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=== Construction details === | === Construction details === | ||
| − | p5E-UAS was made by conventionally subcloning into p5E-MCS a HindIII-EcoRI fragment from | + | p5E-UAS was made by conventionally subcloning into p5E-MCS a HindIII-EcoRI fragment from pBUAS-E1b-RFP from Reinhard Köster, containing multimerized Gal4 upstream activating sequence (UAS) elements, followed by an adenovirus E1b TATA box 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While the source construct nominally contained a 14x UAS element (originally made by Pernilla Rorth), sequencing shows that p5E-UAS only contains 10x UAS elements.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 | While the source construct nominally contained a 14x UAS element (originally made by Pernilla Rorth), sequencing shows that p5E-UAS only contains 10x UAS elements.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. | ||
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Functional tests of constructs using this element show strong activation by Gal4. | Functional tests of constructs using this element show strong activation by Gal4. | ||
| − | === | + | === Sequence === |
| + | Annotated sequence, Genbank format:<br> | ||
| + | [[p5E-UAS Genbank]] | ||
| + | |||
FASTA file with the full-length sequence as well as sequences of individual components:<br> | FASTA file with the full-length sequence as well as sequences of individual components:<br> | ||
[[p5E-UAS sequence]] | [[p5E-UAS sequence]] | ||
Latest revision as of 03:43, 19 December 2007
Construction details
p5E-UAS was made by conventionally subcloning into p5E-MCS a HindIII-EcoRI fragment from pBUAS-E1b-RFP from Reinhard Köster, containing multimerized Gal4 upstream activating sequence (UAS) elements, followed by an adenovirus E1b TATA box 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While the source construct nominally contained a 14x UAS element (originally made by Pernilla Rorth), sequencing shows that p5E-UAS only contains 10x UAS elements.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.
Sequence
Annotated sequence, Genbank format:
p5E-UAS Genbank
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:
