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Part:BBa_K342002:Design
Phasin-Phasin Intein (for silver standard fusion)
- 10INCOMPATIBLE WITH RFC[10]Unknown
- 12INCOMPATIBLE WITH RFC[12]Unknown
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 1693
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1279
- 1000COMPATIBLE 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 sequencing of this part is difficult because of the repetition of phasin's sequence.
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.
Source
Ligation of two synthetized parts : Phasin and Phasin-Intein