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AICSDP-22:CETN2-mTagRFP-T
(Plasmid #101781)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 101781 Standard format: Plasmid sent in bacteria as agar stab 1 $65

This material is available to academics and nonprofits only.

Backbone

Growth in Bacteria

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

Gene/Insert

  • Gene/Insert name
    CETN2 Homology Arms with mTagRFPT-linker
  • Alt name
    CETN2
  • Alt name
    CEN2
  • Species
    H. sapiens (human)
  • Insert Size (bp)
    2744
  • Mutation
    homology arms contain point mutations to disrupt crRNA binding sites used
  • Entrez Gene
    CETN2 (a.k.a. CALT, CEN2)
  • Tag / Fusion Protein
    • mTagRFPT-linker (N terminal on backbone)

Cloning Information

Resource Information

Depositor Comments

This plasmid has been used with locus-specific CRISPR/Cas9 to add a mTagRFP-T tag to the N-terminus of human CETN2 in WTC human induced pluripotent stem cells by the Allen Institute for Cell Science. Linker (AA) sequence: SGLRS. After protein tagging using this donor template plasmid and CRISPR/Cas9 reagents, transfected cells may exhibit varying intensity levels of fluorescence, likely due to editing precision. To obtain cells of uniform intensity levels, see our protocol for fluorescence-assisted cell sorting and subcloning of transfected cells (https://www.allencell.org/instructional-videos-and-tutorials-for-cell-methods.html). Further, we recommend PCR-based assays for identifying precisely edited clones as previously described (https://www.molbiolcell.org/doi/abs/10.1091/mbc.e17-03-0209). For more information on the entire plasmid collection, please see https://www.addgene.org/allen-institute-cell-science/ .

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:

    AICSDP-22:CETN2-mTagRFP-T was a gift from The Allen Institute for Cell Science (Addgene plasmid # 101781 ; http://n2t.net/addgene:101781 ; RRID:Addgene_101781)