Difference between revisions of "Part:BBa K4806103"

 
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<partinfo>BBa_K4806103 short</partinfo>
 
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===Usage and Biology===
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<span class='h3bb'>Sequence and Features</span>
 
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<partinfo>BBa_K4806103 SequenceAndFeatures</partinfo>
 
  
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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 CYP3A4 (<a href=" https://parts.igem.org/Part:BBa_K4806000">BBa_K4806000</a>), mStop (<a href=" https://parts.igem.org/Part:BBa_K4806009">BBa_K4806009</a>) 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>
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<h2>Construct</h2>
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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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  <div class="unterschrift"><b>Fig.1 Construct design</b><br>
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  This construct was designed using the modular cloning system (MoClo).</div>
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<p><br></p>
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<h2>Sequence and Features</h2>
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<partinfo>BBa_K4806103 SequenceAndFeatures</partinfo>
  
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===Functional Parameters===
 
 
<partinfo>BBa_K4806103 parameters</partinfo>
 
<partinfo>BBa_K4806103 parameters</partinfo>
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<h2>Results</h2>
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<p>We confirmed that this construct is built correctly via agarose gel electrophoresis.</p>
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<p>
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  <img class="agarose" src="https://static.igem.wiki/teams/4806/wiki/registry/level2/2d6-ha.png">
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  <div class="unterschrift"><b>Fig.2 Test digest of CYP3A4 level 1 with mStop</b><br>
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We digested this level 2 MoClo part with the restriction enzymes <i>Not</i>I and <i>Bam</i>HI.</div></p>
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<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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<p><br></p>
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<h2>Contribution</h2>
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<p>The <sup>*</sup> marked parts were not created by us. Our results can be found on the experience page of each part.</p>
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</html>

Revision as of 10:06, 2 October 2023


CYP3A4 gene with mStop for Chlamydomonas reinhardtii (Phytobrick)


This level 1 composite part contains the AβSAP(i)-promotor (BBa_K4806013), the coding sequence of CYP3A4 (BBa_K4806000), 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 1846
    Illegal PstI site found at 2168
    Illegal PstI site found at 2228
    Illegal PstI site found at 2700
    Illegal PstI site found at 2769
    Illegal PstI site found at 2873
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 249
    Illegal PstI site found at 1846
    Illegal PstI site found at 2168
    Illegal PstI site found at 2228
    Illegal PstI site found at 2700
    Illegal PstI site found at 2769
    Illegal PstI site found at 2873
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 3180
    Illegal XhoI site found at 530
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 1846
    Illegal PstI site found at 2168
    Illegal PstI site found at 2228
    Illegal PstI site found at 2700
    Illegal PstI site found at 2769
    Illegal PstI site found at 2873
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 1846
    Illegal PstI site found at 2168
    Illegal PstI site found at 2228
    Illegal PstI site found at 2700
    Illegal PstI site found at 2769
    Illegal PstI site found at 2873
    Illegal NgoMIV site found at 2090
    Illegal NgoMIV site found at 3513
    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 CYP3A4 level 1 with mStop
We digested this level 2 MoClo part with the restriction enzymes NotI and BamHI.

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.