pRS318 (ATCC® 77158)

Applications: YC-type (centromeric) shuttle vectorplasmid shufflingvector containing primer sites useful for sequencingvector permitting RNA synthesis in vitrovector permitting production of single-stranded DNAvector permitting visual detection of recombinants  /  Depositors: P Hieter

Designations pRS318
Depositors P Hieter
Biosafety Level 1
Vector Information
Size (kb): 7.4000000953674320
Vector: pRS318 (phagemid)
Promoters: Promoter T3
Construction: pRSS56 [pBluescript KS+, pBS(+)]
Marker(s):LEU2,ampR,CYH2
Construct size (kb): 7.400000095367432
Features: insert detection: lacZ'
marker(s): ampR, LEU2, CYH2
promoter: lac, T3, T7
replicon: pMB1, f1, ARSH4
centromere: CEN6
Applications
YC-type (centromeric) shuttle vector
plasmid shuffling
vector containing primer sites useful for sequencing
vector permitting RNA synthesis in vitro
vector permitting production of single-stranded DNA
vector permitting visual detection of recombinants
Comments
Restriction digests of the clone give the following sizes (kb): HindIII--7.4; XbaI--7.4; PstI--7.4; BamHI--7.4.
One of a series of pBluescript-based centromere vectors (ATCC 77142-77145, 77157-77158) differing in the yeast selectable marker gene.
YC-type centromere vector permitting visual detection of recombinants and production of ssDNA in E. coli. Contains promoters for in vitro RNA synthesis, priming sites useful for sequencing, and encodes the lacZ alpha (lacZ') peptide.
Useful in plasmid shuffle experiments.
pRSS56, constructed by ligating a PvuI fragment (bp 498-2412) of pBluescript KS+ to a PvuI fragment (bp 2850-730) of pBS(+), contains the KS MCS from pBluescript KS+ and the unique NdeI and AatII sites between bla and f1 origin of pBS(+).
The order of the major features in this plasmid is: LEU2 - CYH2 - f1 ori - T7 promoter - lacZ'/MCS - T3 promoter - pMB1 ori - bla - CEN6 - ARSH4.
Media Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin
Growth Conditions
Temperature: 37.0°C
References

Sikorski RS, Boeke JD. In vitro mutagenesis and plasmid shuffling: from cloned gene to mutant gene. Methods Enzymol. 194: 302-318, 1991. PubMed: 2005795

Sikorski RS, Hieter P. A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. Genetics 122: 19-27, 1989. PubMed: 2659436

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