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ID   PCN38      preliminary; circular DNA; SYN; 4100 BP.
AC   ATCC77095;
DT   01-JUL-1993 (Rel. 7, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
DE   Pseudomonas plasmid vector pCN38 - 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).
RN   [2]
RC   from pLUC2
RC   pCP3 from pBR328 & pIAA1, iaaMH operon
RA   Yamada T., Palm C.J., Brooks B., Kosuge T.;
RT   "Nucleotide sequence s of the Pseudomonas savastanoi indoleacetic
RT   acid genes show homology with Agrobacterium tumefaciens";
RL   Proc. Natl. Acad. Sci. U.S.A. 82:6522-6526(1985).
RN   [3]
RC   pIAA1, pIAA2, pIAA3 from Pseudomonas savastanoi, iaaMH operon
RC   pLUC1 from pIAA1 & RSF1010
RA   Kosuge T., Palm C.J., Hutchenson S.V., Louise N., Glass E., Yamada T.;
RT   "pIAA a virulent plasmid in Pseudomonas savastanoi";
RL   Plasmids in Bacteria 0:807-813(1985).
RL   Helinski D.R., Cohen S.N., Clewell D.B., Jackson D.A., Hollaender A.;
RL   Plenum, New York.
RN   [4]
RC   pLUC2 from pBR328 & pIAA1, iaaMH operon
RA   Comai L., Kosuge T.;
RT   "Cloning characterization of iaaM, a virulence determinant of
RT   Pseudomonas savastanoi";
RL   J. Bacteriol. 149:40-46(1982).
CC   Derived from the ligation product of a 7.6 kb PstI fragment of pPS10
CC   and a 4.2 kb PstI fragment of pKT231 which was then shortened by
CC   deletion. A 1.8 kb region of pPS10 was retained (EMBL accession
CC   number X58896) which encodes replication functions.
CC   Compatible with broad-host-range plasmids of the incompatibility
CC   groups P1, P4/Q and W, and maintained at a copy number of 15-20 in
CC   P. savastanoi.
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   General purpose vector. (ATCC staff)
CC   Restriction digests of the clone give the following sizes (kb):
CC   EcoRI--3.2, 0.95; HindIII--4.2; BamHI/KpnI--2.2, 2.0. (ATCC staff)
CC   The order of the major features in this plasmid is:PvuII - BamHI -
CC   kanR - HindIII - KpnI - repA.
CC   Medium is 1236 LB plus kanamycin.
CC   NM (pCN38)
CC   CM (no)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   HO (Pseudomonas aeruginosa PAO1024 [ATCC 47053])(Pseudomonas)
CC   HO (P.aeruginosa PAO1162)(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. P.savastanoi
FT                   -> pPS10 10000bp
FT                   1. E.coli, IncFII plasmids
FT                   -> R6-5
FT                   1. R6-5, kan gene
FT                   -> pKT105
FT                   1. RSF1010 8684bp
FT                   2. pKT105, kan gene
FT                   -> pKT231
FT                   1. pPS10 PstI-PstI 7600bp, ori
FT                   2. pKT231 PstI-PstI 4200bp, kan gene
FT                   -> plasmid 11800bp
FT                   1. plasmid remove,
FT                   -> pCN38 4100bp [1800bp ori, kan]"
FT   misc_binding    0..0
FT                   /note="SIT unique PvuII-BamHI-KpnI"
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 Pseudomonas kanamycin resistance gene
FT                   (kan)"
SQ   Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;