Part:BBa_K4830027
mCherry_C-terminus stop codon_Clover
The mCherry_C-terminus stop codon_Clover is protein coding sequence containing the mCherry fluorescent protein with a premature stop codon installed in the C-terminus before the Clover fluorescent protein. The presence of the premature stop codon prevents the expression of coding regions found downstream. This is particularly useful in assays such as the Traffic Light Reporter (TLR) assay.
Usage and Biology
The mCherry_C-terminus stop codon is utilized in the Traffic Light Reporter (TLR) assay, where a Clover fluorescent protein (commonly known as GFP) is found downstream. In this TLR assay, the Clover is not expressed due to the presence of the stop codon before the GFP. As such, the TLR assay is used to screen for DNA editing activity that swaps the stop codon into a functional amino acid, restoring the expression of Clover.
The entire coding sequence can be cloned into suitable plasmids and subsequently transfected into mammalian cells for in vitro DNA editing activity assay.
Sequence and Features
The sequence shows the mCherry fluorescent protein with a stop codon on the C-terminus. This is followed with a Clover fluorescent protein downstream.
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 352
- 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 352
- 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 352
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 352
- 1000COMPATIBLE WITH RFC[1000]
//cds/reporter/rfp
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