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ID PDO193 preliminary; circular DNA; SYN; 6760 BP.
DT 01-FEB-1995 (Rel. 11, Created)
DT 01-JUL-1995 (Rel. 12, Last updated, Version 1)
DE E. coli cosmid pDO193 - incomplete.
KW cloning vector.
OS Cloning vector
OC Artificial sequences; Cloning vehicles.
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";
RL Gene 141:71-73(1994).
RC pDO193 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).
CC Restriction digests of the clone give the following sizes (kb):
CC EcoRI--7.0; HindIII--7.0; NdeI--7.0; BglI--4.8, 2.2. (ATCC staff)
CC Plasmid contains the following restriction sites (approximate kb from
CC nt 1): BamHI--3.10; BglI--3.29, 5.37; EcoRI--3.06; HindIII--3.14;
CC NdeI--4.19; PstI--0.65, 5.50; PvuI--3.26, 5.62; SacI--3.11;
CC SmaI--3.11; XbaI--2.11. (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'. 
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. pDO193 contains 2 groups of sites recognized by
CC 8 bp cutters separated by a cluster of common restriction sites in the
CC Growth: 1065 plus ampicillin (50 ug/ml) plus kanamycin (20 ug/ml) 30C
CC Deposited by: Ahmed A.L.
CC NM (pDO193)
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 ()
FH Key Location/Qualifiers
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 -> pDO193 6760bp"
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-BssHII-NotI-SalI-KpnI-Acc65I-BamHI-
SQ Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;