Part:BBa_K2201321
functional GFP-streptavidin fusion protein with medium gly-gly-ser linker containing an amber codon
This fusion protein containing GFP and streptavidin connected by a Gly-Gly-Ser-linker containing an amber codon is used to show the properties and usages of the ncAA 2-NPA. The streptavidin of this fusion protein can bind to biotin, such that the protein could be immobilized on any biotinylated surface. The GFP is fluorescent, such that the binding of the protein can easily be detected. The 2-NPA in the linker induces a cleavage of the peptide backbone when radiated with light at ʎ = 365 nm. After the radiation, the GFP can be removed by simply washing it off the surface, so that the following loss of fluorescence shows the usage of this ncAA. To get the fusion proteins encoded in the parts K2201220 and K2201221, a BioBrick assembly with a suitable promoter is needed to achieve sufficient protein expression (e.g. T7 promoter and RBS of K525998). This alone worked for the fusion protein without the amber codon of the part K2201320. To get the fusion protein containing the 2-NPA (K2201321) a co-transformation of the fusion protein and the RS-Part needed.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 825
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 867
Illegal AgeI site found at 918 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 682
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