Part:BBa_K2455008

YddG-AroG construct with an E. coli RBS between (BBa_K2455004 & BBa_K2455000)

This is a composite part consisting of the two YddG (BBa_K2455004) and AroG (BBa_K2455000) biobricks with an RBS in between.

This biobrick has been designed in order to over-express and secrete tryptophan, tyrosine and phenylalanine in E. coli.

YddG is an aromatic amino acid exporter that is responsible for the secretion of Tryptophan, Tyrosine and Phenylalanine. Previous work by Wang et al. have shown that over-expression of YddG in E.coli increases the accumulation of Tryptophan in the culture medium.

AroG gene from E. coli strain MG1655 carrying an P150L point mutation (to avoid negative feedback inhibition) and a hexa histidine-tag attached by a short linker sequence. AroG is the first enzyme of the shikimate pathway leading to synthesis of chorismate, which subsequently can be converted to tryptophan, tyrosine, and phenylalanine. Being the first enzyme in the pathway, it determines the carbon flow towards amino acid synthesis and thus the production. Previous work (Gu et al., 2012) have shown that overexpression of AroG leads to increased tryptophan production in E. coli.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal XhoI site found at 1847
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 577
  Illegal AgeI site found at 639
  Illegal AgeI site found at 756
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI.rc site found at 1154