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ID   YRP14TRP1D preliminary; circular DNA; SYN; 8400 BP.
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AC   ATCC77148;
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DT   01-JUL-1993 (Rel. 7, Created)
DT   01-APR-1995 (Rel. 11, Last updated, Version 1)
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DE   Saccharomyces/E.coli plasmid vector YRp14-trp1delta63 - 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   YRp17-Sc4903his3-delta3' or pNN277 from YRp17 & GAL4903 & HIS3 gene
RC   [YIp5-Sc3103 from YIp5 & yeast DNA]
RC   YIp5-Sc4131ahis3-delta3' or pNN275 from YIp5-Sc3103 & HIS3 gene
RC   YRp14-Sc2605his3-delta200 or pNN214 from YRp14-Sc2605
RC   [YRp15-Sc4914 from YRp15 & yeast DNA]
RC   YRp15-Sc4914his3-delta5' or pNN274 from YRp15-Sc4914 & his3-delta5
RA   Fasullo M.T., Davis R.W.;
RT   "Direction of chromosome rearrangements in Saccharomyces
RT   cerevisiae by use of his3 recombinational substrates";
RL   Mol. Cell. Biol. 8:4370-4380(1988).
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CC   A marker addition/transplacement vector.
CC   Constructed by inserting the equivalent of 2 genomic EcoRI/HindIII
CC   fragments (1.5 kb, centromere proximal and 0.8 kb, 3' end) from the
CC   TRP1 region into YRp14.
CC   Represents the deletion of a 0.6 kb EcoRI/HindIII fragment containing
CC   the promoter and the 5' portion of the gene.
CC   Non-integrants grow poorly selecting for Ura3+ (pinpoints at 5 days)
CC   while integrants grow well because the SUP11 copy number is maintained
CC   at one. If there are no integrants on a transformation plate, streak
CC   onto -URA plates and pick papillae. (personal communication)
CC   Restriction digests of the clone give the following sizes (kb):
CC   EcoRI/HindIII--6.0, 1.6, 0.6; HindIII--8.4; BamHI--8.4; PstI--3.7,
CC   3.4, 1.3; BglII--7.4, 1.1. (ATCC staff)
CC   Medium is 1227 LB plus ampicillin.
CC   NM (YRp14-trp1delta63)
CC   CM (no)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
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. Yrp14 EcoRI
FT                   2. yeast EcoRI-HindIII 1500bp, centromere/TRP1
FT                   3. yeast EcoRI-HindIII 800bp, 3' end of TRP1
FT                   -> plasmid
FT                   -> plasmid remove EcoRI-HindIII 600bp, TRP1 promoter
FT                   -> YRp14-trp1delta63"
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 yeast ARS1"
FT   CDS             0..0
FT                   /note="ANT yeast URA3 gene"
FT   CDS             0..0
FT                   /note="ANT yeast SUP11 gene"
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SQ   Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;
     n
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