Difference between revisions of "Part:BBa K4815011"

 
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<partinfo>BBa_K4815011 short</partinfo>
 
<partinfo>BBa_K4815011 short</partinfo>
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===Description===
 
The functional promoter sequence (about 223 bp) generated by our AI model Pymaker is inserted into the dual-fluorescence reporter system for characterization and we describe it as pDualPH1. Random DNA yields diverse expression levels in a yeast promoter library, while we can use the episomal dual reporter system expressing a constitutive red fluorescent protein (RFP) and a variable yellow fluorescent protein (YFP) to quantify promoter activity. By adpoting the log(YFP/RFP) measured using flow cytometry, we can detect the expression rate of our synthesized promoters. The advantage of this system is its ability to eliminate the influence of plasmid copy number and the growth status of the bacterial host, thereby providing a more direct measurement of the relative expression strength of the synthesized promoters by the Pymaker.
 
The functional promoter sequence (about 223 bp) generated by our AI model Pymaker is inserted into the dual-fluorescence reporter system for characterization and we describe it as pDualPH1. Random DNA yields diverse expression levels in a yeast promoter library, while we can use the episomal dual reporter system expressing a constitutive red fluorescent protein (RFP) and a variable yellow fluorescent protein (YFP) to quantify promoter activity. By adpoting the log(YFP/RFP) measured using flow cytometry, we can detect the expression rate of our synthesized promoters. The advantage of this system is its ability to eliminate the influence of plasmid copy number and the growth status of the bacterial host, thereby providing a more direct measurement of the relative expression strength of the synthesized promoters by the Pymaker.
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===Loci===
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pDualPH1 consists two parts: the synthesized core promoter and the pDual reporter scaffold. The synthesized core promoter is an 80 bp sequence generated by the Pymaker and is seated at approximately -170 to -90 upstream to the codon (which is the presumed transcription start site-TSS and is where most transcription factors binding sites lie).
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The he pDual reporter scaffold 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. 
  
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<!-- Add more about the biology of this part here
 
 
===Usage and Biology===
 
===Usage and Biology===
  

Revision as of 02:15, 12 October 2023


pDualPH1-pDual mcherry-yeGFP High 1

Description

The functional promoter sequence (about 223 bp) generated by our AI model Pymaker is inserted into the dual-fluorescence reporter system for characterization and we describe it as pDualPH1. Random DNA yields diverse expression levels in a yeast promoter library, while we can use the episomal dual reporter system expressing a constitutive red fluorescent protein (RFP) and a variable yellow fluorescent protein (YFP) to quantify promoter activity. By adpoting the log(YFP/RFP) measured using flow cytometry, we can detect the expression rate of our synthesized promoters. The advantage of this system is its ability to eliminate the influence of plasmid copy number and the growth status of the bacterial host, thereby providing a more direct measurement of the relative expression strength of the synthesized promoters by the Pymaker.

Loci

pDualPH1 consists two parts: the synthesized core promoter and the pDual reporter scaffold. The synthesized core promoter is an 80 bp sequence generated by the Pymaker and is seated at approximately -170 to -90 upstream to the codon (which is the presumed transcription start site-TSS and is where most transcription factors binding sites lie). The he pDual reporter scaffold 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.

Usage and Biology

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 396
    Illegal XbaI site found at 3552
    Illegal PstI site found at 964
    Illegal PstI site found at 2196
  • 12
    INCOMPATIBLE 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
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 396
    Illegal BglII site found at 1927
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 396
    Illegal XbaI site found at 3552
    Illegal PstI site found at 964
    Illegal PstI site found at 2196
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 396
    Illegal XbaI site found at 3552
    Illegal PstI site found at 964
    Illegal PstI site found at 2196
    Illegal NgoMIV site found at 1826
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 2886
    Illegal BsaI site found at 3484
    Illegal BsaI.rc site found at 4192
    Illegal SapI site found at 4683
    Illegal SapI site found at 5283
    Illegal SapI.rc site found at 2733