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pcDNA5/FRT/TO-Venus-Flag-Gateway (1124)
(Plasmid #40999)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 40999 Standard format: Plasmid sent in bacteria as agar stab 1 $75

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pcDNA5/FRT/TO
  • Backbone manufacturer
    Invitrogen
  • Modifications to backbone
    Venus-3xFlag added (C-terminal), converted to Gateway destination vector (see notes below)
  • Vector type
    Mammalian Expression
  • Promoter CMV
  • Selectable markers
    Hygromycin
  • Tags / Fusion Proteins
    • Venus YFP (C terminal on insert)
    • 3xFLAG (DYKDHDGDYKDHDIDYKDDDDK) (C terminal on insert)

Growth in Bacteria

  • Bacterial Resistance(s)
    Chloramphenicol and Ampicillin
  • Growth Temperature
    37°C
  • Growth Strain(s)
    ccdB Survival
  • Copy number
    Unknown

Cloning Information

Resource Information

  • Terms and Licenses
  • Industry Terms
    • Not Available to Industry
  • Articles Citing this Plasmid

Depositor Comments

Venus YFP-3xFlag was inserted via BstXI-XhoI sites (to create pcDNA5/FRT/TO-Venus-Flag, Addgene Plasmid# 40998).

The plasmid was then converted to a Gateway destination vector by inserting Gateway cassette C1, containing recombination sites attR1 and attR2, CmR, and ccdB, into the Afl II site (blunt ended).

This plasmid can be used to place genes into the pcDNA5/FRT/TO-Venus-Flag construct by Gateway reaction, to make tetracycline-inducible stable cell lines.

For subcloning, the following restriction sites are present:
Between the CMV promoter and the Gateway cassette: PmeI (non-unique)
Between the Gateway cassette and Venus: HindIII, Asp718I, KpnI, BamHI (non-unique, also in GW casette), and BstXI
Between Venus and BGHpA: XhoI, Eco0109I, ApaI, and PmeI (non-unique)

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:

    pcDNA5/FRT/TO-Venus-Flag-Gateway (1124) was a gift from Jonathon Pines (Addgene plasmid # 40999 ; http://n2t.net/addgene:40999 ; RRID:Addgene_40999)