Part:BBa_K3059621

csgEFG, lacks an RBS before csgE

csgEFG operon isolated from the E. coli (MG1655) genome via PCR. Lacks RBS or any region upstream of csgE, so an additional RBS is necessary when assembling transcriptional units with this part. The native curli operon contains two divergent operons: csgDEFG as well as csgBAC. To create a synthetic curli operon, both operons are necessary (for example, though csgA is 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, csgBAC, 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).

Proposed transcriptional unit with this part. For a 3G-compatible version of this part (and more information about assembling a complete synthetic curli operon), see BBa_K3059630.

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

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 382
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 382
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 382
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 382
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 382
  Illegal AgeI site found at 679
  
• 1000
  COMPATIBLE WITH RFC[1000]