Composite

Part:BBa_K4121051:Design

Designed by: Ziyu Li   Group: iGEM22_NEFU_China   (2022-10-05)


Expression cassette of TmCrtE with constitutive promoter


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 240
    Illegal XbaI site found at 727
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 240
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 240
    Illegal BglII site found at 589
    Illegal BglII site found at 730
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 240
    Illegal XbaI site found at 727
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 240
    Illegal XbaI site found at 727
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

We used the "Goldengate" method for the ligation of the basic parts, We introduced recognition sites and cleavage sites for the type IIs restriction endonuclease BsmBI at both ends of the Basic Parts to ensure that the Basic Parts could be ligated successfully, and changed the location of the recognition sites and cleavage sites so that the BsmBI would not cut repeatedly. To avoid homologous recombination in Saccharomyces cerevisiae, we substitute different promoters and terminators. Please refer to" Design" part of the Wiki for detailed experimental design.


Source

We get sequences of the promoter from Eukaryotic promoter database(EPD), the sequences of CDS and the terminator from National Center for Biotechnology Information(NCBI).

References

None