Part:BBa_K4830029

eGFP_complex premature stop codon

The eGFP_complex premature stop codon is a derivative of the eGFP_simple premature stop codon. Similar to the eGFP_simple premature stop codon, the eGFP_complex premature stop codon can be utilized in a Traffic Light Reporter (TLR) assay.

However, the edit required is complex as it involves the truncation in a particular region from 42bp to 31bp. Specific pegRNA and ngRNA targeting the premature stop codon have been designed - TLR-76 pegRNA and 777BFP ngRNA.

Usage and Biology

The eGFP_complex premature stop codon is a coding sequence used in the Traffic Light Reporter (TLR) assay. It contains a particular region that requires a complex edit to truncate from 42bp to 31bp, before restoring the wild-type eGFP.

Sequence and Features

The coding sequence spans the eGFP with an inherent premature stop codon to be edited.

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal SpeI site found at 192
  Illegal PstI site found at 160
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal SpeI site found at 192
  Illegal PstI site found at 160
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal SpeI site found at 192
  Illegal PstI site found at 160
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal SpeI site found at 192
  Illegal PstI site found at 160
  Illegal NgoMIV site found at 734
  
• 1000
  COMPATIBLE WITH RFC[1000]