Part:BBa_K2540015
Orthogonal RBS for expression across bacteria
This orthogonal RBS was designed using RBS calculator for orthogonal 16S rRNA predicted to function across a variety of bacterial strains. Prediction was done by algorithm written by Rice iGEM 2018 team based on previous work by Chubiz & Rao.
Usage and Biology
The orthogonal RBS contains an altered Shine-Dalgarno sequence, which prevents binding of the wild-type 16S rRNA subunit. Only 16S rRNA subunits containing a complementary orthogonal anti-Shine Dalgarno sequence may bind to the orthogonal RBS. This RBS along with its corresponding 16S rRNA may be used when orthogonal translation is desired in bacteria.
The part was characterized in four E. coli strains, S. oneidensis , and P. putida using mKate2 fluorescent protein. Figure 1 shows fluorescence levels after cells transformed with a plasmid containing mKate2 under the control of the broad host range regulatory element were grown in LB for 24 hours.
Figure 1: Fluorescence levels of mKate2 under the control of the broad host range element (strength 7).
Sequence and Features
- 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 3
Illegal BsaI.rc site found at 58
References
Chubiz, L. M., & Rao, C. V. (2008). Computational design of orthogonal ribosomes. Nucleic Acids Research, 36(12), 4038–4046.
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