Generator

Part:BBa_K1896017

Designed by: Bob Van Hove, Maarten Van Brempt   Group: iGEM16_UGent_Belgium   (2016-10-13)


INP_RC-streptavidin generator

This part contains the INPRC-streptavidin fusion protein controlled by a weak to medium constitutive promoter and RBS.

Usage and Biology

INPRC is a truncated Ice Nucleating Protein from which the N-terminal domain has been removed. The truncated INP is still capable of catalysing the formation of ice crystals, but is no longer localised to the outer cell membrane [1]. The biotin-binding protein Streptavidin has been fused to INPRC for use in protein adhesion experiments performed by the UGent Belgium 2016 iGEM team. This part appears to be toxic and accrues mutations, possibly due to the tetrameric nature of Streptavidin. To remedy this, a version with a monomeric Streptavidin variant was constructed (BBa_K1896018).

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 203
    Illegal NgoMIV site found at 1715
    Illegal NgoMIV site found at 1859
    Illegal NgoMIV site found at 2123
    Illegal NgoMIV site found at 2267
    Illegal NgoMIV site found at 2315
    Illegal NgoMIV site found at 2435
    Illegal AgeI site found at 1379
    Illegal AgeI site found at 3254
    Illegal AgeI site found at 3305
  • 1000
    COMPATIBLE WITH RFC[1000]


References

  1. Green, R. L., Corotto, L. V., & Warren, G. J. (1988). Deletion mutagenesis of the ice nucleation gene from Pseudomonas syringae S203. Molecular and General Genetics MGG, 215(1), 165-172.
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