|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||45826||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
- Backbone size (bp) 6878
Modifications to backboneThe 2.0 kb NdeI/NotI region of pDN19X containing the traJ' allele that allowed efficient replication and/or transfer in M. extorquens AM1 was transferred into the corresponding region of pTJS75 (Schmidhauser & Helinski, 1985), a parent vector of pDN19 (Nunn et al., 1990), to create pCM48. The plasmid pCM48 was then cut with MunI and BlnI, and the DNA ends were blunted and self-ligated to produce pCM50. A 5.3 kb HindIII/XmnI region of pCM50 was ligated with the blunted 1.9 kb AatII/AvaII region of pUC19(Yanisch-Perron et al., 1985) to make pCM62. During the construction of pCM62 an additional 356 bp inward from the site of XmnI cleavage of pCM50 was lost, presumably caused by either non-specific cleavage or by the exonuclease activity of T4 DNA polymerase.
Vector typebroad host range cloning vector
- Promoter lac
Growth in Bacteria
Copy numberHigh Copy
- Cloning method Unknown
- 5′ sequencing primer M13-F20
- 3′ sequencing primer M13R (Common Sequencing Primers)
Terms and Licenses
- Not Available to Industry
Vector is a broad host range vector (bhr) and is used in the Methylobacterium extorquens AM1. Also can be maintained in the a-proteo-bacteria Agrobacterium tumefaciens, Methylobacterium strains CM4 and DM4, and Rhodobacter sphaeroides, the b-proteobacterium Ralstonia eutropha and the c-proteobacteria Methylococcus capsulatus Bath and Pseudomonas aeruginosa (see linked article).
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
For your Materials & Methods section:pCM62 was a gift from Mary Lidstrom (Addgene plasmid # 45826 ; http://n2t.net/addgene:45826 ; RRID:Addgene_45826)
For your References section:Development of improved versatile broad-host-range vectors for use in methylotrophs and other Gram-negative bacteria. Marx CJ, Lidstrom ME. Microbiology. 2001 Aug;147(Pt 8):2065-75. 10.1099/00221287-147-8-2065 PubMed 11495985