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=== Construction details === | === Construction details === | ||
| − | pME-Gal4VP16 was made by PCR amplification of Gal4VP16 ( | + | pME-Gal4VP16 was made by PCR amplification of Gal4VP16 from clones made by Reinhard Koester (described in Koester and Fraser, Dev Biol. 2001), using primers that add att sites, followed by a BP reaction. Sequencing finds a single silent mutation, G1114T. |
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| + | In this construct, the Gal4 DNA binding domain is fused to VP16[413-470], a partially-truncated VP16 transactivation domain. This fusion has reduced toxicity compared to the original Gal4VP16[413-490] from Sadowski et al. 1988. See Ogura et al., Dev Dyn 238:641, 2009 for comparisons of different Gal4VP16 fusions. | ||
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| + | === Sequence === | ||
| + | Annotated sequence, Genbank format:<br> | ||
| + | [[pME-Gal4VP16 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> | ||
[[pME-Gal4VP16 sequence]] | [[pME-Gal4VP16 sequence]] | ||
Latest revision as of 17:24, 7 March 2010
Construction details
pME-Gal4VP16 was made by PCR amplification of Gal4VP16 from clones made by Reinhard Koester (described in Koester and Fraser, Dev Biol. 2001), using primers that add att sites, followed by a BP reaction. Sequencing finds a single silent mutation, G1114T.
In this construct, the Gal4 DNA binding domain is fused to VP16[413-470], a partially-truncated VP16 transactivation domain. This fusion has reduced toxicity compared to the original Gal4VP16[413-490] from Sadowski et al. 1988. See Ogura et al., Dev Dyn 238:641, 2009 for comparisons of different Gal4VP16 fusions.
Sequence
Annotated sequence, Genbank format:
pME-Gal4VP16 Genbank
FASTA file with the full-length sequence as well as sequences of individual components:
pME-Gal4VP16 sequence
Crude map
Screenshot from Sequencher showing locations of components:
