Coding

Part:BBa_K4830027

Designed by: Ong Jie Hao   Group: iGEM23_NTU-Singapore   (2023-10-11)
Revision as of 17:26, 11 October 2023 by Jong154 (Talk | contribs) (Usage and Biology)


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


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 352
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 352
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 352
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 352
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
//cds/reporter/gfp
//cds/reporter/rfp
Parameters
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