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ID   PFN476     preliminary; circular DNA; SYN; 4200 BP.
AC   ATCC86962;
DT   01-FEB-1995 (Rel. 11, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
DE   E. coli plasmid pFN476 - incomplete.
KW   cloning vector.
OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
RN   [1]
RC   pFN476 from pSC101 & pGEM-3Z
RC   NOTE: see J. Bacteriol. 176:939-939(1994)
RA   Sankar P.M., Hutton M.E., VanBogelen R.A., Clark R.L., Neidhardt F.C.;
RT   "Expression analysis of cloned chromosomal segments of Escherichia
RT   coli";
RL   J. Bacteriol. 175:5145-5152(1993).
CC   Restriction digests of the clone give the following sizes (kb):
CC   EcoRI--4.2; HindIII--4.2; PstI--3.1, 1.1. (ATCC staff)
CC   The order of the major features of the plasmid is:pSC101 rep > - BamHI
CC   - < 3' lacZ - T7 promoter > - EcoRI/MCS(pGEM-3Z)/HindIII - < SP6
CC   promoter - 5' lacZ - BamHI - tnpR > - ampR > - par - pSC101 ori.
CC   (personal communication)
CC   Low copy number vector to express transcripts in vivo from the T7
CC   promoter and to minimize toxic gene effects. Maintained at 1-5 copies
CC   per cell.  At this level, the effect of uninduced expression of T7 RNA
CC   polymerase from the UV5 promoter resulting in transcripts of
CC   potentially toxic inserts is minimized. T7 RNA polymerase can also be
CC   provided by infection of F' hosts with M13mGP1-2 (ATCC 40303). [1]
CC   Growth: LB plus ampicillin (ATCC medium number 1227) 37C
CC   Deposited by: Sankar P.M.
CC   NM (pFN476)
CC   CM (no)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   HO (E.coli)(E.coli RC104)
CC   CP (low)
CC   FN (cloning up to 20000bp)(expression)(RNA synthesis in vitro)
CC   SE (color blue)
CC   PA ()
CC   BR ()
CC   OF ()
CC   OR ()
FH   Key             Location/Qualifiers
FT   misc_feature    0..0
FT                   /note="1. pGEM-3Z 2867bp, amp/MCS/lacUV5
FT                   2. pSC101 1300bp, ori
FT                   -> pFN476 4200bp"
FT   rep_origin      0..0
FT                   /note="ORI E. coli pSC101"
FT   CDS             0..0
FT                   /note="ANT E. coli amp gene"
FT   misc_binding    0..0
FT                   /note="MCS EcoRI-SacI-KpnI-AvaI-SmaI-BamHI-XbaI-SalI
FT                   AccI-HincII-PstI-SphI-HindIII"
FT   misc_binding    0..0
FT                   /note="MCS unique EcoRI-KpnI-SmaI-XbaI-SalI
FT                   AccI-HindIII"
SQ   Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;