pIgnaviCas9
(Plasmid #127595)

• Purpose: Expresses IgnaviCas9 in E. coli
• Depositing Lab: Stephen Quake
• Publication: Schmidt et al Proc Natl Acad Sci U S A. 2019 Oct 28. pii: 1904273116. doi: 10.1073/pnas.1904273116.

Backbone

• Vector backbone: pMJ806
• Backbone manufacturer: Addgene plasmid # 39312
• Backbone size w/o insert (bp): 6545
• Total vector size (bp): 10266
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: Use BL21 bacterial cells for protein expression.
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: IgnaviCas9
• Species: Bacterium from Ignavibacteriae phylum
• Insert Size (bp): 3720
• Promoter: T7 promoter
• Tags / Fusion Proteins:
  • 6xHis Tag (N terminal on backbone)
  • Maltose binding protein (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: SpeI (not destroyed)
• 3′ cloning site: AgeI (not destroyed)
• 5′ sequencing primer: acaacactagtgaaaacctgtatttccagggagcagcctcgATGAAAAAAGTATTAGGATTAGATCTTGGAGTATCTTCAAT
• 3′ sequencing primer: ttaaccggtatcaacttttcgtttctttttaggatcctcaacttttcgttttttcttaggtgacccTTTATTATTAGTAATAATTTCTCCCAAACAATTAATTAA

Resource Information

• Supplemental Documents:
  • pMJ806_KANR EC HIS-MBP-TEV-IgnaviCas9-2xNLS.gbk
  • sgRNA Transcription Template

Terms and Licenses

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