Part:BBa_K1900000

pBAD+strong RBS+E. coli tolC signal sequence+K. pneumoniae tolC

Combination of part BBa_K1406000 (BBa_K206000 + BBa_B0034) the E. coli tolC signaling sequence and the Klebsiella pnuemoniae 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 Klebsiella pnuemoniae TolC trimers form a outer-membrane alpha/beta barrel pore can interface with a variety of molecule pumps to form a transperiplasmic pump.
Team WLC-Milwaukee 2019
Authors: Laura Sanford, Ben Bateman The tolC gene from Klebsiella pneumoniae (part BBa_K1900000) was cloned into a plasmid containing the pTrc promoter. This plasmid was transformed into an E. coli strain lacking a chromosomal copy of tolC. Disk sensitivity assays were performed to determine how well the K. pneumoniae protein would work in an E. coli cell.
<img src="File:T--WLC-Milwaukee--figure1new.JPG"/> Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 1603
  Illegal NheI site found at 1624
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 1543
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 1104