Ccas9LacZ
(Plasmid #113083)

• Purpose: (Empty Backbone) Enhanced C. elegans Cas9 vector. Ccas9LacZ contains a lacZ coding sequence, two Cas9 and two type-IIs restriction enzyme recognition sites next to the U6 promoter and gRNA sequences.
• Depositing Lab: Tibor Vellai
• Publication: Sturm et al Nucleic Acids Res. 2018 Jun 19. pii: 5040046. doi: 10.1093/nar/gky516.

Backbone

• Vector backbone: pDD162 (Peft-3::Cas9 + Empty sgRNA, Addgene no.: #47549)
• Modifications to backbone: The original C. elegans Cas9 vector does not include any restriction enzyme cleavage site between the U6 promoter and gRNA sequence that would allow the spacer sequence to be efficiently modified. The redesigned vector (Ccas9lacZ) contains two Cas9 and two type-IIs restriction enzyme recognition sites next to the U6 promoter and gRNA sequences which allow the spacer sequence to be easily modified with restriction digestion coupled linker ligation and homologous recombination-based cloning too. The new vector also contains a lacZ gene that allows blue-white selection after cloning.
• Vector type: Worm Expression, CRISPR

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Cloning Information

• Cloning method: Gibson Cloning

Terms and Licenses

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

Depositor Comments

For plasmid usage, please see the associated publication: Sturm et al.
Highly efficient RNAi and Cas9-based auto-cloning systems for C. elegans research. Nucleic Acids Research, gky516, https://doi.org/10.1093/nar/gky516