Part:BBa_K4139021

Truncated PbEL04

This construct starts with a His-tag, this assists with the purification of the protein. After the His-tag we added thioredoxin to further assist with the purification. Thioredoxin helps with the folding of the protein and acts as a solubility tag as well. This is followed by a T.E.V cut site, which allows us to remove the thioredoxin and His-tag once purified. TEV cut site is cleaved off by TEV protease which is a member of the PA clan of chymotrypsin-like proteases. After the TEV cut site is our truncated version of PbEL04. This is just the target epitope of COAP-FP (BBa_K4139015). The use of this construct is to make a version of PbEL04 that is purified easily. We did end up being able to purify this construct. We first made this coding region as a basic part (BBa_K4139011) 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 1391
  Illegal PstI site found at 1517
  Illegal PstI site found at 1640
  Illegal PstI site found at 1763
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 27
  Illegal NheI site found at 50
  Illegal PstI site found at 1391
  Illegal PstI site found at 1517
  Illegal PstI site found at 1640
  Illegal PstI site found at 1763
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 1391
  Illegal PstI site found at 1517
  Illegal PstI site found at 1640
  Illegal PstI site found at 1763
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 1391
  Illegal PstI site found at 1517
  Illegal PstI site found at 1640
  Illegal PstI site found at 1763
  Illegal AgeI site found at 529
  
• 1000
  COMPATIBLE WITH RFC[1000]