Part:BBa_K4773005:Design
RBS-QsrR
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
When designing the sequence of this part, we considered the following aspects: - We chose a strong RBS (BBa_B0034) to ensure the efficient expression of the QsrR gene. - We chose a QsrR gene from a common skin pathogen, to ensure the high activity and stability of QsrR protein. - We added a KpnI restriction site after the QsrR sequence, to facilitate its connection with other parts. KpnI is a restriction enzyme that can recognize and cut the GGTACC sequence. We avoided this sequence in the rsrr or QsrR gene, to prevent it from being wrongly cut. - We did not add a terminator at the end of the QsrR gene, because we wanted to splice it with other genes into a multifunctional fusion protein. If we added a terminator, it would block the expression of downstream genes. - We also considered some factors that affect prokaryotic expression, such as codon bias, RNA secondary structure, protein folding, etc. We tried to choose codons that match the host bacteria, avoid forming stable RNA secondary structures, and increase protein folding helper factors.
Source
This part consists of a strong bacterial ribosome binding site (RBS) (BBa_B0034) and the QsrR gene. The RBS can enhance the transcription and translation of the QsrR gene, increasing the expression level of QsrR protein. The QsrR gene originates from a common skin pathogen Staphylococcus aureus, which has a high QsrR activity and stability. QsrR protein can work at room temperature and neutral pH, recognizing and binding to specific DNA sequences. This part can be expressed in different host cells, enabling them to sense and respond to quinone compounds.