Part:BBa_K648011

Standard 25-Ready Xyle Reporter

This is a mutated version of the Xyle reporter gene which encodes for the enzyme catechol-2,3-dioxygenase (metapyrocatechase), which converts catechol to the bright yellow product 2-hydroxy-cis,cis-muconic semialdehyde. This version of Xyle has been made compatible with standard 25 assembly methods by removing three restriction sites (two NgoMIV sites: at bp 315 and 486, as well as one AgeI site: at bp 837). These mutations were made synonymous with the original sequence and codon optimized for E. Coli.

Because of the synonymous mutations, this gene can be easily used to create fusion protein parts. For more information on the Xyle gene and its uses as a reporter see part BBa_J33204.

Sequence and Features

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]

This part was used by the Penn State team to construct variations of their improved fast-acting reporters. Proof of its compatibility with assembly 25 methods is shown below by its sequence show here attached to a small fusion linker peptide (BBa_K648007).

Sequence of J23100+B0034+K648011+K648007 (with reverse primer):

The sequence here shows the Xyle gene in green, followed by the assembly 25 scar (ACCGGC) shown highlighted in yellow. The short sequence highlighted in blue is the small linker.