Part:BBa_K2201320
functional GFP-streptavidin fusion protein with medium gly-gly-ser linker and T7-promoter and RBS
This fusion protein containing GFP and streptavidin connected by a Gly-Gly-Ser-linker is used to show the properties and uses of the ncAA 2-NPA. The streptavidin of this fusion protein is able to bind 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. This version of the fusion protein is needed to validate the binding and detection properties without the incorporation of the ncAA 2-NPA compared to the part K2201221. Experiments with this protein can deliver the reference values of binding efficiency and fluorescence signal undisturbed by the ncAA. When compared to the values of the amber variant, it can be determined if there is any loss or change in the binding or in the fluorescence properties caused by the 2-NPA or the synthetase. 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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