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pVEGFR2_TEV(N)_NES_TCS(Q'L)_HA-dCas9(N)_P2A_Puro-WPRE
(Plasmid #101104)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 101104 Standard format: Plasmid sent in bacteria as agar stab 1 $65

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pTMt_NES_TCS(Q’G)_HA-dCas9(N)_P2A-Puro-WPRE
  • Modifications to backbone
    A sequence containing the VEGFR2 (KDR) leader peptide, extracellular domain and transmembrane domain were PCR amplified from plasmid pDONR223-KDR (gift from William Hahn & David Root, Addgene plasmid #23925) and used to replace the TMt in pTMt_NES_TCS(Q’G)_HA-dCas9(N)_P2A-Puro-WPRE. The N-terminal TEV fragment was then amplified from full length TEV protease as previously described (Wehr et al., 2006) and fused to the C-terminus of the VEGFR2 transmembrane domain. In addition, a weak TEV cleavage site (ENLYFQL) was inserted instead of the TCS(Q’G).
  • Vector type
    Mammalian Expression, CRISPR, Synthetic Biology
  • Selectable markers
    Puromycin

Growth in Bacteria

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

Gene/Insert

  • Gene/Insert name
    VEGFR2-dCas9(N)
  • Tag / Fusion Protein
    • HA

Resource Information

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:

    pVEGFR2_TEV(N)_NES_TCS(Q'L)_HA-dCas9(N)_P2A_Puro-WPRE was a gift from Tudor Fulga (Addgene plasmid # 101104 ; http://n2t.net/addgene:101104 ; RRID:Addgene_101104)
  • 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