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p5E-bactin2 was made by PCR of the ''bactin2'' upstream genomic sequence (using primers to add att sites), | p5E-bactin2 was made by PCR of the ''bactin2'' upstream genomic sequence (using primers to add att sites), | ||
| − | followed by a BP reaction. Upstream sequence includes 3.8 kb upstream of transcriptional start, a small 5 | + | followed by a BP reaction. Upstream sequence includes 3.8 kb upstream of transcriptional start, a small 5' UTR exon, a 1.4 kb intron, and then 38 bp of 5' UTR, |
terminating immediately before the native start codon. | terminating immediately before the native start codon. | ||
Sequencing confirms that the ends of the insert are correct. | Sequencing confirms that the ends of the insert are correct. | ||
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=== Sequence === | === Sequence === | ||
| − | Annotated sequence, Genbank format:< | + | Annotated sequence, Genbank format:<br> |
[[p5E-bactin2 Genbank]] | [[p5E-bactin2 Genbank]] | ||
Latest revision as of 17:34, 22 August 2008
Construction details
bactin2 genomic sequence was derived from a 5.3 kb clone (gift of Ken Poss), which was derived in turn from a 10 kb upstream construct from Shin-ichi Higashijima.
p5E-bactin2 was made by PCR of the bactin2 upstream genomic sequence (using primers to add att sites), followed by a BP reaction. Upstream sequence includes 3.8 kb upstream of transcriptional start, a small 5' UTR exon, a 1.4 kb intron, and then 38 bp of 5' UTR, terminating immediately before the native start codon. Sequencing confirms that the ends of the insert are correct.
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."
Sequence
Annotated sequence, Genbank format:
p5E-bactin2 Genbank
FASTA file with the full-length sequence as well as sequences of individual components:
p5E-bactin2 sequence
Crude map
Screenshot from Sequencher showing locations of components:
