Difference between revisions of "Part:BBa K4806006"

 
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<partinfo>BBa_K4806006 short</partinfo>
 
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===Usage and Biology===
 
  
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<p>This basic part contains the coding sequence of mNeonGreen (B5). This part is codon-optimized for <i>Chlamydomonas reinhardtii</i> and was built as part of the CYPurify Collection. In combination with a promotor 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 detection of your expressed target protein like CYP3A4 (<a href=" https://parts.igem.org/Part:BBa_K4806000">BBa_K4806000</a>). </p>
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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K4806006 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/mstop-cons.png">
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  <div class="unterschrift"><b>Fig.1 Construct design</b><br>
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  We designed 4 level 2 constructs containing mStop using the modular cloning system (MoClo).
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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. The POR gene with mStop for <i>Chlamydomonas reinhardtii</i> (Phytobrick) (<a href=" https://parts.igem.org/Part:BBa_K4806211">BBa_K4806211</a>)</li>
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<li>2. CYP3A4 gene with mStop for <i>Chlamydomonas reinhardtii</i> (Phytobrick) (<a href=" https://parts.igem.org/Part:BBa_K4806202">BBa_K4806202</a>)</li>
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<li>3. 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>4. CYP81A10V7 gene with mStop for <i>Chlamydomonas reinhardtii</i> (Phytobrick) (<a href=" https://parts.igem.org/Part:BBa_K4806220">BBa_K4806220</a>)</li>
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</ul>
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  These constructs were transformed into <i>Chlamydomonas reinhardtii</i>. Besides mStop the constructs contain the CAβSAP(i)-promotor (<a href=" https://parts.igem.org/Part:BBa_K4806013">BBa_K4806013</a>), either the POR (<a href=" https://parts.igem.org/Part:BBa_K4806003">BBa_K4806003</a>), CYP3A3 (<a href=" https://parts.igem.org/Part:BBa_K4806000">BBa_K4806000</a>), CYP9Q3 (<a href=" https://parts.igem.org/Part:BBa_K4806004">BBa_K4806004</a>) or the CYP81A10V7 coding sequence (<a href=" https://parts.igem.org/Part:BBa_K4806005">BBa_K4806005</a>) and the tRPL23-terminator (<a href=" https://parts.igem.org/Part:BBa_K3002006">BBa_K3002006</a>)<sup>*</sup>. The resistance cassette for hygromycin or spectinomycin is already built in the level 2 vector pMBS810/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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</p>
  
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<h2>Sequence and Features</h2>
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<partinfo>BBa_K4806009 SequenceAndFeatures</partinfo>
  
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<partinfo>BBa_K4806009 parameters</partinfo>
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===Functional Parameters===
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<partinfo>BBa_K4806006 parameters</partinfo>
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<h2>Results</h2>

Revision as of 15:29, 20 September 2023


mNeonGreen-tag for Chlamydomonas reinhardtii (Phytobrick)

This basic part contains the coding sequence of mNeonGreen (B5). This part is codon-optimized for Chlamydomonas reinhardtii and was built as part of the CYPurify Collection. In combination with a promotor like AβSAP(i) (BBa_K4806013) and a terminator like tRPL23 (BBa_K3002006)*, this level 0 part leads to detection of your expressed target protein like CYP3A4 (BBa_K4806000).


Constructs

Fig.1 Construct design
We designed 4 level 2 constructs containing mStop using the modular cloning system (MoClo).


Here are the links to the built constructs:

  • 1. The POR gene with mStop for Chlamydomonas reinhardtii (Phytobrick) (BBa_K4806211)
  • 2. CYP3A4 gene with mStop for Chlamydomonas reinhardtii (Phytobrick) (BBa_K4806202)
  • 3. CYP9Q3 gene with mStop for Chlamydomonas reinhardtii (Phytobrick) (BBa_K4806223)
  • 4. CYP81A10V7 gene with mStop for Chlamydomonas reinhardtii (Phytobrick) (BBa_K4806220)

These constructs were transformed into Chlamydomonas reinhardtii. Besides mStop the constructs contain the CAβSAP(i)-promotor (BBa_K4806013), either the POR (BBa_K4806003), CYP3A3 (BBa_K4806000), CYP9Q3 (BBa_K4806004) or the CYP81A10V7 coding sequence (BBa_K4806005) and the tRPL23-terminator (BBa_K3002006)*. The resistance cassette for hygromycin or spectinomycin is already built in the level 2 vector pMBS810/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
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 14
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Results