• 3′ cloning site: SalI (not destroyed)
• 5′ sequencing primer: 5'-GCGGATCCATGTTGAATAGTTCAAGAAAATATGCTTGTCGTTCCCTATTCAGACAAGCGAACGTC -3'
• 3′ sequencing primer: 3'- TATGTCGACGTACAATTCATCAATACCATGGGTAATACCAGCAGCAGTAACAAATTCTAACAAGACCA -5'

Resource Information

• Supplemental Documents:
  • Support file with highlighted parts of mitoT insert

Terms and Licenses

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

Depositor Comments

Please note: The plasmid contains a T72A mutation in Sco2p and L64F, T65G, S72A, & Y106C mutations in yeGFP compared to the depositor's provided sequence. The depositor noted that these differences do not affect the plasmid's function.