back Return to this vector's summary.
ID   PECV25     preliminary; circular DNA; SYN; 11317 BP.
XX
AC   ATCC77187;
XX
DT   01-JUL-1993 (Rel. 7, Created)
DT   01-APR-1995 (Rel. 11, Last updated, Version 1)
XX
DE   Vertebrate/E.coli plasmid vector pECV25 - incomplete.
XX
KW   cloning vector.
XX
OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
XX
RN   [1]
RC   pECVZAP1 from pECV25 & lambda ZAP
RA   Belt P.B., Jongmans W., De Wit J., Hoeijmakers J.H., Van De Putte P.,
RA   Backendorf C.;
RT   "Efficient cDNA cloning by direct phenotypic correction of a
RT   mutant human cell line (HPRT-) using an Epstein-Barr
RT   virus-derived cDNA expression vector";
RL   Nucleic Acids Res. 19:4861-4866(1991).
XX
RN   [2]
RC   from pECV25
RA   Belt P.B., Van Oosterwijk M.F., Odijk H., Hoeijmakers J.H.,
RA   Backendorf C.;
RT   "Induction of a mutant phenotype in human repair proficient cells
RT   after overexpression of a mutated human DNA repair gene";
RL   Nucleic Acids Res. 19:5633-5637(1991).
XX
RN   [3]
RC   pUCIBG from pUC19 & pECV1
RC   pSV328neo from pSV328A & pAG60
RC   pECV2 from pUCIBG & pSV328neo
RC   pEVxylE from pECV1 & P.putida TOL plasmid xylE gene
RC   p225 from p205 & p220.2
RC   pRSVxylE from pEVxylE & pRSVneo
RC   pBL21 from p201 & pRSVxylE
RC   pUCneo from pUC19 & pECV2
RC   pECV21 from pECV2 & pBL21
RC   pECV23 from pECV21 & p225
RC   pECV24 from pECV23 & pUCneo
RC   pECV25 from pECV24
RC   pECV5 from pECV2 & pECV21
RC   pECV6 from pECV5 & pECV25
RA   Belt P.B., Groeneveld H., Teubel W.J., Van De Putte P., Backendorf C.;
RT   "Construction and properties of an Epstein-Barr-virus-derived cDNA
RT   expression vector for human cells";
RL   Gene 84:407-417(1989).
XX
RN   [4]
RC   plasmid from pBR329 & linker & pSV2-glo
RC   pECV1 from plasmid & pBR327/RchR-beta-IIIC4, beta-globin gene
RA   Kartasova T., Cornelissen B.J., Belt P., Van De Putte P.;
RT   "Effects of UV, 4-NQO and TPA on expression of genes in human
RT   epidermal keratinocytes";
RL   Nucleic Acids Res. 15:5945-5962(1987).
XX
RN   [5]
RC   pBR328A from pBR328
RC   pSValpha2i from pSValpha2 & rabbit beta-globin gene
RC   pSValpha2j from pBR328 & pSValpha2, SV40 promoter/IFN gene
RC   pSValpha4j from pBR328A & pSValpha4, SV40 promoter/IFN gene
RC   plasmid from pSV328A & MT promoter
RC   pSVMTalpha1 from plasmid & pSValpha1
RC   pSVMTsalpha1 from pSVMTalpha1
RC   pSVGMTalpha1 from pSV2-gpt & pSVMTalpha1
RC   pSVGMTsalpha1 from pSV2-gpt & pSVMTsalpha1
RC   pSVLMOalpha1 from pSValpha1 & pMV5, MoMLV LTR
RC   p32.3 from p32.2 & linker
RC   pPyalpha1 from pSValpha1 & p32.3
RC   pPyMTalpha1 from pSVMTalpha1 & p32.3
RC   pPyLMOalpha1 from pSVLMOalpha1 & p32.2
RA   Van Heuvel M., Bosveld I.J., Luyten W., Trapman J., Zwarthoff E.C.;
RT   "Transient expression of murine interferon-alpha genes in mouse
RT   and monkey cells";
RL   Gene 45:159-165(1986).
XX
RN   [6]
RC   pFG5 from pBR322 & HSV-1 tk gene
RC   pAG0 from pBR322 & pFG5
RC   pAG1 from pBR322 & pFG5
RA   Colbere-Garapin F., Chousterman S., Horodniceanu F., Kourilsky P.,
RA   Garapin A.C.;
RT   "Cloning of the active thymidine kinase gene of herpes simplex
RT   virus type 1 in Escherichia coli K-12";
RL   Proc. Natl. Acad. Sci. U.S.A. 76:3755-3759(1979).
