Part:BBa_K3842006

pPICZαA-Gas6(plasmid)

BBa_K3842000 is a pichia pastoris plasmid which contains AOX1 promoter, Gas6 coding sequence, AOX1 terminator, N-terminal secretion signal from S. cerevisiae alpha-factor, BleoR, high-copy-number ColE1/pMB1/pBR322/pUC origin of replication, transcription terminator for CYC1, promoter for EF-1α, and synthetic bacterial promoter EM7. It can give host cell zeocin resisitance and it has ability to generate mRNA of Gas6 sequence when host cell is induced by methanol. In our project, this plasmid was designed to transformed into pichia pastoris and enable the pichia pastoris to produce Gas6 protein under certain

environment which contain specific concentration of methanol.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 1215
  Illegal EcoRI site found at 4786
  Illegal XbaI site found at 1235
  Illegal PstI site found at 5731
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 1215
  Illegal EcoRI site found at 4786
  Illegal PstI site found at 5731
  Illegal NotI site found at 1221
  Illegal NotI site found at 6847
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 1215
  Illegal EcoRI site found at 4786
  Illegal BglII site found at 1
  Illegal BglII site found at 3553
  Illegal BglII site found at 5297
  Illegal BamHI site found at 1641
  Illegal XhoI site found at 1191
  Illegal XhoI site found at 4754
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 1215
  Illegal EcoRI site found at 4786
  Illegal XbaI site found at 1235
  Illegal PstI site found at 5731
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 1215
  Illegal EcoRI site found at 4786
  Illegal XbaI site found at 1235
  Illegal PstI site found at 5731
  Illegal NgoMIV site found at 2399
  Illegal NgoMIV site found at 2460
  Illegal AgeI site found at 1360
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 1624
  Illegal BsaI.rc site found at 1823
  Illegal BsaI.rc site found at 2821