Part:BBa_K581010

promoter(BBa_J23106)-RBS-luxU-RBS-cqsS-RBS-luxO

Regulatory proteins for receiving signaling of CAI-1.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal SpeI site found at 4021
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 7
  Illegal NheI site found at 30
  Illegal SpeI site found at 4021
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 3096
  Illegal BamHI site found at 404
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal SpeI site found at 4021
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal SpeI site found at 4021
  Illegal AgeI site found at 2374
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 964
  Illegal SapI site found at 3284
  

Design Notes

▪ A silent mutation (A->G) at base 1086 was made to remove PstI site in the region of cqsS gene

▪ A silent mutation (A->G) at base 1650 was made to remove EcoRI site in the region of cqsS gene

▪ A silent mutation (G->A) at base 3234 was made to remove PstI site in the region of luxU gene

▪ cqsS and luxO were amplified by PCR from the genome DNA of Vibrio cholera

▪ promoter Bba_J23106 + luxU was amplified by DNAWorks

Source

▪ We received the genome DNA of Vibrio cholera from Chinese Center for Disease Control and Prevention

▪ Although our system includes several genes from Vibrio cholera, these genes are not directly involved with virulence or antibiotic resistance in any case.

References

• [http://nar.oxfordjournals.org/content/30/10/e43.full.pdf+html] David M. Hoover, Jacek Lubkowski. DNAWorks: an automated method for designing oligonucleotides for PCR-based gene synthesis. Nucl. Acids Res 30 (10): e4 (2002).