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ID   PASZ11     preliminary; circular DNA; SYN; 5762 BP.
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AC   M59824; M32013; ATCC77128;
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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 pASZ11 - 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   pFL39-ADE2 from pFL39 & ADE2 gene
RC   pTZ19B from pTZ19R & linker
RC   pASZ5 from pTZ19B & ARS
RC   pASZ6 from pUC7 & pFL39-ADE2
RC   pASZ7 from pTZ19B & pASZ6
RC   pASZ8 from pASZ7 & pFL39
RC   pASZ9 from pASZ8 & pTZ19B
RC   pASZ11 from pFL39 & pASZ9
RC   pASZ10 from pTZ19B & pASZ9
RC   pEMBLYi32-ADE2 from pASZ8 & pEMBLYi32
RA   Stotz A., Linder P.;
RT   "The ADE2 gene from Saccharomyces cerevisiae: sequence and new
RT   vectors";
RL   Gene 95:91-98(1990).
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CC   The ADE2 segment contains HindIII sites approximately 0.9 and 2.1 kb
CC   from the 5' BglII site.
CC   and the naturally-occurring BglII site at the 3' end (nt 2512) was
CC   used to create a BglII fragment containing the complete coding
CC   sequence.
CC   The naturally-occurring BglII site within ADE2 was destroyed by
CC   replacing the A at nt 1243 of the reported sequence with a G. A BglII
CC   linker was added to the 5' end.
CC   Constructed by replacing the TRP1-containing BglII fragment of pFL39
CC   with a 2.2 kb fragment (Ars- Ade2+).
CC   In E.coli, permits visual detection of recombinants. Encodes the
CC   lacZalpha (lacZ') peptide.
CC   A YC-type shuttle expression vector containing priming sites useful
CC   for sequencing and readily purifiable elements for construction
CC   experiments.
CC   pFL39 was derived from pUC19 by inserting a ClaI fragment containing
CC   ARS-CEN at nt 747 and BglII-linkered fragment containing TRP1 at nt
CC   629.
CC   The order of the major features in this plasmid is:lacZ'/MCS -
CC   pMB1 ori - ADE2 - ARS-CEN.
CC   Restriction digests of the clone give the following sizes (kb):
CC   BglII--3.5, 2.3; EcoRI--5.8; HindIII--3.5, 1.25, 1.15; BamHI--5.8.
CC   (ATCC staff)
CC   Medium is 1227 LB plus ampicillin.
CC   NM (pASZ11)
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   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. pFL39 BamHI 4354bp 418..418
FT                   2. yeast NsiI-NsiI 5200bp, ADE2 gene
FT                   -> pFL39-ADE2 9554bp
FT                   1. pTZ19R 2863bp 416..416
FT                   BglII linker 6bp agatct
FT                   -> pTZ19B 2869bp
FT                   1. pUC7 PstI 2674bp 256..256
FT                   2. pFL39-ADE2 NsiI-NsiI 5200bp, ADE2 gene
FT                   -> pASZ6 7874bp
FT                   1. pTZ19B SalI 2869bp 193..193
FT                   2. pASZ6 SalI-SalI 5200bp, ADE2 gene/pUC7 246..-..258
FT                   -> pASZ7 8100bp
FT                   1. pASZ7 BamHI-KpnI 5200bp, ADE2 gene
FT                   exonuclease III
FT                   2. pFL39
FT                   -> pASZ8
FT                   1. pASZ8 EcoRI-SalI 2500bp, ADE2#0 gene
FT                   blunt end:blunt end
FT                   2. pTZ19B SmaI 2869bp 178..178
FT                   -> pASZ9
FT                   1. pFL39 BglII 4354bp 633..633
FT                   2. pASZ9 BglII, ADE2#1
FT                   mutagenesis
FT                   EcoRI-EcoRI 2500bp, ADE2#1
FT                   Klenow:Klenow
FT                   BglII linker 10bp nnagatctnn:BglII linker 10bp
FT                   \ nnagatctnn
FT                   -> pASZ11 5762bp"
FT   misc_binding    0..0
FT                   /note="MCS EcoRI-BanII-SacI-Asp718-KpnI-AvaI-SmaI-
FT                   XmaI-BamHI-XbaI-AccI-HincII-SalI-PstI-SphI-HindIII"
FT   misc_binding    0..0
FT                   /note="SIT unique EcoRI-BanII-SacI-Asp718-KpnI-AvaI-
FT                   SmaI-XmaI-BamHI-XbaI-AccI-SalI-PstI-SphI"
FT   rep_origin      0..0
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT   rep_origin      0..0
FT                   /note="ORI yeast ARS"
FT   rep_origin      0..0
FT                   /note="ORI bacteriophage f1"
FT   promoter        0..0
FT                   /note="PRO E. coli lac gene"
FT   CDS             0..0
FT                   /note="GEN E. coli beta-galactosidase gene (lacZ');
FT                   reporter gene"
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 ADE2 gene"
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SQ   Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;
     n
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