Part:BBa_K4806108

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

This level 1 composite part contains the AβSAP(i)-promotor (BBa_K4806013), the coding sequence of CYPCamC (BBa_K4806002), 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 1023
  Illegal PstI site found at 1295
  Illegal PstI site found at 2019
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 249
  Illegal PstI site found at 1023
  Illegal PstI site found at 1295
  Illegal PstI site found at 2019
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal XhoI site found at 530
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 1023
  Illegal PstI site found at 1295
  Illegal PstI site found at 2019
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 1023
  Illegal PstI site found at 1295
  Illegal PstI site found at 2019
  Illegal NgoMIV site found at 3040
  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 CYPCamC 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.