Coding

Part:BBa_K4821004

Designed by: Madeline Ho   Group: iGEM23_Washington   (2023-10-12)
Revision as of 14:26, 12 October 2023 by Lnjc (Talk | contribs)


RdhANp_M132P_K12

This sequence codes for a modified RdhANp that has been docked to the para chlorine position in PCB 132 and codon-optimized for expression in E. coli K-12. This enzyme contains the following mutations: F294I K295L E298L K310A R312T P314Y M315F S423T S425A M426L N455L A456M H457D V556I A563G. It was designed as an improvement on part BBa_K4821002, the codon-optimized wild-type RdhANp dehalogenase to possess higher catalytic activity against PCB132.

Usage and Biology

This enzyme was redesigned with the intended purpose of dechlorinating the para chlorine of PCB132 to potentially better facilitate the breakdown of PCB132 into acetyl coA via the biphenyl degradation pathway (bph gene cluster)

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1628
    Illegal BamHI site found at 312
    Illegal BamHI site found at 419
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
None