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ID   PGI21      preliminary; circular DNA; SYN; 1 BP.
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AC   IG5072;
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DT   01-OCT-1993 (Rel. 7, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
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DE   E. coli plasmid vector pGI21 - 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   pGI6130 from pLK69, linker
RC   pGI6131 from pGI6130
RC   pGI6132 from pGI6131 & linker
RC   pGI6133 from pLK59, linker
RC   pGI6134 from pGI6133
RC   pGI6135 from pGI6134 & pGI6132
RC   pGI6136 from pGI6135
RC   pGI6146 from pGI6136
RC   pGI6147 from pGI6136
RC   pGI6137 from pGI6136 & pGI4010, ery gene
RC   pGI6138 from pGI6137 & pNK2805
RC   pGI10 from pHT1030 & pGC2
RC   pGI21 from pGI10 & pGI6138
RC   pGI210 from pGI21
RA   Mahillon J., Kleckner N.;
RT   "New IS10 transposition vectors based on a gram-positive replication
RT   origin";
RL   Gene 116:69-74(1992).
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RN   [2]
RC   pGI4010 from ery gene
RA   Josson K., Scheirlinck T., Michiels F., Patteeuw C., Stanssens P.,
RA   Joos H., Dhaese P., Zabeau M., Mahillon J.;
RT   "Characterization of a Gram-positive broad host range plasmid
RT   isolated from Lactobacillus hilgardii";
RL   Plasmid 21:9-20(1989).
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RN   [3]
RC   lambda NK370 from lambda & Tn10
RC   pNK81 from pBR322 & Tn10
RC   pNK2805 from Tn10
RC   pNK2859 from pBR322 & Tn10
RC   pNK2881 from pBR322 & Tn10
RC   pNK2882 from pACYC184 & Tn10
RC   pNK2883 from pBR322 & Tn10
RC   pNK2884 from pBR322 & Tn10
RC   pNK2885 from pBR322 & Tn10
RC   pNK2887 from pBR322 & Tn10
RC   pNK474 from pBR322 & Tn10
RC   pNK1759 from pBR322 & Tn10 & piAN7
RC   pNK2804 from pBR322 & Tn10
RC   pNK1207 from pBR322 & Tn10
RC   pNK2809 from pBR322 & Tn10
RC   pNK2811 from pBR322 & Tn10
RA   Kleckner N., Bender J., Gottesman S.;
RT   "Uses of transposons, with emphasis on Tn10";
RL   Meth. Enzymol. 204:139-180(1991).
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CC   MCS. not original sequence.
CC   NM (pGI21)
CC   CM (no)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   SP ()
CC   HO (E.coli)
CC   CP ()
CC   FN (cloning)
CC   SE ()
CC   PA (pHT1030)(pGI6136)(pGC2)
CC   BR ()
CC   OF ()
CC   OR ()
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FH   Key             Location/Qualifiers
FH
FT   misc_feature    0..0
FT                   /note="1. pLK69 Asp718 3000bp, in linker
FT                   fill in
FT                   -> pGI6130 3000bp
FT                   1. pGI6130 SacI 3000bp
FT                   Klenow
FT                   AvrII linker 6bp cctagg
FT                   -> pGI6131 3000bp
FT                   1. pGI6131 EcoRV 3000bp
FT                   NotI linker 8bp gcggccgc
FT                   -> pGI6132 3000bp
FT                   1. pLK59 HindIII 2700bp, in linker
FT                   fill in, to make NheI
FT                   -> pGI6133 2700bp
FT                   1. pGI6133 SacII 2700bp
FT                   Klenow
FT                   SpeI linker 6bp actagt
FT                   -> pGI6134 2700bp
FT                   1. pGI6134 BamHI-XbaI
FT                   2. pGI6132 remove BamHI-XbaI
FT                   -> pGI6135
FT                   1. pGI6135 SmaI
FT                   BamHI linker 6bp ggatcc
FT                   -> pGI6136 [RCP1 is BamHI-BamHI 129bp]
FT                   1. ery gene
FT                   -> pGI4010
FT                   1. pGI6136 StuI, in RCP1
FT                   2. pGI4010 KpnI-SacI, ery gene
FT                   Klenow
FT                   -> pGI6137
FT                   1. Tn10 BglII-BglII 2775bp 628..3402, tet gene/#J01830
FT                   -> pNK2805
FT                   1. pGI6137 BamHI-BamHI 1000bp, RCP1/ery gene
FT                   2. pNK2805 BamHI, in Tn10 tet gene
FT                   -> pGI6138
FT                   1. pHT1030 EcoRI-BamHI, Gram positive ori
FT                   2. pGC2 remove small EcoRI-BamHI
FT                   -> pGI10
FT                   1. pGI10 ClaI-BamHI
FT                   Klenow
FT                   2. pGI6138 EcoRI-EcoRI, IS10 transposase/ery
FT                   Klenow
FT                   -> pGI21 [2 EcoRI around pGC2]"
FT   rep_origin      0..0
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322)"
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SQ   Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;
     n
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