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ID   PDO192     preliminary; circular DNA; SYN; 6740 BP.
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AC   ATCC87043;
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DT   01-FEB-1995 (Rel. 11, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
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DE   E. coli cosmid pDO192 - 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   pDO2 from pUC19 & lambda::Tn5 & pNEB193
RC   pDO6 from pDO2
RC   pDO17 from pDO2 & pBR322
RC   pDO19 from pDO17
RC   pDO184 from pK184 & pBR322
RA   Ahmed A.;
RT   "Double origin vectors for isolating bidirectional deletions for DNA
RT   sequence analysis";
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RN   [2]
RC   pDO192 from pBR322 & lambda & kan gene
RA   Ahmed A.L.;
RT   "Isolation of nested, bidirectional deletions by the use of
RT   exonuclease III on double-origin vectors";
RL   Gene 151:97-101(1994).
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CC   Restriction digests of the clone give the following sizes (kb):
CC   EcoRI--6.8; HindIII--6.8; XbaI--5.8, 0.9. (ATCC staff)
CC   Plasmid contains the following restriction sites (approximate kb from
CC   nt 1):  BamHI--3.08; BglI--3.26, 5.34; EcoRI--3.06; KpnI--3.07;
CC   HindIII--3.11; PstI--0.65, 5.47; PvuI--3.23, 5.59; SacI--3.07;
CC   SmaI--3.08; XbaI--2.11, 3.09. (GenBank/EMBL/DDBJ)
CC   Cosmid vector containing two compatible origins of replication,
CC   for generating bidirectional deletions of large inserts to facilitate
CC   sequencing. A series of overlapping deletion segments can be generated
CC   by cutting at the Cos site with lambda terminase, then filling in ends
CC   with a alpha phosphorothioate nucleotides, cutting with a restriction
CC   enzyme that cuts on one side of the insert, and then digesting with
CC   Exo III for various intervals of time. Resulting fragments are blunted
CC   and circularized by ligation, and transformants are selected for
CC   either ampicillin or kanamycin resistance. AmpR transformants can be
CC   sequenced in the clockwise direction using the cosL primer
CC   5'-TCATAAATAGCGAAAACC-3'.  KanR transformants can be sequenced in the
CC   counterclockwise direction using the cosR primer
CC   5'-ACTTTACGGGTCCTTTCCG-3'. [2]
CC   The order of the major features in the
CC   plasmid is:  3' lacZ - HindIII/MCS/EcoRI - 5' lacZ - ori 15 - kanR -
CC   cosR/L - ampR - oriP. pDO192 contains 12 unique restriction sites in
CC   the polylinker but no restiction sites for any 8 bp cutters. [2]
CC   Growth: 1065 plus ampicillin (50 ug/ml) plus kanamycin (20 ug/ml) 30C
CC   Deposited by: Ahmed A.L.
CC   NM (pDO192)
CC   CM (no)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (cosmid)
CC   SP (ATCC)
CC   HO (E.coli)(E.coli AA102)
CC   CP ()
CC   FN (cloning)(sequencing)
CC   SE (color)
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. lambda, ori
FT                   3. E. coli, kan gene
FT                   4. p15A, ori
FT                   -> pDO192 6740bp"
FT   rep_origin      0..0
FT                   /note="ORI E. coli oriP"
FT   rep_origin      0..0
FT                   /note="ORI E. coli p15A"
FT   CDS             0..0
FT                   /note="ANT E. coli amp gene"
FT   CDS             0..0
FT                   /note="ANT E. coli kan gene"
FT   misc_binding    0..0
FT                   /note="MCS EcoRI-SacI-Ecl136II-KpnI-AvaI-SmaI-
FT                   XmaI-BamHI-SphI-HindIII"
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SQ   Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;
     n
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