Composite
WT TP

Part:BBa_K2918019

Designed by: TUDelft2019   Group: iGEM19_TUDelft   (2019-10-04)
Revision as of 22:13, 6 December 2019 by Weronika77 (Talk | contribs) (→‎Toxicity)


WT T7 promoter - Universal RBS - Φ29 TP - T7 terminator

A level 1 construct with a Wild-type T7 promoter, a universal promoter, TP and a T7 terminator

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 237
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 557
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 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: Normalized maximum growth rate of phi29 TP under different promoter strengths (weak, medium, Wild-Type) with no IPTG induction
  • Figure 3B: Normalized maximum growth rate of phi29 TP under different promoter strengths (weak, medium, Wild-Type) with 1 mM IPTG induction
  • Figure 3C: Normalized maximum growth rate of phi29 TP under different promoter strengths (weak, medium, Wild-Type) with 10 mM IPTG induction


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Parameters
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