Part:BBa_K3589105

Mutant laccase from Botrytis aclada for Escherichia coli

This basic part contains the coding sequence of the mutant form L499F of the laccase from the ascomycete Botrytis aclada (hereafter referred to as BaLac) (B3). This part is codon-optimized for Chlamydomonas reinhardtii. Combined with the second part of BaLac (XXXXX), a promoter and a terminator, this level 0 construct should mediate the oxidation of a wide variety of substrates including phenolic compounds and aromatic amines. As this part contains the introns 1 and 2 of RBCS2, it perfectly matches the part BBa_K3002027 (pAR promoter A1-B2), resulting in a high expression (Eichler-Stahlberg et al., 2009 ADD LINK). To detect the target protein, a tag of the Kaiser Collection, introduced by TU Kaiserslautern's 2019 iGEM team like, BBa_K3002017 HA-tag) is recommended. This part was designed to allow synthesis due to a high GC-level. It is only functional in combination with the second part (XXXXX) of the laccase. (Visit the page of the part (XXXX) to gain a deeper understanding of the functionality of BaLac.)

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 775
  Illegal PstI site found at 463
  Illegal PstI site found at 550
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 775
  Illegal PstI site found at 463
  Illegal PstI site found at 550
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 775
  Illegal BamHI site found at 920
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 775
  Illegal PstI site found at 463
  Illegal PstI site found at 550
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 775
  Illegal PstI site found at 463
  Illegal PstI site found at 550
  Illegal NgoMIV site found at 612
  Illegal AgeI site found at 136
  Illegal AgeI site found at 976
  Illegal AgeI site found at 1059
  Illegal AgeI site found at 1315
  
• 1000
  COMPATIBLE WITH RFC[1000]