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ID   PDHFR32    preliminary; circular DNA; SYN; 6400 BP.
AC   ATCC37166;
DT   01-JUL-1993 (Rel. 7, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
DE   Vertebrate/E.coli plasmid vector pdhfr3.2 - incomplete.
KW   cloning vector.
OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
RN   [1]
RC   lambda rva-dhfr3.4 from lambda Ch4ADHFR121 & lambda rva
RC   lambda rvb-DHFR11 from lambda rvb & pDHFR11
RC   lambda dhfr 2.9 from lambda rva-dhfr3.4 & lambda rvb-DHFR11
RC   lambda dhfr 3.2 from lambda rva-dhfr3.4 & lambda rvb-DHFR11
RC   pdhfr2.9 from pBR327 & lambda dhfr 2.9
RC   pdhfr3.2 from pBR327 & lambda dhfr 3.2
RA   Crouse G.F., McEwan R.N., Pearson M.L.;
RT   "Expression and amplification of engineered mouse dihydrofolate
RT   reductase minigenes";
RL   Mol. Cell. Biol. 3:257-266(1983).
CC   Vector which expresses mouse dihydrofolate reductase in eukaryotic
CC   cell lines. The Dhfr minigene is selectable in dhfr- cell lines.
CC   (ATCC staff)
CC   Medium is 1227 LB plus ampicillin.
CC   NM (pdhfr3.2)
CC   CM (no)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   HO (E.coli HB101)(vertebrate cells)(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. lambda
FT                   deletion b2 EcoRI-HindIII, 0.44 to 0.566 on map
FT                   substitute red3 for red
FT                   -> lambda b2 red3
FT                   1. lambda imm21 [P+]
FT                   deletion nin5, for N gene independence
FT                   substitute shn6o
FT                   -> lambda 540 [lambda imm21 P+ nin5 shn6o]
FT                   1. lambda imm434
FT                   deletion b538
FT                   substitute shn6o
FT                   -> lambda 590 [lambda b538 imm434 shn6o]
FT                   1. lambda [with pR & oR, so imm lambda]
FT                   long bio substitution spi84, 0.573 to 0.75 on map
FT                   deletion nin5, for N gene independence
FT                   -> lambda spi84 imm lambda nin5
FT                   1. lambda spi84 imm lambda nin5
FT                   2. lambda 540, shn6o
FT                   -> phage
FT                   1. phage
FT                   2. lambda b2 red3
FT                   -> phage2
FT                   1. phage2 remove HindIII-HindIII, immunity region
FT                   -> lambda rva [lambda b2 spi84 imm lambda nin5 shn6o]
FT                   1. lambda 590
FT                   2. lambda b2 imm lambda
FT                   -> phage3 [lambda b2 imm434 shn6o]
FT                   1. phage3
FT                   2. lambda 540
FT                   -> lambda rvb [lambda b2 imm434 nin5 shn6o]
FT                   1. Charon 4A EcoRI
FT                   2. mouse EcoRI-EcoRI, dhfr gene 16600bp
FT                   -> lambda Ch4ADHFR121
FT                   1. pBR322 4361bp
FT                   2. mouse DHFR gene
FT                   -> pDHFR11
FT                   1. lambda Ch4ADHFR121 HindIII-HindIII 3400bp,dhfr gene
FT                   2. lambda rva HindIII
FT                   -> lambda rva-dhfr3.4
FT                   1. lambda rvb HindIII
FT                   2. pDHFR11 PstI-PstI 1500bp, dhfr gene
FT                   T4 DNA polymerase:T4 DNA polymerase
FT                   HindIII linker 6bp aagctt:HindIII linker 6bp aagctt
FT                   -> lambda rvb-DHFR11
FT                   1. lambda rvb-DHFR11, right arm
FT                   recombination
FT                   2. lambda rva-dhfr3.4, left arm
FT                   -> lambda dhfr 3.2
FT                   1. pBR327 HindIII 3273bp 30..30
FT                   2. lambda dhfr 3.2 HindIII-HindIII 3200bp, dhfr gene
FT                   -> pdhfr3.2 6400bp"
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 mammal methotrexate resistance gene (mtx)"
SQ   Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;