Difference between revisions of "Part:BBa K342002:Design"

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The intein is from the self cleaving protein sequence delta.I-CM mini intein from mycobacterium tuberculosis described in <a href="http://www.faqs.org/patents/app/20090098611"><font color= black><i><u>this patent</i></u></font></a> . <br/>
 
The intein is from the self cleaving protein sequence delta.I-CM mini intein from mycobacterium tuberculosis described in <a href="http://www.faqs.org/patents/app/20090098611"><font color= black><i><u>this patent</i></u></font></a> . <br/>
 
The in between are the same as the one described in <a href="http://2010.igem.org/Team:INSA-Lyon/Project/References#et1"><font color= black><i><u>Banki et al.</i></u></font></a>
 
The in between are the same as the one described in <a href="http://2010.igem.org/Team:INSA-Lyon/Project/References#et1"><font color= black><i><u>Banki et al.</i></u></font></a>
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<p>The sequencing of this part is difficult because of the repetition of phasin's sequence.</p>
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<p>This part should had been synthesized by Mr gene in once but due to technical issue we had to redesign two separates fusion biobrick and ligate them.</p>
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<p>The sequencing of this part confirm the expected sequence.</p>
 
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Revision as of 16:52, 25 October 2010

Phasin-Phasin Intein (for silver standard fusion)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 1693
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1279
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

This part is a self cleavable fusion protein. It has been designed thanks to the results of the publication Banki et al.

The phasin protein bind to the granule membrane. The second phasin makes the binding stronger and then increases the rate of purified protein during the step of granules extraction. The intein is a self cleavable protein which breaks under PH=6 and allows the release od the fused protein at the end of the sequence

The prefix biobrick standard used contains a perfect RBS.
The suffix is a silver standard fusion suffix : 5' ACTAGT A GCGGCCG CTGCAG 3'.
The phasin sequence is the same as the biobrick BBa_K208001 from the Utah 2009 team.
The intein is from the self cleaving protein sequence delta.I-CM mini intein from mycobacterium tuberculosis described in this patent .
The in between are the same as the one described in Banki et al.

This part should had been synthesized by Mr gene in once but due to technical issue we had to redesign two separates fusion biobrick and ligate them.

The sequencing of this part confirm the expected sequence.

Source

Ligation of two synthetized parts : Phasin and Phasin-Intein

References