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ID   PPGKNEO    preliminary; circular DNA; SYN; 1 BP.
AC   IG8010;
DT   01-DEC-1994 (Rel. 10, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
DE   Saccharomyces/E.coli plasmid vector pPGKNeo - incomplete.
KW   cloning vector.
OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
RN   [1]
RC   plasmid from pBS+ & lacI gene & SV40 T-antigen oligo
RC   pPGKLacIN from pPGKNeo & plasmid
RC   pPGKHyg from pPGKNeo & hyg gene
RC   pPGKCAT from pPGKNeo & cat gene
RC   pPN502, pPN504, pPN505, pPN506, pPN507 from pPGKNeo & lacO
RC   pPN503 from pPN502
RC   pPN508 from pPGKNeo & lacO
RA   Hannan G.N., Lehnert S.A., MacAvoy E.S., Jennings P.A., Molloy P.L.;
RT   "An engineered PGK promoter and lac operator-repressor system for
RT   the regulation of gene expression in mammalian cells";
RL   Gene 130:233-239(1993).
RN   [2]
RC   pPGKNeo from mouse Pgk-I gene promoter & pMC1neo, neo gene
RA   McBurney M.W., Sutherland L.C., Adra C.N., Leclair B.,
RA   Rudnicki M.A., Jardine K.;
RT   "The mouse Pgk-I gene promoter contains an upstream activator
RT   sequence";
RL   Nucleic Acids Res. 19:5755-5761(1991).
RN   [3]
RC   [pKM109/90 from R46 or pKM101]
RC   pdeltaMo+PyF101 from PyF101 enhancer
RC   31, 39 from pKM109/90 & pMC1Neo & pdeltaMo+PyF101, PyF101 enhancer
RC   pIT219 from pIT213 & PGK promoter
RC   59A-, 61A+, 61A- from pLG89 & pIT219
RC   pPGK-hyg from PGK promoter/polyA & hyg gene
RA   Riele H.T., Maandag E.R., Clarke A., Hooper M., Berns A.;
RT   "Consecutive inactivation of both alleles of the pim-1 proto-oncogene
RT   by homologous recombination in embryonic stem cells";
RL   Nature 348:649-651(1990).
CC   The promoter and terminator sequences were derived from the
CC   phosphoglycerate kinase gene.
CC   Saccharomyces cerevisiae containing a single copy of the neomycin
CC   phosphotransferase gene are sensitive to G418; resistance to 400 ug/ml
CC   is expressed if there are several copies per cell.
CC   The G418-resistance segment includes a 0.7 kb region of the PGK
CC   promoter and a 1.5 kb BglII/BamHI fragment from Tn5.  The BamHI site
CC   is between the G418R and the PGK terminator.
CC   Restriction digests of the clone give the following sizes (kb):
CC   BamHI--15.0; BglII--15.0; XhoI--9.4, 6.0; EcoRI--13.0, 2.1;
CC   KpnI--13.0, 2.2. (ATCC staff)
CC   Medium is 1227 LB plus ampicillin.
CC   NM (pPGKNeo)
CC   CM (no)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   HO (E.coli JA221)(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. yeast 2 micron
FT                   2. pYe(Leu2)10 SalI-XhoI 2200bp, LEU2 gene
FT                   3. mouse 700bp, PGK-I gene promoter
FT                   4. pMC1neo BglII-BamHI 1541bp 1516..3057,
FT                   \ Tn5 neo gene
FT                   -> pPGKNeo"
FT   misc_binding    0..0
FT                   /note="SIT unique 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 E. coli PGK 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 LEU2 gene"
FT   CDS             0..0
FT                   /note="ANT E. coli Tn5 neo gene"
SQ   Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;