Part:BBa_K1683004

pBAD+Strong RBS+E. coli tolC signal sequence+P. aeruginosa oprM

Combination of part BBa_K1406000 (BBa_K206000 + BBa_B0034) the E. coli tolC signaling sequence and the Pseudomonas aeruginosa PA01 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 Pseudomonas aeruginosa PA01. 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 Pseudomonas' oprM gene. The Pseudomonas oprM failed to restore 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 253
  
• 1000
  COMPATIBLE WITH RFC[1000]