Plasmid

Part:BBa_K165062

Designed by: Aaron Glieberman   Group: iGEM08_BrownTwo   (2008-10-29)
Revision as of 00:47, 30 October 2008 by Aaron Glieberman (Talk | contribs)

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


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 1116
    Illegal EcoRI site found at 3178
    Illegal XbaI site found at 3148
    Illegal SpeI site found at 3154
    Illegal PstI site found at 3172
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1116
    Illegal EcoRI site found at 3178
    Illegal SpeI site found at 3154
    Illegal PstI site found at 3172
    Illegal NotI site found at 3140
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1116
    Illegal EcoRI site found at 3178
    Illegal BamHI site found at 3160
    Illegal XhoI site found at 3211
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1116
    Illegal EcoRI site found at 3178
    Illegal XbaI site found at 3148
    Illegal SpeI site found at 3154
    Illegal PstI site found at 3172
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1116
    Illegal EcoRI site found at 3178
    Illegal XbaI site found at 3148
    Illegal SpeI site found at 3154
    Illegal PstI site found at 3172
    Illegal NgoMIV site found at 2799
    Illegal AgeI site found at 1503
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 4584
    Illegal SapI site found at 3501


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