pSPneoHHgRNAH
(Plasmid #63557)

• Purpose: Expresses gRNA in Leishmania
• Depositing Lab: Greg Matlashewski
• Publication: Zhang et al MBio. 2015 Jul 21;6(4). pii: e00861-15. doi: 10.1128/mBio.00861-15.

Backbone

• Vector backbone: pSPneo
• Total vector size (bp): 3740
• Vector type: Leishmania Expression
• 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: guide sequence
• gRNA/shRNA sequence: Leishmania
• Species: Leishmania

Cloning Information

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

Resource Information

• A portion of this plasmid was derived from a plasmid made by: The gRNA coding sequence was derived from pX330 plasmid distributed by Addgene ( Dr Feng Zhang, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA).

Terms and Licenses

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

Depositor Comments

The guide sequence will be cloned into the two Bbs I sites (See attached map). This plasmid contains secondary structure and needs to be sequenced with a modified protocol.