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ID   PNKY70     preliminary; circular DNA; SYN; 4400 BP.
AC   ATCC77183;
DT   01-JUL-1993 (Rel. 7, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
DE   Saccharomyces/E.coli plasmid vector pNKY70 - incomplete.
KW   cloning vector.
OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
RN   [1]
RC   pNKY2003 from YEp24
RC   pNKY2005 from pNKY2003 & B54
RC   pNKY2010 from pNKY2005 & YEp13
RC   pNKY2009 from pNKY2010 & YEp24
RC   pNKY2022 from pNKY2009 & linker
RC   pNKY2024 from pNKY2022
RC   B184 from B115 & yeast TD50
RC   pNKY2028 from pNKY2024 & B184
RC   pNKY2029 from pNKY2024 & linker
RC   plasmid from pNKY2029 & YRp7
RC   pNKY54 from plasmid & pBR322
RC   pNKY47 from pGC1 & linker
RC   pNKY48 from pNKY2029 & pNKY47
RC   pNKY59, pNKY60 from pNKY48 & pUR290
RC   pNKY70 from pNKY48 & linker
RC   pNKY71 from pNKY70 & pRY116
RC   pNKY1069 from pNKY71 & YRp7
RA   Alani E., Kleckner N.;
RT   "A new type of fusion analysis applicable to many organisms:
RT   protein fusions to the URA3 gene of yeast";
RL   Genetics 117:5-12(1987).
CC   Contains a HIS4C/URA3 fusion cassette that can be used to make fusions
CC   to other proteins of interest.  Also includes a M13 sequencing primer
CC   approximately 300 bp from the HIS4C/URA3 fusion junction.
CC   The order of the major features in this plasmid is: M13 primer - ClaI
CC   - XhoI - SalI - XbaI (stop) - BamHI - EcoRI - HIS4C bridge/URA3 -
CC   BamHI - HindIII - BglII - NcoI - EcoRI - M13 ori - pMB1 ori - ampR.
CC   Encodes 93 amino acids of HIS4C and the complete 267 amino acids of
CC   URA3.
CC   Constructed from pNKY48 (ATCC 77182) by inserting an EcoRI 8-mer at
CC   the HindIII site in the polylinker.
CC   Restriction digests of the clone give the following sizes (kb):
CC   BglII--4.4; EcoRI--2.9, 1.5; PstI--4.4; BamHI--3.2, 1.2; XbaI--4.4.
CC   (ATCC staff)
CC   Can be used for promoter/enhancer trap screens. (personal
CC   communication)
CC   Medium is 1227 LB plus ampicillin.
CC   NM (pNKY70)
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   HO (Saccharomyces cerevisiae DBY745)(Saccharomyces cerevisiae SK1)
CC   HO (Saccharomyces cerevisiae S288C)
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. pNKY48 HindIII 4300bp, nearest to primer
FT                   EcoRI linker 8bp ngaattcn
FT                   -> pNKY70 4400bp"
FT   misc_binding    0..0
FT                   /note="MCS XbaI-BamHI-EcoRI"
FT   misc_binding    0..0
FT                   /note="SIT unique XbaI-BglII-NcoI-SalI"
FT   rep_origin      0..0
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT   rep_origin      0..0
FT                   /note="ORI bacteriophage M13"
FT   CDS             0..0
FT                   /note="ANT E. coli beta-lactamase gene (bla)
FT                   ampicillin resistance gene (apr/amp)"
SQ   Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;