Part:BBa_K5091012:Design
pmsC-pmsB Expression Construct
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 1351
Illegal PstI site found at 1732
Illegal PstI site found at 2110
Illegal PstI site found at 2529 - 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1205
Illegal PstI site found at 1351
Illegal PstI site found at 1732
Illegal PstI site found at 2110
Illegal PstI site found at 2529 - 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1144
Illegal BamHI site found at 1893
Illegal BamHI site found at 2091 - 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 1351
Illegal PstI site found at 1732
Illegal PstI site found at 2110
Illegal PstI site found at 2529 - 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 1351
Illegal PstI site found at 1732
Illegal PstI site found at 2110
Illegal PstI site found at 2529
Illegal AgeI site found at 979 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 961
Design Notes
When designing the pmsC-pmsB expression construct, several important factors should be considered. One of the main considerations was selecting the appropriate plasmid backbone. The plasmid copy number plays a key role in balancing gene expression levels to avoid overproduction of salicylic acid (SA), which can have negative off-target effects on both the host and the environment. A low to medium copy number plasmid would help mitigate these risks while ensuring sufficient SA production.
Additionally, the potential toxicity of excess salicylic acid led to the consideration of incorporating a kill switch into the plasmid. This would act as a safety mechanism to prevent uncontrolled SA production in the event of system failure, protecting both the host and surrounding environment from the adverse effects of high SA concentrations.
Source
This is a composite part including the pmsC protein coding region encoded in part BBa_5091008 and the pmsB protein coding region in part Baa_K5091009.