Difference between revisions of "Part:BBa K4815016:Design"
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<partinfo>BBa_K4815016 SequenceAndFeatures</partinfo> | <partinfo>BBa_K4815016 SequenceAndFeatures</partinfo> | ||
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===Design Notes=== | ===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. | |
| − | + | <html> | |
| − | + | <figure><center> | |
| + | <img alt="" | ||
| + | src="https://static.igem.wiki/teams/4815/wiki/fannei.png" | ||
| + | width="400" | ||
| + | title=""> | ||
| + | <figcaption>pA-core PYPH/PYPL-pT </figcaption> | ||
| + | </figure> | ||
| + | </html> | ||
===Source=== | ===Source=== | ||
| − | + | Originated from the framework yeast_DualReporter(AddGene: 127546) | |
===References=== | ===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. | ||
Latest revision as of 06:42, 12 October 2023
pDualPH6-pDual mcherry-yeGFP High 6
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 396
Illegal XbaI site found at 3645
Illegal PstI site found at 964
Illegal PstI site found at 2196 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 396
Illegal NheI site found at 3407
Illegal PstI site found at 964
Illegal PstI site found at 2196 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 396
Illegal BglII site found at 1927
Illegal BamHI site found at 3608 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 396
Illegal XbaI site found at 3645
Illegal PstI site found at 964
Illegal PstI site found at 2196 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 396
Illegal XbaI site found at 3645
Illegal PstI site found at 964
Illegal PstI site found at 2196
Illegal NgoMIV site found at 1826 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 2886
Illegal BsaI.rc site found at 4285
Illegal SapI site found at 4776
Illegal SapI site found at 5376
Illegal SapI.rc site found at 2733
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.