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pCMV-T7-SpRY-HF1-P2A-EGFP (RTW5008)
(Plasmid #139997)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 139997 Standard format: Plasmid sent in bacteria as agar stab 1 $85
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Backbone

  • Vector backbone
    RTW3027
  • Backbone manufacturer
    Kleinstiver Lab (Addgene ID 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
    human codon optimized SpCas9 variant named SpRY-HF1 with BPNLS-3xFLAG-P2A-EGFP
  • Alt name
    RTW5008
  • Species
    Synthetic
  • Mutation
    SpRY=A61R/L1111R/D1135L/S1136W/G1218K/E1219Q/N1317R/A1322R/R1333P/R1335Q/T1337R; HF1=N497A/R661A/Q695A/Q926A
  • Promoter CMV and T7
  • Tag / Fusion Protein
    • BPNLS-3xFLAG-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-SpRY-HF1-P2A-EGFP (RTW5008) was a gift from Benjamin Kleinstiver (Addgene plasmid # 139997 ; http://n2t.net/addgene:139997 ; RRID:Addgene_139997)

  • For your References section:

    Unconstrained genome targeting with near-PAMless engineered CRISPR-Cas9 variants. Walton RT, Christie KA, Whittaker MN, Kleinstiver BP. Science. 2020 Apr 17;368(6488):290-296. doi: 10.1126/science.aba8853. Epub 2020 Mar 26. 10.1126/science.aba8853 PubMed 32217751
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