Difference between revisions of "Part:BBa K4806104"
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− | <p>This level 1 composite part contains the AβSAP(i)-promotor (<a href=" https://parts.igem.org/Part:BBa_K4806013">BBa_K4806013</a>), the coding sequence of CYP2D6 (<a href=" https://parts.igem.org/Part:BBa_K4806001">BBa_K4806001</a>), the HA-tag (<a href=" https://parts.igem.org/Part:BBa_K3002017">BBa_K3002017</a>)<sup>*</sup> for detection and the tRPL23-terminator (<a href="https://parts.igem.org/Part:BBa_K3002006">BBa_K3002006</a>)<sup>*</sup>. This part is codon-optimized for <i>Chlamydomonas reinhardtii</i> and was built as part of the CYPurify Collection | + | <p>This level 1 composite part contains the AβSAP(i)-promotor (<a href=" https://parts.igem.org/Part:BBa_K4806013">BBa_K4806013</a>), the coding sequence of CYP2D6 (<a href=" https://parts.igem.org/Part:BBa_K4806001">BBa_K4806001</a>), the HA-tag (<a href=" https://parts.igem.org/Part:BBa_K3002017">BBa_K3002017</a>)<sup>*</sup> for detection and the tRPL23-terminator (<a href="https://parts.igem.org/Part:BBa_K3002006">BBa_K3002006</a>)<sup>*</sup>. This part is codon-optimized for <i>Chlamydomonas reinhardtii</i> and was built as part of the CYPurify Collection.</p> |
<br> | <br> | ||
<h2>Construct</h2> | <h2>Construct</h2> | ||
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This construct was designed using the modular cloning system (MoClo).</div> | This construct was designed using the modular cloning system (MoClo).</div> | ||
</p> | </p> | ||
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<p><br></p> | <p><br></p> | ||
<h2>Sequence and Features</h2> | <h2>Sequence and Features</h2> | ||
</html> | </html> | ||
− | <partinfo> | + | <partinfo>BBa_K4806104 SequenceAndFeatures</partinfo> |
− | <partinfo> | + | <partinfo>BBa_K4806104 parameters</partinfo> |
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<img class="agarose" src="https://static.igem.wiki/teams/4806/wiki/registry/level2/2d6-ha.png"> | <img class="agarose" src="https://static.igem.wiki/teams/4806/wiki/registry/level2/2d6-ha.png"> | ||
<div class="unterschrift"><b>Fig.2 Test digest of CYP2D6 level 2 with HA-tag</b><br> | <div class="unterschrift"><b>Fig.2 Test digest of CYP2D6 level 2 with HA-tag</b><br> | ||
− | We digested this level 2 MoClo part with the restriction enzymes <i> | + | We digested this level 2 MoClo part with the restriction enzymes <i>Not</i>I and <i>Bste</i>II.</div></p> |
<p>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.</p> | <p>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.</p> | ||
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</html> | </html> |
Revision as of 09:51, 2 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).
This construct was designed using the modular cloning system (MoClo).
Sequence and Features
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 1098
Illegal PstI site found at 2454
Illegal PstI site found at 2887 - 12INCOMPATIBLE 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 - 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 530
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 1098
Illegal PstI site found at 2454
Illegal PstI site found at 2887 - 25INCOMPATIBLE 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 - 1000COMPATIBLE WITH RFC[1000]
Results
We confirmed that this construct is built correctly via agarose gel electrophoresis.
Fig.2 Test digest of CYP2D6 level 2 with HA-tag
We digested this level 2 MoClo part with the restriction enzymes NotI and BsteII.
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.