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pET22b-ecDHFR-D27S/Y100F
(Plasmid #110827)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 110827 Standard format: Plasmid sent in bacteria as agar stab 1 $85
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Backbone

  • Vector backbone
    pET22b
  • Backbone manufacturer
    Novagen
  • Backbone size w/o insert (bp) 5493
  • Vector type
    Bacterial Expression

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin, 100 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    Low Copy

Gene/Insert

  • Gene/Insert name
    ecDHFR
  • Alt name
    dihydrofolate reductase
  • Alt name
    folA
  • Species
    E.coli
  • Insert Size (bp)
    480
  • Mutation
    D27 mutated to S27, Y100 mutated to F100
  • Promoter T7

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site NdeI (unknown if destroyed)
  • 3′ cloning site BamHI (unknown if destroyed)
  • 5′ sequencing primer TAATACGACTCACTATAGGG
    • (Common Sequencing Primers)

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:

    pET22b-ecDHFR-D27S/Y100F was a gift from Stephen Benkovic (Addgene plasmid # 110827 ; http://n2t.net/addgene:110827 ; RRID:Addgene_110827)

  • For your References section:

    Escherichia coli dihydrofolate reductase catalyzed proton and hydride transfers: temporal order and the roles of Asp27 and Tyr100. Liu CT, Francis K, Layfield JP, Huang X, Hammes-Schiffer S, Kohen A, Benkovic SJ. Proc Natl Acad Sci U S A. 2014 Dec 23;111(51):18231-6. doi: 10.1073/pnas.1415940111. Epub 2014 Dec 1. 10.1073/pnas.1415940111 PubMed 25453098
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