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ID PE5LVP0 preliminary; circular DNA; SYN; 4800 BP.
DT 01-JUL-1993 (Rel. 7, Created)
DT 01-JUL-1995 (Rel. 12, Last updated, Version 1)
DE E. coli plasmid vector pE5LVP0 - incomplete.
KW cloning vector.
OS Cloning vector
OC Artificial sequences; Cloning vehicles.
RC pE5LVP0 from pBR322 & Sp6 & T7 & encephalomyocarditis virus
RL Unpublished (1990).
RL U.S. Patent No. 4,937,190 dated June 26, 1990.
CC Contains DNA corresponding to nt 260 through 2004 of
CC encephalomyocarditis virus RNA, including the 5' non-coding region &
CC coding regions for L, 1A, and IB.
CC Permits increased expression for in vitro transcription translation
CC systems by providing a translational enhancer downstream of the T7
CC Digestion of the plasmid with XbaI plus BalI generates a fragment
CC (approximately 4 kb) that has no EMC coding regions, but retains the
CC translational enhancer. This can be used for cloning.
CC If a potential insert contains transcription terminators, then the
CC BalI site can be used for cloning.
CC For in vitro transcription, the plasmid should be linearized with
CC The order of the major features in this plasmid is:T7 promoter - 5'
CC non-coding region of EMC - BalI - partial coding regions of EMC - XbaI
CC - MCS - pSPT18.
CC Restriction digests of the clone give the following sizes (kb):
CC EcoRI--4.8; HindIII--3.28, 1.54; XbaI--4.8. (personal communication)
CC Medium is 1227 LB plus ampicillin.
CC NM (pE5LVP0)
CC CM (no)
CC NA (ds-DNA)
CC TP (circular)
CC ST ()
CC TY (plasmid)
CC SP (ATCC)
CC HO (E.coli C600)(E.coli)(E.coli HB101)
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. pBR322, ori/amp gene
FT 2. Sp6, promoter
FT 3. T7, promoter
FT 4. encephalomyocarditis virus
FT -> pE5LVP0 4800bp"
FT misc_binding 0..0
FT /note="SIT unique BalI"
FT rep_origin 0..0
FT /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT promoter 0..0
FT /note="PRO bacteriophage SP6"
FT promoter 0..0
FT /note="PRO bacteriophage T7"
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;