Difference between revisions of "Part:BBa K4121052"
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The composite part is called "pFBA1-TmCrtE-tTDH1". We use pFBA1 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 "pFBA1-TmCrtE-tTDH1". We use pFBA1 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). | ||
| + | |||
| + | __NOTOC__ | ||
| + | <partinfo>BBa_K4121052 short</partinfo> | ||
| + | |||
| + | <partinfo>BBa_K4121052 SequenceAndFeatures</partinfo> | ||
| + | |||
| + | |||
| + | ===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). | ||
| + | |||
Revision as of 09:38, 12 October 2022
Expression cassette of TmCrtE with constitutive promoter
The composite part is called "pFBA1-TmCrtE-tTDH1". We use pFBA1 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).
Expression cassette of TmCrtE with constitutive promoter
- 10INCOMPATIBLE WITH RFC[10]Illegal XbaI site found at 728
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 590
Illegal BglII site found at 731 - 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 728
- 25INCOMPATIBLE WITH RFC[25]Illegal XbaI site found at 728
- 1000COMPATIBLE 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).
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal XbaI site found at 728
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 590
Illegal BglII site found at 731 - 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 728
- 25INCOMPATIBLE WITH RFC[25]Illegal XbaI site found at 728
- 1000COMPATIBLE WITH RFC[1000]