Part:BBa_K597102
VioA-AviTag [AviTagged violacein enzyme A]
VioA-AviTag codes for violacein enzyme A with an appended biotinylation marker. When attached to a promoter, it can be 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 culture enhances this mechanism. This enzyme can then be used in the violacein biosynthetic pathway with the added functionality of biotin-streptavidin binding.
The VioA-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 enzymatic activity of VioA.
The violacein biochemical pathway is composed of 5 enzymes, VioA-E. The synthesis of this violet pigment involves five enzymes (VioA, VioB, VioC, VioD, VioE) and begins with L-tryptophan as the initial substrate. However, only three of the five enzymes (VioA, VioB, and VioE) are required to make a colored product. Even with the exclusion of VioC and VioD from the synthesis pathway, L-tryptophan is converted into prodeoxyviolacein, a green pigment.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1012
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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