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ID   MTCHI4     preliminary; circular DNA; SYN; 10700 BP.
AC   ATCC39276;
DT   01-JUL-1993 (Rel. 7, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
DE   Saccharomyces/E.coli plasmid vector Mtchi4 - incomplete.
KW   cloning vector.
OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
RN   [1]
RC   Mtchi4 from YEp13 & DHFR gene
RA   ;
RT   ;
RL   Unpublished (1987).
RL   U.S. Patent No. 4,670,399 dated June 2, 1987.
CC   This vector contains the following sites separated by (bp):
CC   Sau3A I - 255 - BamHI - 280 - SalI - 3700 - EcoRI - 2230 - EcoRI -
CC   860 - SalI - 3250 - HindIII - 130 - Sau3A I.
CC   DHRF transcription runs from HindIII toward the BamHI site.  This
CC   plasmid does NOT confer methotrexate resistance in yeast but does
CC   confer trimethoprim resistance in E.coli.
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.
CC   An episomal Saccharomyces cerevisiae/E.coli shuttle vector.
CC   Restriction digests of the clone give the following sizes (kb):
CC   HindIII--10.5; BamHI--10.5; PstI--5.8, 4.4. (ATCC staff)
CC   Medium is 1227 LB plus ampicillin.
CC   NM (Mtchi4)
CC   CM (no)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   HO (E.coli LE392)(Saccharomyces cerevisiae)(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 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 HindIII-BamHI 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                   -> Mtchi4 10700bp"
FT   misc_binding    0..0
FT                   /note="SIT unique HindIII-BamHI"
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 LEU2 gene"
SQ   Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;