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pTMt_TCS(Q’G)_NLS-HA-dCas9(C)-VP64_T2A_MCP-P65-HSF1
(Plasmid #101102)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 101102 Standard format: Plasmid sent in bacteria as agar stab 1 $75

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pX856
  • Modifications to backbone
    The pX856 vector (gift from Feng Zhang, Addgene plasmid #62888) was modified as follows: the U6 promoter/sgRNA scaffold/U6 terminator cassette was removed; the dCas9(C)VP64 N-terminal NLS+FKBP were also removed; a transmembrane tether (TMt) containing the Igκ signal sequence, (GGGS)2 linker, myc epitope tag, PDGF receptor transmembrane domain and the XTEN linker, was synthesized as a gBlock (IDT). This transmembrane tether was then fused to the N-terminus of HA-dCas9(C)VP64 via a TEV cleavage site (ENLYFQG) and NLS. The MCP-P65-HSF1 from plasmid MS2-P65-HSF1_GFP (gift from Feng Zhang, Addgene plasmid #61423) was fused to the C-terminus of dCas9(C)VP64 via at T2A site.
  • Vector type
    Mammalian Expression, CRISPR, Synthetic Biology

Growth in Bacteria

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

Gene/Insert

  • Gene/Insert name
    TMt-dCas9(C)-VP64
  • Tags / Fusion Proteins
    • Myc (N terminal on insert)
    • HA (N terminal on insert)

Resource Information

  • Supplemental Documents
  • A portion of this plasmid was derived from a plasmid made by
    pX856 vector (gift from Feng Zhang, Addgene plasmid #62888); plasmid MS2-P65-HSF1_GFP (gift from Feng Zhang, Addgene plasmid #61423)
  • Terms and Licenses
  • Industry Terms
    • Not Available to Industry
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:

    pTMt_TCS(Q’G)_NLS-HA-dCas9(C)-VP64_T2A_MCP-P65-HSF1 was a gift from Tudor Fulga (Addgene plasmid # 101102 ; http://n2t.net/addgene:101102 ; RRID:Addgene_101102)
  • For your References section:

    Engineering Synthetic Signaling Pathways with Programmable dCas9-Based Chimeric Receptors. Baeumler TA, Ahmed AA, Fulga TA. Cell Rep. 2017 Sep 12;20(11):2639-2653. doi: 10.1016/j.celrep.2017.08.044. 10.1016/j.celrep.2017.08.044 PubMed 28903044