Part:BBa_K2141000

pET28a(+) expression plasmid

pET28a(+) is a nonviral expression plasmid used in bacteria strains for protein production purposes. It has kanamycin resistance gene as a selection marker. Plasmid sequence consists of T7 promoter and terminator sequences as well as lac operon sequence and lacI promoter for protein expression induction. It has both N- and C- terminal HisTag sequences and N- terminal thrombin cleavage site.
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Additional Information

Team: IISERPUNE-INDIA-2020

• Pet28a(+) is widely used for expression purpose. But unnecessary components often interfere in cloning strategies. So we altered the part into a modified version that satisfies RFC10 criteria, compatible with Gibson's cloning and all the unnecessary components removed: BBa_K358500.

Please see the experience page for more information.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 192
  Illegal XbaI site found at 335
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 192
  Illegal NheI site found at 231
  Illegal NotI site found at 165
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 192
  Illegal BglII site found at 401
  Illegal BamHI site found at 198
  Illegal XhoI site found at 158
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 192
  Illegal XbaI site found at 335
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 192
  Illegal XbaI site found at 335
  Illegal NgoMIV site found at 433
  Illegal NgoMIV site found at 2021
  Illegal NgoMIV site found at 2181
  Illegal NgoMIV site found at 5228
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI site found at 3101