Part:BBa_K648020

Fast-Fusion GFP-Xyle Reporter (RecA cleaveage with small linker)

This is a fast-acting reporter using the Xyle gene (BBa_K648011) fused to GFP. In its normal un-cleaved state the polymerization ability necessary for enzymatic action is inactivated. Once the fusion protein is cleaved by the RecA, which is activated by damaged DNA, the catechol-2,3-dioxygenase enzyme encoded by the Xyle gene is able to convert catechol to the bright yellow product 2-hydroxy-cis,cis-muconic semialdehyde. This reporter is able to produce a visible color response in a matter of minutes.

This version uses the RecA cleavage site (BBa_K648009) followed by the smallest of the three flexible fusion protein linkers (BBa_K648005). The linker is attached to the N-terminus of the Xyle gene.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 7
  Illegal NheI site found at 30
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 704