Coding
Part:BBa_K4830028
Designed by: Ong Jie Hao Group: iGEM23_NTU-Singapore (2023-10-11)
eGFP_simple premature stop codon
eGFP_premature stop codon is a protein coding sequence that contains the eGFP with a premature stop codon. This coding sequence is particularly useful for Traffic Light Reporter (TLR) assays that screen for DNA editing activity. This sequence is used in a TLR assay for prime editing where pegRNAs and ngRNAs targeting the premature stop codon were designed to swap the premature stop codon into a functional amino acid, restoring the wild-type eGFP.
When cloned into a suitable plasmid and subsequently transfected into mammalian cells for in vitro DNA editing, successfully edited cells will fluorescence green which can then be analysed with flow cytometry.
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
[edit]
Categories
Parameters
//cds/reporter/gfp
color | Green |
emission | 509nm |
excitation | 488nm |