Part:BBa_J100335
Attempt at the Creation of a Strong RBS
The rClone sequence developed is a modified sequence using point mutational analysis. To develop the original sequence before we inserted point mutational analysis, we used an RBS calculator created by Salis Lab at Penn State University (link below). We inserted a pre sequence, which was 10 bases before our RBS and the entire protein coding sequence. The calculator then does probabilities for the best possible sequences with the given information. After it runs 200 times it will give you a sequence, and then it will run that given sequence another 200 times and give a new updated sequence. We ran our sequence 2000 times. After having our set sequence we referred to other rClone sequences to try and determine common base pairs that resulted in strong a strong RBS. We noticed the combinations of CA, AA, and GA nucleotides were highly conserved. We kept nucleotide combinations in the sequence and then chose nine nucleotides to make "N" for mutational analysis. Every N will have a different nucleotide inserted to increase the possibility of finding the right sequence for a strong RBS.
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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