pmKaxxte
(Plasmid #113630)

• Purpose: to test Cas9 gRNA efficiency or HR-capacity
• Depositing Lab: Ken Mills
• Publication: Liberante et al Sci Rep. 2019 Feb 25;9(1):2678. doi: 10.1038/s41598-019-39591-7.

Backbone

• Vector backbone: unknown
• Vector type: Mammalian Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): NEB Stable
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: mKaxxte2.5
• Alt name: mKate2.5
• Promoter: CMV

Cloning Information

• Cloning method: Unknown
• 5′ sequencing primer: unknown

Resource Information

• Supplemental Documents:
  • pmKaxxte
• A portion of this plasmid was derived from a plasmid made by: mKate2.5 from Michael Davidson (Addgene plasmid # 54828)

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Evrogen Limited Use Label License for FPs
• Industry Terms:
  • Not Available to Industry

Depositor Comments

One clones a fragment of DNA sequence at the MCS, then uses a targeted nuclease, such as Cas9 to trigger a double-strand break. This is then repaired by homologous recombination resulting in restoration of an intact mKate2.5 fluorescent protein (cloned from Addgene #54828). It can be used to test Cas9 gRNA efficiency, as it was in the paper, or HR-capacity in different cells.