Part:BBa_K4361020
BlcR-binding oligo, 51 bp, Cro repressor variant
BlcR is a transcription factor originating from the bacterium Agrobacterium tumefaciens (Part:BBa_K4361100). Cro is a repressor protein found in bacteriophage λ. Both proteins form homodimers in vivo to create a dimer with a single DNA-binding domain, though the DNA sequences they each bind differ greatly. As described in Usage and Biology below, one of the core principles of our project was to increase the binding strength between BlcR and DNA. One approach to achieve this was to create a chimera protein which combines the DNA-binding and dimerization domain of Cro with the GHB-recognition and tetramerization domain of BlcR. While its application would be similar to those of the previous parts (Part:BBa_K4361000 through Part:BBa_K4361019), it would require a different binding oligo due to its differing DNA-binding domain.
This part has been designed to test the binding strength between Cro-BlcR and DNA. To create this part, the nucleotides originally designated as belonging to IR1 and IR2 in Part:BBa_K4361001 have been exchanged for the consensus sequence which the Cro repressor binds ('TATCACCgcgGGTGATA', Cro IR1) and its reverse complement ('TATCACCcgcGGTGATA', Cro IR2). As the other nucleotides are identical to those in the wildtype oligo, the BlcR-binding domain of this part thus consists of Cro IR1-tca-Cro IR2, where tca is the original 3 nt linker sequence between IRs. The layout of this oligo presumes that other than the DNA-binding domain, the overall structure of Cro-BlcR is identical to that of wildtype BlcR, including its orientation to the DNA, and it retains its ability to form tetramers when bound to DNA.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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