Difference between revisions of "Part:BBa K4121051"
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The composite part is called "pCCW12-TmCrtE-tTDH1". We use pCCW12 as the promoter, tTDH1 as the terminator and the CDS of this part is TmCrtE(from Taxus media). The three basic parts above make up a new transcription unit, it has the fuction to catalyze the conversion of farnesyl pyrophosphate (FPP) to heterologous geranylgeranyl diphosphate(GGPP).  
 
The composite part is called "pCCW12-TmCrtE-tTDH1". We use pCCW12 as the promoter, tTDH1 as the terminator and the CDS of this part is TmCrtE(from Taxus media). The three basic parts above make up a new transcription unit, it has the fuction to catalyze the conversion of farnesyl pyrophosphate (FPP) to heterologous geranylgeranyl diphosphate(GGPP).  
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===Design Notes===
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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.
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To avoid homologous recombination in Saccharomyces cerevisiae, we substitute different promoters and terminators.
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Please refer to" Design" part of the Wiki for detailed experimental design.
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===Source===
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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).
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===References===
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None
  
  

Revision as of 09:46, 12 October 2022


Expression cassette of TmCrtE with constitutive promoter

Expression cassette of TmCrtE with constitutive promoter

The composite part is called "pCCW12-TmCrtE-tTDH1". We use pCCW12 as the promoter, tTDH1 as the terminator and the CDS of this part is TmCrtE(from Taxus media). The three basic parts above make up a new transcription unit, it has the fuction to catalyze the conversion of farnesyl pyrophosphate (FPP) to heterologous geranylgeranyl diphosphate(GGPP).

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 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]