Part:BBa_I20284

Bacteriophage Bxb1 Integrase-Freiburg standard

Bxb1 integrase is a recombinase enzyme from the serine recombinase family. Bxb1 naturally catalyze integration of the phage genome into the GroEL1 gene of M.segmentis, which contains the attB site [2]. Minimal attB, attP,[2],[3] attL and attR [4] have been characterized. Excisionase has been identified, is relatively big (28kDa) for a RDF, and is efficient both in vivo and in vitro [4]. In vitro recombination is very efficient ranging a 100% efficiency in 1hour for integration [2] and for excision [4]. Interestingly the excisionase subunit does not bind DNA and acts stoechiometrically on Bxb1 integrase [4]. The enzyme does not require supercoiled DNA, and can also works with linear products [2]. The enzyme also works in human, rat, and mouse cells [5, 6]. When recombination sites are placed in the same DNA molecule, recombination outcome is inversion of the sequence they surround.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 198
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 472
  Illegal XhoI site found at 559
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 4
  Illegal AgeI site found at 1507
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 1306