Difference between revisions of "Part:BBa K2984009"
 
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This part is a part of the <a href="https://2019.igem.org/Team:Humboldt_Berlin/Part_Collection">Chlamy-HUB-Collection</a>. Another strong constitutive promoter is the AR promoter that is composed of HSP70A and RBCS2 building a tandem promoter. Therefore the HSP70A (A) promoter is fused upstream of another promoter resulting in a stronger transgene expression (Schroda et al. 2000). The combination of the HSP70A (A) promoter and the RBCS2 (R) promoter building <sub>P</sub>AR has the positive effect that transcriptional gene silencing (TGS) is strongly reduced making this promoter popular for transgene expression (Schroda et al. 2002). This part was designed for <i>C. reinhardtii</i>
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This part is a part of the <a href="https://2019.igem.org/Team:Humboldt_Berlin/Part_Collection">Chlamy-HUB-Collection</a>. Another strong constitutive promoter is the AR promoter that is composed of HSP70A and RBCS2 building a tandem promoter. Therefore the HSP70A (A) promoter is fused upstream of another promoter resulting in a stronger transgene expression (Schroda et al. 2000). The combination of the HSP70A (A) promoter and the RBCS2 (R) promoter building <sub>P</sub>AR has the positive effect that transcriptional gene silencing (TGS) is strongly reduced making this promoter popular for transgene expression (Schroda et al. 2002). This part was designed for <i>C. reinhardtii</i>.
 
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This part was designed to be used with the MoClo standard and has A1-A3 overhangs.
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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_K2984009 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K2984009 SequenceAndFeatures</partinfo>
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== Characterization ==  
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==Characterization==
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The AR Promotor was used for two different paromomycin constructs. The first carries the registry part <a href="https://parts.igem.org/Part:BBa_K2703008"> BBa_K2703008 </a> which we wanted to compare to our <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K2984006"> improved </a>
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paromomycin construct which also feautres this promoter. The improvement was done as part of the 2019 <a href="https://2019.igem.org/Judging/Medals"> gold criteria.</a>
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<img src="https://2019.igem.org/wiki/images/4/4a/T--Humboldt_Berlin--AR_Paro.jpeg
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<img src="https://static.igem.org/mediawiki/parts/d/d2/T--Humboldt_Berlin--AR_A1-A3.png" alt="Plate_L0-AR_A1-A3_E.coli" width="500">
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<figcaption>Fig.1 - Successful transformation of L0-AR_A1-A3 into <i>E. coli</i>.</figcaption>
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<figcaption>Fig.1 - TAP-agar plates containing paromomycin. The <i> C.Reinhardtii </i> colonies growing on the plate were tranformed with our improved <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K2984006"> paromomycin construct </a> which is flanked by this AR promoter and the <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K2984018"> Rbcs2 </a> terminator </figcaption>
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<partinfo>BBa_K2984009 parameters</partinfo>
 
<partinfo>BBa_K2984009 parameters</partinfo>
 
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==References==
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Schroda, M., Blöcker, D., & Beck, C. F. (2000). The HSP70A promoter as a tool for the improved expression of transgenes in Chlamydomonas. The plant journal, 21(2), 121-131.
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Schroda, M., et al. (2002). "Sequence elements within an HSP70 promoter counteract transcriptional transgene silencing in Chlamydomonas." Plant J 31(4): 445-455.
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Latest revision as of 21:19, 21 October 2019


AR Promoter A1-A3

This part is a part of the Chlamy-HUB-Collection. Another strong constitutive promoter is the AR promoter that is composed of HSP70A and RBCS2 building a tandem promoter. Therefore the HSP70A (A) promoter is fused upstream of another promoter resulting in a stronger transgene expression (Schroda et al. 2000). The combination of the HSP70A (A) promoter and the RBCS2 (R) promoter building PAR has the positive effect that transcriptional gene silencing (TGS) is strongly reduced making this promoter popular for transgene expression (Schroda et al. 2002). This part was designed for C. reinhardtii.

This part was designed to be used with the MoClo standard and has A1-A3 overhangs.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 282
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 685
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

Characterization

Plate_L0-AR_A1-A3_E.coli
Fig.1 - Successful transformation of L0-AR_A1-A3 into E. coli.


References

  1. Schroda, M., Blöcker, D., & Beck, C. F. (2000). The HSP70A promoter as a tool for the improved expression of transgenes in Chlamydomonas. The plant journal, 21(2), 121-131.
  2. Schroda, M., et al. (2002). "Sequence elements within an HSP70 promoter counteract transcriptional transgene silencing in Chlamydomonas." Plant J 31(4): 445-455.