Part:BBa_K1441001

PGAPz alpha C + Linker

This plasmid is PGAPzα C + modified Multiple Cloning Site(MCS) (Linker) fixed according to iGEM standards. We replaced old multiple cloning cite with new modified multiple cloning site, which has new cutting sites according to iGEM standard. The original MCS has ECORI, KpnI, NotI and XbaI restriction enzyme binding sites compared to the new MCS we designed in which there are ECORI, NotI, XbaI, SpeI and PstI restriction enzyme cutting sites. PGAPzα C + Linker has α-secretion factor before MCS which allows yeast cells and some Eukaryotic cells to secrete the protein into the outside of the cell membrane. Also after MCS there are MYC epitope and 6x histag that are used to tag the protein so it can be easily identified by westernblot technique.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 765
  Illegal XbaI site found at 788
  Illegal SpeI site found at 803
  Illegal PstI site found at 825
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 765
  Illegal SpeI site found at 803
  Illegal PstI site found at 825
  Illegal NotI site found at 776
  Illegal NotI site found at 814
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 765
  Illegal BglII site found at 1
  Illegal BamHI site found at 1243
  Illegal XhoI site found at 736
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 765
  Illegal XbaI site found at 788
  Illegal SpeI site found at 803
  Illegal PstI site found at 825
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 765
  Illegal XbaI site found at 788
  Illegal SpeI site found at 803
  Illegal PstI site found at 825
  Illegal NgoMIV site found at 2000
  Illegal NgoMIV site found at 2061
  Illegal AgeI site found at 962
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal BsaI.rc site found at 1226
  Illegal BsaI.rc site found at 1425
  Illegal BsaI.rc site found at 2422
  Illegal SapI site found at 244