Part:BBa_K1683000

Safety Flag

The iGEM Safety and Security Committee has placed a Red Flag on this part. This part presents safety risks beyond what is normal for the Registry. Researchers who plan to acquire and use this part should take special care to ensure they use it safely and responsibly. Contact safety [AT] igem [DOT] org with any questions.

Reason: outer membrane channel protein

If you are an iGEM team, you must submit a Check-In before acquiring and using this part! See the 2021 Safety Page for more information.

pBAD+Strong RBS+E. coli tolC signal sequence+Y. pestis tolC

Combination of part BBa_K1406000 (BBa_K206000 + BBa_B0034) the E. coli tolC signaling sequence and the Yersenia pestis Angola tolC gene. Transcription is controlled by the arabinose-induced pBAD promoter (BBa_K206000). A strong E. coli RBS (BBa_B0034) controls translation. The signaling sequence is a cleavable N-terminal signaling sequence from E. coli's tolC gene. This is required for trimerization of the TolC monomers as well as insertion into the outer membrane; it forms a chimeric polypeptide with the tolC gene from Yersenia pestis Angola. TolC trimers form a outer-membrane alpha/beta barrel pore can interface with a variety of molecule pumps to form a transperiplasmic pump. Kirby-Bauer Assay of E. coli, knockout tolC E. coli, and knockout tolC E. coli with Yersenia pestis' tolC gene. The Yersenia pestis tolC restored the efflux function of the bacteria, reducing the zones of clearing to that of the WT E. coli.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 125
  Illegal NheI site found at 146
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 65
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 944
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 344