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HuEV-A
(Plasmid #68342)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 68342 Standard format: Plasmid sent in bacteria as agar stab 1 $75

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pCEP4
  • Backbone manufacturer
    Invitrogen
  • Backbone size (bp) 10200
  • Modifications to backbone
    The HuEV-A vector was constructed from a one-step ligation reaction with two inserts and a vector fragment. The backbone for the HuEV-A vector was the previously described pCEP4-Puro plasmid (5). The first insert was a DNA fragment containing the chloramphenicol resistance gene, ccdB gene, and att-R2 site, and was constructed by joining a NotI/KpnI segment of the Gateway Reading frame A cloning vector (Life Technologies, Cat #11828-029) containing the chloramphenicol resistance gene and the att-R2 site to a second synthetic DNA fragment (UEV) containing the complementary att-R1 site along with a series of epitope tags, recombination sites, and the Venus YFP sequence with BsiWI (compatible with BsrGI) and EagI (compatible with NotI) termini. The pCEP Puro backbone vector was digested with KpnI and BsrGI, and the inserts were ligated into the backbone using T4 DNA ligase in ligase buffer (Enzymatics). Vectors were transformed into E. coli and selected on ampicillin/chloramphenicol plates.
  • Vector type
    Mammalian Expression, Cre/Lox
  • Promoter TET-CMV
  • Selectable markers
    Puromycin
  • Tag / Fusion Protein
    • 3xFLAG, V5 and YFP (N terminal on backbone)

Growth in Bacteria

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

Cloning Information

  • Cloning method Gateway Cloning

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:

    HuEV-A was a gift from Jef Boeke (Addgene plasmid # 68342 ; http://n2t.net/addgene:68342 ; RRID:Addgene_68342)
  • For your References section:

    Fluorescence ImmunoPrecipitation (FLIP): a Novel Assay for High-Throughput IP. Mita P, Lhakhang T, Li D, Eichinger DJ, Fenyo D, Boeke JD. Biol Proced Online. 2016 Aug 15;18:16. doi: 10.1186/s12575-016-0046-x. eCollection 2016. 46 [pii] PubMed 27528826