Difference between revisions of "Part:BBa K4806104"

Revision as of 11:10, 6 October 2023

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]
Illegal XhoI site found at 530
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 NcbI and HindIII.

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.

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