Part:BBa_K4830028

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. pegRNA and ngRNA targeting this particular sequence have already been designed - PETLR pegRNA 1 and PETLR ngRNA 1

Usage and Biology

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

The sequence contains the eGFP with a premature stop codon.

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]