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ID   PUN30      preliminary; circular DNA; SYN; 5300 BP.
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AC   ATCC77265;
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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 pUN30 - 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   YC-type shuttle vector permitting visual detection of recombinants by
CC   lacZ alpha complementation and production of ssDNA (in the same sense
CC   as lacZ mRNA).
CC   The order of the major features in this plasmid is: ampR - pMB1 ori -
CC   lacZ'/MCS - CEN4 - M13+ - ARS1 - TRP1.
CC   Restriction digests of the clone give the following sizes (kb):
CC   EcoRI--5.4; KpnI--5.4; HindIII--3.7, 1.7. (ATCC staff)
CC   Medium is 1227 LB plus ampicillin.
CC   NM (pUN30)
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. pBR322 AatII-EcoRI 71bp 4289..4360
FT                   Klenow: , 57bp
FT                   2. pUC18 NdeI-AatII 2437bp 185..2622,
FT                   \ ori/amp gene [2434bp]
FT                   :Klenow
FT                   3. yeast ScaI-EcoRI 1517bp, CEN4
FT                   :Klenow
FT                   4. YRp7 EcoRI-EcoRI 1454bp 2..1456, ARS1/TRP1 gene
FT                   Klenow:Klenow
FT                   -> pUN0 5464bp
FT                   1. pUN0 HindIII-AatII 1400bp,TRP1 gene/pUC18 400..2622
FT                   Klenow:
FT                   2. yeast EcoRI 1456..1456, TRP1 gene
FT                   Klenow
FT                   BglII-HindIII 883bp 604..1487, TRP1 gene
FT                   Klenow:
FT                   -> pUN1 5400bp
FT                   1. pUN1 remove small HpaI-NaeI
FT                   2. M13 HpaI-NaeI 500bp ?..5616, ori
FT                   -> pUN30 5300bp"
FT   misc_binding    0..0
FT                   /note="MCS EcoRI-SacI-KpnI-SmaI-BamHI-XbaI-SalI-
FT                   PstI-SphI-HindIII"
FT   misc_binding    0..0
FT                   /note="SIT unique EcoRI-SacI-KpnI-SmaI-BamHI-SalI-
FT                   SphI-HindIII"
FT   rep_origin      0..0
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT   CDS             0..0
FT                   /note="ANT E. coli beta-lactamase gene (bla)
FT                   ampicillin resistance gene (apr/amp)"
FT   rep_origin      0..0
FT                   /note="ORI bacteriophage M13"
FT   rep_origin      0..0
FT                   /note="ORI yeast ARS1"
FT   misc_feature    0..0
FT                   /note="yeast centromere CEN4"
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 yeast TRP1 gene"
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SQ   Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;
     n
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