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ID   PORF5      preliminary; circular DNA; SYN; 6250 BP.
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AC   ATCC37221;
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DT   01-JUL-1993 (Rel. 7, Created)
DT   01-APR-1995 (Rel. 11, Last updated, Version 1)
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DE   E. coli plasmid vector pORF5 - 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   [pMLB1031 from pMC871 & pBR322]
RC   pMLB1034 from pMC871 & pBR322
RC   pORF5 from pMLB1034 & pBH20
RC   [lambda spc2 from lambda]
RC   pNF3041, pNF3033 from lambda spc2 & pUC8
RC   pNF3042 from pNF3033
RC   pNF3043 from pNF3041
RC   pJS600 from pNF3041
RC   pJS1800 from pNF3033
RC   pJS2200 from pNF3033
RC   pJS1000 from pNF3041 & pORF5
RA   Shultz J., Silhavy T.J., Berman M.L., Fiil N., Emr S.D.;
RT   "A previously unidentified gene in the spc operon of Escherichia
RT   coli K12 specifies a component of the protein export machinery";
RL   Cell 31:227-235(1982).
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RN   [2]
RC   pMLB1034 from pMC1871 & pBR322
RC   pORF5 from pMLB1034 & pBH20
RA   Weinstock G., Berman M., Silhavy T.;
RT   "Chimeric genetics with beta-galactosidase";
RL   Gene Amplif. Anal. 3:27-64(1983).
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RN   [3]
RC   pBH10 from pBR322 & lambda plac5, lacZ gene
RC   pBH20 from pBH10
RC   pSomI from pBH20 & somatostatin gene
RC   pSomII from pSomI & pBR322
RC   pSomII-3, pSomII-5, pSomII-6 from pSomII & lambda plac5, lacZ gene
RC   pSomII-7, pSomII-2, pSomII-4 from pSomII & lambda plac5, lacZ gene
RA   Itakura K., Hirose T., Crea R., Riggs A.D., Heyneuek H.L.,
RA   Bolivar F., Boyer H.W.;
RT   "Expression in Escherichia coli of a chemically synthesized gene
RT   for the hormone somatostatin";
RL   Science 198:1056-1063(1977).
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CC   Open reading frame vector.
CC   E.coli MC1000 = F- araD139 delta[ara,leu]7697 delta[lac]chi74
CC   galK- galU- rpsL.
CC   Medium is 1227 LB plus ampicillin.
CC   NM (pORF5)
CC   CM (no)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   SP (ATCC)
CC   HO (E.coli MC1000)(E.coli)(E.coli HB101)
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. pMC871 EcoRI-AvaI 3317bp, lacZ gene
FT                   2. pBR322 remove small EcoRI-AvaI, tet gene
FT                   -> pMLB1034 [no lacZ, no lacY]
FT                   1. pBR322 EcoRI
FT                   T4 DNA polymerase fill in
FT                   2. lambda plac HaeIII-HaeIII 203bp, lacPO
FT                   -> pBH10
FT                   1. pBH10 EcoRI, 5' to lacPO
FT                   S1 nuclease
FT                   -> pBH20
FT                   1. pMLB1034 remove small EcoRI-PstI
FT                   2. pBH20 EcoRI-PstI, lacPO promoter
FT                   -> pORF5"
FT   misc_binding    0..0
FT                   /note="MCS unique EcoRI-SmaI-BamHI"
FT   misc_binding    0..0
FT                   /note="SIT unique SalI"
FT   misc_binding    0..0
FT                   /note="SIT unique PstI"
FT   misc_binding    0..0
FT                   /note="SIT unique HindIII"
FT   promoter        0..0
FT                   /note="PRO E. coli lac gene"
FT   CDS             0..0
FT                   /note="GEN E. coli beta-galactosidase gene (lacZ');
FT                   reporter gene"
FT   rep_origin      0..0
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT   CDS             0..0
FT                   /note="ANT E. coli beta-lactamase gene (bla)
FT                   ampicillin resistance gene (apr/amp)"
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SQ   Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;
     n
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