Coding

Part:BBa_K3002037

Designed by: Dorothée Klein   Group: iGEM19_TU_Kaiserslautern   (2019-10-16)
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Wildtype MHETase for Chlamydomonas reinhardtii (Phytobrick)

This basic part contains the coding sequence of the wildtype MHETase (B3-B4). This part is codon-optimized for Chlamydomonas reinhardtii. It is a fusion of the parts <a href="https://parts.igem.org/Part:BBa_K3002029">BBa_K3002029</a> and <a href="https://parts.igem.org/Part:BBa_K3002005">BBa_K3002005</a>. In combination with a promoter and a terminator, this level 0 construct mediates MHET degradation ability. As this part contains the introns 1 and two times intron 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>). To detect or purify the target protein a tag of the Kaiser Collection like <a href="https://parts.igem.org/Part:BBa_K3002010">BBa_K3002010</a> (sp20 HA-tag), <a href="https://parts.igem.org/Part:BBa_K3002017">BBa_K3002017</a> (HA-tag), <a href="https://parts.igem.org/Part:BBa_K3002018">BBa_K3002018</a> (sp20 His-tag), <a href="https://parts.igem.org/Part:BBa_K3002028">BBa_K3002028</a> (His-tag) is recommended. A secretion signal (<a href="https://parts.igem.org/Part:BBa_K3002007">BBa_K3002007</a> (cCA), <a href="https://parts.igem.org/Part:BBa_K3002008">BBa_K3002008</a> (GLE) or <a href="https://parts.igem.org/Part:BBa_K3002009">BBa_K3002009</a> (ARS)) can be added, when using the part <a href="https://parts.igem.org/Part:BBa_K3002003">BBa_K3002003</a> (pAR promoter (A1-A3)) as a promoter.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 765
    Illegal PstI site found at 1089
    Illegal PstI site found at 1432
    Illegal PstI site found at 2242
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 765
    Illegal PstI site found at 1089
    Illegal PstI site found at 1432
    Illegal PstI site found at 2242
    Illegal NotI site found at 1100
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 2010
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 765
    Illegal PstI site found at 1089
    Illegal PstI site found at 1432
    Illegal PstI site found at 2242
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 765
    Illegal PstI site found at 1089
    Illegal PstI site found at 1432
    Illegal PstI site found at 2242
    Illegal NgoMIV site found at 698
    Illegal NgoMIV site found at 1159
    Illegal NgoMIV site found at 1189
    Illegal NgoMIV site found at 1504
    Illegal NgoMIV site found at 1522
    Illegal NgoMIV site found at 1558
    Illegal NgoMIV site found at 2201
  • 1000
    COMPATIBLE WITH RFC[1000]


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