Difference between revisions of "Team:iTesla-SoundBio/Parts"
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| − | </ | + | <p>We didn’t successfully complete any BioBricks, but we had the construction of several planned and in progress.</p> |
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| + | <li>This would be the coding sequence for Limulus Clotting Factor C.</li> | ||
| + | <li>To create this, we would first acquire the <a href="https://www.uniprot.org/uniprot/P28175">amino acid sequence</a> from UniProt (UniProtKB - P28175 (LFC_TACTR)), then codon optimize for B. subtilis using JCat and add the BioBrick prefix and suffix.</li> | ||
| + | <li>The Factor C sequence is just over the IDT length limit for free synthesis--the limit is 3kb, while the sequence was 3,133 bases long--so we would need to synthesize it as two separate parts. We would independently digest and ligate these fragments into two separate plasmid backbones, then PCR the plasmids, digest the fragments out, ligate them together, and finally ligate them into the backbone pSB1C3. </li> | ||
| + | <li>To design those fragments, we would have them overlap at a BioBrick-compatible restriction site in the middle of the Factor C sequence. This would be used to ligate the fragments together once we had purified samples of each.</li> | ||
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Revision as of 20:18, 14 October 2018
Parts
We didn’t successfully complete any BioBricks, but we had the construction of several planned and in progress.
- This would be the coding sequence for Limulus Clotting Factor C.
- To create this, we would first acquire the amino acid sequence from UniProt (UniProtKB - P28175 (LFC_TACTR)), then codon optimize for B. subtilis using JCat and add the BioBrick prefix and suffix.
- The Factor C sequence is just over the IDT length limit for free synthesis--the limit is 3kb, while the sequence was 3,133 bases long--so we would need to synthesize it as two separate parts. We would independently digest and ligate these fragments into two separate plasmid backbones, then PCR the plasmids, digest the fragments out, ligate them together, and finally ligate them into the backbone pSB1C3.
- To design those fragments, we would have them overlap at a BioBrick-compatible restriction site in the middle of the Factor C sequence. This would be used to ligate the fragments together once we had purified samples of each.