Size (kb): 4.895
Vector: pRS316 (phagemid)
Promoters: Promoter for in vitro transcription T7
Construction: pRSS56 [pBluescript KS+, pBS(+)]
Construct size (kb): 4.895
Features: insert detection: lacZ'
promoter for in vitro transcription: T3
promoter for in vitro transcription: T7
YC-type (centromeric) shuttle vector
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
Restriction digests of the clone give the following sizes (kb): EcoRI--5.0; BamHI--5.0, PvuII--4.4, 0.5.
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(+).
A fragment (1.112 kb) containing the URA3 gene was inserted into the NdeI site and a cassette containing CEN6 and the ARS associated with histone 4 (ARSH4) was inserted into the AatII site of pRSS56. All ends were blunted.
The order of the major features in this plasmid is: URA3 - f1 ori (NaeI) - T7 promoter - lacZ'/MCS - T3 promoter - pMB1 ori - bla - CEN6 - ARSH4.
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
Frozen glycerol stock of E. coli containing the phagemid.