Composite

Part:BBa_K635001:Design

Designed by: Arjun Athreya   Group: iGEM11_Virginia   (2011-09-27)

Composite coding region for PDGF-β in yeast


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 135
    Illegal BglII site found at 288
    Illegal BglII site found at 351
    Illegal BglII site found at 474
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The PDGF cDNA sequence could not simply be uploaded to the registry because the gene is not expressible in yeast without the addition of a poly-A region. In addition, the gene was constructed for compatibility with the silencing siRNA ([BBa_K635000]) if necessary. Therefore a composite part was designed and uploaded to the registry for efficient complete expression of PDGF within the yeast host chassis.

Source

The E.coli cDNA sequence was obtained from Sinobiological Inc. The cDNA sequence was codon-optimized for yeast expression. The rest of the sequence contains the biobrick kozak sequence [BBa_J63003],the biobrick transcriptional terminator [BBa_K392003], a randomly generated buffer region preceding the kozak sequence (GGAGTTGTGGGCACCTGCTA) which is reverse complementary to part of the siRNA sequence ([BBa_K635000]), and finally a customized poly-A region necessary for expression in yeast.

References