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ID   PLD25      preliminary; circular DNA; SYN; 11000 BP.
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AC   ATCC20687; ATCC39464;
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DT   01-JUL-1993 (Rel. 7, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
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DE   Yarrowia/E.coli plasmid vector pLD25 - 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   pLD25 from pBR322 & LEU2 gene
RA   ;
RT   ;
RL   Unpublished (1989).
RL   U.S. Patent No. 4,880,741 dated Nov. 14, 1989.
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CC   Constructed by a inserting a 6.6 kb Sau3AI fragment (partial)
CC   containing LEU2 into the BamHI site of pBR322.  The gene is contained
CC   in an EcoRI fragment (partial) of approximately 2.4 kb.
CC   Replicates autonomously in E.coli and integrates but does not
CC   replicate autonomously in Y. lipolytica.
CC   The Y. lipolytica portion of the plasmid contains the following
CC   restriction sites: EcoRI (5); SphI (1); SalI (2); AvaI (8); KpnI (1);
CC   EcoRI (4); XhoI (2), BglII (2).  It lacks HindIII, ClaI, BamHI, and
CC   NruI sites.
CC   Medium is 1245 YEPD.
CC   Medium is 1227 LB plus ampicillin.
CC   NM (pLD25)
CC   CM (no)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   SP (ATCC)
CC   HO (Yarrowia lipolytica PC-30827 [ATCC 20688])
CC   HO (Yarrowia lipolytica)(E.coli)(E.coli JC-355)
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 BamHI 4361bp 376..376
FT                   2. yeast Sau3AI/MboI-Sau3AI/MboI 6600bp, LEU2 gene
FT                   -> pLD25 11000bp"
FT   misc_binding    0..0
FT                   /note="SIT unique HindIII"
FT   rep_origin      0..0
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322)"
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"
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SQ   Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;
     n
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