Difference between revisions of "Part:BBa K165062"

Latest revision as of 19:37, 31 October 2008

pRS305 yeast shuttle vector, LEU2 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 LEU2 gene. To be used in conjunction with leucine drop-out media for positive selection of transformed cells. Transformation using this vector requires linearization of the plasmid by cutting with BstEII.

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

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

Vector: NotI, PstI

Prefix: NotI,SpeI

Suffix: XbaI, PstI

Original Sikorski vector was described in R. S. [http://www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pubmed&pubmedid=2659436 Sikorski and P. Hieter, Genetics, 122: 19-27 (1989)].

Vector NTI annotated sequence: File:PRS305.gb

Yeast transformation protocol: R Daniel Gietz & Robert H Schiestl. [http://www.natureprotocols.com/2007/01/31/highefficiency_yeast_transform.php High-efficiency yeast transformation using the LiAc/SS carrier DNA/PEG method]. Nature protocols (2007)

Sequence and Features