pLdCN2
(Plasmid #125186)

• Purpose: Expresses Streptococcus pyogenes Cas9 (SpCas9) and two gRNAs in Leishmania
• Depositing Lab: Greg Matlashewski
• Publication: Zhang et al Methods Mol Biol. 2020;2116:199-224. doi: 10.1007/978-1-0716-0294-2_14.

Backbone

• Vector backbone: pLdCN
• Vector type: CRISPR ; Leishmania
• Selectable markers: Neomycin (select with G418)

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: gRNA and Cas9
• Species: Leishmania

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: Bbs I (unknown if destroyed)
• 3′ cloning site: Bbs I (unknown if destroyed)
• 5′ sequencing primer: SP6 promoter

Resource Information

• A portion of this plasmid was derived from a plasmid made by: The gene sequence was derived from plasmid pX330 from Dr Feng Zhang

Terms and Licenses

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

Depositor Comments

This Leishmania CRISPR vector pLdCN2 was specially designed to facilitate cloning two gRNA guide coding sequences into the vector with a single-step digestion-ligation cloning protocol.