pKSB-610 U6-sgRNA2 GGAA AGAG
(Plasmid #173672)

• Purpose: (Empty Backbone) To clone and express a gRNA in Anopheles
• Depositing Lab: Eric Marois
• Publication: Klug et al PLoS Pathog. 2022 Oct 12;18(10):e1010881. doi: 10.1371/journal.ppat.1010881. eCollection 2022 Oct.

Backbone

• Vector backbone: pBluescript II KS +
• Backbone manufacturer: Stratagene
• Backbone size (bp): 3000
• Modifications to backbone: BsaI restriction site was mutated. U6 promoter from Anopheles gambiae AGAP013557 drives expression of a classical gRNA template (Jinek et al., 2012) in which a specific gRNA protospacer sequence can be cloned using two BbsI restrictions sites.
• Vector type: Insect 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: Restriction Enzyme
• 5′ sequencing primer: M13R
• 3′ sequencing primer: M13F

Resource Information

• Supplemental Documents:
  • 610 pKSB-sgRNA2 GGAA AGAG.gb

Terms and Licenses

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

Depositor Comments

U6 promoter from Anopheles gambiae AGAP013557 drives expression of a classical gRNA template (Jinek et al., 2012) in which a specific gRNA protospacer sequence can be cloned using two BbsI restrictions sites.
This plasmid is part of a kit to assemble a multiple-gRNA expression vector by Golden Gate Cloning for mutagenesis in Anopheles mosquitoes.