Part:BBa_K2429035
phEF1a 3 Exon mKate-HBG Reporter
This part splits the red fluorescent mKate reporter into two exons, with human beta globin (HBG) intron 2 in between The first mKate exon and an exon that contains a stop codon. HBG intron 1 is placed between the exon with a stop codon and the second mKate exon. We used this split mKate construct as a reporter construct that we used to determine if our dCas13 or Ms2 systems were successful in controlling the inclusion of an exon. We used HBG Introns because it is a standard intron used in reporter constructs
In the absence of our system, the following mRNA transcript would be made. The presence of the stop codon would lead to a truncated protein, and there would be a knock down of fluorescence.
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, REST codon, and HBG intron 1 to be spliced out, and would result in more fluorescence than if our system wasn't present.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 2944
Illegal XbaI site found at 2349
Illegal PstI site found at 315
Illegal PstI site found at 820 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 2944
Illegal PstI site found at 315
Illegal PstI site found at 820 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 2944
Illegal BglII site found at 569
Illegal BamHI site found at 1183
Illegal XhoI site found at 968 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 2944
Illegal XbaI site found at 2349
Illegal PstI site found at 315
Illegal PstI site found at 820 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 2944
Illegal XbaI site found at 2349
Illegal PstI site found at 315
Illegal PstI site found at 820
Illegal NgoMIV site found at 703
Illegal AgeI site found at 81 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 1237
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