Map:

Vector IG Sequence Link :

General : cosmid ds-DNA 21000 BP

Functions : (cloning)

Selection : ()

Copy Number :

Hosts : (E.coli HB101)(broad host range)(E.coli)

Suppliers : (ATCC)

Misc.Comments : A broad host range plasmid vector. This is a cosmid with a broad host range that can be used to characterize the genome of C1-utilizing microorganisms. [2] This cosmid can be mobilized for transfer by conjugation between E.coli and other strains in the presence of helper plasmid pRK2013. [2] The BglII and PstI sites interrupt the kanamycin resistance gene; the SalI site interrupts the tetracycline resistance gene. [2] This vector transfers efficiently to a broad host range including E.coli HB101, Methylobacterium organophilum DSM 761, Pseudomonas AM1, Pseudomonas putida, and Pseudomonas aeruginosa. [2] DNA inserts, 25-40 kb in size, can be cloned into the BglII site and mobilized from E.coli to M. organophilum.(personal communication) Restriction digests of the clone give the following sizes (kb): PstI--21.0; HindIII--21.0; BglII--21.0; EcoRI--17.0, 4.0; (ATCC staff) Medium is 1273 LB plus tetracycline.

Parents : ()

Siblings : ()

Descendents : ()

ID  PLA2917    preliminary; circularDNA; 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RL  Unpublished (1989).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 XhoI21000bp

FT                   BAL31 nuclease

FT                   T4 DNA polymerase:T4 DNApolymerase

FT                   2. pMF517 HindIII 7100bp,pBR322 30..30

FT                   BAL31 nuclease

FT                   T4 DNA polymerase:T4 DNApolymerase

FT                  PvuII-PvuII 400bp,lambda cos

FT                   -> pLA2917 21000bp"

FT  misc_binding    0..0

FT                   /note="SIT uniqueBstEII-HpaI-HindIII-SalI-PstI-BglII"

FT  rep_origin      0..0

FT                   /note="ORI E. coliRK2"

FT  CDS             0..0

FT                   /note="ANTMethylobacterium tetracycline resistance

FT                   gene (tet)"

FT  CDS             0..0

FT                   /note="ANTMethylobacterium kanamycin resistance gene

FT                   (kan)"

FT  rep_origin      0..0

FT                   /note="ORIMethylobacterium oriV"

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SQ  Sequence 1 BP; 0 A; 0 C; 0 G; 0 T; 1 other;

   n

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