Coding

Part:BBa_K3002004

Designed by: Dorothée Klein   Group: iGEM19_TU_Kaiserslautern   (2019-10-12)
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Wildtype PETase for Chlamydomonas reinhardtii (Phytobrick)

This basic part contains the coding sequence of the wildtype PETase (B3-B4) and was built as a part of the Kaiser Collection. This part is codon-optimized for Chlamydomonas reinhardtii. Combined with a promoter and a terminator, this level 0 construct mediates PET degradation ability. As this part contains the introns 1-3 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 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 507
    Illegal PstI site found at 1480
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 507
    Illegal PstI site found at 1480
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 507
    Illegal PstI site found at 1480
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 507
    Illegal PstI site found at 1480
    Illegal NgoMIV site found at 242
    Illegal NgoMIV site found at 269
  • 1000
    COMPATIBLE WITH RFC[1000]


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