Part:BBa_K3634004
4-DG Pathway (E.coli Optimised)
The following part describes the constitutive expression of the first 3/4 enzymes of the shinorine production pathway, optimised for use in E.coli. Within the composite part, biobricks BBa_K3634000 (DHQS), BBa_K3634001 (O-MT) and BBa_K3634002 (ATPG) are responsible for converting sedoheptulose 7-phosphate to mycosporine glycine. Once produced, mycosporine glycine will then be converted by BBa_K3634003 (NRPS), present on plasmid A, to the final product of the pathway, shinorine. The shinorine production pathway is separated in this way as a biosafety mechanism so that maximum UV resistance is not conferred in a bacteria which has lost or gained plasmid B alone. As NRPS is determined to be 'rate-limiting' with respect to the pathway, plasmid A will ideally be placed at a higher copy number to plasmid B to ensure 1:1 stoichiometry of reactants.
For our bacteria to provide maximum expression of the UV absorbing compound shinorine, the strong constitutive promoter BBa_J23119 was selected from the Anderson promoter catalogue alongside RBS BBa_B0034 inserted between each ORF. These parts are a preliminary choice based off previous experimental expression data (Berkeley iGEM, 2006). The commonly used double terminator BBa_B0015 will ensure reliable release of the new mRNA strand.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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