Part:BBa_S04615
B0034:K592001
Construction intermediate
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 624
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 2270
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1288
- 1000COMPATIBLE WITH RFC[1000]
Our team has optimized the BBa_S04615 part. In 2018, the SDU-CHINA team optimized the K592001 part to reduce its leakage under the red light. We have improved its promoters this year, which has further bettered the performance of this component.
The component BBa_K2627000 proved the progress of the optimized CcaS by sufficient characterization, so we directly replaced BBa_K592001 in BBa_S04615 with the optimized K2627000 when characterizing this component. In 2014, Schmidl S R , Sheth R U, Wu A, et al. optimized CcaS by constructing an RBS library. This document has fully proven that the performance of components with optimized RBS is better than S04615. In 2019, our team SDU-CHINA has further improved the components based on them. We have introduced site-directed mutagenesis to the improved promoter, further bettering the performance based on all previous studies.
Results
Characaterization of BBa_K2627000
The plasmid carrying our RBS is called 5-10#10. The improved part is BBa_K3127995.
There is less fluorescence intensity than #10 under light of 640-650 nm wavelength.
The following figure shows the relative fluorescence intensity induced by the light of 640-650 nm wavelength(green light) and the light of 530-535 nm wavelength(red light).
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