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.

<iframe width="420" height="315" src="http://www.youtube.com/embed/BLLyUHrrcV4" frameborder="0" allowfullscreen></iframe>

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