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ID   PBP81      preliminary; circular DNA; SYN; 6200 BP.
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AC   ATCC77086;
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DT   01-JUL-1993 (Rel. 7, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
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DE   Saccharomyces/E.coli plasmid vector pBP81 - 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   plasmid from YTCA-1x & pSP73
RC   pBP103 from plasmid & pRS303
RC   pBP108, pBP109 from pBP103 & BLUR8
RC   pBP110, pBP111 from pBP103 & pL1.1A
RC   pBP81 from pBP63A & YTCA-1x
RA   Pavan W.J., Hieter P., Sears D., Burkhoff A., Reeves R.H.;
RT   "High-efficiency yeast artificial chromosome fragmentation
RT   vectors";
RL   Gene 106:125-127(1991).
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RN   [2]
RC   pBP62 from pRS303 & pJS89, yeast Y' sequence
RC   pBP63A, pBP63B from pBP62 & BLUR8, human Alu
RA   Pavan W.J., Hieter P., Reeves R.H.;
RT   "Generation of deletion derivatives by targeted transformation of
RT   human-derived yeast artificial chromosomes";
RL   Proc. Natl. Acad. Sci. U.S.A. 87:1300-1304(1990).
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RN   [3]
RC   pJS71 from yeast YAC
RC   pJS73 from yeast YAC
RC   pJS89 from yeast Y' sequence
RA   Shero J., Hieter P.;
RT   "A suppressor of a centromere DNA mutation encodes a putative
RT   protein kinase (MCK1)";
RL   Genes Dev. 5:549-560(1990).
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CC   Constructed by replacing the telomere-adjacent sequence from pBP63A
CC   with a 0.25 kb SalI/SphI fragment containing the YTCA-1x telomere and
CC   CEN6 sequences. [1]
CC   Linearize with SalI (site adjacent to the telomere sequence) before
CC   transformation. [1]
CC   Contains 2 head to tail Alu sequences with the 5' end closer to the
CC   SalI site. [1]
CC   Centric chromosome fragmentation vector targeting Alu sequences. [1]
CC   Restriction digests of the clone give the following sizes (kb):
CC   BamHI--5.6, 0.35, 0.25; PstI--4.4, 1.4, 0.4; SstI--5.4, 0.8;
CC   SalI/SphI--6.2. (ATCC staff)
CC   The order of the major features in this plasmid is: bla - HIS3 -
CC   3'Alu5' - 3'Alu5' - SalI - TEL - CEN6. [1]
CC   Medium is 1227 LB plus ampicillin.
CC   NM (pBP81)
CC   CM (no)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   SP (ATCC)
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 ()
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FH   Key             Location/Qualifiers
FH
FT   misc_feature    0..0
FT                   /note="1. yeast 2250bp, Y' sequence
FT                   -> pJS89
FT                   1. pRS303 PvuII 4443bp 1953..1953, may be 2398..2398
FT                   2. pJS89 2250bp, yeast Y' sequence
FT                   blunt end:blunt end
FT                   -> pBP62 6700bp [ori, amp, HIS3, lacZ, Y']
FT                   1. pBR322 4361bp, ori/amp
FT                   2. human 300bp, Alu repeat
FT                   linker
FT                   -> BLUR8 4700bp
FT                   1. pBP62 BamHI 6700bp, pRS303 2110..2110
FT                   2. BLUR8 BamHI-BamHI 300bp, human Alu repeats
FT                   -> pBP63A 7300bp
FT                   1. pBR322 4361bp
FT                   2. yeast 250bp, telomere
FT                   -> YTCA-1x 4600bp
FT                   1. pBP63A remove SalI-SphI 1300bp,
FT                   \ yeast Y' sequence/6000bp
FT                   2. YTCA-1x SalI-SphI 250bp, telomere/CEN6
FT                   -> pBP81 6200bp"
FT   rep_origin      0..0
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT   misc_feature    0..0
FT                   /note="yeast centromere CEN6"
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"
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SQ   Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;
     n
//