Part:BBa_K2407014
a modified CUP1 promoter obtained by Error-Prone PCR EP5
Design
This part was improved from BBa_K2407000 and the source can be tracked back to BBa_K2165004.
In our experiment, we noticed that CUP1 promoter still has a certain degree of leakage expression. To make a better biosensor, we planned to reduce the leakage expression and increase the sensitivity. To reach this goal, we took the fluorescence intensity at both induction or not into evaluation indexes.
The technology of error-prone PCR was taken into our experiment. Although there are many methods to introduce genetic diversity into a parent sequence, error-prone PCR is the most common way of creating a combinatorial library based on a single sequence. By adding some heavy metal ions into the PCR buffer and preparing dNTPs with different composition, new mutants were introduced into CUP1 promoter.
This is one of the mutants from CUP1 promoter, which was named with EP-5. The changes in sequence were uploaded in the "sequence and feature" part.
Characterization
We tested the leakage expression and response rages of each selected mutants, and results were shown below.
Visit BBa_K2407013, BBa_K2407015, and BBa_K2407016 to see other mutants.
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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