(New page: === Construction details === p5E-UAS was made by conventionally subcloning into p5E-MCS a HindIII-EcoRI fragment from a UAS-E1b construct from Reinhard Köster, containing multimerized Gal...) |
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| − | p5E-UAS was made by conventionally subcloning into p5E-MCS a HindIII-EcoRI fragment from a UAS-E1b construct from Reinhard Köster, containing multimerized Gal4 upstream activating sequence (UAS) elements, followed by what | + | p5E-UAS was made by conventionally subcloning into p5E-MCS a HindIII-EcoRI fragment from a UAS-E1b construct from Reinhard Köster, containing multimerized Gal4 upstream activating sequence (UAS) elements, followed by what is presumably a carp beta-actin minimal promoter and then by a piece of the carp beta-actin 5' UTR. |
While the source construct nominally contained a 14x UAS element (originally made by Pernilla Rorth), sequencing shows that p5E-UAS only contains 10x UAS elements. Presumably there was an internal rearrangement that shed 4 UAS elements; it is not clear whether had already happened in the UAS-E1b construct, or whether it occurred in the Chien lab. | While the source construct nominally contained a 14x UAS element (originally made by Pernilla Rorth), sequencing shows that p5E-UAS only contains 10x UAS elements. Presumably there was an internal rearrangement that shed 4 UAS elements; it is not clear whether had already happened in the UAS-E1b construct, or whether it occurred in the Chien lab. | ||
Revision as of 03:53, 23 March 2007
Construction details
p5E-UAS was made by conventionally subcloning into p5E-MCS a HindIII-EcoRI fragment from a UAS-E1b construct from Reinhard Köster, containing multimerized Gal4 upstream activating sequence (UAS) elements, followed by what is presumably a carp beta-actin minimal promoter and then by a piece of the carp beta-actin 5' UTR.
While the source construct nominally contained a 14x UAS element (originally made by Pernilla Rorth), sequencing shows that p5E-UAS only contains 10x UAS elements. Presumably there was an internal rearrangement that shed 4 UAS elements; it is not clear whether had already happened in the UAS-E1b construct, or whether it occurred in the Chien lab.
Functional tests of constructs using this element show strong activation by Gal4.
Crudely annotated sequence
FASTA file with the full-length sequence as well as sequences of individual components:
p5E-UAS sequence
Crude map
Screenshot from Sequencher showing locations of components:
