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pKK-mClover3-TEV
(Plasmid #105778)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 105778 Standard format: Plasmid sent in bacteria as agar stab 1 $65

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pcDNA5/FRT/TO
  • Backbone manufacturer
    Invitrogen
  • Vector type
    Mammalian Expression ; Flp-In competent
  • Selectable markers
    Hygromycin
  • Tag / Fusion Protein
    • mClover3-TEV (N terminal on 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
    None
  • Tag / Fusion Protein
    • mClover3-TEV (N terminal on backbone)

Resource Information

Depositor Comments

This vector is a member of the pKK vector family. Its cloning site was optimized for the SLIC approach, however, a traditional restriction and ligation based approach is also possible (see Szczesny RJ et al. for detailed protocols; bioRxiv, doi: https://doi.org/10.1101/160101). This vector is a derivate of the pcDNA5/FRT/TO plasmid (Invitrogen) and is suitable for stable cell line generation using the Flp-In system. Transcription of the gene of interest is driven by a tetracycline responsive CMV promoter. A given coding sequence can be cloned into all pKK vectors using just three universal PCR primers and a ligation-independent DNA cloning method. mClover3 is a brighter derivative of mEGFP. The mClover3-mRuby3 pair is superior to EGFP-mCherry in FRET analysis. At the time of publication (PMID: 26879144) mClover3 was the brightest jellyfish-derived GFP in mammalian cells. mClover3 coding sequence was cloned from Addgene ID: 74252.

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:

    pKK-mClover3-TEV was a gift from Andrzej Dziembowski (Addgene plasmid # 105778 ; http://n2t.net/addgene:105778 ; RRID:Addgene_105778)
  • For your References section:

    Versatile approach for functional analysis of human proteins and efficient stable cell line generation using FLP-mediated recombination system. Szczesny RJ, Kowalska K, Klosowska-Kosicka K, Chlebowski A, Owczarek EP, Warkocki Z, Kulinski TM, Adamska D, Affek K, Jedroszkowiak A, Kotrys AV, Tomecki R, Krawczyk PS, Borowski LS, Dziembowski A. PLoS One. 2018 Mar 28;13(3):e0194887. doi: 10.1371/journal.pone.0194887. eCollection 2018. 10.1371/journal.pone.0194887 PubMed 29590189