back Return to this vector's summary.
ID   PBP96      preliminary; circular DNA; SYN; 4600 BP.
AC   ATCC77084;
DT   01-JUL-1993 (Rel. 7, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
DE   Saccharomyces/E.coli plasmid vector pBP96 - incomplete.
KW   cloning vector.
OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
RN   [1]
RC   plasmid from pBluescript series & HIS3 gene
RC   pBP96 from BLUR8 & plasmid
RC   pBP97 from BLUR8 & plasmid
RC   pBP90 from HIS3 gene & pL1.1A, LINE
RA   Pavan W.J., Reeves R.H.;
RT   "Integrative selection of human chromosome-specific yeast
RT   artificial chromosomes";
RL   Proc. Natl. Acad. Sci. U.S.A. 88:7788-7791(1991).
RN   [2]
RC   from pBP62
RC   from pYAC3
RC   from pYAC4
RC   from pYAC55
RC   from pJS97
RC   from pJS98
RC   from pYACneo
RC   from pYAC-RC
RC   from pCGS966
RC   from pBP81
RC   from pBP100 series
RC   pBP90 from human LINE repeats
RC   pBP96 from human Alu repeats
RC   pBP97 from human Alu repeats
RC   pBP47 from human Alu repeats & HIS3 & SV40 promoter & neo
RA   Reeves R.H., Pavan W.J., Hieter P.;
RT   "Yeast artificial chromosome modification and manipulation";
RL   Meth. Enzymol. 216:584-603(1992).
CC   YI-type shuttle integrative vector targeting human Alu sequences in
CC   YAC libraries derived from somatic cell hybrids. [1]
CC   Constructed by inserting a blunt-ended 1.2 kb fragment containing S.
CC   cerevisiae HIS3 into the blunted MscI within the Alu sequence of
CC   BLUR8. [1]
CC   Digestion with BamHI removes the plasmid sequences from the Alu ends
CC   for targeting integration. [1]
CC   Transformation into pools from a YAC library derived from a
CC   human-mouse somatic cell hybrid, and selection for His+, resulted in
CC   an approximate 50-fold enrichment for Alu-containing YACs. [1]
CC   Larger YACs were preferentially selected.  Some YACs demonstrated loss
CC   of Alu-containing restriction fragments suggesting internal deletions
CC   resulting when the vector targeted two Alu sequences in the YAC. [1]
CC   Restriction digests of the clone give the following sizes (kb):
CC   BamHI--3.0, 1.6; EcoRI--4.6; PstI--3.3, 1.3. (ATCC staff)
CC   The order of the major features in this plasmid is: bla - BamHI - Alu
CC   - HIS3 - Alu - BamHI. (personal communication)
CC   Medium is 1227 LB plus ampicillin.
CC   NM (pBP96)
CC   CM (no)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   HO (E.coli HB101)(Saccharomyces cerevisiae)(E.coli)
CC   HO (Saccharomyces cerevisiae YPH252)
CC   HO (Saccharomyces cerevisiae CGY2516 [ATCC 74013])
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. pBluescript EcoRI 2746bp 424..424, MCS
FT                   Klenow
FT                   2. yeast, -296 of HIS3 gene/pYAC4 5839..5839
FT                   \ pYAC4 1768bp 4238..6006/CDS = complement(4884..5543)
FT                   EcoRI linker 6bp gaattc
FT                   XhoI, end of HIS3 gene/pYAC4 4677..4677
FT                   BamHI linker 6bp ggatcc
FT                   EcoRI-BamHI 1184bp, HIS3 gene/4671..5845
FT                   Klenow:Klenow
FT                   -> plasmid 3950bp
FT                   1. pBR322, ori/amp
FT                   2. human, Alu repeat
FT                   linker
FT                   -> BLUR8 3400bp
FT                   1. BLUR8 MscI, ori/amp/human Alu repeat 3400bp
FT                   \ pBR322 1447..1447
FT                   blunt end:blunt end
FT                   2. plasmid SmaI-EcoRV 1202bp,
FT                   \ pBluescript 424..-..424..442
FT                   blunt end:blunt end
FT                   -> pBP96 4600bp"
FT   rep_origin      0..0
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322)"
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 yeast HIS3 gene"
SQ   Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;