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ID   PNF2Y      preliminary; circular DNA; SYN; 10000 BP.
AC   ATCC37187;
DT   01-JUL-1993 (Rel. 7, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
DE   Saccharomyces/E.coli plasmid vector pNF2Y - incomplete.
KW   cloning vector.
OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
RN   [1]
RC   pNF1 from YEp24 & pMC874
RC   pNF2 from pNF1
RC   pNF3 from pNF2
RC   pNF4 from pNF3
RC   pNF1000 from pNF1 & yeast DNA
RC   pNF2000 from pNF2 & yeast DNA
RC   pNF3000 from pNF3 & yeast DNA
RA   Naumovski L., Friedberg E.C.;
RT   "Construction of plasmid vectors that facilitate subcloning and
RT   recovery of yeast and Escherichia coli DNA fragments";
RL   Gene 22:203-209(1983).
CC   pNF2 can be used to create genomic libraries by inserting DNA from
CC   Sau3AI partial digests into the BamHI site. The inserts can be excised
CC   readily by the SalI sites that flank the BamHI site. [1]
CC   pNF2 was constructed from pNF1 by inserting a BamHI linker into the
CC   filled-in SalI site. pNF1 was constructed from pYE24 (ATCC 37051) by
CC   inserting a kanR-containing fragment from pMC874 into the BamHI site.
CC   [2]
CC   This plasmid confers resistance to G418 in yeast. [1]
CC   An episomal shuttle vector for subcloning and recovery of DNA from
CC   Saccharomyces cerevisiae. [1]
CC   Medium is 1227 LB plus ampicillin.
CC   NM (pNF2Y)
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. ColE1 6646bp
FT                   2. E. coli, lacZ gene
FT                   -> pMC81
FT                   1. pMC81
FT                   2. pACYC177 3941bp, kan gene [HaeII-HaeII 1595..3025]
FT                   -> pMC275
FT                   1. pMC275
FT                   -> pMC874 [kan, unique BamHI in lacZ]
FT                   1. YEp24 BamHI 7769bp 3785..3785
FT                   calf alkaline phosphatase
FT                   2. pMC874 Sau3A-Sau3A 2600bp, kan gene
FT                   \ pACYC177 2534bp 787..3321
FT                   -> pNF1 10400bp
FT                   1. pNF1 SalI 10400bp 4061..4061
FT                   T4 DNA polymerase
FT                   BamHI linker 6bp ggatcc:BamHI linker 6bp ggatcc
FT                   BamHI-BamHI 10400bp
FT                   -> pNF2 10000bp [SalI-BamHI-SalI 3' to kan]"
FT   misc_binding    0..0
FT                   /note="SIT unique BamHI"
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 Saccharomyces G418 resistance gene (G418)"
FT   CDS             0..0
FT                   /note="ANT yeast URA3 gene"
SQ   Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;