Part:BBa_K802004

Shuttle vector for E. coli and B. subtilis

The shuttle vector of 6,5kb is Ampicillin resistant in E. coli and Erythromycin resistant in B. subtilis. It is a high plasmid copy number in both B. subtilis and E.coli. It has a polylinker containing all 4 of the iGEM sites.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal prefix found in sequence at 6506
  Illegal suffix found in sequence at 1
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 6506
  Illegal NheI site found at 3377
  Illegal SpeI site found at 2
  Illegal PstI site found at 16
  Illegal NotI site found at 9
  Illegal NotI site found at 6512
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 6506
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal prefix found in sequence at 6506
  Illegal suffix found in sequence at 2
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal prefix found in sequence at 6506
  Illegal XbaI site found at 6521
  Illegal SpeI site found at 2
  Illegal PstI site found at 16
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 3203
  Illegal BsaI.rc site found at 5190
  

Characterization

Transformation yield

In the NM522 E.coli strain the average yield was 3,3XE06 cells/µg.The transformed cells were selected on LB media supplemented with Ampicillin at a concentration of 100µg/mL.

In the BS168 B.subtilis strain the maximum yield was 70 cells/µg. The transformed cells were selected on LB media supplemented with Erythromycin at a concentration of 15µg/mL.