Difference between revisions of "Part:BBa K165062"
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+ | 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. | 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. |
Revision as of 00:45, 30 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
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
- 10INCOMPATIBLE 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 - 12INCOMPATIBLE 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 - 21INCOMPATIBLE 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 - 23INCOMPATIBLE 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 - 25INCOMPATIBLE 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 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 4584
Illegal SapI site found at 3501