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ID   PET9C      preliminary; circular DNA; SYN; 1 BP.
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AC   IG2024;
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DT   01-JUN-1993 (Rel. 7, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
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DE   E. coli plasmid vector pET9c - complete.
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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   pLysE, pLysS from pACYC184 & T7 lysozyme gene
RC   phage CE6 from lambda
RC   plasmid from pBR322 & lacI gene
RC   pET-9 from kan gene pET-7 & pET-3
RC   pET-10 from pET-7 & lac operator & plasmid
RC   pET-11 from pET-7 & lac operator & plasmid & pET-3
RC   pET-3d from pET-3
RC   pET-8, pET-8c from pET series
RA   Studier F.W., Rosenberg A.H., Dunn J.J., Dubendorff J.W.;
RT   "Use of T7 RNA polymerase to direct expression of cloned genes";
RL   Meth. Enzymol. 185:60-89(1990).
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CC   NM (pET9c)
CC   CM (yes)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   SP (NEB)
CC   HO (E.coli)
CC   CP ()
CC   FN (expression)
CC   SE ()
CC   PA (pBR322)
CC   BR ()
CC   OF ()
CC   OR ()
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FH   Key             Location/Qualifiers
FH
FT   misc_feature    0..0
FT                   /note="1. pBR322 4361bp
FT                   2. Tn5 XhoI-XhoI 2347bp 486..2833,
FT                   \ kan/neo gene with BglII
FT                   -> pKP772 6708bp
FT                   1. pKP772 6708bp
FT                   linker
FT                   -> pFB70 6708bp
FT                   1. pUC4K EcoRI-EcoRI 2632bp 1679..3914..397
FT                   2. linker EcoRI-BamHI-SacI-SmaI/XmaI-HindIII-XhoI
FT                   3. pFB70 HindIII-XhoI 1637bp 1196..2833,
FT                   \ Tn5 kan/neo gene
FT                   4. linker XhoI-HindIII-SmaI/XmaI-SacI-BamHI-EcoRI
FT                   -> pUC4KISS 4200bp
FT                   1. pBR322 BamHI 4361bp 376..376, tet
FT                   2. T7 HhaI-HhaI 122bp 24106..24228 -104 to +19,
FT                   \ Tphi term
FT                   -> pAR939 4483bp
FT                   1. pAR939 BamHI 4483bp, pBR322 376..376
FT                   Klenow
FT                   BglII linker 8bp gagatctc
FT                   -> pAR2516 4483bp
FT                   1. pBR322 BamHI 4361bp 376..376, tet
FT                   2. T7 TaqI-XbaI 48bp 22880..22928, phi10
FT                   BamHI linker 10bp cgggatcccg:BamHI linker 10bp
FT                   \ cgggatcccg
FT                   -> pAR1959 4429bp
FT                   1. pAR1959 BamHI 4429bp
FT                   Klenow
FT                   BglII linker 8bp gagatctc
FT                   -> pET-2 4430bp
FT                   1. pET-2 PstI-BamHI 1400bp, phi10
FT                   2. pAR2516 BamHI-PstI 3238bp, pBR322 376..3614
FT                   -> pET-3 4600bp
FT                   1. pET-3 remove small BspHI-EcoRI 50bp, amp gene
FT                   \ 4600bp
FT                   \ pBR322 4309..4360 or 4204..4360
FT                   2. pUC4KISS BspHI-EcoRI 867bp, kan gene
FT                   \ pUC4K BspHI = 1582 2755 3763 3868
FT                   \ pUC4K EcoRI = 397 1679
FT                   -> pET-9 5500bp
FT                   1. pET-9 BglII 5500bp, 3' to tphi
FT                   EcoRV linker 6bp gatatc
FT                   -> pET-9c 5500bp"
FT   misc_binding    23..23
FT                   /note="SIT unique ClaI"
FT   misc_binding    29..29
FT                   /note="SIT unique HindIII"
FT   promoter        complement(185..461)
FT                   /note="PRO bacteriophage T7"
FT   promoter        185..461
FT                   /note="MCS"
FT   terminator      194..316
FT                   /note="PRO bacteriophage T7 phi10 gene"
FT   promoter        403..427
FT                   /note="PRO E. coli lac operator"
FT   misc_feature    429..429
FT                   /note="bacteriophage T7 transcription initiation
FT                   site"
FT   promoter        430..446
FT                   /note="PRO bacteriophage T7 phi10 gene"
FT   misc_binding    461..461
FT                   /note="SIT unique BglI"
FT   misc_binding    654..654
FT                   /note="SIT unique SphI"
FT   misc_binding    714..714
FT                   /note="SIT unique EcoNI"
FT   CDS             833..1915
FT                   /note="REP E. coli lacIq gene"
FT   misc_binding    904..904
FT                   /note="SIT unique DrdII"
FT   misc_binding    1183..1183
FT                   /note="SIT unique MluI"
FT   misc_binding    1197..1197
FT                   /note="SIT unique BclI"
FT   misc_binding    1364..1364
FT                   /note="SIT unique BstEII"
FT   misc_binding    1390..1390
FT                   /note="SIT unique ApaI"
FT   misc_binding    1594..1594
FT                   /note="SIT unique BssHII"
FT   misc_binding    2024..2024
FT                   /note="SIT unique PshAI"
FT   misc_binding    2251..2251
FT                   /note="SIT unique EagI"
FT   misc_binding    2284..2284
FT                   /note="SIT unique NruI"
FT   misc_binding    2375..2375
FT                   /note="SIT unique BspMI"
FT   misc_binding    2665..2665
FT                   /note="SIT unique BsmI"
FT   misc_binding    2737..2737
FT                   /note="SIT unique AvaI"
FT   misc_binding    2756..2756
FT                   /note="SIT unique MscI"
FT   misc_binding    2892..2892
FT                   /note="SIT unique Bpu10I"
FT   CDS             >3000..<3558
FT                   /note="REP E. coli ROP gene"
FT   misc_binding    3531..3531
FT                   /note="SIT unique Tth111I"
FT   misc_binding    3539..3539
FT                   /note="SIT unique BsaAI"
FT   misc_binding    3558..3558
FT                   /note="SIT unique AccI"
FT   misc_binding    3666..3666
FT                   /note="SIT unique SapI"
FT   rep_origin      complement(>3666..<4400)
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT   rep_origin      complement(3850..3850)
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322) start"
FT   misc_binding    4200..4200
FT                   /note="SIT unique AlwNI"
FT   CDS             complement(4619..5469)
FT                   /note="ANT E. coli beta-lactamase gene (bla)
FT                   ampicillin resistance gene (apr/amp)"
FT   misc_binding    4677..4677
FT                   /note="SIT unique Eam1105I"
FT   misc_binding    4749..4749
FT                   /note="SIT unique BsaI"
FT   misc_binding    4923..4923
FT                   /note="SIT unique PstI"
FT   misc_binding    5049..5049
FT                   /note="SIT unique PvuI"
FT   misc_binding    5160..5160
FT                   /note="SIT unique ScaI"
FT   misc_binding    5484..5484
FT                   /note="SIT unique SspI"
FT   misc_binding    5600..5600
FT                   /note="SIT unique AatII"
FT   misc_binding    5675..5675
FT                   /note="SIT unique EcoRI"
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SQ   Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;
     n
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