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Addgene

pEM-Cas9HF1-D1135E
(Plasmid #154924)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 154924 Standard format: Plasmid sent in bacteria as agar stab 1 $85
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Backbone

  • Vector backbone
    p15A
  • Vector type
    Bacterial Expression, CRISPR, Synthetic Biology

Growth in Bacteria

  • Bacterial Resistance(s)
    Chloramphenicol, 25 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    Low Copy

Gene/Insert

  • Gene/Insert name
    Cas9HF1-D1135E
  • Insert Size (bp)
    4104
  • Mutation
    N497A/R661A/Q695A/Q926A/D1135E and stop codon on the Tet repressor (see depositor comments)
  • Promoter pTet
  • Tag / Fusion Protein
    • ssrA (C terminal on insert)

Cloning Information

Terms and Licenses

  • Academic/Nonprofit Terms
  • Industry Terms
    • Not Available to Industry
Trademarks:
  • Zeocin® is an InvivoGen trademark.

Depositor Comments

Addgene NGS had a stop codon on the Tet repressor. The depositing lab has not found that this difference impacts the function of the plasmid but it is possible that the expression of SpCas9HF1 is not well controlled by Tetracycline or its analogs.

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:

    pEM-Cas9HF1-D1135E was a gift from Michael Lynch (Addgene plasmid # 154924 ; http://n2t.net/addgene:154924 ; RRID:Addgene_154924)

  • For your References section:

    CRISPR-Cas "Non-Target" Sites Inhibit On-Target Cutting Rates. Moreb EA, Hutmacher M, Lynch MD. CRISPR J. 2020 Dec;3(6):550-561. doi: 10.1089/crispr.2020.0065. 10.1089/crispr.2020.0065 PubMed 33346713
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