Part:BBa_K5492101

pETDUET-1

pETDUET-1 plasmid used for expression of Histamine-N-Methyltransferase BBa_K5492400, BBa_K5492401 and Diamine Oxidase BBa_K5492200 in multiple e.coli chassis.

Sequence and Features

As seen upon the map below, pETDUET-1 contains two lac operons, an AmpR gene, and contains multiple restriction sites. Our team has utilised the EcoRI site starting in position 112, and the BglII site starting in position 305. Additionally, we have utilised an RBS in position 58, and a start codon is located before the multiple cloning site, in position 71. A 6xHis tag is also found at position 83, which we have utilised in our gene designs in order to ease the protein purification process.

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 112
  Illegal XbaI site found at 30
  Illegal PstI site found at 131
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 112
  Illegal PstI site found at 131
  Illegal NotI site found at 149
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 112
  Illegal BglII site found at 305
  Illegal BamHI site found at 106
  Illegal XhoI site found at 354
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 112
  Illegal XbaI site found at 30
  Illegal PstI site found at 131
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 112
  Illegal XbaI site found at 30
  Illegal PstI site found at 131
  Illegal NgoMIV site found at 324
  Illegal NgoMIV site found at 671
  Illegal NgoMIV site found at 5348
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 1256
  Illegal SapI.rc site found at 2916