Composite

Part:BBa_K1638036

Designed by: Jens Sivk�r Pettersen   Group: iGEM15_SDU-Denmark   (2015-09-14)

Sce VMA Intein with promoter

This part consist of an intein from Saccharomyces cerevisiae VMA1 gene including a chitin-binding domain (CBD)-tag. This intein is a thiol-induced self-cleavable protein that enables the release of a C-terminal fused protein. The chitin-binding domain is an affinity tag that enables affinity purification on a chitin column. First the target protein fused to intein is loaded and washed on the chitin column. Using a thiol reagent, like dithiothreitol (DTT), an on-column cleavage is induced and the target protein is released [1].

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 897
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 563
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1440
  • 1000
    COMPATIBLE WITH RFC[1000]

References

[1] IMPACT™ Kit. 2015. New England Biolabs; [accessed 2015 Sep 11]. https://www.neb.com/products/e6901-impact-kit


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