Part:BBa_M36888:Design
Sequence for sigma 32 transcription factor
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 601
- 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 601
- 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 601
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 601
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 303
Illegal SapI.rc site found at 652
Design Notes
Anyone incorporating this part into a feedback amplification loop must beware of likely rpoH-deletion mutants. This is due to overproduction of sigma 32 under leaky promoters such as htpG (BBa_J45405). Less deletion mutants in E. coli are noted when plasmids containing rpoH are cloned at higher temperature.
Consider two facts before using this part. First, rpoH transcript activity is reduced under excess heat shock protein production. As such, feedback loops using rpoH should employ weaker bicistronic ribosome binding sites so as to optimize heat-shock response. You might also consider using the wild-type RBS (BBa_K1895001). Second, sigma 32 is turned over rapidly, with a half-life under 1 minute.
Source
The particular part described here is specific to the genome of W3110 strain E. coli.
http://ecoliwiki.net/colipedia/index.php/rpoH:Gene
References
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC211317/?page=5
http://www.researchgate.net/publication/20461625_The_activity_of_sigma_32_is_reduced_under_conditions_of_excess_heat_shock_protein_production_in_Escherichia_coli