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ID   PAS2       preliminary; circular DNA; SYN; 8500 BP.
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AC   IG9002; ATCC87008;
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DT   01-DEC-1994 (Rel. 10, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
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DE   Yeast/E.coli phagemid vector pAS2 - 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   pACT, pACT2 from LEU2 gene
RA   Durfee T., Becherer K., Chen P.L., Yeh S.H., Yang Y., Kilburn A.E.,
RA   Lee W.H., Elledge S.J.;
RT   "The retinoblastoma protein associates with the protein
RT   phosphatase type 1 catalytic subunit";
RL   Genes Dev. 7:555-569(1993).
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RN   [2]
RC   pAS2 from CYH2 gene & pAS1
RC   from pAS2
RC   pCMV-CIP1 from pCMVbeta
RA   Harper J.W., Adami G.R., Wei N., Keyomarsi K., Elledge S.J.;
RT   "The p21 Cdk-interacting protein Cip1 is a potent inhibitor of G1
RT   cyclin-dependent kinases";
RL   Cell 75:805-816(1993).
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CC   Restriction digests of the clone give the following sizes (kb):
CC   BamHI--8.6; SalI--8.6; SmaI--8.6. (ATCC staff)
CC   Shuttle expression vector used to create fusion proteins
CC   consisting of the nuclear localization sequence from SV40 T antigen,
CC   the GAL4 DNA-binding domain (aa 1-147), and a HA (hemagglutinin)
CC   epitope tag in frame with the activation domain. [1]
CC   The order of the major features in this plasmid is:
CC   ADC1 (ADH) promoter -> - GAL4 DNA binding domain - HA -
CC   NdeI/MCS/SalI - ADC1 terminator - pMB1 ori - ampR -
CC   2 micron ori - TRP1 -> - f1 ori - <- CYH2. [2]
CC   Growth: LB plus ampicillin (ATCC medium number 1227) 37C
CC   Deposited by: Elledge S.J.
CC   NM (pAS2)
CC   CM (no)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (phagemid)
CC   SP (CLONTECH)(ATCC)
CC   HO (E.coli)(yeast)(Saccharomyces cerevisiae)
CC   CP ()
CC   FN (cloning)(expression)(shuttle - episomal)(DNA library
CC   SE (CYH2 ribosomal protein L29)
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. pAS1 remove small BglII-SacI, 7000bp
FT                   2. yeast BamHI-SacI 1500bp, CYH2 gene
FT                   -> pAS2 8500bp"
FT   rep_origin      0..0
FT                   /note="ORI yeast 2 micron"
FT   misc_binding    0..0
FT                   /note="SIT XbaI"
FT   CDS             0..0
FT                   /note="YEA yeast TRP1 gene"
FT   misc_binding    0..0
FT                   /note="SIT XbaI"
FT   misc_binding    0..0
FT                   /note="SIT EcoRV"
FT   rep_origin      0..0
FT                   /note="ORI phage f1"
FT   misc_binding    0..0
FT                   /note="SIT NaeI"
FT   misc_binding    0..0
FT                   /note="SIT SacI"
FT   CDS             0..0
FT                   /note="YEA yeast CYH2 gene"
FT   misc_binding    0..0
FT                   /note="SIT EcoRV"
FT   misc_binding    0..0
FT                   /note="SIT EcoRI"
FT   misc_binding    0..0
FT                   /note="SIT BglII"
FT   misc_binding    0..0
FT                   /note="SIT XbaI"
FT   misc_binding    0..0
FT                   /note="SIT EcoRV"
FT   promoter        0..0
FT                   /note="PRO yeast ADH1 gene"
FT   promoter        0..0
FT                   /note="PRO yeast GAL4 bd"
FT   misc_feature    0..0
FT                   /note="HA sequences"
FT   misc_binding    0..0
FT                   /note="SIT EcoRI"
FT   misc_binding    0..0
FT                   /note="MCS unique NdeI-SfiI-NcoI-SmaI-BamHI-SalI"
FT   terminator      0..0
FT                   /note="TER yeast ADH1 gene"
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/amr)"
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SQ   Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;
     n
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