Coding

Part:BBa_K4821005

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


RdhANp_M132O_K12

This sequence codes for a modified RdhANp that has been docked to the ortho chlorine position in PCB 132 and codon-optimized for expression in E. coli K-12. This enzyme contains the following mutations: F294Y K295L E298L K310A P314Y M315L S423A S425V M426L N455I 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 PCB 132.

Usage and Biology

This enzyme was redesigned with the intended purpose of dechlorinating the chlorine at the ortho position of PCB132 to potentially better facilitate the breakdown of PCB132 into acetyl CoA by 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