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ID   PCYM03     preliminary; circular DNA; SYN; 9591 BP.
AC   ATCC37827;
DT   01-JUL-1993 (Rel. 7, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
DE   Saccharomyces/vertebrate/E.coli plasmid vector pCYM03 - incomplete.
KW   cloning vector.
OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
RN   [1]
RC   pCYM0 from pSV2-cat & pUT332
RC   pCYM1 from pCYM0
RC   pCYM2 from pCYM0 & YRp7
RC   pCYM3 from pCYM2 & pTG2856
RC   pCYM00 from pCYM0 & pMC1403
RC   pCYM01 from pCYM1 & pMC1403
RC   pCYM02 from pCYM2 & pMC1403
RC   pCYM03 from pCYM3 & pMC1403
RC   pCYM1-11 from pCYM1 & M13mp11
RC   pCYM3-11 from pCYM3 & M13mp11
RA   Camonis J.H., Cassan M., Rousset J.P.;
RT   "Of mice and yeast: versatile vectors which permit gene expression
RT   in both budding yeast and higher eukaryotic cells";
RL   Gene 86:263-268(1990).
CC   The EcoRI, SmaI, and BamHI sites are 3' to the unique available
CC   in-phase initiation codon under control of the SV40 early promoter.
CC   Expresses beta-galactosidase activity in E.coli, S. cerevisiae, and
CC   mouse cells, and thus can be used as a control to calibrate
CC   transformation efficency.  Expression can be regulated by using medium
CC   composition to modulate copy number.
CC   Constructed by inserting a 3.048 kb EcoRI/DraI fragment from pMC1403
CC   into pCYM3 cut with BamHI (nt 427), linkered to add an EcoRI site, and
CC   cut with BalI (nt 702).
CC   URA3d is a modified URA3 gene which is not efficiently expressed and
CC   requires a large number of plasmids to complement a ura3 mutation.
CC   Restriction digests of the clone give the following sizes (kb):
CC   BamHI/ClaI--4.9, 3.5, 0.76, 0.48; EcoRI--9.4; SmaI--9.4. (ATCC staff)
CC   Shuttle expression vector which includes a reporter group. (ATCC
CC   staff)
CC   Also permits blue/white detection in E.coli.
CC   Medium is 1227 LB plus ampicillin.
CC   NM (pCYM03)
CC   CM (no)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   HO (E.coli CM5alpha)(Saccharomyces cerevisiae)(vertebrate cells)
CC   HO (E.coli)(Saccharomyces cerevisiae W303-1B/D)(mouse C11D cell line)
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. pCYM3 BamHI 6543bp, pSV2-cat 3370..3370
FT                   mung bean nuclease
FT                   EcoRI linker 12bp nnngaattcnnn
FT                   BalI 6465bp,
FT                   \ yeast 2 micron 267..267/pSV2-cat 4489..4489
FT                   2. pMC1403 EcoRI-DraI 3126bp 3640..6766, lacZ gene
FT                   \ [3048bp]
FT                   -> pCYM03 9591bp"
FT   misc_binding    0..0
FT                   /note="MCS EcoRI-SmaI-BamHI"
FT   misc_binding    0..0
FT                   /note="SIT unique EcoRI-SmaI-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 SV40 early genes"
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 URA3d gene"
FT   CDS             0..0
FT                   /note="GEN E. coli beta-galactosidase gene (lacZ');
FT                   reporter gene"
SQ   Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;