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ID   YPGX265GAL preliminary; circular DNA; SYN; 12300 BP.
AC   ATCC67233;
DT   03-FEB-1994 (Rel. 8, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
DE   Saccharomyces/E.coli plasmid vector YpGX265GAL4 - incomplete.
KW   cloning vector.
OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
RN   [1]
RC   YpGX265GAL4 from GAL1/10 prom & MFalpha1 & GAL4 gene & LEU2d gene
RA   ;
RT   ;
RL   Unpublished (1991).
RL   U.S. Patent No. 5,013,652 dated May 7, 1991.
CC   High copy number YE-type shuttle expression vector. [1]
CC   The promoter consists of nt -610 to -157 of the GAL 1-10 upstream
CC   activation site,fused to the transcription initiation site
CC   (nt -158 to -1) of the alpha mating factor gene MFalpha1. [1]
CC   The 3.65 kb HindIII fragment encodes the positive regulator protein
CC   GAL4, necessary for transcription from the GAL1-10 promoter. [1]
CC   The PHO5 signal sequence (51 nt) was modified to use preferred
CC   Saccharomyces codons. [1]
CC   The plasmid is maintained at approximately 200 copies per cell in
CC   haploid S. cerevisiae cells. [1]
CC   The order of the major features in the plasmid is:  HindIII - GAL4 -
CC   HindIII - Gal1-10 UAS - MFalpha1 transcription initiation sequence -
CC   PHO5 signal sequence - EcoRV - BamHI - GAPDH terminator - ampR -
CC   LEU2-d - 2 micron ori. [1]
CC   Restriction digests of the clone give the following sizes (kb):
CC   EcoRV--5.1, 4.2, 3.0; BamHI--8.1, 4.2; HindIII--8.5, 3.8. (ATCC staff)
CC   Medium is 1227 LB plus ampicillin.
CC   NM (YpGX265GAL4)
CC   CM (no)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   HO (E.coli)(Saccharomyces cerevisiae)(S.cerevisiae 442)
CC   HO (S.cerevisiae SHY1)
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. pBR322 4361bp, ori/amp gene
FT                   2. yeast 453bp, GAL1-GAL10 promoter -610 to -157
FT                   3. yeast 157bp, MFalpha1 -158 to -1
FT                   4. yeast HindIII-HindIII 3650bp, GAL4 gene
FT                   5. pYe(Leu2)10 SalI-XhoI 2200bp, LEU2 gene
FT                   6. yeast 1500bp, PHO5 signal sequence
FT                   -> YpGX265GAL4 12300bp"
FT   misc_binding    0..0
FT                   /note="MCS unique EcoRV-BamHI"
FT   rep_origin      0..0
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT   rep_origin      0..0
FT                   /note="ORI yeast 2 micron"
FT   promoter        0..0
FT                   /note="PRO yeast GAL1-10/MFalpha1"
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 LEU2-d gene"
SQ   Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;