Difference between revisions of "Part:BBa K2984048"

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<partinfo>BBa_K2984048 short</partinfo>
 
<partinfo>BBa_K2984048 short</partinfo>
  
This part is composed of the PsaD promoter (BBa_K2984008), bleomycin resistance with self cleaving peptide scp (BBa_K2984045), the ARS secretion signal (BBa_K2984000), the PETase enzyme (), the secretion enhancing SP20 glycomodule (BBa_K2984015), and the Rbcs2 terminator (BBa_K2984018).  
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This vector is a part of the <a href="https://2019.igem.org/Team:Humboldt_Berlin/Part_Collection">Chlamy-HUB-Collection</a>. This part is composed of the <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K2984008">PsaD promoter</a>, <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K2984045">bleomycin resistance</a> with self cleaving peptide scp, the <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K2984000">ARS</a> secretion signal, the <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K2984011">PETase enzyme</a>, the secretion enhancing <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K2984016">SP20</a> glycomodule, and the <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K2984018">Rbcs2 terminator</a>.  
  
 
The part can be used to express and secrete the PETase enzyme in C. reinhardtii. The bleomycin resistance leads to a higher expression of the enzyme, due to its working mechanism. For every bleomycin moleucle, the bleomycin resistance gene must be expressed once. The resistance works in a ratio of 1:1 to the antibiotic. By having the resistance in the same open reading frame, the enzyme is also expressed, leading to a higher expression of the enzyme.  
 
The part can be used to express and secrete the PETase enzyme in C. reinhardtii. The bleomycin resistance leads to a higher expression of the enzyme, due to its working mechanism. For every bleomycin moleucle, the bleomycin resistance gene must be expressed once. The resistance works in a ratio of 1:1 to the antibiotic. By having the resistance in the same open reading frame, the enzyme is also expressed, leading to a higher expression of the enzyme.  
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<!-- Add more about the biology of this part here
 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K2984048 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K2984048 SequenceAndFeatures</partinfo>
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==Characterization==
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<figure>
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<img src="https://2019.igem.org/wiki/images/e/e8/T--Humboldt_Berlin--Konstruk_14.jpg" alt="colonies_total" width="500">
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<figcaption>Fig.1 - Image of a successful transformation in E.coli after ligation of the construct. The construct can then be isolated from E. coli to be transformed in C. reinhardtii</figcaption>
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Revision as of 10:53, 17 October 2019


L1c-PsaD-bleRscp-ARS-PETase-SP20-RbcS2


This vector is a part of the Chlamy-HUB-Collection. This part is composed of the PsaD promoter, bleomycin resistance with self cleaving peptide scp, the ARS secretion signal, the PETase enzyme, the secretion enhancing SP20 glycomodule, and the Rbcs2 terminator. The part can be used to express and secrete the PETase enzyme in C. reinhardtii. The bleomycin resistance leads to a higher expression of the enzyme, due to its working mechanism. For every bleomycin moleucle, the bleomycin resistance gene must be expressed once. The resistance works in a ratio of 1:1 to the antibiotic. By having the resistance in the same open reading frame, the enzyme is also expressed, leading to a higher expression of the enzyme.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 2249
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 4
    Illegal XhoI site found at 1973
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1607
  • 1000
    COMPATIBLE WITH RFC[1000]


Characterization

colonies_total
Fig.1 - Image of a successful transformation in E.coli after ligation of the construct. The construct can then be isolated from E. coli to be transformed in C. reinhardtii