Part:BBa_K2179003:Design
Chlorite Dismutase CDS+RBS from I. dechloratans with His-tag
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 807
Illegal BsaI.rc site found at 6
Design Notes
Chlorite Dismutase is the second enzyme utilized in a number of bacterial species' perchlorate reducing pathway. In their original work, Thorell et al. cloned, sequenced, and functionally expressed the natural coding sequence for Chlorite Dismutase from Ideonella dechloratans. They determined that the gene was capable of converting ClO2- ions to O2 gas and Cl- ions and that its activity was localized to the periplasm, even though its putative signal peptide had not been cleaved. We designed this Cld(-SP) gene to lack this putative signal sequence. The purpose of this modification was so that once translation was complete, the Chlorite Dismutase enzyme would remain in the cytoplasm of our E. coli. chassis. A C-terminal histidine tag was also added to this translational unit in order to purify the enzymes in the event that O2 production from living cells proved inefficient. Flanking these BsaI sites were XbaI (5’) and the BBa_ std 1 suffix (3’) to facilitate parts creation for the iGEM Registry. We note that the natural coding sequence contained none of the standard BBa_10 restrictions sites, but did contain a single internal BsaI site which we eliminated with a single base silent substitution
Source
Derived from the soil bacterium Ideonella dechloratans
References
Thorell, H.D, Karlsson, J., Portelius, E.and Nilsson, T. Biochimica et Biophysica Acta 1577 (2002) 445–451 Samant,S., Gupta, G., Karthikeyan, S., Haq, S., Sambasivam, N.G., and Sukumaran, S. J Ind Microbiol Biotechnol (2014) 41:1435–1442