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ID   PNKY48     preliminary; circular DNA; SYN; 4300 BP.
AC   ATCC77182;
DT   01-JUL-1993 (Rel. 7, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
DE   Saccharomyces/E.coli plasmid vector pNKY48 - 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).
RN   [2]
RC   pRY110, pRY111 from pSc4816 & linker
RC   pRY112 from pRY110 & linker
RC   pRY114 from pRY112
RC   pRY116 from pRY114 & linker
RC   pRY117 from pRY114 & linker
RC   pRY113 from pRY111
RC   pRY115 from pRY113
RC   pRY118 from pRY115 & linker
RC   pRY119 from pRY115 & linker
RC   pRY121 from pRY116 & pLG669
RC   pRY131 from pRY121 & linker
RC   pRY140 from pRY121
RC   pRY123 from pRY118 & pLG669
RC   pRY133 from pRY123 & linker
RC   pRY124 from pRY119 & pSZ211
RC   pRY134 from pRY124 & linker
RC   pRY151 from pRY134 & pRY131
RC   pRY161 from pRY151 & YCp19
RC   pRY181 from pRY121 & YIp32
RC   pRY183 from pRY123 & YIp32
RC   pLR1delta3 from pRY131 & linker
RC   pRY171 from pLR1delta3
RC   pRY173 from pLR1delta3
RA   Yocum R., Hanley S., West R., Ptashne M.;
RT   "Use of lacZ fusions to delimit regulatory elements of the
RT   inducible divergent GAL1-GAL10 promoter in Saccharomyces cerevisiae";
RL   Mol. Cell. Biol. 4:1985-1998(1984).
RN   [3]
RC   B54 from HIS4 gene
RC   B115 from pBR322 & URA3/HIS4
RA   Donahue T.;
RT   ;
RL   Unpublished (1987).
RN   [4]
RC   pSc4816 from pBR325 & Gal1-Gal10 & linker
RA   St John T.;
RT   ;
RL   Unpublished (1984).
RN   [5]
RC   pSZ211 from YIp32
RA   Szostak J.;
RT   ;
RL   Unpublished (1984).
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   Encodes 93 amino acids of HIS4C and the complete 267 amino acids of
CC   URA3.
CC   Can be used for promoter/enhancer trap screens. (personal
CC   communication)
CC   Constructed by inserting a 1.4 kb HindIII fragment from pNKY2029
CC   containing the HIS4/URA3 fusion into pNKY47,which is a derivative of
CC   pGC-1 with the original HindIII site destroyed & a new one created at
CC   the BamHI site of the MCS.
CC   The order of the major features in this plasmid is: M13 primer - ClaI
CC   - XhoI - SalI - XbaI (stop) - BamHI - HindIII - 90 aa HIS4C
CC   bridge/URA3 - BamHI - HindIII - BamHI - BglII - NcoI - EcoRI - M13 ori
CC   - pMB1 ori - ampR.
CC   Restriction digests of the clone give the following sizes (kb):
CC   PstI--4.4; BglII--4.4; HindIII--3.1, 1.3; EcoRI--4.4; XbaI--4.4.
CC   (ATCC staff)
CC   Medium is 1227 LB plus ampicillin.
CC   NM (pNKY48)
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 SK-1)
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. yeast, HIS4 gene
FT                   -> B54
FT                   1. YEp24 remove HindIII-HindIII 1166bp 2273..3439,
FT                   \ URA3 gene/6603bp
FT                   -> pNKY2003 6603bp
FT                   1. pNKY2003 SphI 6603bp 3976..3976
FT                   2. B54 SstI
FT                   SphI linker 6bp gcatgc
FT                   SphI-SphI 4000bp, HIS4 gene
FT                   -> pNKY2005 10600bp
FT                   1. pNKY2005 BamHI 10600bp 3785..3785
FT                   2. YEp13 BglII-BglII 2500bp, LEU2 gene
FT                   -> pNKY2010 13100bp
FT                   1. pNKY2010 BglII 13100bp
FT                   2. YEp24 BglII-BamHI 1500bp ?..3785, URA3 gene
FT                   -> pNKY2009 14600bp
FT                   1. pNKY2009 BglII 14600bp
FT                   BAL31 nuclease
FT                   BglII linker 6bp agatct
FT                   -> pNKY2022 14600bp
FT                   1. pNKY2022 BglII 14600bp
FT                   BAL31 nuclease
FT                   -> pNKY2024 14600bp
FT                   1. pNKY2024 remove small SmaI-BstEII, 14000bp
FT                   BglII linker 8bp nagatctn
FT                   -> pNKY2029 14000bp
FT                   1. pGC1 HindIII 2900bp, pBR322 30..30
FT                   T4 DNA polymerase
FT                   BamHI 2900bp, MCS/pBR322 376..376
FT                   T4 DNA polymerase
FT                   HindIII linker 6bp aagctt
FT                   -> pNKY47 2900bp [BamHI]
FT                   1. pNKY2029 HindIII-HindIII 1400bp, HIS4/URA3
FT                   2. pNKY47 HindIII 2900bp, pBR322 30..30
FT                   -> pNKY48 4300bp"
FT   misc_binding    0..0
FT                   /note="MCS XbaI-BamHI-HindIII"
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   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   CDS             0..0
FT                   /note="ANT yeast URA3 gene"
SQ   Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;