Difference between revisions of "Part:BBa K346000:Design"

 
 
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__NOTOC__
 
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<partinfo>BBa_K346000 short</partinfo>
 
<partinfo>BBa_K346000 short</partinfo>
  
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<partinfo>BBa_K346000 SequenceAndFeatures</partinfo>
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<html><a href="https://static.igem.org/mediawiki/parts/1/14/Design1.jpg" target="blank"><img src="https://static.igem.org/mediawiki/parts/1/14/Design1.jpg"></a></html>
  
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T3 RNA polymerase has a dramatically high transcription rating; therefore a strong RBS is not necessary as a medium RBS (BBa_B0032) will be enough to reach the maximum of transcription activation at T3 promoters.
  
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===Design Notes===
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What’s more, we observed that maximum transcription rating might result in protein misfolding sometimes and the apparent transcription activation fold will decrease instead. Therefore we strongly recommend a low expression intensity of T3 polymerase in the case of genetic circuit design, or the combination of T3 polymerase with a low-strength T3 promoter, such as promoter phi 1.3 or 4.3. You can find more info about the T3 promoter intensity in the following Figure
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We cloned this part from plasmid NR1, which was a gift from Anne O. Summers.
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<html><a href="https://static.igem.org/mediawiki/parts/a/ae/T3POLfig4.png" target="blank"><img src="https://static.igem.org/mediawiki/parts/a/ae/T3POLfig4.png"></a></html>
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'''Figure 5: The strength of different T3 promoters under the same expression level of T3 polymerase. BBa_E0840 was combined with 11 T3 promoters from the genome of T3 phage. The x axis denotes strains that only differ in the T3 promoter strength, in accordance with the order from strong to weak. The Y axis denotes the GFP intensity normalized by OD600. We can see that these T3 promoters can be divided into 3 subgroups: weak, medium and strong. (You can find more details about the promoter intensity characterization on the Experience Page)'''
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<partinfo>BBa_K346000 SequenceAndFeatures</partinfo>
  
  
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===Source===
 
===Source===
  
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Tn21
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Enterobacteria phage T3 (Bacteriophage T3).
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----
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We used the following primers to standardize this part:
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T3pol_For: ccg gaattc gcggccgc t tctag ATGAACATCATCGAAAACATCGAA
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T3pol_Rev: aaa ctgcag cggccgc t actagt a TTATGCAAAGGCAAAGTCAGAC
  
 
===References===
 
===References===

Latest revision as of 07:11, 25 October 2010

RBS(B0032)+T3 DNA-directed RNA polymerase

T3 RNA polymerase has a dramatically high transcription rating; therefore a strong RBS is not necessary as a medium RBS (BBa_B0032) will be enough to reach the maximum of transcription activation at T3 promoters.

What’s more, we observed that maximum transcription rating might result in protein misfolding sometimes and the apparent transcription activation fold will decrease instead. Therefore we strongly recommend a low expression intensity of T3 polymerase in the case of genetic circuit design, or the combination of T3 polymerase with a low-strength T3 promoter, such as promoter phi 1.3 or 4.3. You can find more info about the T3 promoter intensity in the following Figure

Figure 5: The strength of different T3 promoters under the same expression level of T3 polymerase. BBa_E0840 was combined with 11 T3 promoters from the genome of T3 phage. The x axis denotes strains that only differ in the T3 promoter strength, in accordance with the order from strong to weak. The Y axis denotes the GFP intensity normalized by OD600. We can see that these T3 promoters can be divided into 3 subgroups: weak, medium and strong. (You can find more details about the promoter intensity characterization on the Experience Page)



Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 2585


Source

Enterobacteria phage T3 (Bacteriophage T3).

We used the following primers to standardize this part: T3pol_For: ccg gaattc gcggccgc t tctag ATGAACATCATCGAAAACATCGAA T3pol_Rev: aaa ctgcag cggccgc t actagt a TTATGCAAAGGCAAAGTCAGAC

References