Part:BBa_K1441002

PGAPz alpha A + 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 760
  Illegal XbaI site found at 783
  Illegal SpeI site found at 798
  Illegal PstI site found at 820
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 760
  Illegal SpeI site found at 798
  Illegal PstI site found at 820
  Illegal NotI site found at 771
  Illegal NotI site found at 809
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 760
  Illegal BglII site found at 1
  Illegal BamHI site found at 1238
  Illegal XhoI site found at 736
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 760
  Illegal XbaI site found at 783
  Illegal SpeI site found at 798
  Illegal PstI site found at 820
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 760
  Illegal XbaI site found at 783
  Illegal SpeI site found at 798
  Illegal PstI site found at 820
  Illegal NgoMIV site found at 1995
  Illegal NgoMIV site found at 2056
  Illegal AgeI site found at 957
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal BsaI.rc site found at 1221
  Illegal BsaI.rc site found at 1420
  Illegal BsaI.rc site found at 2417
  Illegal SapI site found at 244