XX
RN   [7]
RC   pHyg from pKan2 & pLG89, hyg B gene
RC   pdeltaSstII-3 from EBV oriP
RC   pHEBo from pHyg & pdeltaSstII-3, EBV oriP
RA   Sugden B., Marsh K., Yates J.L.;
RT   "A vector that replicates as a plasmid and can be efficiently
RT   selected in B-lymphoblasts transformed by Epstein-Barr virus";
RL   Mol. Cell. Biol. 5:410-413(1985).
XX
RN   [8]
RC   lambda gt10 mouse ERCC-1 from lambda gt10 & mouse brain DNA
RC   pM4a-1, pM4a-2 from pTZ19R & lambda gt10 mouse ERCC-1
RC   plasmid from pM4a-2 & pcD-X, SV40 early gene promoter
RC   pTIME from plasmid & pM4a-1
RC   pSPSma from pSP65 & pcDE series, human ERCC-1 gene
RC   plasmid2 from pSPSma & linker
RC   pcDEMP-1, pcDEMP-2 from plasmid2 & pcDE series
RC   pcDEBg1 from pcDE series
RC   pcDEdeltaStu from pcDE series
RC   pcDEASt from pcDEdeltaStu & pcDE series
RA   Van Duin M., Van Den Tol J., Warmerdam P., Odijk H., Meijer D.,
RA   Westerveld A., Bootsma D., Hoeijmakers J.H.;
RT   "Evolution and mutagenesis of the mammalian excision repair gene
RT   ERCC-1";
RL   Nucleic Acids Res. 16:5305-5322(1988).
XX
RN   [9]
RC   pKan2 from pBR322 & Tn5, kan gene
RA   Skopek T.R., Liber H.L., Penman B.W., Thilly W.G.;
RT   "Isolation of a human lymphoblastoid line heterozygous at the
RT   thymidine kinase locus: possibility for a rapid human cell mutation
RT   assay";
RL   Biochem. Biophys. Res. Commun. 84:411-416(1978).
XX
RN   [10]
RC   pBR327/RchR-beta-IIIC4 from pBR327 & beta-globin gene
RA   Diercks P.;
RT   ;
RL   Unpublished (1987).
XX
RN   [11]
RC   pcDE72, pcDE series from human ERCC-1 gene
RA   Van Duin M., De Wit J., Odijk H., Westerveld A., Yasui A.,
RA   Koken M.H., Hoeijmakers J.H., Bootsma D.;
RT   "Molecular characterization of the human excision repair gene
RT   ERCC-1: cDNA cloning and amino acid homology with the yeast DNA
RT   repair gene RAD10";
RL   Cell 44:913-923(1986).
XX
RN   [12]
RC   pSV328A from pBR328A & SV40 promoter
RC   pSValpha1 from pSV328A & IFN gene
RC   pSValpha2 from pSV328A & IFN gene
RC   pSValpha4 from pSV328A & IFN gene
RA   Van Heuvel M., Bosveld I.J., Mooren A.T., Trapman J., Zwarthoff E.C.;
RT   "Properties of natural and hybrid murine interferons";
RL   J. Gen. Virol. 67:2215-2222(1986).
XX
RN   [13]
RC   pMV5 from MoMLV LTR & mouse DNA
RA   Luciw P.;
RT   ;
RL   Unpublished (1986).
XX
RN   [14]
RC   p32.2 from polyoma virus
RC   p48.19 from polyoma virus
RA   Veldman G.M.;
RT   ;
RL   Unpublished (1986).
XX
CC   The order of the major features in this plasmid is:bla (ampR) - hph
CC   (hygR) - RSV LTR - HindIII - rabbit beta-globin intron splicing
CC   signals - NotI - agpt (G418R) - XhoI - rabbit beta-globin
CC   polyadenylation signal - SalI - oriP - SacI - EBNA-1 - ClaI. [3]
CC   Shuttle expression vector maintained episomally in human cells at a
CC   copy number of 25 - 50.  Shows high transfection efficiency and can be
CC   introduced as supercoiled plasmids. [3]
CC   The EBNA-1 gene contains an in-frame deletion of a repetitive region
CC   of 700 +/- 20 bp.  This deletion increases the number of stably
CC   maintained plasmid molecules per cell. [3]
CC   Vector can be rescued from transfected cells with no detectable
CC   rearrangements. [3]
CC   Restriction digests of the clone give the following sizes (kb):
CC   HindIII/SacI--5.7 (doublet); SalI--11.5; XhoI--11.5. (ATCC staff)
CC   Medium is 1227 LB plus ampicillin.
