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3xFLAG-dCas9/pCMV-7.1
(Plasmid #47948)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 47948 Standard format: Plasmid sent in bacteria as agar stab 1 $65

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pCMV-7.1
  • Backbone manufacturer
    Sigma-Aldrich
  • Backbone size w/o insert (bp) 4636
  • Total vector size (bp) 8848
  • Vector type
    Mammalian Expression, CRISPR

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin
  • Growth Temperature
    30°C
  • Growth Strain(s)
    NEB Stable
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    3xFLAG-dCas9
  • Insert Size (bp)
    4212
  • Mutation
    human codon-optimized, D10A + H840A (catalytically inactive)
  • Promoter CMV
  • Tags / Fusion Proteins
    • 3xFLAG (N terminal on insert)
    • NLS (C terminal on insert)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site Hind III (destroyed during cloning)
  • 3′ cloning site EcoR V (destroyed during cloning)
  • 5′ sequencing primer AAT GTC GTA ATA ACC CCG CCC CGT TGA CGC
  • 3′ sequencing primer TAT TAG GAC AAG GCT GGT GGG CAC
  • (Common Sequencing Primers)

Resource Information

Depositor Comments

This plasmid can be used to isolate specific genomic regions of interest using a catalytically inactive Cas9 fused with a tag(s).

Purify: locus-specific chromatin immunoprecipitation (enChIP)
This system is compatible with gRNA_cloning_vector (www.addgene.org/41824) from the Church lab.

Additional information and protocols can be found at:
http://www.med.hirosaki-u.ac.jp/~bgb/iChIP_protocols/index_e.html

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

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:

    3xFLAG-dCas9/pCMV-7.1 was a gift from Hodaka Fujii (Addgene plasmid # 47948 ; http://n2t.net/addgene:47948 ; RRID:Addgene_47948)
  • For your References section:

    Efficient isolation of specific genomic regions and identification of associated proteins by engineered DNA-binding molecule-mediated chromatin immunoprecipitation (enChIP) using CRISPR. Fujita T, Fujii H. Biochem Biophys Res Commun. 2013 Sep 13;439(1):132-6. doi: 10.1016/j.bbrc.2013.08.013. Epub 2013 Aug 11. 10.1016/j.bbrc.2013.08.013 PubMed 23942116