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Addgene

pE5n
(Plasmid #17459)

Ordering

This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 17459 Standard format: Plasmid sent in bacteria as agar stab 1 $85

Backbone

  • Vector backbone
    pENTR1A
  • Backbone manufacturer
    Invitrogen
  • Backbone size (bp) 3030
  • Modifications to backbone
    A short cloning site region and stop codon replaced ccdB gene in pENTR1A. Inserted ECFP tag
  • Vector type
    Mammalian Expression, Bacterial Expression, Yeast Expression, Insect Expression ; modified Gateway Entry vector

Growth in Bacteria

  • Bacterial Resistance(s)
    Kanamycin, 50 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    None
  • Tag / Fusion Protein
    • ECFP (N terminal on backbone)

Cloning Information

  • Cloning method Gateway Cloning
  • 5′ sequencing primer pENTR-F (5'-CTACAAACTCTTCCTGTTAGTTAG-3')
  • 3′ sequencing primer pENTR-R (5'-ATGGCTCATAACACCCCTTG-3')
  • (Common Sequencing Primers)

Terms and Licenses

  • Academic/Nonprofit Terms
  • Industry Terms
    • Not Available to Industry
Trademarks:
  • Zeocin® is an InvivoGen trademark.

Depositor Comments

Part of a modified Gateway Entry vector system for generating epitope tagged constructs. Clone your gene of interest into this vector using the BamHI and NotI cloning sites to obtain a N-terminal ECFP tagged Entry vector. The resulting Entry vector can be recombined into Gateway Destination vectors for bacteria, insect mammalian, plant or yeast expression via the Gateway LR reaction.

The tag is present in the Entry cassette to avoid the presence of att-derived protein linker sequences between the tag and gene or interest corresponding to the attB recombination site. A stop codon is already present within the vector (see plasmid map).

GenBank accession number: EU344820

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:

    pE5n was a gift from Giovanna Benvenuto (Addgene plasmid # 17459 ; http://n2t.net/addgene:17459 ; RRID:Addgene_17459)
  • For your References section:

    A modified Gateway cloning strategy for overexpressing tagged proteins in plants. Dubin MJ, Bowler C, Benvenuto G. Plant Methods. 2008 Jan 22. 4(1):3. 10.1186/1746-4811-4-3 PubMed 18211686
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