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ID   PMB        preliminary; circular DNA; SYN; 5590 BP.
AC   ATCC77385;
DT   03-FEB-1994 (Rel. 8, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
DE   Vertebrate/E.coli plasmid vector pMB - incomplete.
KW   cloning vector.
OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
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).
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   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 ()
FH   Key             Location/Qualifiers
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)"
SQ   Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;