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ID   PCN50      preliminary; circular DNA; SYN; 8500 BP.
AC   ATCC77099;
DT   01-JUL-1993 (Rel. 7, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
DE   Pseudomonas/E.coli plasmid vector pCN50 - incomplete.
KW   cloning vector.
OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
RN   [1]
RC   pPS10 from P.savastanoi
RC   pCN38 from pPS10 & pKT231
RC   pCN39 from pCN38 & pUC7
RC   pCN40 from pCN39 & pKT231
RC   pCN41 from pCN40
RC   pCN50 from pCN38 & pBR322
RC   pCN51 from pCN50
RC   pCN60 from pUC18 & pCN40
RC   pCN61 from pCN60
RC   pNSV1 from pCN38 & pCP3
RA   Nieto C., Fernandez-Tresguerres E., Sanchez N., Vicente M., Diaz R.;
RT   "Cloning vectors, derived from a naturally occurring plasmid of
RT   Pseudomonas savastanoi, specifically tailored for genetic
RT   manipulations in Pseudomonas";
RL   Gene 87:145-149(1990).
CC   Multiple selectible markers permits insertional activation to screen
CC   for recombinants (PstI site in ampR).
CC   Constructed by ligating pCN38 (ATCC 77095) and pBR322 which had been
CC   linearized with PvuII.
CC   Contains the basic replicon of pPS10 which has been sequenced (EMBL
CC   accession number X58896).  The repA portion is flanked by EcoRI sites
CC   (nt 536 & 1500), with the site at nt 536 between the repA promoter and
CC   coding sequences. (personal communication)
CC   Retains the mobilization site of pBR322-derived vectors and can be
CC   mobilized if suitable functions are provided in trans.
CC   Restriction digests of the clone give the following sizes (kb):
CC   BamHI--6.2, 3.1; HindIII/PstI--4.5, 3.4, 0.8; PvuII--4.4 (doublet);
CC   PvuII/KpnI--4.4, 2.5, 1.6. (ATCC staff)
CC   The order of the major features in this plasmid is:PvuII - pBR322
CC   (pMB1 ori - bla - tetR) - PvuII - BamHI - kanR - HindIII - repA.
CC   Medium is 1227 LB plus ampicillin.
CC   NM (pCN50)
CC   CM (no)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   HO (E.coli HB101)(Pseudomonas aeruginosa)(Pseudomonas putida)(E.coli)
CC   HO (P.aeruginosa PAO1162)(P.aeruginosa PAO1024)
CC   HO (P.putida KT2440 [ATCC 47054])
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. pCN38 PvuII 4100bp, RSF1010 1824 1951
FT                   2. pBR322 PvuII 4361bp 2067..2067
FT                   -> pCN50 8500bp"
FT   misc_binding    0..0
FT                   /note="SIT unique PstI-KpnI-BamHI"
FT   rep_origin      0..0
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT   rep_origin      0..0
FT                   /note="ORI Pseudomonas pPS10; incompatibility groups
FT                   P1, P4/Q and WrepA"
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 E. coli tetracycline resistance gene (tet)"
FT   CDS             0..0
FT                   /note="ANT Pseudomonas kanamycin resistance gene
FT                   (str)"
SQ   Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;