Difference between revisions of "Part:BBa K1189018:Design"
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<partinfo>BBa_K1189018 SequenceAndFeatures</partinfo> | <partinfo>BBa_K1189018 SequenceAndFeatures</partinfo> | ||
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| − | + | <h1>Design notes</h1> | |
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| + | <p>This part was created by fusing the heavy chain (<a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K1189025">BBa_K1189025</a>, <a href="http://www.uniprot.org/uniprot/P02794">P02794 [UniParc]</a>) and light chains (<a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K1189024">BBa_K1189024</a>, <a href="http://www.uniprot.org/uniprot/P02792">P02792 [UniParc]</a>) of human ferritin together. It is expressed under the LacI promoter (<a href="https://parts.igem.org/Part:BBa_J04500">BBa_J04500</a>) and has a his-tag for protein purification. An E-coil (<a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K1189011">BBa_K1189011</a>) is included in order to allow binding of parts containing the respective K-coil (<a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K1189010">BBa_K1189010</a>). This particular part is expressed under a strong RBS and the LacI promoter. Note that this part is identical to <a href="https://parts.igem.org/Part:BBa_K1189037">BBa_K1189037</a>, say for the exclusion of a hexahistidine purification tag. Please see <a href="https://parts.igem.org/Part:BBa_K1189037">BBa_K1189037</a> for full characterization of this part.</p> | ||
| + | <h1>Source</h1> | ||
| + | <p>We synthesized heavy ferrtin <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K1189019">(BBa_K1189019)</a> and light ferritin <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K1189020">(BBa_K1189020)</a> from a commerical synthesis company. These parts were optimized for expression in <i>E. coli K12</i>. These two elements were extracted via PCR, the pSB1C3 backbone was isolated with PCR, and these linear fragments were assembled into BBa_K1189018 using isothermal assembly.</p> | ||
| − | + | <h1>References</h1> | |
| − | + | <li>Chasteen, N. D., & Harrison, P. M. (1999). Mineralization in ferritin: an efficient means of iron storage. Journal of structural biology, 126(3), 182-194.</li> | |
| − | + | <li>Dehal, P. K., Livingston, C. F., Dunn, C. G., Buick, R., Luxton, R., & Pritchard, D. J. (2010). Magnetizable antibody‐like proteins. Biotechnology journal, 5(6), 596-604.</li> | |
Latest revision as of 02:20, 1 November 2013
Human ferritin di-subunit with E coil w/ LacI promoter
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1289
Design notes
This part was created by fusing the heavy chain (BBa_K1189025, P02794 [UniParc]) and light chains (BBa_K1189024, P02792 [UniParc]) of human ferritin together. It is expressed under the LacI promoter (BBa_J04500) and has a his-tag for protein purification. An E-coil (BBa_K1189011) is included in order to allow binding of parts containing the respective K-coil (BBa_K1189010). This particular part is expressed under a strong RBS and the LacI promoter. Note that this part is identical to BBa_K1189037, say for the exclusion of a hexahistidine purification tag. Please see BBa_K1189037 for full characterization of this part.
Source
We synthesized heavy ferrtin (BBa_K1189019) and light ferritin (BBa_K1189020) from a commerical synthesis company. These parts were optimized for expression in E. coli K12. These two elements were extracted via PCR, the pSB1C3 backbone was isolated with PCR, and these linear fragments were assembled into BBa_K1189018 using isothermal assembly.