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ID   PAB366     preliminary; circular DNA; SYN; 4200 BP.
AC   ATCC77134;
DT   01-JUL-1993 (Rel. 7, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
DE   Aspergillus/Neurospora/E.coli plasmid vector pAB366 - incomplete.
KW   cloning vector.
OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
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).
RN   [2]
RC   pRMH200 from pBR322 & Tn5, ble gene
RC   pRMH201, pRMH207, pRMH209 from pRMH200
RC   pRMH213 from pRMH207
RC   pRMH215, pRMH219 from pRMH213
RC   pRMH214 from pRMH215
RA   Collis C.M., Hall R.M.;
RT   "Identification of a Tn5 determinant conferring resistance to
RT   phleomycins, bleomycins and tallisomycins";
RL   Plasmid 14:143-151(1985).
RN   [3]
RC   clone 10 from lambda L47 & Neurospora
RC   [pAm200 from pUC8 & Neurospora crassa am gene]
RA   Kinnaird J.H., Keighren M.A., Kinsey J.A., Eaton M., Finscham J.R.;
RT   "Cloning of the am (glutamate dehydrogenase) gene of Neurospora
RT   crassa through the use of a synthetic DNA probe";
RL   Gene 20:387-396(1982).
RN   [4]
RC   pBT3 from pUC12 & N. crassa beta-tubulin gene
RC   [pBT6 from pUC12 & N. crassa beta-tubulin mutant gene (ben gene)]
RA   Orbach M.J., Porro E.B., Yanofsky C.;
RT   "Cloning and characterization of the gene doe beta-tubulin from a
RT   benomyl-resistant mutant of Neurospora crassa and its use as a
RT   dominant selectable marker";
RL   Mol. Cell. Biol. 6:2452-2461(1986).
RN   [5]
RC   pMOO6 from pUC19 & pMT201, A.nidulans argB gene
RA   Upshall A.;
RT   "Genetic and molecular characterization of argB transformants
RT   of Apergillus nidulans";
RL   Curr. Genet. 10:593-599(1986).
RN   [6]
RC   lambda L47 from lambda
RA   Loenen W.A., Brammar W.J.;
RT   "A bacteriophage lambda vector for cloning large DNA fragments made
RT   with several restriction enzymes";
RL   Gene 10:249-259(1980).
RN   [7]
RC   pMT201 from pBR329 & A.nidulans argB gene
RA   Timberlake W.E.;
RT   ;
RL   Unpublished (1986).
CC   not original sequence. fusion regions.
CC   Shuttle vector conferring bleomycin and phleomycin resistance.
CC   The order of the major features in this plasmid is:pUC8 - EcoRI -
CC   BamHI - am promoter - ble - XhoI - am 3' region - HindIII - pUC8.
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   Restriction digests of the clone give the following sizes (kb):
CC   BamHI/HindIII--2.7,1.5; BamHI/XhoI--3.4, 0.8; BglI--3.0, 1.2. (ATCC
CC   staff)
CC   A (GT)n repeat. (personal communication)
CC   Medium is 1227 LB plus ampicillin.
CC   NM (pAB366)
CC   CM (no)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
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 ()
FH   Key             Location/Qualifiers
FT   misc_feature    0..0
FT                   /note="1. pUC8 BamHI 2665bp 241..241
FT                   2. Neurospora crassa BamHI-BamHI 2700bp, am gene
FT                   -> JR2 5365bp
FT                   1. JR2 remove KpnI-PstI, Neurospora to pUC8 PstI 257
FT                   -> pAm200
FT                   1. pBR322 4361bp, in tet gene
FT                   transposition
FT                   2. Tn5, bleo gene/#U00004
FT                   -> pRMH200
FT                   1. pRMH200 AsuII-XhoI, Tn5 bleo gene
FT                   2. pAm200 ClaI-XhoI, N.crassa am gene
FT                   -> pAB366 4200bp"
FT   misc_binding    0..0
FT                   /note="SIT unique BamHI-HindIII"
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)"
SQ   Sequence 50 BP; 14 A; 20 C; 5 G; 11 T; 0 other;
     ctcatccatc ttgccctccc accaagcaca actttatcga aatgaccgac