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ID   PASZ10     preliminary; circular DNA; SYN; 5396 BP.
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AC   M59824; M32013; ATCC77127;
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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 phagemid vector pASZ10 - 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 1.17 kb and
CC   2.36 kb from the 5' BglII site.
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   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   Constructed by inserting a 2.5 kb fragment (Ars+ Ade2+) into the BglII
CC   site of pTZ19B.
CC   A YR-type shuttle expression vector containing promoters for in vitro
CC   RNA synthesis and priming sites useful for sequencing. Contains
CC   readily purifiable elements for construction experiments.
CC   In E.coli, permits visual detection of recombinants and production of
CC   ssDNA. Encodes the lacZalpha (lacZ') peptide.
CC   pTZ19B was derived from pTZ19R by adding a BglII linker at coordinate
CC   416.
CC   The order of the major features in this plasmid is:lacZ'/MCS - T7
CC   promoter - ADE2 - pMB1 ori - ampR - f1 ori.
CC   Restriction digests of the clone give the following sizes (kb):
CC   BglII--2.9, 2.6; EcoRI--5.6; HindIII--2.9, 1.3, 1.1, 0.41; BamHI--5.6.
CC   (ATCC staff)
CC   Medium is 1227 LB plus ampicillin.
CC   NM (pASZ10)
CC   CM (no)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (phagemid)
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 KpnI-BamHI 4354bp 413..418
FT                   -> pASZ8 9500bp
FT                   1. pASZ8 EcoRI-SalI 2500bp, ADE2#0 gene
FT                   \ pFL39 397..-..430
FT                   blunt end:blunt end
FT                   2. pTZ19B SmaI 2869bp 178..178
FT                   -> pASZ9 5400bp
FT                   1. pTZ19B BglII 2869bp 416..416
FT                   2. pASZ9 BglII, ADE2#0
FT                   mutagenesis
FT                   EcoRI-EcoRI 2500bp, ADE2#0
FT                   Klenow:Klenow
FT                   BglII linker 10bp nnagatctnn:BglII linker 10bp
FT                   \ nnagatctnn
FT                   -> pASZ10 5396bp"
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-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   promoter        0..0
FT                   /note="PRO bacteriophage T7"
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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