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ID   PJEL144    preliminary; circular DNA; SYN; 12700 BP.
AC   X05629; ATCC37526;
DT   01-JUL-1993 (Rel. 7, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
DE   E. coli plasmid vector pJEL144 - incomplete.
KW   cloning vector.
OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
RN   [1]
RC   pVH7 from pBR322 & deoCABD genes
RC   pJEL153 from pJEL75 & pPR633
RC   pJEL154 from pJEL153 & pBR322
RC   pJEL170 from pJEL126 & pJEL154
RC   pJEL144 from pJEL134 & pJEL170
RC   pJEL147 from pVH7 & pJEL170
RC   pJEL141, pJEL143, pJEL145, pJEL146 from pVH7 & pJEL144
RA   Larsen J.E., Albrechtsen B., Valentin-Hansen P.;
RT   "Analysis of the terminator region after the deoCABD operon of
RT   Escherichia coli K-12 using a new class of single copy number
RT   operon-fusion vectors";
RL   Nucleic Acids Res. 15:5125-5140(1987).
RN   [2]
RC   pPR95 from R1, parB gene
RC   pPR171 from pBR322 & R1
RC   pPR633 from pBR322 & R1, parB gene
RC   pPR122 from pOU491 & pPR95
RC   pPR633 from R1, parB gene
RC   pPR634, pPR341 from pPR633
RC   pKG171 from pPR171 & pOU82
RC   pKG341 from pPR341 & pOU82
RC   pKG634 from pPR634 & pOU82
RC   pKG633 from pOU82 & pPR633
RC   pF634 from F factor
RA   Gerdes K., Rasmussen P.B., Molin S.;
RT   "Unique type of plasmid maintenance function: postsegregational
RT   killing of plasmid-free cells";
RL   Proc. Natl. Acad. Sci. U.S.A. 83:3116-3120(1986).
RN   [3]
RC   lambda cIts857 xis am6 Sam7 nin5 b515 b519 from lambda
RA   Schrenk W.J, Weisberg R.A.;
RT   "A simple method for making new transducing lines of coliphage
RT   lambda";
RL   Mol. Gen. Genet. 137:101-107(1975).
RN   [4]
RC   pJEL134 from deoP2 promoter
RA   Valentin-Hansen P.;
RT   ;
RL   Gene Manipulation and Expression 0:273-288(1985).
RL   Glass R.E., Spizek J., eds., Croom Helm, London.
RN   [5]
RC   lambda c (int)delta h80 from lambda
RA   Shimada K., Weisberg R.A., Gottesman M.;
RT   "Prophage lambda at unusual chromosomal locations. I. Location of
RT   the secondary attachment sites and the properties of the lysogens";
RL   J. Mol. Biol. 80:297-314(1972).
CC   Terminator-testing plasmid.  Termination signal-containing sequences
CC   cloned into the BamHI site may reduce expression of
CC   beta-galactosidase activity regulated by the deoP2 promoter.
CC   Below 37C, there is one copy per genome equivalent stabilized by the
CC   parB locus.  At 42C replication is uncontrolled because induction of
CC   the cI857-regulated PR promoter permits expression of the copB, copA,
CC   and repA genes.
CC   Restriction digests of the clone give the following sizes (kb):
CC   BamHI--13.6; EcoRI--13.6. (ATCC staff)
CC   Constructed by inserting a 0.22 kb EcoRI/BamHI fragment containing the
CC   deoP2 regulatory region and the first 16 codons of deoC from pJEL134
CC   into pJEL170.
CC   Transcription from deoP2 is negatively regulated by the DeoR and CytR
CC   repressors and depends on the cAMP/CRP complex.
CC   Expression of beta-galactosidase from this vector can be increased by
CC   the use of hosts containing repressor mutations such as S0929 (cytR),
CC   S0930 (deoR) and S0931 (cytR, deoR).
CC   Medium is 1227 LB plus ampicillin.
CC   NM (pJEL144)
CC   CM (no)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   HO (E.coli CSH50)(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. pBR322 remove EcoRI-BamHI 377bp
FT                   \ 4360..4361..376, 3984bp
FT                   2. R1 BamHI-EcoRI 580bp, 0 to +580 parB gene
FT                   -> pPR633 4564bp
FT                   1. R1
FT                   2. lambda, pR/cI857 gene
FT                   -> pOU71
FT                   1. pOU71 remove HindIII-HindIII
FT                   -> pJEL75
FT                   1. pJEL75 NdeI-BamHI
FT                   Klenow:
FT                   2. pPR633 EcoRI-BamHI 580bp, 0 to +580 parB gene
FT                   Klenow:
FT                   -> pJEL153
FT                   1. pJEL153 remove small BamHI-ClaI
FT                   2. pBR322 XhoII-ClaI 1269bp 3117..4361..25, amp gene
FT                   -> pJEL154
FT                   1. lambda, cI857 gene
FT                   2. pMC903, lac gene
FT                   3. R1, parA gene
FT                   -> pJEL126
FT                   1. pJEL126 EcoRI-HindIII, lac gene/cI857
FT                   2. pJEL154 large EcoRI-HindIII
FT                   -> pJEL170
FT                   1. E. coli EcoRI-BamHI 220bp, deoP2 promoter
FT                   -> pJEL134
FT                   1. pJEL134 EcoRI-BamHI 220bp, deoP2 promoter
FT                   2. pJEL170 large EcoRI-BamHI, 12500bp
FT                   -> pJEL144 12700bp"
FT   misc_binding    0..0
FT                   /note="SIT unique EcoRI-BamHI"
FT   rep_origin      0..0
FT                   /note="ORI E. coli R1"
FT   promoter        0..0
FT                   /note="PRO bacteriophage lambda deoP2 gene"
FT   CDS             0..0
FT                   /note="GEN E. coli beta-galactosidase gene (lacZ');
FT                   reporter gene"
FT   CDS             0..0
FT                   /note="REP bacteriophage lambda cI857 repressor gene"
FT   CDS             0..0
FT                   /note="ANT E. coli beta-lactamase gene (bla)
FT                   ampicillin resistance gene (apr/amp)"
SQ   Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;