Difference between revisions of "Part:BBa K2918030"

 
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<partinfo>BBa_K2918030 short</partinfo>
 
<partinfo>BBa_K2918030 short</partinfo>
  
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A level 1 assembly of Wild type T7 promoter, a universal RBS, GFP from the registry and T7 terminator
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A level 1 assembly of Wild type T7 promoter, a universal RBS, GFP from the registry and T7 terminator.
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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_K2918030 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K2918030 SequenceAndFeatures</partinfo>
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The construct is confirmed by sequencing and there are no mutations.
  
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===Toxicity===
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Our Sci-Phi 29 tool is based on four components of the Φ29 bacteriophage: DNAP, TP, p5 and p6. However, overexpression of these proteins are toxic for the cell. In order to determine the optimal expression levels of the proteins in live cells, we carried out viability assays in <i>E. coli</i> BL21(DE3) pLysS. The results are shown in the graphs below.
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<div><ul>
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    <li style="display: inline-block;"> [[File:T--TUDelft--WTGFP-nomM.png|thumb|none|444px|<b>Figure 3A:</b> The growth curve of GFP under a WT T7 promoter in hours with no IPTG induction]] </li>
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    <li style="display: inline-block;"> [[File:T--TUDelft--WTGFP-1mM.png|thumb|none|444px|<b>Figure 3B:</b> The growth curve of GFP under a WT T7 promoter in hours with 1 mM IPTG induction]] </li>
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    <li> [[File:T--TUDelft--WTGFP-10mM.png|thumb|center|444px|<b>Figure 3C:</b> The growth curve of GFP under a WT T7 promoter in hours with 10 mM IPTG induction]] </li>
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    </ul></div>
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<!-- Add more about the biology of this part here
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===Usage and Biology===
  
 
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Latest revision as of 22:18, 6 December 2019


WT T7 promoter - Universal RBS - GFP - T7 terminator

A level 1 assembly of Wild type T7 promoter, a universal RBS, GFP from the registry and T7 terminator.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 465
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 465
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 465
    Illegal BamHI site found at 674
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 465
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 465
  • 1000
    COMPATIBLE WITH RFC[1000]

The construct is confirmed by sequencing and there are no mutations.

Toxicity

Our Sci-Phi 29 tool is based on four components of the Φ29 bacteriophage: DNAP, TP, p5 and p6. However, overexpression of these proteins are toxic for the cell. In order to determine the optimal expression levels of the proteins in live cells, we carried out viability assays in E. coli BL21(DE3) pLysS. The results are shown in the graphs below.

  • Figure 3A: The growth curve of GFP under a WT T7 promoter in hours with no IPTG induction
  • Figure 3B: The growth curve of GFP under a WT T7 promoter in hours with 1 mM IPTG induction
  • Figure 3C: The growth curve of GFP under a WT T7 promoter in hours with 10 mM IPTG induction