Part:BBa_K597101
GFP-AviTag [AviTagged green fluorescent protein]
GFP-AviTag codes for green fluorescent protein with an appended biotinylation marker. When attached to a promoter, it can be used as a fluorescent reporter, in addition to getting biotinylated by Escherichia coli. E. coli has a natural mechanism encoded by the birA gene to produce biotin and biotinylate proteins. Adding biotin to the bacterial cultures enhances this mechanism. This enzyme can then be used in biotin-streptavidin binding applications.
The GFP-AviTag fusion protein was constructed to have the AviTag sequence on the C-terminus, followed by a stop codon. The resulting protein, once biotinylated, is capable of a ligand reaction with surface-coated avidin, while retaining the fluorescence of GFP. This molecule was used to test the efficacy of the binding interaction between the functionalized protein and a streptavidin-coated surface.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 715
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 644
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