Skip to main content
This website uses cookies to ensure you get the best experience. By continuing to use this site, you agree to the use of cookies.

Please note: Your browser does not support the features used on Addgene's website. You may not be able to create an account or request plasmids through this website until you upgrade your browser. Learn more

Please note: Your browser does not fully support some of the features used on Addgene's website. If you run into any problems registering, depositing, or ordering please contact us at [email protected]. Learn more

pMB67
(Plasmid #47947)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 47947 Standard format: Plasmid sent in bacteria as agar stab 1 $85

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pBluescript SK+
  • Backbone manufacturer
    Stratagene
  • Backbone size w/o insert (bp) 2958
  • Total vector size (bp) 8127
  • Vector type
    Worm Expression, CRISPR

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin, 100 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    EPI400
  • Growth instructions
    Plasmid isolation from standard DH5alpha results in low yield. The plasmid may need to first be retransformed into BL21 cells to obtain normal DNA yields. Requires extra growth time.
  • Copy number
    Unknown

Gene/Insert

  • Gene/Insert name
    Cas9
  • Species
    Synthetic
  • Insert Size (bp)
    4254
  • Promoter hsp-16.48
  • Tags / Fusion Proteins
    • 3xFLAG (N terminal on insert)
    • SV40 NLS (N terminal on insert)
    • egl-13 NLS (C terminal on insert)

Cloning Information

Resource Information

Terms and Licenses

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

Depositor Comments

To generate the unique SpeI site, a short, 37 bp, linker was inserted into an existing NdeI site. This linker seems to have been inserted 2 or more times, creating a short repeat region that we have been unable to sequence across. Sequencing with the M13 reverse primer will fail.

This plasmid is provided in EPI400 bacteria which may need to be induced to obtain a high copy number. Please refer to this protocol: http://www.epibio.com/docs/default-source/protocols/copycutter-epi400-electrocompetent-e-coli-copycutter-epi400-chemically-competent-e-coli-copycutter-induction-solution.pdf?sfvrsn=10 for more information.

For more information on Boxem Lab CRISPR Plasmids please refer to: http://www.addgene.org/crispr/boxem/

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:

    pMB67 was a gift from Mike Boxem (Addgene plasmid # 47947 ; http://n2t.net/addgene:47947 ; RRID:Addgene_47947)
  • For your References section:

    CRISPR/Cas9-Targeted Mutagenesis in Caenorhabditis elegans. Waaijers S, Portegijs V, Kerver J, Lemmens BB, Tijsterman M, van den Heuvel S, Boxem M. Genetics. 2013 Aug 26. 10.1534/genetics.113.156299 PubMed 23979586