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Addgene

pEG584
(Plasmid #40191)

Full plasmid sequence is not available for this item.

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 40191 Standard format: Plasmid sent in bacteria as agar stab 1 $85

Backbone

  • Vector backbone
    pCG150
  • Backbone manufacturer
    Addgene plasmid 17247
  • Backbone size w/o insert (bp) 6351
  • Modifications to backbone
    See comment below
  • Vector type
    Worm Expression
  • Selectable markers
    unc-119 rescuing fragment

Growth in Bacteria

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

Gene/Insert

  • Gene/Insert name
    Mex-5 (RNAi resistant)
  • Alt name
    Mex-5
  • Species
    C. elegans (nematode)
  • Mutation
    Exon 2 recoded to be RNAi-resistant
  • GenBank ID
    NM_070165.4 NP_502566.1
  • Entrez Gene
    mex-5 (a.k.a. CELE_W02A2.7)
  • Promoter 4.4 kb mex-5 promoter fragment
  • Tags / Fusion Proteins
    • Dendra2 PAFP (sequence optimized for worm expression; three synthetic introns inserted) (N terminal on backbone)
    • TEV protease cleavage site (N terminal on backbone)
    • S-peptide (N terminal on backbone)

Cloning Information

Terms and Licenses

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

Depositor Comments

Dendra::MEX-5 constructs were constructed as follows. A 4.4 kb mex-5 promoter fragment based on Tenlen et al. (2008) was cloned into pDONRP4P1R (Invitrogen). Dendra2/TEV/S-peptide (Gallo et al., 2010) was cloned into pDONR201. A MEX-5 genomic fragment from the start ATG through 648 bp of 3'UTR was cloned into pDONRP2RP3. Exon 2 of mex-5 was recoded to be RNAi-resistant (Gen-Script) so as to allow depletion of endogenous MEX-5/6 without depletion of the transgene. These constructs were assembled into pCG150 using three-way Gateway system (LR reaction) (Invitrogen) (Merritt et al., 2008). Mutations were made by recombinant PCR and all inserts were sequenced verified.

Alternate plasmid name: mex-5 prom::Dendra2/TEV/Speptide::MEX-5RR - mex-5 3'UTR

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:

    pEG584 was a gift from Geraldine Seydoux (Addgene plasmid # 40191 ; http://n2t.net/addgene:40191 ; RRID:Addgene_40191)
  • For your References section:

    Regulation of the MEX-5 gradient by a spatially segregated kinase/phosphatase cycle. Griffin EE, Odde DJ, Seydoux G. Cell. 2011 Sep 16;146(6):955-68. 10.1016/j.cell.2011.08.012 PubMed 21925318