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Permits
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These permits may be required for shipping this product:
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Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
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Vector Information
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Size (kb): 5.6999998092651370 Vector: pUN65 (plasmid) Promoters: Promoter lac Construction: pUC18, pBluescript KS+ Marker(s):URA3,ampR,SUP11 Construct size (kb): 5.699999809265137 Features: insert detection: lacZ'
marker(s): SUP11
marker(s): URA3
marker(s): ampR
promoter: lac
promoter for in vitro transcription: T3
promoter for in vitro transcription: T7
replicon: ARS1
replicon: pMB1
MCS: KpnI...SacI
centromere: CEN4
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Applications
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YC-type (centromeric) shuttle vector mutation detection shuttle vector vector permitting RNA synthesis in vitro vector permitting visual detection of recombinants beta-galactosidase beta-D-galactosidase
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Comments
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Restriction digests of the clone give the following sizes (kb): XhoI--5.8; EcoRI--5.8; NotI--5.8. SUP11 in the vector suppresses the red colony phenotype of ade2-101 ochre allele host strains, grown on limiting adenine media. In the absence of plasmid selection, plasmids are lost at a frequency of 1 to 3%, giving rise to red/white sectoring. One strategy for mutation scanning is to clone the normal allele of a gene into a SUP11 vector and transform into an ade2-101 host with a null mutation in the same gene, thus selecting for plasmid maintenance and producing white colonies. Vectors pUN20 ( ATCC 77263), pUN25 ( ATCC 77264), pUN60 ( ATCC 77266) and pUN65 ( ATCC 77267) contain SUP11. A second, potentially mutated copy of the gene can then be cloned into a second centromeric shuttle vector (pUN30, ATCC 77265; pUN70, ATCC 77268; pUN90, ATCC 77269; pUN100, ATCC 77270) for transformation into the SUP11 plasmid containing host. A normal gene in plasmid #2 can relieve the selective pressure on the SUP11 plasmid, restoring the red/white sectoring; a null allele in plasmid #2 will result in SUP11 plasmid maintenance and white colonies. YC-type shuttle vector useful for the sectoring-shuffle mutagenesis assay. Contains promoters for in vitro RNA synthesis and permits visual detection of recombinants by lacZalpha complementation.
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References
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Elledge SJ, Davis RW. A family of versatile centromeric vectors designed for use in the sectoring-shuffle mutagenesis assay in Saccharomyces cerevisiae. Gene 70: 303-312, 1988. PubMed: 3063604
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