Difference between revisions of "Part:BBa K4806004"

 
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<partinfo>BBa_K4806004 short</partinfo>
 
<partinfo>BBa_K4806004 short</partinfo>
  
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===Usage and Biology===
 
  
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<p>This basic part contains the coding sequence of CYP9Q3 (B3-B4). This part is codon-optimized for <i>Chlamydomonas reinhardtii</i> and was built as part of the CYPurify Collection. In combination with a promoter like AβSAP(i) (<a href=" https://parts.igem.org/Part:BBa_K4806013">BBa_K4806013</a>) and a terminator like tRPL23 (<a href="https://parts.igem.org/Part:BBa_K3002006">BBa_K3002006</a>)<sup>*</sup>, this level 0 part leads to expression and potential detoxification of specific chemicals (Ohkawa & Inui, 2015). To detect the target protein a tag like HA-tag (<a href=" https://parts.igem.org/Part:BBa_K3002017">BBa_K3002017</a>)<sup>*</sup> is recommended. </p>
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<span class='h3bb'>Sequence and Features</span>
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<br>
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<partinfo>BBa_K4806004 SequenceAndFeatures</partinfo>
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<h2>Constructs</h2>
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<p>
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  <img class="bild" src="https://static.igem.wiki/teams/4806/wiki/registry/level-0/cyp9q3-constructs.png"/">
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  <div class="unterschrift"><b>Fig.1 Construct design</b><br>
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  We designed 3 level 2 constructs containing CYP9Q3 using the modular cloning system (MoClo).
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  </div>  
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<p><br></p>
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<p>
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    Here are the links to the built constructs:<br>
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<ul>
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<li>1. CYP9Q3 gene with HA-tag for <i>Chlamydomonas reinhardtii</i> (Phytobrick) (<a href=" https://parts.igem.org/Part:BBa_K4806222">BBa_K4806222</a>)</li>
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<li>2. CYP9Q3 gene with mStop for <i>Chlamydomonas reinhardtii</i> (Phytobrick) (<a href=" https://parts.igem.org/Part:BBa_K4806223">BBa_K4806223</a>)</li>
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<li>3. CYP9Q3 gene with mNeonGreen for <i>Chlamydomonas reinhardtii</i> (Phytobrick) (<a href=" https://parts.igem.org/Part:BBa_K4806224">BBa_K4806224</a>)</li>
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</ul>
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</p>
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<p>
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  These constructs were transformed into <i>Chlamydomonas reinhardtii</i>. Besides the CYP9Q3 coding sequence the constructs contain the AβSAP(i)-promotor (<a href=" https://parts.igem.org/Part:BBa_K4806013">BBa_K4806013</a>), either the HA-tag (<a href=" https://parts.igem.org/Part:BBa_K3002017">BBa_K3002017</a>)<sup>*</sup> or mNeonGreen (<a href=" https://parts.igem.org/Part:BBa_K4806006">BBa_K4806006</a>) for detection or 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>. The resistance cassette for spectinomycin is already built in the level 2 vector pMBS807 we are using. The usage of this vector allows the direct assembly of level 0 parts to level 2 constructs, facilitating the cloning time (Niemeyer & Schroda, 2022).
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<h2>Sequence and Features</h2>
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===Functional Parameters===
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<partinfo>BBa_K4806004 parameters</partinfo>
 
<partinfo>BBa_K4806004 parameters</partinfo>
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<h2>Results</h2>

Revision as of 11:45, 20 September 2023


CYP9Q3 gene for Chlamydomonas reinhardtii (Phytobrick)

This basic part contains the coding sequence of CYP9Q3 (B3-B4). This part is codon-optimized for Chlamydomonas reinhardtii and was built as part of the CYPurify Collection. In combination with a promoter like AβSAP(i) (BBa_K4806013) and a terminator like tRPL23 (BBa_K3002006)*, this level 0 part leads to expression and potential detoxification of specific chemicals (Ohkawa & Inui, 2015). To detect the target protein a tag like HA-tag (BBa_K3002017)* is recommended.


Constructs

Fig.1 Construct design
We designed 3 level 2 constructs containing CYP9Q3 using the modular cloning system (MoClo).


Here are the links to the built constructs:

  • 1. CYP9Q3 gene with HA-tag for Chlamydomonas reinhardtii (Phytobrick) (BBa_K4806222)
  • 2. CYP9Q3 gene with mStop for Chlamydomonas reinhardtii (Phytobrick) (BBa_K4806223)
  • 3. CYP9Q3 gene with mNeonGreen for Chlamydomonas reinhardtii (Phytobrick) (BBa_K4806224)

These constructs were transformed into Chlamydomonas reinhardtii. Besides the CYP9Q3 coding sequence the constructs contain the AβSAP(i)-promotor (BBa_K4806013), either the HA-tag (BBa_K3002017)* or mNeonGreen (BBa_K4806006) for detection or mStop (BBa_K4806009) and the tRPL23-terminator (BBa_K3002006)*. The resistance cassette for spectinomycin is already built in the level 2 vector pMBS807 we are using. The usage of this vector allows the direct assembly of level 0 parts to level 2 constructs, facilitating the cloning time (Niemeyer & Schroda, 2022).

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 1249
    Illegal PstI site found at 1429
    Illegal PstI site found at 1935
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 1249
    Illegal PstI site found at 1429
    Illegal PstI site found at 1935
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1791
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 1249
    Illegal PstI site found at 1429
    Illegal PstI site found at 1935
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 1249
    Illegal PstI site found at 1429
    Illegal PstI site found at 1935
    Illegal NgoMIV site found at 2206
  • 1000
    COMPATIBLE WITH RFC[1000]


Results