Plasmid

Part:BBa_K4815017:Design

Designed by: Chen Xi   Group: iGEM23_NJU-China   (2023-10-12)
Revision as of 08:24, 12 October 2023 by ChenXi (Talk | contribs)

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pDualPH7-pDual mcherry-yeGFP High 7


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 396
    Illegal XbaI site found at 3677
    Illegal SpeI site found at 1351
    Illegal PstI site found at 991
    Illegal PstI site found at 2232
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 396
    Illegal NheI site found at 3443
    Illegal SpeI site found at 1351
    Illegal PstI site found at 991
    Illegal PstI site found at 2232
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 396
    Illegal BglII site found at 1963
    Illegal BamHI site found at 3640
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 396
    Illegal XbaI site found at 3677
    Illegal SpeI site found at 1351
    Illegal PstI site found at 991
    Illegal PstI site found at 2232
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 396
    Illegal XbaI site found at 3677
    Illegal SpeI site found at 1351
    Illegal PstI site found at 991
    Illegal PstI site found at 2232
    Illegal NgoMIV site found at 1862
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 2922
    Illegal BsaI.rc site found at 4317
    Illegal SapI site found at 4808
    Illegal SapI site found at 5408
    Illegal SapI.rc site found at 2769

Design Notes

Our Pymaker synthesized core promoter sequence is extracted through PCR, and introducing the restriction sites ofBamH I and Xho I, the same as the scaffold. The scaffold is a preserved sequence in all PYPHs. It is a structure that we learned and utilized from previous research that can link the core promoter with the codon and provide restriction sites of BamH I and Xho I which make it possible for the plasmids with the scaffold to be inserted by various core promoter sequences at ease. The scaffold is already maintained in the plasmid framework yeast_DualReporter(AddGene: 127546),using this assay we create a promoter library that can drive the high expression of YeGFP and be used quickly for the detection of expression rate.

pA-core PYPH/PYPL-pT

Source

Originated from the framework yeast_DualReporter(AddGene: 127546)

References

[1] Liu, X., et al., Dual Fluorescent Protein (yEGFP/DsRed-Express-2) Bioassay System for Rapid Screening for Chemical Mutagens Based on RNR3 Regulation in Saccharomyces Cerevisiae. Biomed Environ Sci, 2021. 34(5): p. 421-424.