Composite
Weak GFP

Part:BBa_K2918031

Designed by: TUDelft2019   Group: iGEM19_TUDelft   (2019-10-04)
Revision as of 22:28, 6 December 2019 by HuyenMy (Talk | contribs) (→‎Toxicity)

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Weak T7 promoter - Universal RBS - GFP - T7 terminator

A level 1 with weak T7 promoter, a universal RBS, GFP and a 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 weak T7 promoter in hours with no IPTG induction
  • Figure 3B: The growth curve of GFP under a weak T7 promoter in hours with 1 mM IPTG induction
  • Figure 3C: The growth curve of GFP under a weak T7 promoter in hours with 10 mM IPTG inductionn


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