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pCMV-T7-TadCBEd-SpCas9(MKRCKS)-P2A-EGFP (RAS3445)
(Plasmid #223130)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 223130 Standard format: Plasmid sent in bacteria as agar stab 1 $85
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Backbone

  • Vector backbone
    pCMV_ABEmax_P2A_GFP
  • Backbone manufacturer
    David Liu, Addgene plasmid #112101
  • Vector type
    Mammalian Expression, CRISPR

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
    human codon optimized TadA-CDd-SpCas9(MKRCKS)-2xUGI-P2A-EGFP
  • Alt name
    RAS3445
  • Alt name
    TadCBEd-SpCas9(MKRCKS)
  • Species
    Synthetic
  • Mutation
    TadA-CDd mutations; nSpCas9(D10A/MKRCKS)=D1135M, S1136K, G1218R, E1219C, R1335K, T1337S
  • Promoter CMV and T7
  • Tags / Fusion Proteins
    • BPNLS-TadA-CDd (N terminal on insert)
    • 2xUGI-BPNLS-P2A-EGFP (C terminal on insert)

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.
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:

    pCMV-T7-TadCBEd-SpCas9(MKRCKS)-P2A-EGFP (RAS3445) was a gift from Benjamin Kleinstiver (Addgene plasmid # 223130 ; http://n2t.net/addgene:223130 ; RRID:Addgene_223130)

  • For your References section:

    Custom CRISPR-Cas9 PAM variants via scalable engineering and machine learning. Silverstein RA, Kim N, Kroell AS, Walton RT, Delano J, Butcher RM, Pacesa M, Smith BK, Christie KA, Ha LL, Meis RJ, Clark AB, Spinner AD, Lazzarotto CR, Li Y, Matsubara A, Urbina EO, Dahl GA, Correia BE, Marks DS, Tsai SQ, Pinello L, De Ravin SS, Liu Q, Kleinstiver BP. Nature. 2025 Apr 22. doi: 10.1038/s41586-025-09021-y. 10.1038/s41586-025-09021-y PubMed 40262634
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