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ID   MTCHI5     preliminary; circular DNA; SYN; 10700 BP.
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AC   ATCC39275;
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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 Mtchi5 - 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   Mtchi5 from YEp13 & DHFR gene
RA   ;
RT   ;
RL   Unpublished (1987).
RL   U.S. Patent No. 4,670,399 dated June 2, 1987.
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CC   This vector contains the following sites separated by (bp):
CC   Sau3A I - 130 - BamHI - 280 - SalI - 3700 - EcoRI - 2230 - EcoRI -
CC   860 - SalI - 3250 - HindIII - 255 - Sau3A I.
CC   A 370 bp HaeIII/AluI fragment encoding dihydrofolate reductase (ends
CC   filled, BamHI & HindIII linkers added) was inserted between the BamHI
CC   and HindIII sites of YEp13.  DHFR transcription runs from BamHI toward
CC   the HindIII site.
CC   An episomal Saccharomyces cerevisiae/E.coli shuttle vector.
CC   Restriction digests of the clone give the following sizes (kb):
CC   BamHI--11.0; HindIII--11.0; EcoRI--11.5. (ATCC staff)
CC   Medium is 1227 LB plus ampicillin.
CC   NM (Mtchi5)
CC   CM (no)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   SP (ATCC)
CC   HO (E.coli LE392)(Saccharomyces cerevisiae)(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. pBR322 EcoRI 4361bp 4360..4360
FT                   2. yeast 2 micron EcoRI-EcoRI 2406bp 2..2408, ori
FT                   -> plasmid 6767bp
FT                   1. plasmid SalI 6767bp 652..652
FT                   2. pYe(Leu2)10 SalI-XhoI 2200bp, LEU2 gene
FT                   -> YEp13 9000bp
FT                   1. YEp13 remove BamHI-HindIII 346bp, pBR322 30..376
FT                   \ 8600bp [385bp]
FT                   2. yeast HaeIII-AluI 2100bp, DHFR gene
FT                   fill in:fill in
FT                   BamHI linker 6bp ggatcc:HindIII linker 6bp aagctt
FT                   -> Mtchi5 10700bp"
FT   misc_binding    0..0
FT                   /note="SIT unique BamHI-HindIII"
FT   rep_origin      0..0
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT   rep_origin      0..0
FT                   /note="ORI yeast 2 micron"
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 DHFR gene"
FT   CDS             0..0
FT                   /note="ANT yeast LEU2 gene"
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SQ   Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;
     n
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