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ID   PRS169     preliminary; circular DNA; SYN; 5800 BP.
AC   ATCC77151;
DT   01-JUL-1993 (Rel. 7, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
DE   Saccharomyces/E.coli phagemid vector pRS169 - incomplete.
KW   cloning vector.
OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
RN   [1]
RC   pRSS56 from pBluescript KS+ & BlueScribe
RC   pRS303 from pRSS56 & HIS3 gene
RC   pRS304 from pRSS56 & TRP1 gene
RC   pRS305 from pRSS56 & LEU2 gene
RC   pRS306 from pRSS56 & URA3 gene
RC   pUC19-CEN6:32 from pUC19 & CEN6
RC   pRSS83 from pUC19-CEN6:32 & pAB9, ARS
RC   pRSS84 from pRSS83
RC   pRS313 from pRS303 & pRSS84
RC   pRS314 from pRS304 & pRSS84
RC   pRS315 from pRS305 & pRSS84
RC   pRS316 from pRS306 & pRSS84
RC   pRSS93 from pRSS84 & pYCF5
RA   Sikorski R.S., Hieter P.;
RT   "A system of shuttle vectors and yeast host strains designed for
RT   efficient manipulation of DNA in Saccharomyces cerevisiae";
RL   Genetics 122:19-27(1989).
CC   pRSS56, constructed by ligating a PvuI fragment (bp 498-2412) of
CC   pBluescript KS+ to a PvuI fragment (bp 2850-730) of pBS(+), contains
CC   the KS MCS from pBluescript KS+ and the unique NdeI and AatII sites
CC   between bla and f1 origin of pBS(+).
CC   pRS169 (ATCC 77151) and pRS173 (ATCC 77152) differ in the orientation
CC   of the GAL1/10 cassette. (personal communication)
CC   The order of the major features in this plasmid is: URA3 - f1 ori
CC   (NaeI) - T7 promoter - lacZ'/MCS - T3 promoter - GAL1 promoter -
CC   GAL10 promoter - pMB1 ori - bla. (personal communication)
CC   Transcription from the GAL1 promoter (no ATG included in the sequence)
CC   procedes through the polylinker. (personal communication)
CC   YX-type expression shuttle vector permitting visual detection of
CC   recombinants and production of ssDNA in E.coli. Contains promoters for
CC   in vitro RNA synthesis and encodes the lacZ alpha (lacZ') peptide.
CC   (personal communication)
CC   Expression is inducible by galactose. (personal communication)
CC   A fragment containing the URA3 gene was inserted into the NdeI site
CC   and a DraIII/EcoRI fragment containing the GAL1 and GAL10 promoters
CC   was inserted into the Asp718 site of the MCS of pRSS56.   All ends
CC   were blunted. (personal communication)
CC   Restriction digests of the clone give the following sizes (kb):
CC   PvuI--3.3 (doublet); SalI--6.6; EcoRI--6.6; SmaI--6.6; BamHI--6.6.
CC   (ATCC staff)
CC   Medium is 1227 LB plus ampicillin.
CC   NM (pRS169)
CC   CM (no)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (phagemid)
CC   HO (E.coli HB101)(Saccharomyces cerevisiae)(E.coli)
CC   CP ()
CC   FN (cloning)
CC   SE ()
CC   PA ()
CC   BR ()
CC   OF ()
CC   OR ()
FH   Key             Location/Qualifiers
FT   misc_feature    0..0
FT                   /note="1. pBluescript KS+ PvuI-PvuI 1914bp 498..2412,
FT                   \ MCS
FT                   2. pBS+ PvuI-PvuI 1084bp 2850..3204..730, amp/f1 ori
FT                   -> pRSS56 2998bp [unique NdeI, unique AatII]
FT                   1. pRSS56 NdeI 2998bp, pBS+ 185..185
FT                   blunt end:blunt end
FT                   2. yeast NdeI-NdeI 2800bp, URA3 gene
FT                   blunt end:blunt end
FT                   -> plasmid 5800bp
FT                   1. plasmid Asp718I/KpnI 5800bp, MCS
FT                   \ pBS+ 898/pBluescript KS+ 760
FT                   blunt end:blunt end
FT                   2. yeast DraIII-EcoRI 500bp, GAL1/GAL10 promoters
FT                   blunt end:blunt end
FT                   -> pRS169 5800bp"
FT   misc_binding    0..0
FT                   /note="MCS KpnI-ApaI-XhoI-SalI-ClaI-HindIII-EcoRV-
FT                   EcoRI-PstI-SmaI-
FT                   BamHI-SpeI-XbaI-EagI-NotI-BstXI-SacII-SacI"
FT   misc_binding    0..0
FT                   /note="SIT unique XhoI-SalI-EcoRI-SmaI-BamHI-NotI-
FT                   SacI"
FT   rep_origin      0..0
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT   rep_origin      0..0
FT                   /note="ORI bacteriophage f1"
FT   promoter        0..0
FT                   /note="PRO E. coli lac gene"
FT   promoter        0..0
FT                   /note="PRO bacteriophage T3"
FT   promoter        0..0
FT                   /note="PRO bacteriophage T7"
FT   promoter        0..0
FT                   /note="PRO yeast GAL1/GAl10 gene"
FT   CDS             0..0
FT                   /note="GEN E. coli beta-galactosidase gene (lacZ');
FT                   reporter gene"
FT   CDS             0..0
FT                   /note="ANT E. coli beta-lactamase gene (bla)
FT                   ampicillin resistance gene (apr/amp)"
FT   CDS             0..0
FT                   /note="ANT yeast URA3 gene"
SQ   Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;