Difference between revisions of "Part:BBa K2271105:Design"
 
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===Design Notes===
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===Part design===
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This part is based on the yeasts PEX5 gene. In the course of a targeted mutagensis approach this PEX5 variant was designed to interact with a non-native PTS1 sequence. We verified that this variant does not detect the wildtype PTS1 but our designed PTS1* and leads to the import of the protein attached to it.
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  This part was designed in the course of a targeted mutagenesis approach. With the predicted three dimensional strucutre of the TPR domains we carried out our workflow in order to obtain the PEX5 variant we wanted.
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We thereby provide a construct which allows full control over the peroxisomes protein contents.
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  <br>
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  After gene synthesis we performed golden gate cloning and made it compatible to the biobrick standard via overhang PCR with the appropriate primers. This product then was ligated into the biobrick backone <b>pSB1C3</b>.
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</p>
  
 
===Source===
 
===Source===
  
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Synthesized by IDT.
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  The original sequence was taken from NCBI (GenBank: KZV12481.1). After mutagenesis we performed the IDT codon optimization for <i>Saccharomyces cerevisiae</i>, removed any restrictions sites that would interfere with the biobrick or golden gate standard and synthesized the gene with appropiate Golden Gate overhangs.
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</p>
  
 
===References===
 
===References===

Latest revision as of 18:53, 1 November 2017


PEX5 variant R15


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 409
  • 1000
    COMPATIBLE WITH RFC[1000]


Part design

This part was designed in the course of a targeted mutagenesis approach. With the predicted three dimensional strucutre of the TPR domains we carried out our workflow in order to obtain the PEX5 variant we wanted.
After gene synthesis we performed golden gate cloning and made it compatible to the biobrick standard via overhang PCR with the appropriate primers. This product then was ligated into the biobrick backone pSB1C3.

Source

The original sequence was taken from NCBI (GenBank: KZV12481.1). After mutagenesis we performed the IDT codon optimization for Saccharomyces cerevisiae, removed any restrictions sites that would interfere with the biobrick or golden gate standard and synthesized the gene with appropiate Golden Gate overhangs.

References