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ID   PAB520     preliminary; circular DNA; SYN; 4100 BP.
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AC   ATCC77137;
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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   Aspergillus/Neurospora/E.coli plasmid vector pAB520 - 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   pAB366 or pAMPh366 from pRMH200 & pAm200
RC   pAB366-1 from pAB366
RC   pAB366-2 from pAB366 & pTZ19R
RC   pAB520 or pAmPh520 from pUC18 & pAB366-1
RC   pAB366Ben from pAB366 & pBT6
RA   Austin B., Hall R.M., Tyler B.M.;
RT   "Optimized vectors and selection for transformation of Neurospora
RT   crassa and Aspergillus nidulans to bleomycin and phleomycin
RT   resistance";
RL   Gene 93:157-162(1990).
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CC   That expression was improved by modifying the 17 bp before the
CC   initiation codon in pAB366 to match the sequence of the am gene.
CC   Shuttle vector conferring bleomycin and phleomycin resistance.
CC   Expression of ble (bleomycin resistance) is regulated by the promoter
CC   and terminator regions of the am gene (amination, glutamate
CC   dehydrogenase) from N. crassa.
CC   The am::ble fusion cassette can be removed by digestion with either
CC   HindIII, SphI, XbaI or BamHI or any combination of them.
CC   The order of the major features in this plasmid is:pUC18 - MCS
CC   (HindIII to BamHI) - am promoter - ble - XbaI - am 3' region - EcoRI
CC   - MCS (HindIII to EcoRI) - pUC18.
CC   Restriction digests of the clone give the following sizes (kb):
CC   HindIII--2.7, 1.4; KpnI--4.1; SmaI--3.1, 1.0. (ATCC staff)
CC   A (GT)n repeat. (personal communication)
CC   Medium is 1227 LB plus ampicillin.
CC   NM (pAB520)
CC   CM (no)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   SP (ATCC)
CC   HO (E.coli HB101)(Aspergillus nidulans)(Neurospora crassa)(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. pAB366 4200bp, bleo gene/#U00004
FT                   mutagenesis to convert 17bp to the am one
FT                   -> pAB366-1 4200bp
FT                   1. pAB366-1 1400bp, am/bleo fusion
FT                   2. pUC18 2686bp, MCS
FT                   -> pAB520 4100bp"
FT   misc_binding    0..0
FT                   /note="MCS HindIII-SphI-PstI-SalI-XbaI-BamHI-
FT                   HindIII-SphI-PstI-SalI-XbaI-BamHI-SmaI-KpnI-SacI-
FT                   EcoRI"
FT   misc_binding    0..0
FT                   /note="SIT unique KpnI-SacI"
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 Aspergillus/Neurospora bleomycin resistance
FT                   gene (ble)"
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SQ   Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;
     n
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