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ID PLA2917 preliminary; circular DNA; SYN; 21000 BP.
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AC ATCC37355; ATCC39840;
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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 Broad host range/E.coli cosmid vector pLA2917 - 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 pLA2901 from pRK290 & pBR322
RC pLA2905 from pLA2901
RC pLA2910, pLA2917 from pLA2901 & pMF517
RA Allen L.N., Hanson R.S.;
RT "Construction of broad-host-range cosmid cloning vectors:
RT identification of genes necessary for growth of Methylobacterium
RT organophilum on methanol";
RL J. Bacteriol. 161:955-962(1985).
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RN [2]
RC pLA2917 from pLA2901 & pMF517
RA ;
RT ;
RL Unpublished (1989).
RL U.S. Patent No. 4,824,786 dated Apr. 25, 1989.
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CC A broad host range plasmid vector.
CC This is a cosmid with a broad host range that can be used to
CC characterize the genome of C1-utilizing microorganisms. [2]
CC This cosmid can be mobilized for transfer by conjugation between
CC E.coli and other strains in the presence of helper plasmid
CC pRK2013. [2]
CC The BglII and PstI sites interrupt the kanamycin resistance gene; the
CC SalI site interrupts the tetracycline resistance gene. [2]
CC This vector transfers efficiently to a broad host range including
CC E.coli HB101, Methylobacterium organophilum DSM 761, Pseudomonas
CC AM1, Pseudomonas putida, and Pseudomonas aeruginosa. [2]
CC DNA inserts, 25-40 kb in size, can be cloned into the BglII site and
CC mobilized from E.coli to M. organophilum.(personal communication)
CC Restriction digests of the clone give the following sizes (kb):
CC PstI--21.0; HindIII--21.0; BglII--21.0; EcoRI--17.0, 4.0; (ATCC staff)
CC Medium is 1273 LB plus tetracycline.
CC NM (pLA2917)
CC CM (no)
CC NA (ds-DNA)
CC TP (circular)
CC ST ()
CC TY (cosmid)
CC SP (ATCC)
CC HO (E.coli HB101)(broad host range)(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. pLA2905 XhoI 21000bp
FT BAL31 nuclease
FT T4 DNA polymerase:T4 DNA polymerase
FT 2. pMF517 HindIII 7100bp, pBR322 30..30
FT BAL31 nuclease
FT T4 DNA polymerase:T4 DNA polymerase
FT PvuII-PvuII 400bp, lambda cos
FT -> pLA2917 21000bp"
FT misc_binding 0..0
FT /note="SIT unique BstEII-HpaI-HindIII-SalI-PstI-BglII"
FT rep_origin 0..0
FT /note="ORI E. coli RK2"
FT CDS 0..0
FT /note="ANT Methylobacterium tetracycline resistance
FT gene (tet)"
FT CDS 0..0
FT /note="ANT Methylobacterium kanamycin resistance gene
FT (kan)"
FT rep_origin 0..0
FT /note="ORI Methylobacterium oriV"
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SQ Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;
n
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