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pDuRCC-N
(Plasmid #81194)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 81194 Standard format: Plasmid sent in bacteria as agar stab 1 $75

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pRS42N
  • Backbone manufacturer
    Euroscarf
  • Vector type
    Yeast Expression, CRISPR
  • Selectable markers
    Nourseothricin

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    High Copy

Gene/Insert 1

  • Gene/Insert name
    Cas9
  • Species
    S. pyogenes
  • Mutation
    WT - codon optimized
  • Promoter ROX3
  • Tag / Fusion Protein
    • NTS (C terminal on backbone)

Cloning Information for Gene/Insert 1

Gene/Insert 2

  • Gene/Insert name
    empty gRNA cassette
  • Species
    Synthetic
  • Mutation
    Please view depositor comments below
  • Promoter SNP52p

Cloning Information for Gene/Insert 2

Gene/Insert 3

  • Gene/Insert name
    empty gRNA cassette
  • Species
    Synthetic
  • Mutation
    Please view depositor comments below
  • Promoter SNP52p

Cloning Information for Gene/Insert 3

Resource Information

Depositor Comments

Please note: Addgene’s quality control sequences found the guanine nucleotide at the start of the structural guide-RNA sequence was missing. The Boles Lab has indicated that this is a “virtual guanine”. If you sequence this plasmid, you will notice this missing guanine. The Boles lab omitted the guanine in order to work with the empty vector in yeast, since once the structural gRNA has this G, it is active and could create problems in the cell. However, when primers are designed to create the CRISPR/Cas9 vector for a specific target, this G should be in the primer, so that the gRNA becomes functional again. If you do not include this G in your primer sequence, the system will not work.

Plasmid contains 2 gRNA cassettes together with the Cas9 overexpression cassette.

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:

    pDuRCC-N was a gift from Eckhard Boles (Addgene plasmid # 81194 ; http://n2t.net/addgene:81194 ; RRID:Addgene_81194)
  • For your References section:

    Simplified CRISPR-Cas genome editing for Saccharomyces cerevisiae. Generoso WC, Gottardi M, Oreb M, Boles E. J Microbiol Methods. 2016 Aug;127:203-5. doi: 10.1016/j.mimet.2016.06.020. Epub 2016 Jun 17. 10.1016/j.mimet.2016.06.020 PubMed 27327211