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ID   PAP12BD    preliminary; circular DNA; SYN; 15000 BP.
AC   ATCC53231;
DT   01-JUL-1993 (Rel. 7, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
DE   Saccharomyces/E.coli plasmid vector pAP12BD - incomplete.
KW   cloning vector.
OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
RN   [1]
RC   pAP12BD from yeast 2 micron & pBR322 & LEU2 gene & neo gene
RA   ;
RT   ;
RL   Unpublished (1990).
RL   U.S. Patent No. 4,935,350 dated June 19, 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 (pAP12BD)
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. pBR322 4361bp, ori/amp gene
FT                   2. yeast 2 micron, ori
FT                   3. yeast, PGK promoter
FT                   4. pYe(Leu2)10 SalI-XhoI 2200bp, LEU2 gene
FT                   5. yeast, G418 gene
FT                   -> pAP12BD 15000bp"
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 Saccharomyces G418 resistance gene (G418)"
SQ   Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;