Difference between revisions of "Part:BBa K165061"

Revision as of 05:52, 30 October 2008

pRS304* yeast shuttle vector, TRP1 selection

Ampicillin resistance when grown in bacteria. This plasmid is not currently Biobrick compatible, but is useful for inserting a final construct into a yeast cell.

Useful for chromosomal integration of a device into yeast strains. This will insert the vector at the specific locus of the TRP13 gene. To be used in conjunction with tryptophan drop-out media for positive selection of transformed cells. Transformation using this vector requires linearization of the plasmid by cutting with PstI.

To incorporate a single Biobrick part into this vector for subsequent transformation into yeast, one should cut both vector and part with EcoRI and SpeI.

Alternatively, one can do the final ligation step between two Biobrick parts into this vector by cutting with the following:

Vector: EcoRI, Not I

Prefix: EcoRI, SpeI

Suffix: XbaI, NotI

Original Sikorski vector was described in [http://www.genetics.org/cgi/reprint/122/1/19.pdf R. S. Sikorski and P. Hieter, Genetics, 122: 19-27 (1989)].

Yeast transformation protocol: R Daniel Gietz & Robert H Schiestl. High-efficiency yeast transformation using the LiAc/SS carrier DNA/PEG method . Nature protocols (2007)

Sequence and Features