Coding

Part:BBa_K3589110

Designed by: Yannik Schermer   Group: iGEM20_TU_Kaiserslautern   (2020-10-23)


Mutant laccase from Botrytis aclada for Chlamydomonas reinhardtii, Part 2 (Phytobrick)

This basic part contains the coding sequence of the laccase from the ascomycete Botrytis aclada mutant L499F (B4). This part is codon-optimized for Chlamydomonas reinhardtii. Combined with the first part of the laccase (BBa_K3589107), a promoter and a terminator, this basic part 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). To detect the target protein a 3xHA-tag from the Kaiser Collection (BBa_K3002017) is recommended. We recommend using the pAR promoter (A1-A3) (BBa_K3002003).
This part was designed to allow synthesis due to a high GC-level. It is only functional in combination with the second part (BBa_K3589110), of the laccase. Visit the page of the part BBa_K3589108 to gain a deeper understanding of the functionality of the laccase

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 57
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 57
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 575
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 57
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 57
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None