Part:BBa_K560013

BsaI Golden Gate Assembly No.2

The second vector of the Golden gate series (adapted from psb1c3) This vector expedites golden gate cloning with BBa_K560012, BBa_K560014 and BBa_K560015. To make this set of vector compatible with the standard biobricks, we have maintained PstI and XbaI recognition sites. Flanking these two sites are the cleavage and recognition sites of BsaI, a kind of Type IIS restriction endonuclease. This kind of restriction endonucleases cleaves outside their recognition sites thus creating different sticky ends. If you want to assemble four biobricks at a time, you can put the biobricks in order into corresponding vector by the digestion of PstI and Xbal. Then mix these assembled vectors and a backbone (PFUS4 from TALEN golden gate kit). Under the effect of ligase and Bsal, these biobricks will have sticky ends nose to tails and ligate in order in a single reaction.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal XbaI site found at 2073
  Illegal PstI site found at 3
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal PstI site found at 3
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal XhoI site found at 1041
  Illegal XhoI site found at 1933
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal XbaI site found at 2073
  Illegal PstI site found at 3
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal XbaI site found at 2073
  Illegal PstI site found at 3
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal BsaI site found at 2062
  Illegal BsaI.rc site found at 14