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ID   PTAG1      preliminary; circular DNA; SYN; 6000 BP.
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AC   ATCC77318;
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DT   03-FEB-1994 (Rel. 8, Created)
DT   01-APR-1995 (Rel. 11, Last updated, Version 1)
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DE   Vertebrate/E.coli phagemid vector pTAG-1 - 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   pBLCAT3.f1 from pBLCAT3 & pUC-f1
RC   pUC.A.1.5 from pUC18
RC   pJFCAT1 from pBLCAT3.f1 & pUC.A.1.5
RC   pGEM2-human-beta-globin from pGEM-2 & human beta-globin gene
RC   pbetaGloXH from pGEM-7 series & pGEM2-human-beta-globin
RC   pbetaGlo4 from pbetaGloXH & mouse tk promoter
RC   pbetaGlo4pA from pbetaGlo4 & pJFCAT1
RC   pTAG-1 from pbetaGlo4pA & pBLCAT3
RA   Fridovich-Keil J.L., Gudas J.M., Bryan I.B., Pardee A.B.;
RT   "Improved expression vectors for eukaryotic promoter/enhancer
RT   studies";
RL   Biotechniques 11:572-579(1991).
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CC   Deposited by: Judith L. Fridovich-Keil
CC   The order of the major features in this plasmid is: SalI - SV40
CC   polyadenylation trimer - XhoI/MCS/HindIII - beta-globin - PstI - SphI
CC   - ApaI - ampR/pGEM-7Zf(+). [1]
CC   Contains a trimer cassette of the SV40 major late polyadenylation
CC   signal to block background readthrough expression of the reporter
CC   gene. [1]
CC   A promoter/enhancer-cloning shuttle vector using human beta-globin as
CC   the reporter gene. Primarily useful for analysis of gene expression at
CC   the RNA level. [1]
CC   The ApaI site can be used for cloning enhancer elements. [1]
CC   Restriction digests of the clone give the following sizes (kb):
CC   PstI--3.8, 2.2; SphI--3.8, 2.2; HindIII--6.0; XhoI--6.0;
CC   BamHI--5.5, 0.55. (ATCC staff)
CC   Medium is 1227 LB plus ampicillin.
CC   NM (pTAG-1)
CC   CM (no)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (phagemid)
CC   SP (ATCC)
CC   HO (E.coli)(E.coli HB101)(vertebrate cells)
CC   CP ()
CC   FN (cloning)
CC   SE ()
CC   PA (pbetaGlo4pA from pbetaGlo, from pbetaGloXH from pGEM7, & pJFCAT1)
CC   PA (pBLCAT3)
CC   BR ()
CC   OF ()
CC   OR ()
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FH   Key             Location/Qualifiers
FH
FT   misc_feature    0..0
FT                   /note="1. pGEM-2 remove small HindIII-XbaI
FT                   2. human beta-globin gene HindIII-XbaI
FT                   -> pGEM2-human-beta-globin
FT                   1. pGEM-7 remove small HindIII-XbaI
FT                   2. pGEM2-human-beta-globin HindIII-XbaI
FT                   -> pbetaGloXH
FT                   1. pbetaGloXH remove small HindIII-NsiI
FT                   2. mouse tk promoter PstI-HindIII
FT                   -> pbetaGlo4
FT                   1. pbetaGlo4 XbaI
FT                   fill in
FT                   2. pJFCAT1 HindIII-HindIII, SV40 major late polyA
FT                   Klenow, fill in
FT                   -> pbetaGlo4pA
FT                   1. pbetaGlo4pA remove small HindIII-XhoI, mouse tk
FT                   2. pBLCAT3 small HindIII-XhoI, MCS
FT                   -> pTAG-1"
FT   misc_binding    0..0
FT                   /note="SIT blunted XbaI site"
FT   polyA_signal    0..0
FT                   /note="PLA SV40 major late gene trimer, from pJFCAT1
FT                   HindIII-HindIII, blunt-ended, insert into blunt XbaI
FT                   of pbetaGlo4pA"
FT   misc_binding    0..0
FT                   /note="MCS pBLCAT3 XhoI-HindIII,
FT                   replaces mouse thymidine kinase promoter PstI-HindIII
FT                   inserted into HindIII-NsiI of pGEM7"
FT   misc_binding    0..0
FT                   /note="MCS XhoI-BglII-BamHI-PstI-SphI-HindIII"
FT   misc_binding    0..0
FT                   /note="SIT unique XhoI"
FT   misc_binding    0..0
FT                   /note="SIT unique BglII"
FT   misc_binding    0..0
FT                   /note="SIT unique HindIII"
FT   CDS             0..0
FT                   /note="GEN human beta-globin gene;
FT                   reporter gene, human beta-globin gene
FT                   HindIII-XbaI inserted in pGEM7 HindIII-XbaI"
FT   misc_binding    0..0
FT                   /note="SIT PstI"
FT   misc_binding    0..0
FT                   /note="SIT SphI"
FT   misc_binding    0..0
FT                   /note="SIT unique ApaI"
FT   rep_origin      0..0
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322),
FT                   from pGEM7"
FT   CDS             0..0
FT                   /note="ANT E. coli beta-lactamase gene (bla)
FT                   ampicillin resistance gene (apr/amp), from pGEM7"
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SQ   Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;
     n
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