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ID   PTACNEO12  preliminary; circular DNA; SYN; 5440 BP.
AC   ATCC37687;
DT   01-JUL-1993 (Rel. 7, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
DE   Vertebrate/E.coli plasmid vector pTacNeo1.2 - incomplete.
KW   cloning vector.
OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
RN   [1]
RC   pTacNeo1.2 from pBR322 & tac promoter & Tn601/Tn5 APH(3')-I gene
RA   Hunter-Cevera J.C.;
RT   ;
RL   Unpublished (1993).
CC   The upstream translation of LacZ is out of frame with APH(3')-I.
CC   (personal communication)
CC   The vector contains pBR322 (EcoRI-SphI), a TAC promoter, the first 8
CC   amino-terminal codons of beta-galactosidase, & a gene encoding a
CC   modified aminoglycoside phosphotransferase (3')-I (APH(3')-I).
CC   (personal communication)
CC   The start of the modified APH(3')-I is followed by the eleventh codon
CC   (serine) of APH(3')-I.  Codons 2-10 have been deleted. (personal
CC   communication)
CC   Sequences from the TAA stop codon of APH(3')-I to the immediate
CC   downstream PvuII site encode a portion of Tn601. Sequences from the
CC   PvuII site to the PstI site are derived from Tn5. (personal
CC   communication)
CC   This is an expression vector capable of producing a
CC   beta-galactosidase: insert: APH(3')-I fusion protein. (personal
CC   communication)
CC   Restriction digests of the clone give the following sizes (kb):
CC   BamHI--5.45; XhoI--5.45; EcoRI--5.45; PvuI--4.0, 1.45; PstI--3.1,
CC   2.35. (ATCC staff)
CC   Medium is 1227 LB plus ampicillin.
CC   NM (pTacNeo1.2)
CC   CM (no)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   HO (E.coli DG101)(vertebrate cells)(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. pBR322 SphI-EcoRI 3793bp 567..4360,
FT                   \ ori/amp gene
FT                   2. E. coli, tac promoter
FT                   3. E. coli, lac gene
FT                   4. Tn601, TAA stop codon of APH(3')-I to PvuII
FT                   5. Tn5 PvuII-PstI 107bp 2548..2655, APH(3')-I gene
FT                   -> pTacNeo1.2 5440bp"
FT   misc_binding    0..0
FT                   /note="MCS XmaIII-BamHI-XhoI"
FT   misc_binding    0..0
FT                   /note="SIT unique BamHI"
FT   rep_origin      0..0
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT   promoter        0..0
FT                   /note="PRO E. coli tac"
FT   CDS             0..0
FT                   /note="ANT E. coli beta-lactamase gene (bla)
FT                   ampicillin resistance gene (apr/amp)"
SQ   Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;