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ID   PSI4       preliminary; circular DNA; SYN; 11900 BP.
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AC   ATCC77171;
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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 pSI4 - 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   pMJ5 from pBR322 & yeast 2 micron B form
RC   pMJ6 from pBR322 & yeast 2 micron B form
RC   pC4 from pBR322 & yeast 2 micron ori & LEU2 gene
RC   pC4B from pC4
RC   pCV7 from pBR322 & yeast 2 micron & LEU2 gene
RC   pCV19 from pBR322 & yeast 2 micron & LEU2 gene
RC   pCV20 from pBR322 & LEU2 gene
RC   pCV21 from pBR322 & yeast 2 micron & LEU2 gene
RC   pSI4 from pJDB219 & pBR322
RC   YEp13 from pBR322 & yeast 2 micron & LEU2 gene
RC   YEp13S from YEp13
RC   YEp6 from pBR322 & yeast 2 micron & HIS3 gene
RC   YEp24 from pBR322 & yeast 2 micron & URA3 gene
RC   YEp16 from pBR322 & yeast 2 micron & URA3 gene
RC   YEp420 or beta72 from YIp5 & yeast 2 micron form B
RC   pJDB207 from pJDB219 & pAT153
RA   Broach J.R.;
RT   "Construction of high copy yeast vectors using 2-micron circle
RT   sequences";
RL   Meth. Enzymol. 101:307-325(1983).
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CC   Consists of the entire 2 micron circle genome cloned, at the EcoRI
CC   site of the small unique region, into the EcoRI site of pBR322.  The
CC   leu2-d sequence (approximately 1.4 kb) is inserted into the PstI site
CC   of the 2 micron circle. [2]
CC   High-copy YE-type shuttle vector. [2]
CC   Restriction digests of the clone give the following sizes (kb):
CC   BamHI--12.0; EcoRI--4.4, 4.1, 2.6, 0.8; HindIII--4.6, 3.4, 2.7, 1.3;
CC   PstI--12.0. (ATCC staff)
CC   Medium is 1227 LB plus ampicillin.
CC   NM (pSI4)
CC   CM (no)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   SP (ATCC)
CC   HO (E.coli)(Saccharomyces cerevisiae)
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. pMB9 EcoRI 5300bp, 5' to tet gene
FT                   2. yeast 2 micron B form EcoRI-EcoRI 2235bp
FT                   \ 2408..3942 & 703..2
FT                   \ UL = 3239bp 703..3942 US = 3079bp 3942..6318..703 A
FT                   \ UL = 3239bp 703..3942 US = 3079bp 703..6318..3942 B
FT                   -> pJBD41 7500bp [tet gene]
FT                   1. pJBD41 PstI 7500bp, amp gene
FT                   \ yeast 2 micron 2657..2657
FT                   dAT tailing
FT                   2. yeast 1200bp, LEU2 gene
FT                   sheared
FT                   dAT tailing
FT                   -> pJDB219 8700bp [HindIII-HindIII 3600bp, ori/leu2]
FT                   1. pJDB219 remove EcoRI-EcoRI 5300bp, pMB9/7500bp
FT                   2. pBR322 EcoRI 4361bp 4360..4360
FT                   -> pSI4 11900bp"
FT   misc_binding    0..0
FT                   /note="SIT unique BamHI-SalI-PstI-HpaI"
FT   rep_origin      0..0
FT                   /note="ORI yeast 2 micron ori"
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 E. coli tetracycline resistance gene (tet)"
FT   CDS             0..0
FT                   /note="ANT yeast LEU2-d gene"
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SQ   Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;
     n
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