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This basic part contains the coding sequence of the laccase from the ascomycete <I>Botrytis aclada</i> mutant  L499F (B3). This part is codon-optimized for <I>Chlamydomonas reinhardtii</I>. Combined with the second part of the laccase (<a href="https://parts.igem.org/Part:BBa_K3589110">BBa_K3589110</a>), 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 1 and 2 of RBCS2, it perfectly matches the part <a href="https://parts.igem.org/Part:BBa_K3002027">BBa_K3002027</a> (pAR promoter A1-B2), resulting in a high expression (<a href="https://www.researchgate.net/publication/23762345_Strategies_to_facilitate_transgene_expression_in_Chlamydomonas_reinhardtii">Eichler-Stahlberg et al., 2009</a>).
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To detect the target protein a 3xHA-tag from the Kaiser Collection (<a href="https://parts.igem.org/Part:BBa_K3002017">BBa_K3002017</a>) is recommended.
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We recommend using the pAR promoter (A1-A3) (<a href="https://parts.igem.org/Part:BBa_K3002003">BBa_K3002003</a>).
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This part was designed to allow synthesis due to a high GC-level. It is only functional in combination with the second part (<a href="https://parts.igem.org/Part:BBa_K3589110">BBa_K3589110</a>), of the laccase. Visit the page of the part <a href="https://parts.igem.org/Part:BBa_K3589108">BBa_K3589108</a> to gain a deeper understanding of the functionality of the laccase</b> </br>
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===Usage and Biology===
 
===Usage and Biology===

Latest revision as of 13:35, 25 October 2020


Mutant laccase from Botrytis aclada for Chlamydomonas reinhardtii, Part 1 (Phytobrick)

This basic part contains the coding sequence of the laccase from the ascomycete Botrytis aclada mutant L499F (B3). This part is codon-optimized for Chlamydomonas reinhardtii. Combined with the second part of the laccase (BBa_K3589110), 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 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). 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 608
    Illegal PstI site found at 695
    Illegal PstI site found at 759
    Illegal PstI site found at 1131
    Illegal PstI site found at 1419
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 608
    Illegal PstI site found at 695
    Illegal PstI site found at 759
    Illegal PstI site found at 1131
    Illegal PstI site found at 1419
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1055
    Illegal BamHI site found at 1394
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 608
    Illegal PstI site found at 695
    Illegal PstI site found at 759
    Illegal PstI site found at 1131
    Illegal PstI site found at 1419
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 608
    Illegal PstI site found at 695
    Illegal PstI site found at 759
    Illegal PstI site found at 1131
    Illegal PstI site found at 1419
  • 1000
    COMPATIBLE WITH RFC[1000]