Part:BBa_K590015

sfGFP_mamK_pGA1C3

This part was made by [http://2011.igem.org/Team:Washington UW iGEM Team 2011] and was contributed to the [http://2011.igem.org/Team:Washington/Magnetosomes/Results Magnetosome Toolkit].

Usage and Biology

This part consists of the mamK gene from Magnetospirillum magneticum strain AMB-1, fused to superfolder gfp on pGA1C3. MamK has been reported to be essential for proper magnetosome formation in magnetotactic bacteria; its bacterial actin-like cytoskeleton protein is required for proper alignment of the magnetosomes in a chain. Beyond magnetosome alignment, MamK filaments could act as a scaffold for protein localization or to alter cell morphology by preventing cells from fully dividing (see images below).

We transformed this part into an E. coli BL-21 lacI^q strain and observed fluorescent filament structures that in some cases, prevented cells from dividing properly. The same construct has been successfully cloned into Gibson assembly vectors pGA1A3 and pGA4C5.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal PstI site found at 1430
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal PstI site found at 1430
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 1134
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 1430
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 1430
  Illegal NgoMIV site found at 1156
  Illegal NgoMIV site found at 1523
  Illegal NgoMIV site found at 1670
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI.rc site found at 80
  Illegal SapI.rc site found at 1802