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ID   PCE54      preliminary; circular DNA; SYN; 14500 BP.
AC   ATCC39019;
DT   01-JUL-1993 (Rel. 7, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
DE   Corynebacterium/Brevibacterium/E.coli plasmid vector pCE54 - incomp.
KW   cloning vector.
OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
RN   [1]
RC   pCG2 from pCG1
RC   pCG4 from pCG2
RC   pCG11 from pCG1 & pCG4
RC   pGA22 from pGA1
RC   pCE54 from pCG2 & pGA22
RA   ;
RT   ;
RL   Unpublished (1987).
RL   U.S. Patent No. 4,710,471 dated Dec. 1, 1987.
RN   [2]
RC   pGA1, pGA2 from Corynebacterium glutamicum LP-6
RC   pEM5.2 from pK19 & pSUP102, RP4 traJ gene/oriT/mob gene
RC   pHSKm1 from pUC18 & pACYC177, Tn903 kan gene
RC   pHS2-1, pHS2-2 from pGA1 & pHSKm1
RC   pDM10, pDM11, pDM12 from pGA1 & pEM5.2
RC   pCC1 from Corynebacterium
RC   pBL1 from Corynebacterium
RC   pHM1519 from Corynebacterium
RA   Sonnen H., Thierbach G., Kautz S., Kalinowski J., Schneider J.,
RA   Puhler A., Kutzner R.;
RT   "Characterization of pGA1, a new plasmid from Corynebacterium
RT   glutamicum LP-6";
RL   Gene 107:69-74(1991).
RN   [3]
RC   pECM1 from pHM1519
RA   Schafer A., Kalinowski J., Seep-Feldhaus A.H., Puhler A.;
RT   "High-frequency conjugal plasmid transfer from Gram-negative
RT   Escherichia coli to various Gram-positive coryneform bacteria";
RL   J. Bacteriol. 172:1663-1666(1990).
RN   [4]
RC   pCC1 from C.callunae
RA   Sandoval H., Aguilar A., Panagua C., Martin J.F.;
RT   "Isolation and physical characterization of plasmid pCC1 from
RT   Corynebacterium callunae and construction of hybrid derivatives";
RL   Appl. Microbiol. Biotechnol. 19:404-413(1984).
RN   [5]
RC   pHM1519 from C.glutamicum
RC   pAM330 from B.lactofermentum
RA   Miwa K., Matsui H., Terabe M., Nakamori S., Sano K., Momose H.;
RT   "Cryptic plasmids in glutamic acid producing bacteria";
RL   Agric. Biol. Chem. 48:2901-2903(1984).
RN   [6]
RC   pBL1 from B.lactofermentum
RA   Santamaria R., Gil J.A., Mesas J.M., Martin J.F.;
RT   "Characterization of an endogenous plasmid and development of cloning
RT   vectors and a transformation system in Brevibacterium lactofermentum";
RL   J. Gen. Microbiol. 130:2237-2246(1984).
RN   [7]
RC   pCC100 from pCC1 & pBR322
RA   Shaw P.C.;
RT   "Transformation of a Corynebacterium callunae-Escherichia coli hybrid
RT   plasmid to an Arthrobacter species";
RL   Agric. Biol. Chem. 53:1717-1719(1989).
RN   [8]
RC   pBL100 from pBL1 & pBR322
RC   pCG100 from pCG1 & pBR322
RA   Shaw P.C., Hartley B.S.;
RT   "A host vector system for an Arthrobacter species";
RL   J. Gen. Microbiol. 134:903-911(1988).
RN   [9]
RC   pWS101 from pBL1
RC   pSR1 from C.glutamicum
RA   Yoshihama M., Higashino K., Rao E.R., Akledo M., Shanabruch W.G.,
RA   Follettie M.T., Walker G., Sinskey A.J.;
RT   "Cloning vector system for Corynebacterium  glutamicum";
RL   J. Bacteriol. 162:591-597(1985).
RN   [10]
RC   pGX1901 from pBL1
RA   Smith M.D., Flickinger J.L., Lieberger P.W., Schmid B.;
RT   "Protoplast transformation in coryneform bacteria and introduction of
RT   an alpha-amylase gene form Bacillus amyloliquefaciens into
RT   Brevibacterium lactofermentum";
RL   Appl. Environ. Microbiol. 51:634-639(1986).
RN   [11]
RC   pX18 from pBL1
RA   Yeh P., Oreglia J., Prevots F., Sicard A.M.;
RT   "A shuttle vector system for Brevibacterium lactofermentum";
RL   Gene 47:301-306(1986).
RN   [12]
RC   pRN3.1 from C.glutamicum
RA   Nutter R.C., Panganiban L.C.;
RT   "Corynebacterium plasmid and vector";
RL   Unpublished (1985).
RL   U.S. Patent No. 4,559,308 dated Dec. 17. 1985.
RN   [13]
RC   pCG1 from Corynebacterium glutamicum strain 225-57
RA   Katsumata R., Furuya A.;
RT   "Plasmid pCG1";
RL   Unpublished (1982).
RL   European Patent Appl. 0058889 dated Sept. 1, 1982.
CC   This was constructed by ligating pCG2 (6.6 kb) linearized with PstI to
CC   pGA22 (7.9 kb) linearized with PstI. [1]
CC   This vector contains the following restriction sites (in approximate
CC   kb from one PstI site): KpnI--0.8, 3.8; BamHI--4.4, 10.7, 13.4;
CC   HpaI--5.8; EcoRI--8.3; SalI--10.3; XhoI--12.5. [1]
CC   The pGA22 portion of the construct contains the drug resistance genes.
CC   cmlR can be insertionally inactivated by cloning into the EcoRI site;
CC   kanR can be inactivated by cloning into the HindIII or XhoI sites. [2]
CC   The original vials from the depositor are being used for distribution.
CC   (ATCC staff)
CC   Growth at 30C in the presence of chloramphenicol, kanamycin, or
CC   tetracycline was verified.  Recommended tetracycline concentration is
CC   1.5 ug/ml on LB agar. (ATCC staff)
CC   Medium is -1 LB plus tetracycline (1.5 ug/ml).
CC   NM (pCE54)
CC   CM (no)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   HO (Corynebacterium glutamicum LA103)
CC   HO (Corynebacterium glutamicum)(Brevibacterium)(E.coli)
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. Corynebacterium glutamicum strain 225-57
FT                   -> pCG1
FT                   1. Corynebacterium glutamicum LP-6
FT                   -> pGA1 4900bp
FT                   1. pGA1 4900bp
FT                   -> pGA22
FT                   1. pCG1
FT                   -> pCG2
FT                   1. pCG2
FT                   2. pGA22
FT                   -> pCE54 14500bp"
FT   misc_binding    0..0
FT                   /note="SIT unique EcoRI-SalI-XhoI"
FT   rep_origin      0..0
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT   rep_origin      0..0
FT                   /note="ORI Corynebacterium pCG2"
FT   CDS             0..0
FT                   /note="ANT E. coli tetracycline resistance gene (tet)"
FT   CDS             0..0
FT                   /note="ANT E. coli chloramphenicol acetyltransferase
FT                   gene (cat); chloramphenicol resistance gene (cmr/cml)"
FT   CDS             0..0
FT                   /note="ANT E. coli kanamycin resistance gene (kan)"
SQ   Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;