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ID   PBNSPNEOAL preliminary; circular DNA; SYN; 5300 BP.
AC   ATCC37803;
DT   01-JUL-1993 (Rel. 7, Created)
DT   01-APR-1995 (Rel. 11, Last updated, Version 1)
DE   Trypanosoma/E.coli phagemid vector pBNsp-Neo-Alpha - incomplete.
KW   cloning vector.
OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
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).
CC   Restriction digests of the clone give the following sizes (kb):
CC   BglIII, EcoRI--3.0,1.6,0.8; SacI--2.9,2.5; EcoRI--5.4; HindIII,
CC   BamHI--2.9,2.5; Hind III--5.4. (ATCC staff)
CC   The listed cloning sites are derived from the multiple cloning site of
CC   pBluescript KS+ and are located 3' to the 3' splice site of the
CC   alpha-tubulin sequence. (personal communication)
CC   The alpha-tubulin sequences are located downstream of the PARP
CC   promoter and were derived from the immediate upstream of the ATG in
CC   the alpha-tubulin gene to the first EcoRI site located in the upstream
CC   region. (personal communication)
CC   This is an unpublished modification of BNsp-Neo-T (ATCC 37802). (ATCC
CC   staff)
CC   Linearizing with MluI before transfection is essential for efficient
CC   integration.
CC   Other restriction sites found in the polylinker region of pBluescript
CC   SK+ (for example: SpeI, NotI, SacII, EagI, BstXI, ApaII, AccI, XhoI,
CC   HindII, etc.) have not yet been tested for their presence in the
CC   insert. (personal communication)
CC   Shuttle vector for expressing foreign DNA polycistronicly from the
CC   procyclic acidic repetitive protein (PARP) promoter. Integrates at the
CC   beta- alpha tubulin tandem array. (ATCC staff)
CC   Medium is 1227 LB plus ampicillin.
CC   NM (pBNsp-Neo-Alpha)
CC   CM (no)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (phagemid)
CC   HO (E.coli HB101)(Trypanosoma brucei)(E.coli)
CC   CP ()
CC   FN (cloning)
CC   SE ()
CC   PA ()
CC   BR ()
CC   OF ()
CC   OR ()
FH   Key             Location/Qualifiers
FT   misc_feature    0..0
FT                   /note="1. BNsp-Neo-T
FT                   2. alpha-tubulin gene
FT                   -> pBNsp-Neo-Alpha"
FT   misc_binding    0..0
FT                   /note="MCS SacI-XbaI-BamHI-HindIII-ClaI-SalI-KpnI"
FT   misc_binding    0..0
FT                   /note="SIT unique SacI-XbaI-BamHI-HindIII-ClaI-SalI-
FT                   KpnI"
FT   rep_origin      0..0
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT   rep_origin      0..0
FT                   /note="ORI bacteriophage f1"
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)"
SQ   Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;