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SaCas9 variant click editor (CE1) - pCMV-T7-PCV2-nSaCas9-EcKlenow (MLE194)
(Plasmid #217801)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 217801 Standard format: Plasmid sent in bacteria as agar stab 1 $85
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Backbone

  • Vector backbone
    pCMV-T7-SpCas9-P2A-EGFP (RTW3027)
  • Backbone manufacturer
    Addgene #139987
  • Vector type
    Mammalian Expression, CRISPR ; in vitro transcription; T7 promoter

Growth in Bacteria

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

Gene/Insert

  • Gene/Insert name
    PCV2-XTEN-nSaCas9-BPNLS-EcKlenow(-exo)-BPNLS
  • Alt name
    MLE194
  • Species
    Synthetic
  • Mutation
    nSaCas9(N580A); EcKlenow(-exo;D355A/E357A)
  • Promoter CMV and T7
  • Tag / Fusion Protein
    • BPNLS (C terminal on backbone)

Cloning Information

  • Cloning method Gibson Cloning
  • 5′ sequencing primer oBK6928-CCAAGTCTCCACCCCATTGACG
  • 3′ sequencing primer oBK219-GGGAGTGGCACCTTCCAGGGTC
    • (Common Sequencing Primers)

Resource Information

Terms and Licenses

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

Depositor Comments

Please visit https://doi.org/10.1101/2023.09.12.557440 for bioRxiv preprint.

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:

    SaCas9 variant click editor (CE1) - pCMV-T7-PCV2-nSaCas9-EcKlenow (MLE194) was a gift from Benjamin Kleinstiver (Addgene plasmid # 217801 ; http://n2t.net/addgene:217801 ; RRID:Addgene_217801)

  • For your References section:

    Click editing enables programmable genome writing using DNA polymerases and HUH endonucleases. Ferreira da Silva J, Tou CJ, King EM, Eller ML, Rufino-Ramos D, Ma L, Cromwell CR, Metovic J, Benning FMC, Chao LH, Eichler FS, Kleinstiver BP. Nat Biotechnol. 2024 Jul 22. doi: 10.1038/s41587-024-02324-x. 10.1038/s41587-024-02324-x PubMed 39039307
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