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pCRIS-PITChv2-dTAG-Puro (BRD4)
(Plasmid #91796)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 91796 Standard format: Plasmid sent in bacteria as agar stab 1 $65

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pCRIS-PITChv2 (addgene #63672)
  • Backbone manufacturer
    Takashi Yamamoto lab
  • Backbone size w/o insert (bp) 4579
  • Total vector size (bp) 5738
  • Modifications to backbone
    A C-terminal dTAG cassette for endogenous protein degradation replaced the EGFP cassette in the original pCRIS-PITChv2 plasmid (addgene #63672)
  • Vector type
    Mammalian Expression, CRISPR
  • Selectable markers
    Hygromycin

Growth in Bacteria

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

Gene/Insert

  • Gene/Insert name
    FKBP_F36V-2xHA-P2A-Puro
  • Species
    Synthetic
  • Mutation
    Phenylalanine 36 to valine in FKBP12
  • Promoter Promotorless

Cloning Information

  • Cloning method Gibson Cloning
  • 5′ sequencing primer tcgcccttaattgtgagcgga
  • 3′ sequencing primer gaaaggacagtgggagtggca
  • (Common Sequencing Primers)

Resource Information

Depositor Comments

See:
Sakuma, T., Nakade, S., Sakane, Y., Suzuki, K.-I. & Yamamoto, T. MMEJ-assisted gene knock-in using TALENs and CRISPR-Cas9 with the PITCh systems. Nat Protoc 11, 118–133 (2016).
for the original backbone and general cloning protocol.

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:

    pCRIS-PITChv2-dTAG-Puro (BRD4) was a gift from James Bradner (Addgene plasmid # 91796 ; http://n2t.net/addgene:91796 ; RRID:Addgene_91796)
  • For your References section:

    The dTAG system for immediate and target-specific protein degradation. Nabet B, Roberts JM, Buckley DL, Paulk J, Dastjerdi S, Yang A, Leggett AL, Erb MA, Lawlor MA, Souza A, Scott TG, Vittori S, Perry JA, Qi J, Winter GE, Wong KK, Gray NS & Bradner JE.. Nature Chemical Biology (2018) 10.1038/s41589-018-0021-8