|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||11573||Standard format: Plasmid sent in bacteria as agar stab||1||$65|
This material is available to academics and nonprofits only.
Backbone manufacturerHajdukiewicz et al 1994
- Backbone size w/o insert (bp) 11073
Vector typePlant Expression
Growth in Bacteria
Growth instructionsPlasmid grows in standard E. coli at 37 degrees, but the four enhancer repeats may be unstable in E. coli and A. tumefaciens if stored at 4'C for extended time. See comments.
Copy numberHigh Copy
Gene/Insert nameCaMV 35S enhancers
- Cloning method Restriction Enzyme
- 5′ cloning site See paper and map. (not destroyed)
- 3′ cloning site See paper and map. (not destroyed)
- 5′ sequencing primer M13 reverse (Common Sequencing Primers)
Used to confirm activation-tagged genes. Contains multiple cloning sites adjacent to the same tetramerized CaMV 35S enhancers as in the activation-tagging vectors from this article.
Kanamycin gene for plant selection on MS media.
Plasmid selection in E. coli and in A. tumefaciens is spectinomycin (100 mg/l).
The four enhancer repeats in the construct may be unstable in E. coli and Agrobacterium if stored at +4'C for extended time. Check by PCR with T7 (or M13-20) oligo and this one derived from the RB sequence: 5' acc cgc caa tat atc ctg 3'. This should give 1.4 kb band. Note that these oligos will not work in a transgenic plant, as the RB sequence is not transfered. Using this test, we found that after 1-2 weeks in a fridge the Agrobacterium strain loses on average one copy of a repeat, and after a month in a fridge there is only one copy left. It is a good idea to use a freshly streaked colony from a good -70'C glycerol stock for every infiltration.
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:pMN20 was a gift from Detlef Weigel (Addgene plasmid # 11573 ; http://n2t.net/addgene:11573 ; RRID:Addgene_11573)
For your References section:Activation tagging in Arabidopsis. Weigel D, Ahn JH, Blazquez MA, Borevitz JO, Christensen SK, Fankhauser C, Ferrandiz C, Kardailsky I, Malancharuvil EJ, Neff MM, Nguyen JT, Sato S, Wang ZY, Xia Y, Dixon RA, Harrison MJ, Lamb CJ, Yanofsky MF, Chory J. Plant Physiol. 2000 Apr . 122(4):1003-13. 10.1104/pp.122.4.1003 PubMed 10759496