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ID   PMB        preliminary; circular DNA; SYN; 5590 BP.
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AC   ATCC77385;
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DT   03-FEB-1994 (Rel. 8, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
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DE   Vertebrate/E.coli plasmid vector pMB - 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   pS1 from pUC19 & oligo
RC   pS1-polyA from pS1 & pMAMneo
RC   pS1-MT from pS1 & pT24
RC   pS1-MMTV from pS1 & pMAMneo
RC   pS3 from pS1
RC   pS5 from pS3
RC   pK from pUC19 & oligo
RC   pM from pK & pS1-polyA & pS1-MMTV, RSV LTR/MMTV LTR
RC   pT from pK & pS1-polyA & pS1-MT, MT gene
RC   pM-hyg from pM & pHT, hyg gene
RC   pMB-hyg from pM-hyg & pS5, lacZ-amb
RC   pMB from pMB-hyg
RC   pT-hyg from pT & pHT, hyg gene
RC   pTB-hyg from pT-hyg & pS5, lacZ-amb
RC   pTB from pTB-hyg
RA   Wang Q., Maher V.M., McCormick J.J.;
RT   "Mammalian expression vectors with modulatable promoters and two
RT   multiple cloning sites";
RL   Gene 119:155-161(1992).
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CC   Deposited by: J. Justin McCormick
CC   The first site was designed for insertion of a marker selectable in
CC   mammalian cells, and the second for the sequence to be expressed. [1]
CC   Both sites permit detection of inserts by alpha complementation, with
CC   discrimination between the two provided by the amber mutation in the
CC   second lacZ. [1]
CC   Expression vector containing two lacZalpha-based multiple cloning
CC   sites, with the second containing an amber mutation and being flanked
CC   by an RSV enhancer-MMTVLTR promoter at one end and splicing and
CC   polyadenylation signals at the other. [1]
CC   The plasmid contains the following restriction sites (bp from
CC   position 0):
CC   NotI--0; EcoRI--9, 3071; ClaI--425, 1560, 1745; Drd--1398, 3583, 5452;
CC   PvuI--1596, 2953, 4741; BglII--1782; SphI--2907, 3110; NsiI--2909;
CC   HindIII--99, 1794, 3112; BsaI--4441. (personal communication)
CC   The order of the major features in this plasmid is: NotI - 5' RSV
CC   enhancer - MMTV LTR 3' - ClaI - 5' lacZ'amb5 - NruI/MCSII''/HindIII -
CC   3' lacZ'amb5 - splicing - polyadenylation - NsiI -
CC   3' lacZ' EcoRI/MCS/HindIII - 5' lacZ'- pMB1 ori - ampR. [1]
CC   Restriction digests of the clone give the following sizes (kb):
CC   PvuI--2.5, 1.8, 1.3; EcoRI--3.0, 2.6; HindIII--2.6, 1.7, 1.3;
CC   BglII--5.6; ClaI--4.3, 1.3. (ATCC staff)
CC   Medium is 1227 LB plus ampicillin.
CC   NM (pMB)
CC   CM (no)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   SP (ATCC)
CC   HO (E.coli)(E.coli HB101)(E.coli JM105)(E.coli JM109)
CC   HO (vertebrate cells)
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. pM BamHI 5169bp, MCS/pUC19 418
FT                   2. pHT BamHI-BamHI 1920bp, hyg gene
FT                   -> pM-hyg 7089bp
FT                   1. pM-hyg NheI-BglII 2000bp, hyg gene
FT                   2. pS5 BglI-EarI 4600bp, lacZ'-amb gene
FT                   -> pMB-hyg 6600bp
FT                   1. pMB-hyg remove BamHI-BamHI 1920bp, hyg gene
FT                   -> pMB 5590bp"
FT   misc_binding    0..0
FT                   /note="MCS NruI-ClaI-NheI-BssHII-SpeI-XhoI-NarI-
FT                   NaeI-MluI-SplI-BglII-HpaI-HindIII; pUC19 MCSI/MSCII''"
FT   misc_binding    0..0
FT                   /note="SIT unique NruI-NheI-BssHII-SpeI-XhoI-
FT                   NaeI-MluI-SplI-BglII-HpaI-HindIII; pUC19 MCSI/MSCII''"
FT   misc_binding    0..0
FT                   /note="SIT unique KpnI-SmaI-BamHI-XbaI-SalI-AccI-PstI"
FT   rep_origin      0..0
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT   promoter        0..0
FT                   /note="PRO E. coli lac"
FT   promoter        0..0
FT                   /note="PRO MMTV LTR"
FT   CDS             0..0
FT                   /note="GEN E. coli beta-galactosidase gene (lacZ');
FT                   reporter gene"
FT   CDS             0..0
FT                   /note="ANT E. coli beta-lactamase gene (bla)
FT                   ampicillin resistance gene (apr/amp)"
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SQ   Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;
     n
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