pRS173


Vector IG Sequence Link :
General : phagemid ds-DNA 5800 BP
Functions : (cloning)
Selection : ()
Copy Number :
Hosts : (E.coli HB101)(Saccharomyces cerevisiae)(E.coli)
Suppliers : (ATCC)
Misc.Comments : YX-type expression shuttle vector permitting visual detection of recombinants and production of ssDNA in E.coli. Contains promoters for in vitro RNA synthesis and encodes the lacZ alpha (lacZ') peptide. (personal communication) Transcription from the GAL10 promoter (ATG is included in the sequence) permits the synthesis of fusion protein starting with 26 aa of GAL10 into the polylinker in the same frame as lacZ'. (personal communication) Expression is inducible by galactose. (personal communication) A fragment containing the URA3 gene was inserted into the NdeI site and a DraIII/EcoRI fragment containing the GAL1 and GAL10 promoters was inserted into the Asp718 site of the MCS of pRSS56. All ends were blunted. (personal communication) pRS169 (ATCC 77151) and pRS173 (ATCC 77152) differ in the orientation of the GAL1/10 cassette. (personal communication) 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: URA3 - f1 ori (NaeI) - T7 promoter - lacZ'/MCS - T3 promoter - GAL10 promoter - GAL1 promoter - pMB1 ori - bla. (personal communication) Restriction digests of the clone give the following sizes (kb): EcoRI--6.6; BamHI--6.6; SalI--6.6. (ATCC staff) Medium is 1227 LB plus ampicillin.
Parents : ()
Siblings : ()
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