Part:BBa_K4121064
Expression cassette of PaCrtB with constitutive promoter
The part is called "pACT1-PaCrtB-tPRM9". We use pACT1 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
- 10INCOMPATIBLE WITH RFC[10]Illegal XbaI site found at 1106
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1456
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1151
Illegal BglII site found at 1367 - 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 1106
- 25INCOMPATIBLE WITH RFC[25]Illegal XbaI site found at 1106
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 34
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