Part:BBa_K3589106

Wildtype laccase from uncultured marine bacteria 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) (B4). This part is codon-optimized for Chlamydomonas reinhardtii. Combined with the first part of BaLac (BBa_K3589105), 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 2 and 3 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 (3xHA-tag) is recommended. This part was designed to allow synthesis due to a high GC-level. It is only functional in combination with the first part (BBa_K3589105) 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 517
  Illegal PstI site found at 22
  Illegal PstI site found at 52
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 517
  Illegal PstI site found at 22
  Illegal PstI site found at 52
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 517
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 517
  Illegal PstI site found at 22
  Illegal PstI site found at 52
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 517
  Illegal PstI site found at 22
  Illegal PstI site found at 52
  Illegal NgoMIV site found at 840
  Illegal AgeI site found at 93
  Illegal AgeI site found at 1105
  
• 1000
  COMPATIBLE WITH RFC[1000]