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ID   PBNSPNEOT  preliminary; circular DNA; SYN; 6200 BP.
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AC   ATCC37802;
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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   Trypanosoma/E.coli plasmid vector pBNsp-Neo-T - 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   pBNsp-Neo-T from pUC18 & neo gene & T.brucei PARP gene promoter
RA   Lee M.G., Van der Ploeg L.H.;
RT   "Homologous recombination and stable transfection in the parasitic
RT   protozoan Trypanosoma brucei";
RL   Science 250:1583-1587(1990).
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RN   [2]
RC   pUC18-PARP from pUC18 & B1 PARP locus
RC   pBNspCAT-0, pBNspCAT-1 from pCOCAT & pUC18-PARP
RC   pdeltaBNspCAT from pBNspCAT-1
RC   pBSsp-CAT from pCOCAT & pUC18-PARP
RC   pBintCAT-0, pBintCAT-1 from pCOCAT & pUC18-PARP
RC   pEMR1, pEMR2, pEMR2.5, pEMR3 from pBluescript series & trypanosome
RC   pEMR5 from pBluescript series & trypanosome
RA   Rudenko G., Le Blancq S.,  Smith J., Lee M.G., Rattray A.,
RA   Van der Ploeg L.H.;
RT   "Procyclic acidic repetitive protein (PARP) genes located in an
RT   unusually small alpha-amanitin-resistant transcription unit: PARP
RT   promoter activity assayed by transient DNA transfection of
RT   Trypanosoma brucei";
RL   Mol. Cell. Biol. 10:3492-3504(1990).
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RN   [3]
RC   pAL91 from pAL61
RC   pJF2 from pAL61 & parpA gene promoter
RC   pJF3 from pAL61 & parpB gene promoter
RC   pJF5 from pJF2
RC   pJF6 from pJF2
RC   pJF7 from pAL61 & parpA gene promoter
RC   pJP9, pJP10, pJP11, pJP13, pJP14A, pJP14B from pJF6
RA   Clayton C.E., Fueri J.P., Itzhaki J.E., Bellofatto V., Sherman D.R.,
RA   Wisdom G.S., Vijayasarathy S., Mowatt M.R.;
RT   "Transcription of the procyclic acidic repetitive protein genes of
RT   Trypanosoma brucei";
RL   Mol. Cell. Biol. 10:3036-3047(1990).
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RN   [4]
RC   pTBR1 from T. brucei parp gene
RC   pTBR3 from T. brucei parp gene
RA   Campbell D.A., Kubo K., Clark C.G., Boothroyd J.C.;
RT   "Precise identification of cleavage sites involved in the unusual
RT   processing of trypanosome ribosomal RNA";
RL   J. Mol. Biol. 196:113-124(1987).
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RN   [5]
RC   pPRO2001 from T. brucei parpBbeta gene
RA   Roditi I., Carrington M., Turner M.;
RT   "Expression of a polypetide containing a dipeptide repeat is
RT   confined to the insect stage of Trypanosoma brucei";
RL   Nature 325:272-274(1987).
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RN   [6]
RC   Gemini 3 from T.brucei
RC   pAL61 from Gemini 3 & L. seymouri mini-exon promoter
RA   Bellofatto V., Cross G.A.;
RT   "Expression of a bacterial gene in a trypanosomal protozoan";
RL   Science 244:1167-1169(1989).
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RN   [7]
RC   pCOCAT from pLW2
RA   Lowndes N.F., Paul J., Wu J., Allan M.;
RT   "C-Ha-ras gene bidirectional promoter expressed in vitro: location
RT   and regulation";
RL   Mol. Cell. Biol. 9:3758-3770(1989).
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CC   Restriction digests of the clone give the following sizes (kb):
CC   HindIII, BamHI--3.5,2.7; EcoRI, BglII--5.6,0.6; BamHI--6.2; MluI--6.2.
CC   (ATCC staff)
CC   Constructed from the PARP promoter and 3' splice site from BNsp-CAT,
CC   neo (BglII/BamHI fragment) from pSV2neo, and a tubulin intergenic
CC   region (639 bp, blunted EcoRI/BglII) inserted into the SmaI site 3'
CC   to the termination codon of neo.
CC   Linearizing with MluI before transfection is essential for efficient
CC   integration. The MluI site is in the center of the tubulin intergenic
CC   sequence.
CC   Shuttle vector for integration by homologous recombination targeted
CC   to the beta- alpha-tubulin intergenic region of Trypanosoma brucei.
CC   (ATCC staff)
CC   Medium is 1227 LB plus ampicillin.
CC   NM (pBNsp-Neo-T)
CC   CM (no)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   SP (ATCC)
CC   HO (E.coli HB101)(Trypanosoma brucei)(E.coli)
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. pUC18
FT                   2. PARP promoter
FT                   3. cat gene
FT                   -> BNsp-CAT
FT                   1. BNsp-CAT remove BglII-BamHI, cat gene
FT                   2. pSV2-neo BglII-BamHI, neo gene
FT                   -> plasmid
FT                   1. plasmid SmaI, 3' to neo gene
FT                   2. EcoRI-BglII 639bp, alpha-tubulin gene IVS
FT                   blunt end:blunt end
FT                   -> pBNsp-Neo-T"
FT   misc_binding    0..0
FT                   /note="SIT unique HindIII-BamHI"
FT   rep_origin      0..0
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT   promoter        0..0
FT                   /note="PRO E. coli PARP gene"
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 Trypanosoma G418 resistance gene (G418)"
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SQ   Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;
     n
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