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

pEDA5_GFPmut3_Y66H
(Plasmid #80085)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 80085 Standard format: Plasmid sent in bacteria as agar stab 1 $85

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pET29b(+)
  • Backbone manufacturer
    Novagen
  • Backbone size w/o insert (bp) 3583
  • Total vector size (bp) 4303
  • Modifications to backbone
    - Swapped antibiotic resistance from KanR to AmpR - Promoter is proB (see doi: 10.1371/journal.pone.0109105)
  • Vector type
    Bacterial Expression

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin, 100 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Growth instructions
    Plasmid must be prepared fresh and avoid multiple freeze/thaw cycles for use as a positive control in Nicking Mutagenesis.
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    GFPmut3_Y66H
  • Species
    Synthetic; Aequorea victoria
  • Insert Size (bp)
    720
  • Mutation
    changed Tyrosine 66 to Histidine
  • Promoter proB
  • Tag / Fusion Protein
    • 6x-His tag (C terminal on backbone)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site NdeI (not destroyed)
  • 3′ cloning site XhoI (not destroyed)
  • 5′ sequencing primer pED_2ND
  • 3′ sequencing primer T7 term
  • (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:

    pEDA5_GFPmut3_Y66H was a gift from Timothy Whitehead (Addgene plasmid # 80085 ; http://n2t.net/addgene:80085 ; RRID:Addgene_80085)
  • For your References section:

    Plasmid-based one-pot saturation mutagenesis. Wrenbeck EE, Klesmith JR, Stapleton JA, Adeniran A, Tyo KE, Whitehead TA. Nat Methods. 2016 Nov;13(11):928-930. doi: 10.1038/nmeth.4029. Epub 2016 Oct 10. 10.1038/nmeth.4029 PubMed 27723752