Composite

Part:BBa_K5143024

Designed by: Perrine Ferdinand and Jess Philippon   Group: iGEM24_UnivLyon1-INSALyon   (2024-08-14)
Revision as of 21:14, 14 August 2024 by Perrine-fdn (Talk | contribs)

Yellow protein and BioGlue protein secreted by Saccharomyces cerevisiae

Protein Description

Description

This composite part corresponds to the association of two composite parts (BBa_K5143023 & BBa_K5143022) linked by a P2A system (BBa_K5143012), all under the control of a single promoter. During translation, this would give two distinct fusion proteins from the same transcriptional unit, which would then be secreted by their respective alpha factors and associate with the cellulose membrane via their CBD domains. This composite part is cloned within the backbone (BBa_K5143005), then digested by XhoI so that the composite part is framed by zones of homology. Following transformation, the whole is integrated by homologous recombination within the yeast Saccharomyces cerevisiae .

Diagram of the composite part BBa_K5143024

Construction

The composite biobricks were optimised for transcription and synthesis in S. cerevisiae . This composite part was first constructed using TEDA cloning by integrating the "yellow" fragment into the backbone (BBa_K5143005) opened by ? . The newly constructed plasmid was checked by sequencing, in order to carry out a new cloning to add the "bioglue" fragment. The plasmid will be opened by PCR and the fragment cloned by HIFI. This construction will be carried out without scarring. Size: 6365 bp

References

METTRE LES REF ICI


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1361
    Illegal PstI site found at 2789
    Illegal PstI site found at 3092
    Illegal PstI site found at 3185
    Illegal PstI site found at 3191
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1361
    Illegal PstI site found at 2789
    Illegal PstI site found at 3092
    Illegal PstI site found at 3185
    Illegal PstI site found at 3191
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1361
    Illegal BamHI site found at 1906
    Illegal BamHI site found at 2620
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1361
    Illegal PstI site found at 2789
    Illegal PstI site found at 3092
    Illegal PstI site found at 3185
    Illegal PstI site found at 3191
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1361
    Illegal PstI site found at 2789
    Illegal PstI site found at 3092
    Illegal PstI site found at 3185
    Illegal PstI site found at 3191
    Illegal NgoMIV site found at 1876
    Illegal AgeI site found at 1915
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None