pET28-SaCas9
(Plasmid #178901)

• Purpose: SaCas9 (Cas9 from Staphylococcus aureus)
• Depositing Lab: Hendrik Milting
• Publication: Flottmann et al STAR Protoc. 2022 Apr 5;3(2):101276. doi: 10.1016/j.xpro.2022.101276. eCollection 2022 Jun 17.

Backbone

• Vector backbone: pET-28b-RfxCas13d-His
• Backbone manufacturer: Addgene, #141322
• Backbone size w/o insert (bp): 5198
• Total vector size (bp): 8502
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: SaCas9
• Alt name: SauCas9
• Species: Staphylococcus aureus
• Insert Size (bp): 3300
• Tags / Fusion Proteins:
  • nuclear localization signal of SV40 (simian virus 40) large T antigen (N terminal on insert)
  • bipartite nuclear localization signal from nucleoplasmin (C terminal on insert)
  • His6-Tag (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NcoI (not destroyed)
• 3′ cloning site: BamHI (not destroyed)
• 5′ sequencing primer: T7
• 3′ sequencing primer: GCTAGTTATTGCTCAGCGG

Resource Information

• A portion of this plasmid was derived from a plasmid made by: SaCas9 was cloned from pX601-AAV-CMV::NLS-SaCas9-NLS-3xHA-bGHpA;U6::BsaI-sgRNA (Addgene, #61591)

Terms and Licenses

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