Composite

Part:BBa_K274111:Design

Designed by: Siming Ma   Group: iGEM09_Cambridge   (2009-10-19)


CrtY under Pbad promoter (I0500)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1205
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1144
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 979
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 961


Design Notes

The colourless precursor, farnesyl pyrophosphate, is naturally produced in E.coli using Non-mevalonate Pathway (from pyruvate and glyceraldehyde-3-phosphate). BBa_K2741x0 series (BBa_K274100, BBa_K274110 and BBa_K274120) contain enzyme cassette CrtEBI that convert farnesyl pyrophosphate to lycopene (red). BBa_K274111 (I0500+CrtY) then converts lycopene (red) to beta-carotene (orange).

Our experiences showed that this Biobrick works better in E.coli strains with up-regulated level of precursors. E.coli strain MG1655 demonostrates higher level of pigment production than strain TOP10.


Source

The (rbs+CrtY) component comes from BBa_K118013. The coding sequence for CrtY comes from Pantoea ananatis (formerly Erwinia uredovora) (Accession number D90087).

The Pbad promoter comes from BBa_I0500.

References