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ID   PARO181    preliminary; circular DNA; SYN; 6070 BP.
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AC   ATCC77125;
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DT   01-JUL-1993 (Rel. 7, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
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DE   Enterobacter/E.coli plasmid vector pARO181 - incomplete.
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KW   cloning vector.
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OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
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RN   [1]
RC   pARO160 from pUC18 & pSUP2021
RC   pARO161 from pARO160 & pBR325, Tn5 kan gene
RC   pARO163 from pUC18 & pARO161
RC   pARO173 from pUC19 & pARO161
RC   pARO190 from pUC19 & pARO173
RC   pARO191 from pARO173
RC   pARO181 from pARO191 & pARO163
RC   pARO180 from pARO190 & pUC18
RA   Parke D.;
RT   "Construction of mobilizable vectors derived from plasmids RP4,
RT   pUC18 and pUC19";
RL   Gene 93:135-137(1990).
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CC   Mobilizable expression vector encoding the lacZalpha (lacZ') peptide
CC   permitting visual detection of recombinants. Contains priming sites
CC   useful for sequencing.
CC   Replicates to high copy number in E.coli and Enterobacter aerogenes.
CC   Contains the basis of mobilization (bom) and origin of transfer (oriT)
CC   sequences from RP4.
CC   The order of the major features in this plasmid is:lacZ'/MCS -
CC   pMB1 ori - kanR - oriT/bom.  pARO181 (ATCC 77125) and pARO191 (77126)
CC   differ in the orientation of the MCS in lacZ'.
CC   Can be used as an expression vector in bacteria of the family
CC   Enterobacteriaceae, or as a mobilizable suicide vector for
CC   site-specific insertional mutagenesis in diverse other bacterial
CC   strains.
CC   Restriction digests of the clone give the following sizes (kb):
CC   HindIII/PvuI--6.0; PvuI--6.0; EcoRI/BglII--3.7,2.3; EcoRI/PvuI--6.0.
CC   (ATCC staff)
CC   Medium is 1236 LB plus kanamycin.
CC   NM (pARO181)
CC   CM (no)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   SP (ATCC)
CC   HO (E.coli DH5alpha)(Enterobacter aerogenes)(E.coli)
CC   HO (E.coli S17-1 [ATCC 47055])
CC   CP ()
CC   FN (cloning)
CC   SE ()
CC   PA ()
CC   BR ()
CC   OF ()
CC   OR ()
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FH   Key             Location/Qualifiers
FH
FT   misc_feature    0..0
FT                   /note="1. pACYC184
FT                   2. RP4, Mob region/oriT
FT                   -> pSUP101
FT                   1. pBR325, ori/amp
FT                   2. pSUP101
FT                   -> pSUP202-1
FT                   1. pUC18 remove HindIII-EcoRI 51bp 400..451,
FT                   \ MCS 2635bp
FT                   2. pSUP202-1 EcoRI-HindIII 3150bp, RP4 oriT
FT                   -> pARO160 5785bp
FT                   1. pARO160 HindIII 5785bp 400..400
FT                   2. Tn5 HindIII-HindIII 3423bp 1196..4619, kan gene
FT                   -> pARO161 9185bp
FT                   1. pUC18 SspI-Cfr101 1960bp 2504..2686..1780,
FT                   \ MCS/lacZ gene
FT                   2. pARO161 SspI-XmaI 4110bp, oriT/kan gene
FT                   -> pARO163 6072bp
FT                   1. pUC19 SspI-Cfr101 1962bp 2504..2686..1780,
FT                   \ MCS/lacZ gene
FT                   2. pARO161 SspI-XmaI 4110bp, oriT/kan gene
FT                   -> pARO173 6072bp
FT                   1. pARO173 HindIII 6072bp
FT                   fill in
FT                   -> pARO191 6072bp
FT                   1. pARO191 NcoI-NdeI 4000bp
FT                   2. pARO163 NcoI-NdeI 2000bp
FT                   -> pARO181 6000bp"
FT   misc_binding    0..0
FT                   /note="MCS HindIII-SphI-PstI-AccI-HincII-SalI-XbaI-
FT                   BamHI-AvaI-SmaI-XmaI-KpnI-BanII-SacI-EcoRI"
FT   misc_binding    0..0
FT                   /note="SIT unique HindIII-SalI-HincII-XbaI-BamHI-
FT                   SmaI-KpnI-SacI-EcoRI"
FT   rep_origin      0..0
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT   promoter        0..0
FT                   /note="PRO E. coli lac gene"
FT   CDS             0..0
FT                   /note="GEN E. coli beta-galactosidase gene (lacZ');
FT                   reporter gene"
FT   CDS             0..0
FT                   /note="ANT E. coli kanamycin resistance gene (kan)"
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SQ   Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;
     n
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