Part:BBa_J18903

pSB1AT3F construction plasmid for protein fusion BioBricks

pSB1AT3F is almost identical to pSB1AT3 but uses the "Freiburg" extension of the standard BioBrick prefix and suffix and features silent mutations in the tetracycline resistance gene. The Freiburg extension adds a new pair of inner restriction enzymes -- NgoMIV (prefix) and AgeI (suffix) -- which allows the in-frame fusion of protein parts. See [http://openwetware.org/wiki/The_BioBricks_Foundation:Standards/Technical/Formats BB formats].

pSB1AT3F is a high copy number plasmid carrying ampicillin and tetracyclin resistance. The replication origin is a pUC19-derived pMB1 (copy number of 100-300 per cell). For the version of this plasmid with a CcdB insert, see BBa_P1010.

The given sequence starts with the Freiburg BB suffix and ends with the Freiburg BB prefix. The complete vector sequence is therefore obtained from: part + vector

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal prefix found in sequence at 3434
  Illegal suffix found in sequence at 10
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 3434
  Illegal NheI site found at 1279
  Illegal SpeI site found at 11
  Illegal PstI site found at 25
  Illegal NotI site found at 18
  Illegal NotI site found at 3440
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 3434
  Illegal BamHI site found at 1425
  Illegal XhoI site found at 1044
  Illegal XhoI site found at 2327
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal prefix found in sequence at 3434
  Illegal suffix found in sequence at 11
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal prefix found in sequence at 3434
  Illegal suffix found in sequence at 1
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 2473