Champion™ pET104 BioEase™ Gateway® Biotinylation System

Catalog number: K10401

Invitrogen™  Related applications: Gateway Cloning | Protein Expression

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The Champion™ pET104-DEST BioEase™ Gateway® Expression System provides an easy, efficient method for expressing, purifying, and detecting biotinylated recombinant proteins. The Champion™ pET104-DEST vector (Figure 1) incorporates a 72 amino acid sequence from K. pneumoniae that directs in vivo biotinylation of a specific lysine residue. This tag is recognized and efficiently biotinylated by the enzyme Biotin Protein Ligase (BPL) encoded by the birA gene in E. coli(1). Proteins produced in the Champion™ pET104-DEST vector are expressed as fusion proteins to this sequence. Biotinylated fusion proteins can be purified on streptavidin agarose or detected on western blots using streptavidin-HRP (Figure 2) or streptavidin-AP conjugates.

In addition to the BioEase™ tag, the Champion™ pET104-DEST vector includes the following features:

attR sites for efficient recombination with any attL-flanked Gateway® entry vector
• The T7lac bacteriophage promoter for high-level expression of the recombinant fusion protein
• An enterokinase cleavage site for removal of the N-terminal BioEase™ fusion tag
For Research Use Only. Not for use in diagnostic procedures.


Expression: E. coli
Promoter: T7 and lacO
Bacterial or Yeast Strain: BL21 Star™(DE3)
Antibiotic Resistance (Bacterial): Ampicillin (AmpR)
Cleavage: EK (Enterokinase) Recognition Site
Cloning Method: Gateway®
Constitutive or Inducible System: Inducible
Expression Mechanism: Cell-Based Expression
Inducing Agent: IPTG
Product Line: BioEase™, Champion™, Gateway®
Product Size: 1 kit
Protein Tag or Fusion: BioEase Tag
Selection Agent (Eukaryotic): None
Vector Type: pET

Contents & storage

The Champion™ pET104-DEST BioEase™ Gateway® Expression Kit includes 6 flg of the Champion™ pET104-DEST vector, 10 µg of the Champion™ pET104/GW/lacZ control vector, twenty-one 50-µl aliquots of BL21 Star™(DE3) One Shot¤ competent cells, S.O.C. medium, and pUC19 control DNA. Store vectors at -20°C. Store competent cells at -80°C. Guaranteed stable for 6 months when properly stored.


Manuals & protocols