Part:BBa_K4121051

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]