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ID   PDHFR29    preliminary; circular DNA; SYN; 6100 BP.
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AC   ATCC37165;
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DT   01-JUL-1993 (Rel. 7, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
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DE   Vertebrate/E.coli plasmid vector pdhfr2.9 - incomplete.
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KW   cloning vector.
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OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
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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).
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RN   [2]
RC   [lambda b2 imm from lambda]
RC   [lambda spi84 imm lambda nin5 from lambda]
RC   lambda rva from lambda spi84 imm lambda nin5 & lambda 540
RC   lambda rvb from lambda 590 & lambda b2 imm
RC   lambda rvc from lambda 598 & lambda b2 imm
RA   Carroll D., Ajioka R.S., Georgopoulos C.;
RT   "Bacteriophage lambda cloning vehicles for studies of genetic
RT   recombination";
RL   Gene 10:261-271(1980).
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RN   [3]
RC   from pDHFR11
RC   from pDHFR26
RC   lambda Ch4ADHFR103 from Charon 4A & mouse DHFR gene
RC   lambda Ch4ADHFR121, lambda Ch4ADHFR84 from Charon 4A & mouse dhfr gene
RC   pDR28 from DHFR gene
RC   pDBR14, pDBR22 from pBR322 & DHFR gene
RC   pD89Bg14 from DHFR gene
RC   pDHB9.8 from DHFR gene
RC   pDPH5 from DHFR gene
RC   pCHOR32 from pBR327 & lambda Ch16ACHOg49
RA   Crouse G.F., Simonsen C.C., McEwan R.N., Schimke R.T.;
RT   "Structure of amplified normal and variant dihydrofolate reductase
RT   genes in mouse sarcoma S180 cells";
RL   J. Biol. Chem. 257:7887-7897(1982).
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RN   [4]
RC   pDHFR7 from pBR322 & mouse DHFR gene
RC   pDHFR10, pDHFR11 from pBR322 & mouse DHFR gene
RC   pDHFR12, pDHFR13 from pBR322 & mouse DHFR gene
RC   pDHFR21, pDHFR22, pDHFR23, pDHFR24 from pBR322 & DHFR gene
RC   pDHFR25, pDHFR26, pDHFR27, pDHFR28, pDHFR29 from pBR322 & DHFR gene
RA   Chang A.C., Nunberg J.H., Kaufman R.J., Erlich H.A., Schimke R.T.,
RA   Cohen S.N.;
RT   "Phenotypic expression in E. coli of a DNA sequence coding for
RT   mouse dihydrofolate reductase";
RL   Nature 275:617-624(1978).
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RN   [5]
RC   lambda NM540 or lambda 540 from lambda imm21
RA   Murray K., Murray N.E.;
RT   "Phage lambda receptor chromosomes for DNA fragments made with
RT   restriction endonuclease III of Haemophilus influenzae and
RT   restriction endonuclease I of Escherichia coli";
RL   J. Mol. Biol. 98:551-564(1975).
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RN   [6]
RC   lambda Ch16ACHOg49 from Charon 16 & CHO gene
RA   Haynes S.R., Toomey T.P., Leinwand L., Jelinek W.R.;
RT   ;
RL   Mol. Cell. Biol. 1:573-583(1981).
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RN   [7]
RC   pSM24, pSM31 from DHFR gene
RA   Cheng S.M.;
RT   ;
RL   Unpublished (1982).
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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 (pdhfr2.9)
CC   CM (no)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   SP (ATCC)
CC   HO (E.coli HB101)(vertebrate cells)(E.coli)
CC   CP ()
CC   FN (cloning)
CC   SE ()
CC   PA ()
CC   BR ()
CC   OF ()
CC   OR ()
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FH   Key             Location/Qualifiers
FH
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 2.9
FT                   1. pBR327 HindIII 3273bp 30..30
FT                   2. lambda dhfr 2.9 HindIII-HindIII 1550bp, dhfr gene
FT                   -> pdhfr2.9 6100bp"
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)"
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SQ   Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;
     n
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