pCas9_sgRNA_0
(Plasmid #70763)

• Purpose: expresses Ustilago maydis codon-optimized Cas9, contains U. maydis U6 promoter, is self-replicating
• Depositing Lab: Regine Kahmann
• Publication: Schuster et al Fungal Genet Biol. 2015 Sep 11. pii: S1087-1845(15)30025-6. doi: 10.1016/j.fgb.2015.09.001.

Backbone

• Vector backbone: pNEBUC
• Backbone manufacturer: Weinzierl et al., 2002, Mol. Microbiol. 45
• Total vector size (bp): 11105
• Vector type: self-replicating in Ustilago maydis, conferring carboxin resistance
• Selectable markers: carboxin resistance conferred by mutated ip gene

Growth in Bacteria

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

Gene/Insert 1

• Gene/Insert name: U6 promoter
• Species: Ustilago maydis
• Insert Size (bp): 826
• Promoter: U6 from Ustilago maydis

Cloning Information for Gene/Insert 1

• Cloning method: Restriction Enzyme
• 5′ cloning site: HindIII (not destroyed)
• 3′ cloning site: Acc651 (not destroyed)
• 5′ sequencing primer: TACGCCAAGCTTTAATACGTTCGTTCCG
• 3′ sequencing primer: GGTACGGGTACCGTTGTAGAATGGAATTTTG

Gene/Insert 2

• Gene/Insert name: cas9
• Species: Synthetic
• Insert Size (bp): 5393
• Mutation: Streptococcus pyogenes gene codon-optimized for Ustilago maydis
• Promoter: synthetic otef promoter (Spellig et al., 1996, Mol. Gen. Genet. 252)
• Tag / Fusion Protein:
  • N-terminal NLS, C-terminal HA-tag +NLS (N terminal on insert)

Cloning Information for Gene/Insert 2

• Cloning method: Restriction Enzyme
• 5′ cloning site: HindIII (not destroyed)
• 3′ cloning site: BamHI (not destroyed)
• 5′ sequencing primer: TACGCCAAGCTTTAATACGTTCGTTCCG
• 3′ sequencing primer: GTAAAACGACGGCCAG

Gene/Insert 3

• Gene/Insert name: tnos terminator
• Species: Agrobacterium tumefaciens
• Insert Size (bp): 291

Resource Information

• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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