One of a series of pBluescript-based integrating vectors (ATCC 77138-77141) differing in the yeast selectable marker gene. This is a YI-type (integrating shuttle vector ) permitting visual detection of recombinants and production of ssDNA in E. coli. It contains promoters for invitro RNA synthesis, priming sites useful for sequencing, and encodes the lacZ alpha (lacZ?) peptide. The pRSS56, base vector was constructed by ligating a PvuI fragment (bp 498-2412) of pBluescript KS+ to a PvuI fragment (bp 2850-730) of pBS(+). It contains the KS MCS from pBluescript KS+ and the unique NdeI and AatII sites between bla and the f1 origin of pBS(+). A fragment (1.112 kb) containing the URA3 gene was inserted into the NdeI 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. - Genetics 122: 19-27, 1989.
The distribution host strain for this plasmid is E. coli HB101.
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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
Characteristics
Comments
Restriction digests of the clone give the following sizes (kb): PstI--2.7, 1.65; PvuI--3.9, 0.40; EcoRI--4.4; HindIII--4.4.
One of a series of pBluescript-based integrating vectors (ATCC 77138-77141) differing in the yeast selectable marker gene.
YI-type integrating shuttle 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.
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 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.
The following restriction sites in the multiple cloning site (MCS) are no longer unique: ApaI EcoRV PstI.
This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use.
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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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