Part:BBa_K3059619

csgBAC + 22 bp upstream

csgBAC operon isolated from the E. coli (MG1655) genome via PCR. Contains 22 basepair upstream region, so an additional RBS is unnecessary when assembling transcriptional units with this part. The native curli operon contains two divergent operons: csgBAC as well as csgDEFG. To create a synthetic curli operon, csgEFG is necessary as well as csgBAC (for example, though csgA constitutes the main subunit of the eventual fiber, csgG is responsible for fiber export out of the cell). csgD regulates csgBAC, and is thus unnecessary in a synthetic operon where both component operons are combined under the control of one promoter.

When successfully assembled in a transcriptional unit with a promoter, csgEFG, an RBS preceding csgEFG, and a terminator, this part results in curli amyloid fiber production. Curli fibers constitute the main proteinaceous component of Enterobacteriaceae species such as E. coli (Barnhart & Chapman, 2006).

Though this part contains a non-Biobrick PstI cut site, it lacks BsaI and SapI cutsites and is thus type IIS-compatible and legal.

For a 3G-compatible version of this part (and information on assembling a complete synthetic curli operon), see BBa_K3059628.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal PstI site found at 889
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal PstI site found at 889
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 889
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 889
  
• 1000
  COMPATIBLE WITH RFC[1000]