3xEGFP-mMeikin(1-391) - hRad21(142-275, E169R, R172E) - mMeikin(387-434)
(Plasmid #174713)

• Purpose: Mammalian expression of mouse Meikin with inserted C-terminal Separase cleavage site inactivated
• Depositing Lab: Iain Cheeseman
• Publication: Maier et al Dev Cell. 2021 Aug 9;56(15):2192-2206.e8. doi: 10.1016/j.devcel.2021.06.019. Epub 2021 Jul 30.

Backbone

• Vector backbone: pCS2+
• Vector type: Mammalian 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: Meikin
• Species: M. musculus (mouse)
• Mutation: mMeikin: 1-391; hRad21: 142-275, E169R, R172E; mMeikin: 387-434
• Tag / Fusion Protein:
  • 3xEGFP (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: Unknown (unknown if destroyed)
• 3′ cloning site: Unknown (unknown if destroyed)

Resource Information

• Supplemental Documents:
  • pNM963.gb

Terms and Licenses

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

Depositor Comments

pNM963, compatible with in vitro transcription from SP6 promoter, C-terminal Separase cleavage site is derived from human Rad21 (aa142-275) with Separase resistant mutations (E169R, R172E), kinetochore localization is maintainted by C-terminal mMeikin fragment (aa387-434) . Please visit https://doi.org/10.1101/2020.11.29.402537 for bioRxiv preprint