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ID   PRS166     preliminary; circular DNA; SYN; 6300 BP.
AC   ATCC77149;
DT   01-JUL-1993 (Rel. 7, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
DE   Saccharomyces/E.coli phagemid vector pRS166 - incomplete.
KW   cloning vector.
OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
RN   [1]
RC   pRSS56 from pBluescript KS+ & pBS+
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   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   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   Fragments containing the URA3 and TRP1 genes were inserted into the
CC   NdeI site and a DraIII/EcoRI fragment containing the GAL1 and GAL10
CC   promoters was inserted into the Asp718 site of the MCS of pRSS56.  All
CC   ends were blunted. (personal communication)
CC   pRS166 (ATCC 77149) and pRS167 (ATCC 77150) differ in the orientation
CC   of the GAL1/10 cassette. (personal communication)
CC   Transcription from the GAL10 promoter (ATG is included in the
CC   sequence) permits the synthesis of fusion protein starting with 26 aa
CC   of GAL10 into the polylinker in the same frame as lacZ'. (personal
CC   communication)
CC   Expression is inducible by galactose. (personal communication)
CC   The order of the major features in this plasmid is:TRP1 - URA 3 -
CC   f1 ori (NaeI) - T7 promoter - lacZ'/MCS - T3 promoter - GAL10 promoter
CC   - GAL1 promoter - pMB1 ori - bla. (personal communication)
CC   Restriction digests of the clone give the following sizes (kb):
CC   PvuI--3.6, 3.0; SalI--6.6; EcoRI--6.6; SmaI--6.6; BamHI--6.6. (ATCC
CC   staff)
CC   Medium is 1227 LB plus ampicillin.
CC   NM (pRS166)
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   HO (Saccharomyces cerevisiae YPH499)(Saccharomyces cerevisiae YPH500)
CC   HO (Saccharomyces cerevisiae YPH501)
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/TRP1 genes
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                   -> pRS166 6300bp"
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 TRP1 gene"
FT   CDS             0..0
FT                   /note="ANT yeast URA3 gene"
SQ   Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;