Composite

Part:BBa_K2455007

Designed by: Jon Fugl   Group: iGEM17_UCopenhagen   (2017-10-27)
Revision as of 18:39, 28 October 2017 by Jon UCPH (Talk | contribs)

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YddG-TrpE construct with an E. coli RBS between (BBa_K2455004 & BBa_K2455001)

This is a composite part consisting of the two YddG (BBa_K2455004) and TrpE (BBa_K2455001) biobricks with an RBS in between.

This biobrick has been designed in order to over-express and secrete tryptophan 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.

TrpE codes for the Anthranilate enzyme responsible for the first step of the conversion of Chorismate into to tryptophan. Previous work (Gu et al., 2012) have shown that over-expression of TrpE leads to increased tryptophan production in E. coli. The trpE gene carries a M293T point mutation (to avoid negative feedback inhibition) and a hexa histidine-tag attached by a short linker sequence.

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 1876
    Illegal BglII site found at 1937
  • 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
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
//cds/biosynthesis
//cds/enzyme
//cds/membrane/transporter
//chassis/prokaryote/ecoli
//direction/forward
Parameters
chassisEscherichia coli
directionForward
functionTransporter and Biosynthesis
originEscherichia coli MG1655 - Codon optimized
proteinEnzyme
targetTryptophan, Tyrosine, Phenylalanine