pML104-KanMx4
(Plasmid #83476)

• Purpose: Expresses Cas9 and contains guide RNA expression cassette with BclI-SwaI cloning sites for guide sequence cloning; Contains KanMx4 marker for yeast transformation.
• Depositing Lab: Patricia Wittkopp
• Publication: Patricia Wittkopp yeast CRISPR plasmids (unpublished)

Backbone

• Vector backbone: pRSII426
• Vector type: Yeast Expression
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): NEB dam-/dcm-
• Growth instructions: Growth in dam- E.coli cells is preferred
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: KanMX4

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: pRS-marker
• 3′ sequencing primer: F1ori-F

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Addgene plasmid #67638 pML104, deposited by John Wyrick
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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

Depositor Comments

This plasmid is similar to pML104 except that the URA3 marker has been switched for KanMX4 marker using Gibson Assembly. Cloning of guide sequence using BclI-SwaII restriction sites requires growth in dam- E. coli cells.