Part:BBa_K283007

sp-eGFP-streptavidin-AIDAc

Autotransporters are synthesized as pre-proproteins with modular organization. A C-terminal membrane-embedded domain promotes secretion of parts of the protein across the outer membrane and is composed of a β-barrel and a α-helix spanning its lumen. The adhesion involved in diffuse adherence (AIDA-I) is a monemeric autotransporter that has been extensively studied. The cleavage of the autotransporter adhesion involve in diffuse adherence (AIDA-I) of Escherichia coli yields a membrane embedded fragments, AIDAc, and an extracellular fragments, the mature AIDA-I adhesion. In this part, AIDAc directs the fusion protein sp-eGFP-streptavidin-AIDAc (sp means signal sequence) to the outer membrane of the bacteria by the AIDA-I autotransporter pathway. The surface localization of sp-eGFP-streptavidin-AIDAc was demonstrated in our wiki website.In addition, the restriction of autotransporters to the pole is dependent on the presence of a complete lipopolysaccharide (LPS), consistent with known effects of LPS composition on membrane fluidity.This part does not pose any biological threat.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 181
  Illegal EcoRI site found at 1675
  Illegal EcoRI site found at 2371
  Illegal PstI site found at 1723
  Illegal PstI site found at 2309
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 181
  Illegal EcoRI site found at 1675
  Illegal EcoRI site found at 2371
  Illegal PstI site found at 1723
  Illegal PstI site found at 2309
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 181
  Illegal EcoRI site found at 1675
  Illegal EcoRI site found at 2371
  Illegal BamHI site found at 17
  Illegal BamHI site found at 949
  Illegal BamHI site found at 2573
  Illegal XhoI site found at 2779
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 181
  Illegal EcoRI site found at 1675
  Illegal EcoRI site found at 2371
  Illegal PstI site found at 1723
  Illegal PstI site found at 2309
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 181
  Illegal EcoRI site found at 1675
  Illegal EcoRI site found at 2371
  Illegal PstI site found at 1723
  Illegal PstI site found at 2309
  Illegal AgeI site found at 991
  Illegal AgeI site found at 1042
  
• 1000
  COMPATIBLE WITH RFC[1000]