pMZ283
(Plasmid #52224)

• Purpose: Expression of sgRNA precursor with CspCI placeholder for target cloning (see comments).
• Depositing Lab: Mikel Zaratiegui
• Publication: Jacobs et al Nat Commun. 2014 Oct 29;5:5344. doi: 10.1038/ncomms6344.

Backbone

• Vector backbone: pUR18
• Backbone size w/o insert (bp): 5672
• Total vector size (bp): 6215
• Vector type: Yeast Expression, CRISPR
• Selectable markers: ura4

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): Top10
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: rrk1-sgRNA
• Species: S. pombe (fission yeast)
• Promoter: rrk1

Cloning Information

• Cloning method: Ligation Independent Cloning
• 5′ sequencing primer: CAG GAA ACA GCT ATG AC
• 3′ sequencing primer: TGT AAA ACG ACG GCC AGT

Resource Information

• Supplemental Documents:
  • gRNA design and cloning for the rrk1 fission yeast CRISPR/Cas9 vectors
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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

Depositor Comments

Please note that some batches of CspCI from New England Biolabs may cut inefficiently and alters the mobility of the DNA, possibly because it remains bound to the target. The two batches that are known to work are 0041303 and 0051407. This may be due to the particular target sequence present in pMZ283 and pMZ374.