Team:Carnegie Mellon/Bio-Submitted

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BBa_K613007: TAATACGACTCACTATAGGGAGAGGAATTGTGAGCGGATAACAA <br>
BBa_K613007: TAATACGACTCACTATAGGGAGAGGAATTGTGAGCGGATAACAA <br>
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Mutant I: TAATGCGACTCACTATAGGACAATTGTGGGCGGACAACAATTCCAA <br>
+
(BBa_K921000) Mutant I: TAATGCGACTCACTATAGGACAATTGTGGGCGGACAACAATTCCAA <br>
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Mutant II: TAATACGACTCACTACAGGGCGGAATTGTGAGCGGATAACAATTCCAA <br>
+
(BBa_K921001) Mutant II: TAATACGACTCACTACAGGGCGGAATTGTGAGCGGATAACAATTCCAA <br>
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Mutant III: CAATCCGACTCACTAAAGAGAGAATTGTGAGCGGATAACAATTCCAA <br>
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(BBa_K921002) Mutant III: CAATCCGACTCACTAAAGAGAGAATTGTGAGCGGATAACAATTCCAA <br>
</p><p class="promoter">
</p><p class="promoter">

Revision as of 21:18, 1 October 2012

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Submitted Parts

We have submitted three T7Lac promoter parts to the registry. The followings show the sequences of these constructs.
BBa_K613007: TAATACGACTCACTATAGGGAGAGGAATTGTGAGCGGATAACAA
(BBa_K921000) Mutant I: TAATGCGACTCACTATAGGACAATTGTGGGCGGACAACAATTCCAA
(BBa_K921001) Mutant II: TAATACGACTCACTACAGGGCGGAATTGTGAGCGGATAACAATTCCAA
(BBa_K921002) 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

RBS used in construct is: CATATG AAGAAGGAGA TATACC

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%
Cheemeng:Need to comment on the results here.

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