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pET28b-SpRY-His
(Plasmid #179320)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 179320 Standard format: Plasmid sent in bacteria as agar stab 1 $85
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Backbone

  • Vector backbone
    pET-28b-Cas9-His (#47327)
  • Backbone manufacturer
    Alex Schier
  • Backbone size w/o insert (bp) 5271
  • Total vector size (bp) 9375
  • Vector type
    Bacterial Expression, CRISPR

Growth in Bacteria

  • Bacterial Resistance(s)
    Kanamycin, 50 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Growth instructions
    For expression and purification of SpRY-Cas9, transform into Rosetta strain.
  • Copy number
    Unknown

Gene/Insert

  • Gene/Insert name
    Human codon-optimized Cas9 variant SpRY
  • Alt name
    SpRY
  • Species
    Synthetic
  • Insert Size (bp)
    4104
  • Mutation
    SpRY=A61R/L1111R/D1135L/S1136W/G1218K/E1219Q/ N1317R/A1322R/R1333P/R1335Q/T1337R
  • Tags / Fusion Proteins
    • SV40-NLS (C terminal on insert)
    • 6xHis (C terminal on insert)

Resource Information

  • Supplemental Documents
  • A portion of this plasmid was derived from a plasmid made by
    gBlock containing the indicated mutations was cloned into pET-28b-Cas9-His (#47327) linearized by PCR using primers Lin_Cas9_pET28b_fwd (5'-CTACCAAGGAGGTCCTGGACGCC-3') and Lin_Cas9_pET28b_rev (5'-GATACTTTCCTTGGAGCCTCCGG-3').

Terms and Licenses

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

Depositor Comments

This plasmid is owned by Moreno-Mateos Lab (CABD, Seville, Spain) and Cerón Lab (IDIBELL, Barcelona, Spain).

This plasmid was used to express a His-tagged SpRY-Cas9 protein in E. coli and purify it by nickel-based affinity purification.

Please visit https://www.biorxiv.org/content/10.1101/2021.06.06.447255v2 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:

    pET28b-SpRY-His was a gift from Julián Cerón & Miguel Angel Moreno-Mateos (Addgene plasmid # 179320 ; http://n2t.net/addgene:179320 ; RRID:Addgene_179320)

  • For your References section:

    Genome editing in animals with minimal PAM CRISPR-Cas9 enzymes. Vicencio J, Sanchez-Bolanos C, Moreno-Sanchez I, Brena D, Vejnar CE, Kukhtar D, Ruiz-Lopez M, Cots-Ponjoan M, Rubio A, Melero NR, Crespo-Cuadrado J, Carolis C, Perez-Pulido AJ, Giraldez AJ, Kleinstiver BP, Ceron J, Moreno-Mateos MA. Nat Commun. 2022 May 12;13(1):2601. doi: 10.1038/s41467-022-30228-4. 10.1038/s41467-022-30228-4 PubMed 35552388
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