Composite

Part:BBa_K3789010:Design

Designed by: MEI YING YI   Group: iGEM21_UM_Macau   (2021-09-28)


PNP enzyme secretion system


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 1448
    Illegal PstI site found at 1023
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 1023
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 2273
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 1448
    Illegal PstI site found at 1023
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 1448
    Illegal PstI site found at 1023
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The pGK1 promoter can efficiently drive high levels of gene expression. Since the purine nucleoside phosphorylase(PNP) needs to be released in the brewing environment to be effective, we choose the common secretion signal sequence “mating factor alpha prepro-leader (MF alpha)” to make the enzyme secretions become possible. Due to these reasons, the system can provide a stable secretable PNP expression.


Source

pGHK1 promoter, Mating factor alpha prepro-leader (MF alpha), and the PNP1 coding sequence come from Saccharomyces cerevisiae S288c. HA-tag sequence comes from the human influenza virus.

References