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ID   PFDTN5     preliminary; circular DNA; SYN; 8400 BP.
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AC   ATCC77330;
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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   E. coli plasmid vector pfd-Tn5 - 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   pfdA31 from pfdA3
RC   pfdA31-Tn5 or pfd-Tn5 from pfdA31 & Ti plasmid, omega fragment Tn5
RA   Metzger M., Bellemann P., Schwartz T., Geider K.;
RT   "Site-directed and transposon-mediated mutagenesis with
RT   pfd-plasmids by electroporation of Erwinia amylovora and
RT   Escherichia coli cells";
RL   Nucleic Acids Res. 20:2265-2270(1992).
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CC   Deposited by: Klaus Geider
CC   Suicide plasmid for transposon-mediated or site-directed mutagenesis
CC   of Erwinia amylovora and Escherichia coli after electroporation. [1]
CC   Suggested conditions for electroporation in these bacterial strains:
CC   E. coli C600Sm, fields strength 20 kV/cm, pulse length 5 to 12 msec;
CC   Erwinia amylovora Ea7/74Sm, 25 kV/cm, 5 msec; E. amylovora Ea1/79Sm,
CC   12.5 kV/cm, 17.5 msec. [1]
CC   To propagate the plasmid, the host must provide the bacteriophage fd
CC   gene 2 product. [1]
CC   The order of the major features in this plasmid is:  EcoRI - ampR -
CC   fd ori - EcoRI - Tn5/kanR (6.5 kb). [1]
CC   Restriction digests of the clone give the following sizes (kb):
CC   EcoRI--6.5, 2.0; BamHI--4.4, 4.0; HindIII--5.0, 3.4; PstI--2.45
CC   (doublet); 1.5, 0.96, 0.84. (ATCC staff)
CC   Medium is 1227 LB plus ampicillin.
CC   NM (pfd-Tn5)
CC   CM (no)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   SP (ATCC)
CC   HO (E.coli)(E.coli C600-2Sm)
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. pBR322, amp gene
FT                   2. phage fd, ori
FT                   -> pfdA3
FT                   1. pfdA3 remove HaeII-HaeII, kan gene
FT                   -> pfdA31
FT                   1. pfdA31 EcoRI
FT                   2. Ti plasmid EcoRI-EcoRI 800bp, Tn5/EcoRI omega frag
FT                   -> pfd-Tn5 8400bp"
FT   rep_origin      0..0
FT                   /note="ORI bacteriophage fd"
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 kanamycin resistance gene (kan)"
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SQ   Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;
     n
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