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pAc5 HA3-eDHFR-T2A-neo
(Plasmid #86396)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 86396 Standard format: Plasmid sent in bacteria as agar stab 1 $65

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pAc5
  • Modifications to backbone
    see Gonzalez et al Scientific Reports 1:75, 2011
  • Vector type
    CRISPR
  • Selectable markers
    Neomycin (select with G418)

Growth in Bacteria

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

Gene/Insert

  • Gene/Insert name
    HA3 eDHFR T2A neo
  • Species
    E coli
  • Mutation
    R12Y, G67S and Y100I mutations in E coli DHFR
  • Tag / Fusion Protein
    • HA3 eDHFR T2A neo (C terminal on insert)

Resource Information

  • A portion of this plasmid was derived from a plasmid made by
    The mutant E coli DHFR gene used in the two plasmids was derived from Cho et al PlosOne 8: e72393 Addgene plasmid 29325.
  • Terms and Licenses

Depositor Comments

pAc5 HA3-eDHFR-T2A-neo was derived from pAc5gRNA Cas9 (Bassett, A.R.,et al Biol Open 3:42, 2014) by replacing the EcoRI-Hind III fragment encoding Cas9 with a PCR fragment encoding HA3-eDHFR amplified from pBMN DHFR(DD)-YFP (Iwamoto eet al Chem Biol 17:981, 2010.) (Addgene plasmid #29325) and by replacing the puro gene with neo using the NheI-XhoI sites
This degron has R12Y, G67S and Y100I mutations in eDHFR

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:

    pAc5 HA3-eDHFR-T2A-neo was a gift from David Bentley (Addgene plasmid # 86396 ; http://n2t.net/addgene:86396 ; RRID:Addgene_86396)
  • For your References section:

    Selectable one-step PCR-mediated integration of a degron for rapid depletion of endogenous human proteins. Sheridan RM, Bentley DL. Biotechniques. 2016 Feb 1;60(2):69-74. doi: 10.2144/000114378. eCollection 2016 Feb. 10.2144/000114378 PubMed 26842351