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pKMW123_SpRY-Cas9_U6-entry
(Plasmid #244820)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 244820 Standard format: Plasmid sent in bacteria as agar stab 1 $94
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DNA 244820-D50 50 μg of DNA in Tris buffer $495
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DNA 244820-D100 100 μg of DNA in Tris buffer $585
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Backbone

  • Vector backbone
    PX458
  • Vector type
    Mammalian Expression, CRISPR

Growth in Bacteria

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

Gene/Insert

  • Gene/Insert name
    SpRY Cas9
  • Species
    Streptococcus pyogenes
  • Mutation
    A61R/L1111R/D1135L/S1136W/G1218K/E1219Q/N1317R/A1322R/R1333P/R1335Q/T1337R
  • Promoter CAG, U6
  • Tags / Fusion Proteins
    • 3xFLAG-SV40 NLS (N terminal on insert)
    • Nucleoplasmin NLS (C terminal on insert)

Cloning Information

  • Cloning method Gibson Cloning

Resource Information

Terms and Licenses

  • Academic/Nonprofit Terms
  • Industry Terms
    • Not Available to Industry

Trademarks:

  • Zeocin® is an InvivoGen trademark.

Depositor Comments

Also expresses single guide RNA driven by U6 promoter.

Purpose

Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.

Delivery

  • Amount 50 μg
  • Concentration 1 μg/μL
  • Pricing $495 USD
  • Storage 4 ℃ (short-term) or -20 ℃ (long-term)

Terms and Licenses

  • Academic/Nonprofit Terms
  • Industry Terms
    • Not Available to Industry

Quality Control

Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.

Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.

Purpose

Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.

Delivery

  • Amount 100 μg
  • Concentration 1 μg/μL
  • Pricing $585 USD
  • Storage 4 ℃ (short-term) or -20 ℃ (long-term)

Terms and Licenses

  • Academic/Nonprofit Terms
  • Industry Terms
    • Not Available to Industry

Quality Control

Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.

Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.

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:

    pKMW123_SpRY-Cas9_U6-entry was a gift from Jennifer Doudna (Addgene plasmid # 244820 ; http://n2t.net/addgene:244820 ; RRID:Addgene_244820)

  • For your References section:

    Rapid two-step target capture ensures efficient CRISPR-Cas9-guided genome editing. Shi H, Al-Sayyad N, Wasko KM, Trinidad MI, Doherty EE, Vohra K, Boger RS, Colognori D, Cofsky JC, Skopintsev P, Bryant Z, Doudna JA. Mol Cell. 2025 May 1;85(9):1730-1742.e9. doi: 10.1016/j.molcel.2025.03.024. Epub 2025 Apr 23. 10.1016/j.molcel.2025.03.024 PubMed 40273916
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