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Addgene

pET28-SaCas9
(Plasmid #178901)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 178901 Standard format: Plasmid sent in bacteria as agar stab 1 $85
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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
    • (Common Sequencing Primers)

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
  • Industry Terms
    • Not Available to Industry
Trademarks:
  • Zeocin® is an InvivoGen trademark.
How to cite this plasmid ( Back to top)

These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.

  • For your Materials & Methods section:

    pET28-SaCas9 was a gift from Hendrik Milting (Addgene plasmid # 178901 ; http://n2t.net/addgene:178901 ; RRID:Addgene_178901)

  • For your References section:

    A detailed protocol for expression, purification, and activity determination of recombinant SaCas9. Flottmann F, Pohl GM, Gummert J, Milting H, Brodehl A. STAR Protoc. 2022 Apr 5;3(2):101276. doi: 10.1016/j.xpro.2022.101276. eCollection 2022 Jun 17. 10.1016/j.xpro.2022.101276 PubMed 35403007
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