Difference between revisions of "Team:East Chapel Hill/Contribution"

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<h1>Contribution</h1>
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Part: BBa_K2290000 </p>
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<p style="font-size:18px;"> <b>Plasmid Backbone:</b> pSB1A3 </p>
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Below is a list of the basic parts that we used in our part K2290000: </p>
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<p style="font-size:18px;"> BBa_J23100 – a constitutively active promoter from the Anderson collection. </p>
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<p style="font-size:18px;"> BBa_K2290008 – The <i>B. cereus</i> riboswitch with an engineered terminator that results in less read through/background. (This part is highly similar to BBa_K911003, but the BBa_K911003 sequence includes both the Lys promoter and the riboswitch). </p>
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<p style="font-size:18px;"> BBa_K2290004 – This is a sequence for a strong RBS that doesn't appear to be in the iGEM repository. </p>
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<p style="font-size:18px;"> BBa_K2290005 – This is an <i>E. coli</i> optimized chloramphenicol resistance gene that we had synthesized. </p>
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<p style="font-size:18px;"> BBa_B0010 – This is the T1 terminator from <i>E. coli</i> rrnB, reportedly a strong terminator </p>
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<p style="font-size:18px;"> We characterized and improved upon part BBa_K911003. First, BBa_K911003 listed as a basic part in the iGEM repository is actually a composite part that consists of a promoter and the <i>B. cereus</i> fluoride riboswitch. We more accurately annotated the <i>B. cereus</i> fluoride riboswitch alone as BBa_K2290008. We then characterized the responsiveness of the the <i>B. cereus</i> riboswitch and found that the switch works best at 100uM fluoride. To our knowledge part BBa_K911003 was not characterized in iGEM. We also used part BBa_J23100 from the Anderson promoter collection to regulate the expression of our operon and the BBa_B0010 terminator to efficiently terminate transcription . Given, that our composite part works these other two parts are functioning as intended in our operon.
  
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Revision as of 01:24, 2 November 2017

Contribution

Part: BBa_K2290000

Plasmid Backbone: pSB1A3

Below is a list of the basic parts that we used in our part K2290000:

BBa_J23100 – a constitutively active promoter from the Anderson collection.

BBa_K2290008 – The B. cereus riboswitch with an engineered terminator that results in less read through/background. (This part is highly similar to BBa_K911003, but the BBa_K911003 sequence includes both the Lys promoter and the riboswitch).

BBa_K2290004 – This is a sequence for a strong RBS that doesn't appear to be in the iGEM repository.

BBa_K2290005 – This is an E. coli optimized chloramphenicol resistance gene that we had synthesized.

BBa_B0010 – This is the T1 terminator from E. coli rrnB, reportedly a strong terminator

We characterized and improved upon part BBa_K911003. First, BBa_K911003 listed as a basic part in the iGEM repository is actually a composite part that consists of a promoter and the B. cereus fluoride riboswitch. We more accurately annotated the B. cereus fluoride riboswitch alone as BBa_K2290008. We then characterized the responsiveness of the the B. cereus riboswitch and found that the switch works best at 100uM fluoride. To our knowledge part BBa_K911003 was not characterized in iGEM. We also used part BBa_J23100 from the Anderson promoter collection to regulate the expression of our operon and the BBa_B0010 terminator to efficiently terminate transcription . Given, that our composite part works these other two parts are functioning as intended in our operon.