pKSB-631 U6-sgRNA3/4 AACA GCTT
(Plasmid #173675)

• Purpose: (Empty Backbone) To clone and express a gRNA in Anopheles
• Depositing Lab: Eric Marois
• Publication: Marois lab kit for CRISPR and transgenesis in Anopheles and Aedes (unpublished)

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 gRNA template improved for efficiency (Dang et al., Genome Biology 2015) 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:
  • pKSB-631 sgRNA3:4 AACA GCTT.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 an improved gRNA template (Dang et al., Genome Biology 2015) 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.