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ID   PFE364     preliminary; circular DNA; SYN; 4000 BP.
AC   K01479;
DT   07-NOV-1984 (Rel. 1, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
DE   E. coli plasmid vector pFE364 - incomplete, methotroxate-resist DHFR.
KW   cloning vector; dihydrofolate reductase; DHFR; methotrexate-resistant.
OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
RN   [1]
RP   1-90
RC   pSV4 from pBR322 & SV40 late early region
RC   pBR1 from pSV4 & linker
RC   pBR2 from pBR1 & SV40 late early region
RC   pKCR from pBR2 & ppCR1/Rchr beta-G-1, rabbit beta-globin
RC   pHG from pKCR & pFE364, DHFR gene
RC   pMCG from pKCR & pFE373, DHFR gene
RA   O'Hare K., Benoist C.O., Breathnach R.;
RT   "Transformation of mouse fibroblasts to methotrexate resistance by a
RT   recombinant plasmid expressing a prokaryotic dihydrofolate
RT   reductase";
RL   Proc. Natl. Acad. Sci. U.S.A. 78:1527-1531(1981).
RN   [2]
RC   pFE504, pFE506 from pBR322 & ColE1::Tn7, R483 Tn7 DHFR I gene
RC   pFE332 from pBR322 & R67, DHFR II gene
RC   pFE364 from pFE332
RC   pFE413 from pSC101 & R67, DHFR II gene
RC   pFE420 from pFE413
RC   pFE425 from pFE413
RC   pFE373 from pBR322 & R388, DHFR II gene
RA   Fling M.E., Elwell L.P.;
RT   "Protein expression in Escherichia coli minicells containing
RT   recombinant plasmids specifying trimethoprim-resistant
RT   dihydrofolate reductases";
RL   J. Bacteriol. 141:779-785(1980).
RN   [3]
RC   ppCR1/Rchr beta-G-1 from pCR1 & rabbit beta-globin gene
RA   Weissman C.;
RT   ;
RL   Unpublished (1981).
CC   A recombinant plasmid expressed inserted DNA sequences coding for
CC   prokaryotic methotrexate-resistant dihydrofolate reductase of
CC   unspecified origin, using an SV40 early promoter and
CC   polyadenylation signals from the rabbit beta-globin gene. The DHFR
CC   insert begins at base 4. The plasmids were grown in mouse
CC   fibroblasts and produced a chimeric mRNA coding for the prokaryotic
CC   enzyme. The mouse fibroblasts became Mtx resistant.
CC   [1] also reports a DHFR sequence in plasmid pFE373.
CC   NCBI gi: 208994
CC   NM (pFE364)
CC   CM (no)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   SP ()
CC   HO (E.coli)(mouse fibroblast cells)
CC   CP ()
CC   FN (cloning)
CC   SE ()
CC   PA ()
CC   BR (pFE373)
CC   OF ()
CC   OR ()
FH   Key             Location/Qualifiers
FT   misc_feature    0..0
FT                   /note="1. E. coli, DHFR II gene
FT                   -> R67
FT                   1. pBR322 EcoRI 4361bp 4360..4360
FT                   2. SV40 HindIII-PvuII 344bp 5172..5243..273,
FT                   \ early promoter
FT                   3. R67 EcoRI-EcoRI 4700bp, DHFR II gene
FT                   4. rabbit, beta-globin gene polyA
FT                   -> pFE332 9100bp
FT                   1. pFE332 remove BamHI-BamHI 4900bp
FT                   -> pFE364 4000bp [trim gene]"
FT   promoter        0..0
FT                   /note="PRO SV40 early gene"
FT   CDS             43..>90
FT                   /note="GEN E. coli dihydrofolate reductase gene
FT                   (DHFR); NCBI gi: 208995"
FT   polyA_signal    0..0
FT                   /note="PLA rabbit beta-globin gene"
SQ   Sequence 90 BP; 33 A; 18 C; 22 G; 17 T; 0 other;
     ggccacatag aacacctaga agttcacaag aaaggttcgg aaatggaacg aagtagcaat
     gaagtcagta atccagtcgc tggcaacttt