pZK12
(Plasmid #248912)

• Purpose: expresses yeast Isc10 fused to maltose binding protein and Cak1 in bacteria
• Depositing Lab: Edward Winter
• Publication: Rimal et al Mol Cell Biol. 2020 Jul 29;40(16). pii: MCB.00097-20. doi: 10.1128/MCB.00097-20. Print 2020 Jul 29.

Backbone

• Vector backbone: pACYCDuet-1 w/NheI site eliminated
• Backbone manufacturer: Novagen
• Backbone size w/o insert (bp): 4008
• Total vector size (bp): 6948
• Modifications to backbone: The NheI site in pACYC-DUET was eliminated from pACYC-DUET
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Chloramphenicol, 25 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert 1

• Gene/Insert name: Isc10-MBP
• Alt name: Isc10, Inhibitor of the Smk1 MAPK
• Species: S. cerevisiae (budding yeast); Sequence is optimized for expression of S. cerevisiae Isc10 sequence in bacteria
• Insert Size (bp): 1945
• Mutation: Nucleotide sequence is optimized for bacterial expression of ISC10.
• GenBank ID: NP_011107.1
• Entrez Gene: ISC10 (a.k.a. YER180C)
• Promoter: T7
• Tag / Fusion Protein:
  • maltose binding protein (N terminal on insert)

Cloning Information for Gene/Insert 1

• Cloning method: Restriction Enzyme
• 5′ cloning site: AflIII (not destroyed)
• 3′ cloning site: XhoI (not destroyed)
• 5′ sequencing primer: standard T7 promoter primer

Gene/Insert 2

• Gene/Insert name: Cak1
• Alt name: CDK activating kinase
• Species: S. cerevisiae (budding yeast)
• Insert Size (bp): 1104
• GenBank ID: NM_001179937.1
• Entrez Gene: CAK1 (a.k.a. YFL029C, CIV1)
• Promoter: T7

Cloning Information for Gene/Insert 2

• Cloning method: Restriction Enzyme
• 5′ cloning site: AflIII (not destroyed)
• 3′ cloning site: XhoI (not destroyed)
• 5′ sequencing primer: standard T7 primer

Terms and Licenses

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