Part:BBa_K533012

safety version of pSB1C3 plasmid

The plasmid has a ccdB suicide gene and a mutation-controller sequence, variant of I-SceI restriction site. It can hence eliminate devious mutations and help to restrain biobricks to only our engineered organisms.

The variant of I-SceI site present in the plasmid allows for elimination of heavily mutated vectors in I-SceI expressing strains.

This year, we tried to construct an E. coli strain expressing ccdB resistance gene and I-SceI endonuclease from its chromosome and wanted to use this new strain as the engineering strain for all the biobricks. Due to the limited time, we did not construct this strain. Any teams interested in enhancing safety level might consider this undertaking.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Plasmid lacks a prefix.
  Illegal EcoRI site found at 2590
  Illegal XbaI site found at 2596
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 2590
  Illegal SpeI site found at 2
  Illegal PstI site found at 16
  Illegal NotI site found at 9
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 2590
  Illegal BamHI site found at 2566
  Illegal XhoI site found at 1033
  Illegal XhoI site found at 1925
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Plasmid lacks a prefix.
  Illegal suffix found at 2
  Illegal EcoRI site found at 2590
  Illegal XbaI site found at 2596
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 2590
  Illegal XbaI site found at 2596
  Illegal SpeI site found at 2
  Illegal PstI site found at 16
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal BsaI.rc site found at 2146