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ID   PCATX      preliminary; circular DNA; SYN; 4986 BP.
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AC   ATCC68669;
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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   E. coli plasmid vector pCAT - 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   pTIF from pCA15 & pPOH14
RC   pNFT from pTIF & murine NF-L promoter
RC   [pCAT from pNFT & mice ova]
RA   Byrne G.W., Ruddle F.H.;
RT   "Multiplex gene regulation: a two-tiered approach to transgene
RT   regulation in transgenic mice";
RL   Proc. Natl. Acad. Sci. U.S.A. 86:5473-5477(1989).
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RN   [2]
RC   pPO22 from pGH12
RC   delta108.CAT from pCatB' & pPO22
RC   p175KCAT.SV.L from p175KCAT & SV40 enhancer
RC   delta108.SV.L from pPO22 & linker & pCatB' & SV40 enhancer
RC   [pPOH14 from ICP4 promoter & cat gene & SV40 splice/polyA]
RC   [pCA15 from HSV-1 VP16 gene]
RA   O'Hare P., Hayward G.S.;
RT   "Comparison of upstream sequence requirements for positive and
RT   negative regulation of a herpes simplex virus immediate-early
RT   gene by three virus-encoded trans-acting factors";
RL   J. Virol. 61:190-199(1987).
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RN   [3]
RC   p175KCAT from pBR322 & HSV-1 IE175K gene & cat gene
RC   pGH12 from pBR322 & HSV-1
RA   O'Hare P., Hayward G.S.;
RT   "Expression of recombinant genes containing herpers simplex virus
RT   delayed-early and immediate-early regulatory regions and trans
RT   activation by herpesvirus infection";
RL   J. Virol. 52:522-531(1984).
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RN   [4]
RC   pXhoI-C from pBR322 & HSV IE175K gene
RC   pIGA-15 from pBR322 & HSV IE110K gene
RC   pGR135 from pBR322 & HSV-2 Vmw65 gene
RC   pGR212 from pBR322 & HSV-1 Vmw65 gene
RA   O'Hare P., Hayward G.S.;
RT   "Three trans-acting regulatory proteins of herpes simplex virus
RT   modulate immediate-early gene expression in a pathway involving
RT   positive and negative feedback regulation";
RL   J. Virol. 56:723-733(1985).
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CC   The component of a multiplex gene regulatory system containing the
CC   target transponding promoter sequence & reporter gene.  The
CC   transactivating gene product is encoded in pNFT (ATCC 68668).
CC   Constructed by inserting the ClaI/SalI (end filled) fragment of pPO2
CC   into the HincII site of the vector.  Transcription from the SP6
CC   promoter gives sense RNA. (personal communication)
CC   The 2.1 kb SmaI/SalI fragment contains the promoter fragment linked to
CC   coding sequences for chloramphenicol acetyltransferase (CAT) along
CC   with SV40 splice and polyadenylation signals.
CC   Restriction digests of the clone give the following sizes (kb):
CC   EcoRI/HindIII--2.9, 1.6, 0.4, 0.3; PvuI/PvuII--1.8, 1.7, 1.5;
CC   SmaI/SalI--5.0; SalI--uncut; SmaI--5.0. (ATCC staff)
CC   Medium is 1227 LB plus ampicillin.
CC   NM (pCAT)
CC   CM (no)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   SP (ATCC)
CC   HO (E.coli DH5alpha)(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. HSV-1 SmaI-BamHI 360bp, ICP4 imm-early prom
FT                   2. pCatB' large SmaI-BamHI, cat gene/SV40 splice/polyA
FT                   -> pPOH14
FT                   1. HSV-1 VP16 gene
FT                   -> pCA15
FT                   1. pCA15 BamHI-AsuII 1700bp, 5'UTR/VP16/polyA
FT                   2. pPOH14 2100bp, imm-early promoter/cat
FT                   -> pTIF
FT                   1. pTIF remove small HindIII-SmaI, 5' to VP16/3100bp
FT                   2. murine HindIII-SmaI 1500bp, NF-L promoter
FT                   -> pNFT 4600bp [NF-L prom/VP16/polyA]
FT                   1. pBR322
FT                   2. HSV-1, immediate early promoter
FT                   -> pGH12
FT                   1. pGH12, immediate early promoter
FT                   -> pPO22
FT                   1. pNFT HincII 4600bp
FT                   2. pPO22 ClaI-SalI 360bp, HSV-1 IE promoter
FT                   fill in:fill in
FT                   -> pCAT 4986bp"
FT   misc_binding    0..0
FT                   /note="MCS EcoRI-SacI-SmaI-BamHI-XbaI-AccI-SalI-
FT                   PstI-HindIII"
FT   rep_origin      0..0
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT   promoter        0..0
FT                   /note="PRO HSV-1 ICP4 gene"
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 E. coli chloramphenicol acetyltransferase
FT                   gene (cat); chloramphenicol resistance gene (cmr/cml)"
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SQ   Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;
     n
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