Part:BBa_K4815017:Design
pDualPH7-pDual mcherry-yeGFP High 7
- 10INCOMPATIBLE 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 - 12INCOMPATIBLE 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 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 396
Illegal BglII site found at 1963
Illegal BamHI site found at 3640 - 23INCOMPATIBLE 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 - 25INCOMPATIBLE 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 - 1000INCOMPATIBLE 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.
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.