P5E-UAS: Difference between revisions
Hggenusrwki (talk | contribs) Created page with "== Construction Details == This template is used to display DNA sequence information in a standardized format. == Crude Map == photo showing locations of components == Sequence == * Annotated Sequence: (Genbank format) link * Full-Length Sequence with individual component sequences included: (FASTA file) link == Reference == This template is used to display DNA sequence information in a standardized format." |
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== Construction Details == | == 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While the source construct nominally contained a 14x UAS element (originally made by Pernilla Rorth), sequencing shows that p5E-UAS only contains 10x UAS elements.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. | |||
== Crude Map == | == Crude Map == | ||
Screenshot from Sequencher showing locations of components: | |||
P5E-UAS.png | |||
== Sequence == | ==Download the Sequence == | ||
* Annotated Sequence | * '''Annotated Sequence'''(Genbank format) and '''Full-Length Sequence with individual component sequences included'''(FASTA file) | ||
: [[Media:233._pME-nlsmCherry.zip|Download p5E-UAS]] | |||
== Reference == | == Reference == | ||
Higashijima S, Okamoto H, Ueno N, Hotta Y, Eguchi G. (1997) Dev Biol.15;192:289-99. "High-frequency generation of transgenic zebrafish which | |||
reliably express GFP in whole muscles or the whole body by using promoters of zebrafish origin." | |||
Revision as of 16:28, 11 March 2025
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While the source construct nominally contained a 14x UAS element (originally made by Pernilla Rorth), sequencing shows that p5E-UAS only contains 10x UAS elements.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.
Crude Map
Screenshot from Sequencher showing locations of components: P5E-UAS.png
Download the Sequence
- Annotated Sequence(Genbank format) and Full-Length Sequence with individual component sequences included(FASTA file)
Reference
Higashijima S, Okamoto H, Ueno N, Hotta Y, Eguchi G. (1997) Dev Biol.15;192:289-99. "High-frequency generation of transgenic zebrafish which reliably express GFP in whole muscles or the whole body by using promoters of zebrafish origin."