Difference between revisions of "Part:BBa K3589110"

Latest revision as of 13:54, 25 October 2020

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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-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 first part of the laccase (<a href="https://parts.igem.org/Part:BBa_K3589107">BBa_K3589107</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 2 and 3 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>).
+This basic part contains the coding sequence of the laccase from the ascomycete <I>Botrytis aclada</i> mutant  L499F (B4). This part is codon-optimized for <I>Chlamydomonas reinhardtii</I>. Combined with the first part of the laccase (<a href="https://parts.igem.org/Part:BBa_K3589107">BBa_K3589107</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 2 and 3 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>).
 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.
 We recommend using the pAR promoter (A1-A3) (<a href="https://parts.igem.org/Part:BBa_K3002003">BBa_K3002003</a>).