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ID   YPGX265GAL preliminary; circular DNA; SYN; 12300 BP.
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AC   ATCC67233;
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DT   03-FEB-1994 (Rel. 8, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
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DE   Saccharomyces/E.coli plasmid vector YpGX265GAL4 - incomplete.
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KW   cloning vector.
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OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
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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RL   Unpublished (1991).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.
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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   SP (ATCC)
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 ()
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FH   Key             Location/Qualifiers
FH
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"
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SQ   Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;
     n
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