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ID   PJA50      preliminary; circular DNA; SYN; 6541 BP.
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AC   ATCC77251;
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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   Saccharomyces/E.coli phagemid vector pJA50 - 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   pUN0 or pNN328 from pBR322 & pUC18 & CEN4 & ARS1/TRP1 gene
RC   pNN328 or pMJ1 from pBR322 & pUC18 & CEN4 & ARS1/TRP1 gene
RC   pUN1 or pNN329 from pUN0 & TRP1 gene
RC   pUN10 or pNN330 from pUN1
RC   pUN20 or pNN331 from pUN1 & SUP11 gene
RC   pUN30 or pNN332, pUN40 or pNN333 from pUN1 & M13, ori
RC   pUN50 or pNN334 from pUN0 & URA3 gene
RC   pUN60 or pNN335 from pUN50 & SUP11 gene
RC   pUN70 or pNN336 from pUN50 & pUN30
RC   pUN80 or pNN337 from pUN50 & pUN40
RC   pUN90 or pNN338 from pUN0 & pNN69, HIS3 gene
RC   pUN100 or pNN339 from pUN0 & pUN90 & YEp13, LEU2 gene
RC   pUN15 or pNN340 from pUN10 & pBluescript KS+
RC   pUN25 or pNN341 from pUN20 & pBluescript KS+
RC   pUN35 or pNN342 from pUN30 & pBluescript KS+
RC   pUN45 or pNN343 from pUN40 & pBluescript KS+
RC   pUN55 or pNN344 from pUN50 & pBluescript KS+
RC   pUN65 or pNN345 from pUN60 & pBluescript KS+
RC   pUN75 or pNN346 from pUN70 & pBluescript KS+
RC   pUN85 or pNN347 from pUN80 & pBluescript KS+
RC   pUN95 or pNN348 from pUN90 & pBluescript KS+
RC   pUN105 or pNN349 from pUN100 & pBluescript KS+
RA   Elledge S.J., Davis R.W.;
RT   "A family of versatile centromeric vectors designed for use in the
RT   sectoring-shuffle mutagenesis assay in Saccharomyces cerevisiae";
RL   Gene 70:303-312(1988).
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CC   Shuttle vector to facilitate construction of gene disruptions in
CC   E. coli.
CC   The gene of interest is cleaved at an appropriate site, made flush,
CC   and ligated to the SmaI fragment from this plasmid that contains kanR
CC   and the yeast selectable marker. (personal communication)
CC   The order of the major features in this plasmid is:kanR - HIS3 -
CC   EcoRI/MCS/HindIII - f1 ori - ampR - pMB1 ori. (personal communication)
CC   The appropriate product is selected for by kanR in E.coli. (personal
CC   communication)
CC   Restriction digests of the clone give the following sizes (kb):
CC   EcoRI--6.54; PstI--5.1, 0.7, 0.7; HindIII--4.1, 1.8, 0.65.(ATCC staff)
CC   Medium is 1227 LB plus ampicillin.
CC   NM (pJA50)
CC   CM (no)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (phagemid)
CC   SP (ATCC)
CC   HO (E.coli HB101)(Saccharomyces cerevisiae)(E.coli)
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. pUC18 2686bp
FT                   2. kan gene
FT                   3. pYAC4 BamHI-BamHI 1768bp 4238..6006, HIS3 gene
FT                   -> pJA50 6541bp"
FT   misc_binding    0..0
FT                   /note="MCS EcoRI-ClaI-EcoRV-XbaI-BglII-XhoI-SacI-
FT                   HindIII"
FT   misc_binding    0..0
FT                   /note="SIT unique EcoRI-EcoRV-XbaI-XhoI-SacI"
FT   rep_origin      0..0
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT   rep_origin      0..0
FT                   /note="ORI bacteriophage f1"
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 beta-lactamase gene (bla)
FT                   ampicillin resistance gene (apr/amp)"
FT   CDS             0..0
FT                   /note="ANT E. coli kanamycin resistance gene (kan)"
FT   CDS             0..0
FT                   /note="ANT yeast HIS3 gene"
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SQ   Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;
     n
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