Part:BBa_K4229021
sfGFP AMBER stop codon at the position of the 4th amino acid
Short Description:
Superfolder GFP (sfGFP) with an amber stop codon mutation at position 4, replacing phenylalanine
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 421
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Short Description:
The G4 variant of sfGFP was created using site-directed mutagenesis, aiming to insert the amber stop codon at position 4. The sfGFP can only be completely synthesized upon successful amber stop codon suppression. Hence, sfGFP G4 can be used as a model to test the efficiency of incorporation of non-canonical amino acids via amber stop codon suppression technology.
Usage:
Besides the well-known 21 canonical amino acids, there is a variety of non-canonical amino acids which can be used to further modify proteins. One way of incorporating non-canonical amino acids is via the amber stop codon suppression technology. This part, sfGFP G4, is a model protein to test non-canonical amino acid incorporation. Without successful incorporation, a non-functional version of sfGFP will be translated, and therefore not give a fluorescence signal. However, after successful incorporation of the non-canonical amino acid, fluorescence will be restored providing an immediate readout of the efficiency of amber stop codon suppression via orthogonal translation systems.
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