Composite

Part:BBa_K4121065

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

Expression cassette of PaCrtB with constitutive promoter

The part is called "pTPI1-PaCrtB-tPRM9". We use pTPI1 as the promoter, tPRM9 as the terminator and the CDS of this part is PaCrtB(Pantoea agglomerans and phytoene desaturase). The three basic parts above make up a new transcription unit,It has the function to catalyze the conversion of heterologous geranylgeranyl diphosphate(GGPP) to phytoene.

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


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 1106
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1456
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1151
    Illegal BglII site found at 1367
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 1106
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 1106
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None