CC   NM (pECV25)
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)(HeLa cells)
CC   CP ()
CC   FN (cloning)
CC   SE ()
CC   PA ()
CC   BR ()
CC   OF ()
CC   OR ()
XX
FH   Key             Location/Qualifiers
FH
FT   misc_feature    0..0
FT                   /note="1. pBR327
FT                   2. beta-globin gene
FT                   -> pBR327/RchR-beta-IIIC4
FT                   1. pSV2
FT                   2. globin gene
FT                   -> pSV2-glo
FT                   1. pBR329 PstI-AvaI
FT                   2. pSV2-glo remove EcoRI-BamHI, SV40
FT                   remove PstI-PvuII
FT                   -> plasmid [no PvuII]
FT                   1. plasmid BglII
FT                   KpnI linker
FT                   remove HindIII-KpnI, beta-globin gene
FT                   2. pBR327/RchR-beta-IIIC4 HindIII-KpnI, globin 2nd int
FT                   -> pECV1
FT                   1. p201 remove 700bp, EBNA-1 repetitive DNA [in frame]
FT                   -> p205
FT                   1. p205 ClaI-SacI, EBNA-1 gene
FT                   2. p220.2 remove ClaI-SacI, EBNA-1 gene
FT                   -> p225
FT                   1. pECV1 remove beta-globin region
FT                   2. P.putida TOL plasmid xylE gene
FT                   -> pEVxylE
FT                   1. pEVxylE HindIII-BamHI, xylE gene/SV40 late polyA
FT                   2. pRSVneo remove small HindIII-BamHI, 3' to RSV LTR
FT                   -> pRSVxylE
FT                   1. p201 NarI
FT                   Klenow fill in
FT                   BglII linker
FT                   2. pRSVxylE NdeI, 5' RSV LTR
FT                   Klenow fill in
FT                   BglII linker
FT                   BglII-BamHI, RSV LTR/xylE gene/SV40 polyA
FT                   -> pBL21
FT                   1. pUC19 remove HindIII-KpnI, MCS
FT                   2. pECV1 HindIII-KpnI, rabbit beta-globin 2nd/polyA
FT                   -> pUCIBG
FT                   1. pSV328A PvuII
FT                   2. pAG60 PvuII-PvuII, agpt gene/HSV1 tk promoter
FT                   -> pSV328neo
FT                   1. pUCIBG HindIII-SacI, rabbit beta-globin 2nd intron
FT                   2. pSV328neo remove small HindIII-SacI
FT                   -> pECV2
FT                   1. pUC19 remove BamHI-SmaI, MCS
FT                   2. pECV2 BglII-SmaI, agpt gene
FT                   -> plasmid
FT                   1. plasmid SmaI
FT                   XhoI linker
FT                   -> pUCneo
FT                   1. pECV2 HindIII-SalI, rabbit beta-globin 2nd/polyA
FT                   2. pBL21 remove HindIII-SalI, xylE gene
FT                   -> pECV21
FT                   1. pECV21 remove ClaI-SalI, SV40
FT                   2. p225 ClaI-SalI
FT                   -> pECV23
FT                   1. pECV23 SacI
FT                   T4 DNA polymerase fill in
FT                   XhoI linker
FT                   -> pECV23Xho
FT                   1. pECV23Xho XhoI
FT                   2. pUCneo SalI-XhoI, agpt gene
FT                   -> pECV24
FT                   1. pECV24 KpnI
FT                   NotI-KpnI linker gcggccgcgtac
FT                   -> pECV25"
FT   misc_binding    0..0
FT                   /note="MCS EcoRI-NotI-XbaI-XhoI-EcoRI"
FT   misc_binding    0..0
FT                   /note="SIT unique NotI-XhoI-SalI"
FT   rep_origin      0..0
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT   rep_origin      0..0
FT                   /note="ORI Epstein-Barr virus (EBV) oriP"
FT   promoter        0..0
FT                   /note="PRO RSV LTR"
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 hygromycin phosphotransferase gene;
FT                   hygromycin resistance gene (hyg)"
FT   CDS             0..0
FT                   /note="ANT mammal G418 resistance gene (G418)"
XX
SQ   Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;
     n
//