Part:BBa_K1896017

INP_RC-streptavidin generator

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

Usage and Biology

INP_RC 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 INP_RC 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.