pHis-parallel1-NLSH2BCas9
(Plasmid #112065)

• Purpose: Expression and purification of NLSH2BCas9 for fungal gene editing
• Depositing Lab: Jeff Coleman
• Publication: Wang et al Fungal Genet Biol. 2018 May 12. pii: S1087-1845(18)30081-1. doi: 10.1016/j.fgb.2018.05.003.

Backbone

• Vector backbone: pHis-parallel1
• Backbone manufacturer: Sheffield et al (1999) Protein Expression Purification 15, 34-39
• Backbone size w/o insert (bp): 5466
• Total vector size (bp): 9765
• Modifications to backbone: None
• Vector type: Bacterial Expression, CRISPR

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: NLS of the Fusarium oxysporum histone H2B fused to SpCas9 that was codon optimized fro Aspergillus niger
• Species: Fusarium oxysporum, Streptococcus pyogenes
• Insert Size (bp): 4299
• Mutation: Cas9 was codon optimized for Aspergillus niger
• Promoter: promoter in vector
• Tag / Fusion Protein:
  • His tag (N terminal on backbone)

Cloning Information

• Cloning method: Unknown
• 5′ sequencing primer: GGATCCGATGGCTCCCAAGGCTGCTG
• 3′ sequencing primer: CCGCAAGCTTTCAGACCTTGCGCTTCTTC

Resource Information

• A portion of this plasmid was derived from a plasmid made by: The A. niger codon optimized Cas9 gene was obtained from Uffe Mortensen and was described in Nodvig et al. (2015) A CRISPR-Cas9 system for genetic engineering of filamentous fungi. PLoS ONE 10(7): e0133085.

Terms and Licenses

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