Part:BBa_K2787000
Constitutive promoter J23100 + orthogonal RBS A2 + GFP
This is a part used for justification and measurement of the orthogonality of the ribosomal 16sRNA and its corresponding ribosome binding site. It is composed of a constant expressing promoter(BBa_J23100), followed by a mutated ribosome binding site that can be specifically recognized by the orthogonal 16s rRNA A2(BBa_K2787004) and a reporter protein GFP(BBa_E0040). The GFP would generate fluorescence only upon the presence of o-16SA2 while showing high orthogonality toward the innate 16s rRNA of E. coli.
Usage and Biology
a plasmid with GFP under control of A2 orthogonal RBS is constructed to test the orthogonality of A2 RBS.
The expression of GFP under control of A2 orthogonal RBS is greatly repressed without A2 orthogonal ribosome, while it is upregulated with the presence of A2 orthogonal 16s rRNA.
With the presence of A2 orthogonal 16s rRNA, the expression of GFP controlled by the A2 RBS is activated and the fluorescence grows up gradually over time. However, GFP is highly repressed without A2 orthogonal 16s, and keep low expression in a period of time
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]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 706
| None |