pRS166


Vector IG Sequence Link :
General : phagemid ds-DNA 6300 BP
Functions : (cloning)
Selection : ()
Copy Number :
Hosts : (E.coli HB101)(Saccharomyces cerevisiae)(E.coli)(Saccharomyces cerevisiae YPH499)(Saccharomyces cerevisiae YPH500)(Saccharomyces cerevisiae YPH501)
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) 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(+). Fragments containing the URA3 and TRP1 genes were 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) pRS166 (ATCC 77149) and pRS167 (ATCC 77150) differ in the orientation of the GAL1/10 cassette. (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) The order of the major features in this plasmid is:TRP1 - URA 3 - 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): PvuI--3.6, 3.0; SalI--6.6; EcoRI--6.6; SmaI--6.6; BamHI--6.6. (ATCC staff) Medium is 1227 LB plus ampicillin.
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