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Team:Carnegie Mellon/Bio-Submitted
From 2012.igem.org
Submitted Parts
We have submitted three T7Lac promoter parts to the registry. The followings show the sequences of these constructs.
BBa_K613007: TAATACGACTCACTATAGGGAGAGGAATTGTGAGCGGATAACAA
Mutant I: TAATGCGACTCACTATAGGACAATTGTGGGCGGACAACAATTCCAA
Mutant II: TAATACGACTCACTACAGGGCGGAATTGTGAGCGGATAACAATTCCAA
Mutant III: CAATCCGACTCACTAAAGAGAGAATTGTGAGCGGATAACAATTCCAA
Predicted strength of the hybrid T7Lac promoters
Expected promoter strength of the mutants (relative to BBa_K613007):Mutant I: <100%
Mutant II: ~100%
Mutant III: ~50%
Expected LacI leaky expression of different mutants:
Mutant I: More than average
Mutant II: Average
Mutant III: Average
Measured strength of the hybrid T7Lac promoters
Using the model we developed in MATLAB, we fitted experimental results of both RNA and protein expression rates and calculated the following properties with respect to the wild-type promoter.| Promoter | Mutant I | Mutant II | Mutant III |
|---|---|---|---|
| Transcription Strength | 97% | 72% | 127% |
| Translational Efficiency | 6% | 6% | 94% |
| RNA degradation constant (assumed) | 100% | 100% | 100% |
| Protein degradation constant (fit) | 4% | 6% | 60% |
