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ID   PSE1076    preliminary; circular DNA; SYN; 9000 BP.
AC   ATCC77271;
DT   01-JUL-1993 (Rel. 7, Created)
DT   01-APR-1995 (Rel. 11, Last updated, Version 1)
DE   Saccharomyces/E.coli plasmid vector pSE1076 - incomplete.
KW   cloning vector.
OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
RN   [1]
RC   pNK294 from pRB91, Salmonella hisOGD region
RC   pNKY3 from YEp24 & linker
RC   pNKY49 from pNK294 & pNKY3
RC   pNKY50 from pNKY49 & pNK294
RC   pNKY51 from pNKY50
RC   pNKY1006 from YRp7 & linker
RC   pNKY1009 from pNKY1006 & pNKY51
RA   Alani E., Cao L., Kleckner N.;
RT   "A method for gene disruption that allows repeated use of URA3
RT   selection in the construction of multiply disrupted yeast
RT   strains";
RL   Genetics 116:541-545(1987).
RN   [2]
RC   lambda 408 from lambda & Tn10
RC   lambda 634 from lambda & IS10
RC   lambda 757, lambda 768, lambda 809 from lambda & Tn10
RC   pBR333 from pBR322
RC   pNK75 from pBR333 & pRB91, Salmonella hisOGD region
RC   pNK81 from pNK75 & F' factor
RC   pNK82, pNK83 from pNK81
RC   pNK217 from pNK81 & Tn10, tet gene
RC   pNK289 from pNK217 & Tn5, kan gene
RA   Foster T.J., Davis M., Roberts D., Takeshita K., Kleckner N.;
RT   "Genetic organization of transposon Tn10";
RL   Cell 23:201-213(1981).
CC   A derivative of pNKY51 with kanR from Tn903 inserted at the SmaI site
CC   of pNKY51. (personal communication)
CC   The order of the major features in this plasmid is: BglII (nt 1) -
CC   hisG - nt 1150 - URA3 - nt 2350 - kanR - nt 3850 - hisG -
CC   BamHI (nt 5000) - pMB1 ori - ampR. (personal communication)
CC   Restriction digests of the clone give the following sizes (kb):
CC   BglII--9.0; BamHI--9.0; BamHI/BglII--5.2, 3.8. (ATCC staff)
CC   A plasmid containing an excisable URA3 cartridge for gene disruption
CC   that permits repeated use of URA3 selection in constructing multiply
CC   disrupted yeast strains.
CC   Used by inserting the excised cassette into a cloned target gene,
CC   isolating a restriction fragment containing the cassette with flanking
CC   target gene sequences, transforming Ura3- strains, & selecting for
CC   stable integrants.
CC   Medium is 1227 LB plus ampicillin.
CC   NM (pSE1076)
CC   CM (no)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   HO (E.coli HB101)(Saccharomyces cerevisiae)(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. pRB91, Salmonella hisOGD region
FT                   -> pNK294
FT                   1. YEp24 remove EcoRI-EcoRI, yeast 2 micron
FT                   2. BglII linker
FT                   -> pNKY3 [unique EcoRI]
FT                   1. pNK294 BglII-BamHI 1100bp, hisG gene
FT                   2. pNKY3 BglII
FT                   -> pNKY49
FT                   1. pNKY49 BamHI
FT                   2. pNK294 BglII-BamHI 1100bp, hisG gene
FT                   -> pNKY50
FT                   1. pNKY50 EcoRI, 5' URA3 gene
FT                   fill in
FT                   -> pNKY51 [hisG-URA3-hisG 3800bp]
FT                   1. pNKY51 SmaI
FT                   2. Tn903 SmaI-SmaI, kan gene
FT                   -> pSE1076"
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   CDS             0..0
FT                   /note="ANT E. coli kanamycin resistance gene (kan)"
FT   CDS             0..0
FT                   /note="ANT yeast URA3 gene"
SQ   Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;