Part:BBa_K4806104

CYP2D6 gene with HA-tag for Chlamydomonas reinhardtii (Phytobrick)

This level 1 composite part contains the AβSAP(i)-promotor (BBa_K4806013), the coding sequence of CYP2D6 (BBa_K4806001), the HA-tag (BBa_K3002017)^* for detection and the tRPL23-terminator (BBa_K3002006)^*. This part is codon-optimized for Chlamydomonas reinhardtii and was built as part of the CYPurify Collection.

Construct

Fig.1 Construct design
This construct was designed using the modular cloning system (MoClo).

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal PstI site found at 1098
  Illegal PstI site found at 2454
  Illegal PstI site found at 2887
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 249
  Illegal PstI site found at 1098
  Illegal PstI site found at 2454
  Illegal PstI site found at 2887
  Illegal NotI site found at 1604
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Unknown
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 1098
  Illegal PstI site found at 2454
  Illegal PstI site found at 2887
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 1098
  Illegal PstI site found at 2454
  Illegal PstI site found at 2887
  Illegal NgoMIV site found at 1741
  Illegal NgoMIV site found at 2090
  Illegal NgoMIV site found at 3431
  Illegal AgeI site found at 268
  
• 1000
  COMPATIBLE WITH RFC[1000]

Results

We confirmed that this construct is built correctly via agarose gel electrophoresis.

Fig.2 Test digest of CYP2D6 level 1 with HA-tag
We digested this level 2 MoClo part with the restriction enzymes NotI and BsteII.

The test digest in Fig.2 was compared to an in-silico digest. Together with the sequencing results we were able to demonstrate that our construct was built correctly.

Contribution

The ^* marked parts were not created by us. Our results can be found on the experience page of each part.