Difference between revisions of "Part:BBa K4806105"
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<h2>Construct</h2>
 
<h2>Construct</h2>
 
<p>
 
<p>
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   <img class="bild" src="https://static.igem.wiki/teams/4806/wiki/registry/level2/cyp2d6-ha-construct.png">
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   <img class="bild" src="https://static.igem.wiki/teams/4806/wiki/registry/level-1/con-2d6-mstop.png">
 
   <div class="unterschrift"><b>Fig.1 Construct design</b><br>
 
   <div class="unterschrift"><b>Fig.1 Construct design</b><br>
 
   This construct was designed using the modular cloning system (MoClo).</div>
 
   This construct was designed using the modular cloning system (MoClo).</div>
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<p>We confirmed that this construct is built correctly via agarose gel electrophoresis.</p>
 
<p>We confirmed that this construct is built correctly via agarose gel electrophoresis.</p>
 
<p>
 
<p>
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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/level-1/2d6-mstop.png">
 
   <div class="unterschrift"><b>Fig.2 Test digest of CYP2D6 level 1 with mStop</b><br>
 
   <div class="unterschrift"><b>Fig.2 Test digest of CYP2D6 level 1 with mStop</b><br>
 
We digested this level 2 MoClo part with the restriction enzymes <i>Nde</i>I and <i>Sap</i>HI.</div></p>
 
We digested this level 2 MoClo part with the restriction enzymes <i>Nde</i>I and <i>Sap</i>HI.</div></p>

Revision as of 10:35, 2 October 2023


CYP2D6 gene with mStop for Chlamydomonas reinhardtii (Phytobrick)


This level 1 composite part contains the AβSAP(i)-promotor (BBa_K4806013), the coding sequence of CYP2D6 (BBa_K4806001), mStop (BBa_K4806009) 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 BamHI site found at 3017
    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 3350
    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 mStop
We digested this level 2 MoClo part with the restriction enzymes NdeI and SapHI.

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.