Part:BBa_K4139027

COAPE-GFP

This construct starts with an OmpA tag and is a signaling peptide that will bring the protein from cytoplasm to the periplasm. This is followed by enhanced green fluorescent protein (eGFP) then our Chimeric Anti-PbEL04 (CAPE). The CAPE proportion is the same as our CAPE protein. Finally we added a His-tag to further help with purification. The function of this protein is to bind to PbEL04 and glow once binded correctly. The reason for using eGFP is to show if proper binding has occurred. If proper binding has occurred, then the protein will glow green when blue light in the 470 nm to 498 nm range. The difference between CAPE-GFP (BBa_K4139013) and COAPE-GFP (BBa_K4139015) is the OmpA tag which was used as a solubility tag. This construct was successfully purified by our team. We first made this coding region as a basic part (BBa_K4139015) which we have made as a composite part with a ribosome binding site, promoter, and terminator.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal PstI site found at 1839
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 27
  Illegal NheI site found at 50
  Illegal PstI site found at 1839
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 1839
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 1839
  Illegal AgeI site found at 529
  
• 1000
  COMPATIBLE WITH RFC[1000]