pSPneogRNA241510+MT
(Plasmid #84292)

• Purpose: Express LdPBK_241510.1 and LdMT targeting gRNAs simutaneously
• Depositing Lab: Greg Matlashewski
• Publication: Zhang et al mSphere. 2017 Jan 18;2(1). pii: e00340-16. doi: 10.1128/mSphere.00340-16. eCollection 2017 Jan-Feb.

Backbone

• Vector backbone: pSP72
• Total vector size (bp): 3900
• Vector type: CRISPR ; Leishmania donovani
• 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: LdBPK_241510.1 targeting gRNA and LdMT targeting gRNA
• gRNA/shRNA sequence: GGTGTCGTTCTGCAGCCA and GTCCAGGCGCAGTGGATGC
• Species: Leishmania donovani
• GenBank ID: LdBPK_241510.1 in TritrypDB LdBPK_131590.1 in TritrypDB
• Promoter: L. donovani ribosome RNA promoter

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 Promoter
• 3′ sequencing primer: T7 promoter

Resource Information

• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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

Depositor Comments

This dual gRNA expression plasmid is used to target both LdBPK_241510.1 and LdMT gene. In the article, it was used to generate the targeted chromosome translocations between chromosome 13 and 24 and to enrich the edited LdBPK_241510.1 mutants by selection with miltefosine. For addgene customers, this dual gRNA expression plasmid will mainly be used as the PCR template to generate new double gRNAs expression vector or co-targeting MT vector. See article for more detail.