Difference between revisions of "Part:BBa K2429019"
 
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In the absence of our system, the following mRNA transcript would be made. The presence of the stop codon in between the eYFP and eBFP proteins would lead to a truncaated protein in which only the conserved region and eYFP sequence would remain, and only yellow fluorescent protein would be produced.
 
In the absence of our system, the following mRNA transcript would be made. The presence of the stop codon in between the eYFP and eBFP proteins would lead to a truncaated protein in which only the conserved region and eYFP sequence would remain, and only yellow fluorescent protein would be produced.
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https://parts.igem.org/File:C-y-b.png
 
  
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https://static.igem.org/mediawiki/parts/c/c5/C-y-b.png
  
  
 
In the presence of our system, the following mRNA transcript would be made. The dCas13a or Ms2 protein would bind to the 3' splice site, and cause HBG intron 2, eYFP exon, and HBG intron 1 to be spliced out, and would result in only blue fluorescent protein to be produced.
 
In the presence of our system, the following mRNA transcript would be made. The dCas13a or Ms2 protein would bind to the 3' splice site, and cause HBG intron 2, eYFP exon, and HBG intron 1 to be spliced out, and would result in only blue fluorescent protein to be produced.
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https://static.igem.org/mediawiki/parts/2/28/Case2.2.1.png
 
https://static.igem.org/mediawiki/parts/2/28/Case2.2.1.png
  

Latest revision as of 22:19, 23 October 2017


3 Exon eYFP Reporter

This part consists of 3 exons: the first exon is made of sequence for a "conserved" region that most fluorescent proteins share. The second intron is made of the The second exon is made up of the sequences unique to YFP, as well as the remaining parts of the fluorescent protein. The third exon is very similar to the second, except instead of sequences unique to yellow fluorescence, it has sequences unique to blue. In between the conserved region and YFP exon is the second human beta globin (HBG) intron, and in between the YFP exon and BFP exon is the first HBG intron.

We used this construct to determine whether our RNA binding proteins (dCas13a or Ms2) were successful in controlling the exclusion of an exon. 3ex_FP.png


In the absence of our system, the following mRNA transcript would be made. The presence of the stop codon in between the eYFP and eBFP proteins would lead to a truncaated protein in which only the conserved region and eYFP sequence would remain, and only yellow fluorescent protein would be produced.

C-y-b.png


In the presence of our system, the following mRNA transcript would be made. The dCas13a or Ms2 protein would bind to the 3' splice site, and cause HBG intron 2, eYFP exon, and HBG intron 1 to be spliced out, and would result in only blue fluorescent protein to be produced.

Case2.2.1.png


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 2282
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 2282
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 2282
    Illegal BamHI site found at 1
    Illegal BamHI site found at 1412
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 2282
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 2282
  • 1000
    COMPATIBLE WITH RFC[1000]