Part:BBa_K3002029

Wildtype MHETase for Chlamydomonas reinhardtii Part 2 (Phytobrick)

This basic part contains the coding sequence of the wildtype MHETase (B4) and was built as a part of the Kaiser Collection. This part is codon-optimized for Chlamydomonas reinhardtii. Combined with the first part of the MHETase (BBa_K3002005), a promoter and a terminator, this level 0 construct mediates PET degradation ability. As this part contains the second intron of RBCS2, it perfectly matches the part BBa_K3002027(pAR promoter A1-B2) resulting in a high expression (Eichler-Stahlberg, 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 57
  Illegal PstI site found at 331
  Illegal PstI site found at 1141
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal PstI site found at 57
  Illegal PstI site found at 331
  Illegal PstI site found at 1141
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 909
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 57
  Illegal PstI site found at 331
  Illegal PstI site found at 1141
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 57
  Illegal PstI site found at 331
  Illegal PstI site found at 1141
  Illegal NgoMIV site found at 403
  Illegal NgoMIV site found at 421
  Illegal NgoMIV site found at 457
  Illegal NgoMIV site found at 1100
  
• 1000
  COMPATIBLE WITH RFC[1000]