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Sa-dCas9-NLS-3xFLAG/pcDNA3.1
(Plasmid #98041)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 98041 Standard format: Plasmid sent in bacteria as agar stab 1 $65

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pcDNA3.1
  • Backbone manufacturer
    Invitrogen
  • Backbone size w/o insert (bp) 5500
  • Total vector size (bp) 8767
  • Vector type
    Mammalian Expression, CRISPR
  • Selectable markers
    Neomycin (select with G418)

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin
  • Growth Temperature
    30°C
  • Growth Strain(s)
    NEB Stable
  • Growth instructions
    High frequency of recombination. Culture at 30°C.
  • Copy number
    Unknown

Gene/Insert

  • Gene/Insert name
    Sa-dCas9-NLS-3xFLAG
  • Species
    Synthetic; Staphylococcus aureus
  • Insert Size (bp)
    3264
  • Mutation
    D10A + H557A
  • Promoter CMV
  • Tags / Fusion Proteins
    • NLS (nuclear localization signal) (C terminal on insert)
    • 3xFLAG tag (C terminal on insert)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site Nhe I (not destroyed)
  • 3′ cloning site Not I (not destroyed)
  • 5′ sequencing primer T7
  • 3′ sequencing primer BGH Reverse
  • (Common Sequencing Primers)

Resource Information

  • A portion of this plasmid was derived from a plasmid made by
    A portion of this plasmid was derived from a plasmid made by Keith Joung (Addgene plasmid 70703)
  • Terms and Licenses

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 BPK2660 (www.addgene.org/70709) from the Joung lab.

Additional information and protocols can be found at:
http://www.biken.osaka-u.ac.jp/lab/microimm/fujii/iChIP_protocols/english.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:

    Sa-dCas9-NLS-3xFLAG/pcDNA3.1 was a gift from Hodaka Fujii (Addgene plasmid # 98041)
  • For your References section:

    enChIP systems using different CRISPR orthologues and epitope tags. Fujita T, Yuno M, Fujii H. BMC Res Notes. 2018 Feb 27;11(1):154. doi: 10.1186/s13104-018-3262-4. 10.1186/s13104-018-3262-4 PubMed 29482606