Difference between revisions of "Part:BBa K1404008:Design"
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__NOTOC__ | __NOTOC__ | ||
<partinfo>BBa_K1404008 short</partinfo> | <partinfo>BBa_K1404008 short</partinfo> | ||
<partinfo>BBa_K1404008 SequenceAndFeatures</partinfo> | <partinfo>BBa_K1404008 SequenceAndFeatures</partinfo> | ||
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===Design Notes=== | ===Design Notes=== | ||
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| − | + | If you intend to use this part, be aware that CsgA might be toxic to <i>E. coli</i> cells in large quantities. We were unable to clone <i>csgA</i> in front of the strong promoter <a href="https://parts.igem.org/Part:BBa_J23119">J23119</a> or the weaker promoter <a href="https://parts.igem.org/Part:BBa_J23114">J23114</a> | |
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===Source=== | ===Source=== | ||
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| + | A <a href="http://2009.igem.org/Team:Paris/Parts_RBS"> strong RBS</a> desgined by team Paris Bettencourt 2009 and the coding region for the <i>csgA</i> gene as found in the genome of the <i>E. coli</i> strain MC4100 were synthesized by Genecust. An illegal restriction site was removed by introducing a silent mutation. A sequence coding for (Gly-Gly-Gly-Gly-His-His-His-His-His-His-Gly-Gly-Gly-Gly-His-His-His-His-His-His) was added at the end of the sequence. This direct synthesis was then cloned downstream of the <a href="http://partsregistry.org/Part:BBa_K342000">K342000</a> promoter by standard assembly. | ||
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===References=== | ===References=== | ||
Latest revision as of 23:56, 17 October 2014
p70-CsgA-His*2, double His-tagged curli generator
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
If you intend to use this part, be aware that CsgA might be toxic to E. coli cells in large quantities. We were unable to clone csgA in front of the strong promoter J23119 or the weaker promoter J23114 ,
Source
A strong RBS desgined by team Paris Bettencourt 2009 and the coding region for the csgA gene as found in the genome of the E. coli strain MC4100 were synthesized by Genecust. An illegal restriction site was removed by introducing a silent mutation. A sequence coding for (Gly-Gly-Gly-Gly-His-His-His-His-His-His-Gly-Gly-Gly-Gly-His-His-His-His-His-His) was added at the end of the sequence. This direct synthesis was then cloned downstream of the K342000 promoter by standard assembly.