Coding

Part:BBa_K3002005

Designed by: Dorothée Klein   Group: iGEM19_TU_Kaiserslautern   (2019-10-12)
Revision as of 20:59, 12 October 2019 by Registry (Talk | contribs)

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Wildtype MHETase for Chlamydomonas reinhardtii Part 1 (Phytobrick)

This basic part contains the coding sequence of the wildtype MHETase (B3) and was built as a part of the Kaiser Collection. This part is codon-optimized for Chlamydomonas reinhardtii. Combined with the second part of the MHETase (BBa_K3002029), a promoter and a terminator, this level 0 construct mediates PET degradation ability. 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 or purify the target protein a tag of the Kaiser Collection like BBa_K3002010 (sp20 HA-tag), BBa_K3002017 (HA-tag), BBa_K3002018 (sp20 His-tag), BBa_K3002028 (His-tag) is recommended. A secretion signal (BBa_K3002007 (cCA), BBa_K3002008 (GLE) or BBa_K3002009 (ARS)) can be added, when using the part BBa_K3002003 (pAR promoter (A1-A3)) as a promoter.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 711
    Illegal PstI site found at 1089
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 711
    Illegal PstI site found at 1089
    Illegal NotI site found at 1100
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 711
    Illegal PstI site found at 1089
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 711
    Illegal PstI site found at 1089
    Illegal NgoMIV site found at 1027
    Illegal NgoMIV site found at 1159
    Illegal NgoMIV site found at 1189
  • 1000
    COMPATIBLE WITH RFC[1000]


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Parameters
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