Plasmid

Part:BBa_K165062

Designed by: Aaron Glieberman   Group: iGEM08_BrownTwo   (2008-10-29)
Revision as of 19:50, 29 October 2008 by Registry (Talk | contribs)

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pRS305 yeast shuttle vector, LEU2 selection

Useful for chromosomal integration of a part into yeast strains. Insertion of a single Biobrick (or biofusion) part requires cutting with XbaI and PstI. Another option for this vector is to insert two Biobrick/biofusion parts by cutting the first part with NotI and SpeI and the second part with XbaI and PstI, while cutting the vector with NotI and PstI. Note that this part does not conform to Standard Assembly requirements and will maintain idempotency. Once a construct of interest has been inserted into the vector, it should be linearized with a digest using BstEII. This vector will supplement a LEU2 auxotrophic yeast strain. The part is then transformed into yeast according to the protocol outlined in this paper:

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


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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