Difference between revisions of "Part:BBa K5207011"

 
 
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This is an expression cassette consisting of a promoter pCCW12, the SmCPS1 gene, and a terminator tSSA1.
 
This is an expression cassette consisting of a promoter pCCW12, the SmCPS1 gene, and a terminator tSSA1.
  
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===Usage and Biology===
 
  
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K5207011 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K5207011 SequenceAndFeatures</partinfo>
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===Functional Parameters===
 
===Functional Parameters===
 
<partinfo>BBa_K5207011 parameters</partinfo>
 
<partinfo>BBa_K5207011 parameters</partinfo>
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===Usage and Biology===
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We combined each Level 0 junction module, promoter, CDS, terminator, and ending module, and constructed them into vector pYTK095 through BsaI digestion and ligation, and this step was the construction of Level 1.
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<html>
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<body>
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<figure>
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<div class = "center">
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<center><img src = "https://static.igem.wiki/teams/5207/parts/4.png" style = "width:600px"></center>
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</div>
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<figcaption><center>Figure 1. Snapgene diagrams of pYTK095</center></figcaption>
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</figure>
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</body>
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</html>
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After transformation in E. coli DH10B, we selected the single bacterial colony (pYTK095) that does not fluoresce under UV light from the Petri dishes. It was used for colony PCR amplification and validation.
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<html>
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<body>
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<figure>
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<div class = "center">
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<center><img src = "https://static.igem.wiki/teams/5207/parts/7.png" style = "width:600px"></center>
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</div>
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<figcaption><center>Figure 2. Fluorescent labeling screening of pYTK095</center></figcaption>
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</figure>
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</body>
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</html>
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<html>
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<body>
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<figure>
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<div class = "center">
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<center><img src = "https://static.igem.wiki/teams/5207/parts/9.png" style = "width:600px"></center>
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</div>
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<figcaption><center>Figure 3. Validation plot of pYTK095 gel electrophoresis</center></figcaption>
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</figure>
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</body>
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</html>
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Latest revision as of 13:21, 30 September 2024


pCCW12-SmCPS1-tSSA1

This is an expression cassette consisting of a promoter pCCW12, the SmCPS1 gene, and a terminator tSSA1.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 2397
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1158
    Illegal BamHI site found at 756
    Illegal XhoI site found at 892
    Illegal XhoI site found at 916
    Illegal XhoI site found at 2209
    Illegal XhoI site found at 2419
    Illegal XhoI site found at 2891
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 561
    Illegal NgoMIV site found at 1978
    Illegal NgoMIV site found at 2542
  • 1000
    COMPATIBLE WITH RFC[1000]


Usage and Biology

We combined each Level 0 junction module, promoter, CDS, terminator, and ending module, and constructed them into vector pYTK095 through BsaI digestion and ligation, and this step was the construction of Level 1.

Figure 1. Snapgene diagrams of pYTK095

After transformation in E. coli DH10B, we selected the single bacterial colony (pYTK095) that does not fluoresce under UV light from the Petri dishes. It was used for colony PCR amplification and validation.

Figure 2. Fluorescent labeling screening of pYTK095

Figure 3. Validation plot of pYTK095 gel electrophoresis