Plasmid

Part:BBa_K165061

Designed by: Aaron Glieberman   Group: iGEM08_BrownTwo   (2008-10-29)
Revision as of 00:46, 30 October 2008 by Aaron Glieberman (Talk | contribs)

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


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


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1942
    Illegal XbaI site found at 550
    Illegal XbaI site found at 1912
    Illegal SpeI site found at 1918
    Illegal PstI site found at 251
    Illegal PstI site found at 1936
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1942
    Illegal SpeI site found at 1918
    Illegal PstI site found at 251
    Illegal PstI site found at 1936
    Illegal NotI site found at 1904
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1942
    Illegal BamHI site found at 1924
    Illegal XhoI site found at 1975
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1942
    Illegal XbaI site found at 550
    Illegal XbaI site found at 1912
    Illegal SpeI site found at 1918
    Illegal PstI site found at 251
    Illegal PstI site found at 1936
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1942
    Illegal XbaI site found at 550
    Illegal XbaI site found at 1912
    Illegal SpeI site found at 1918
    Illegal PstI site found at 251
    Illegal PstI site found at 1936
    Illegal NgoMIV site found at 1563
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 3348
    Illegal SapI site found at 2265


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