Modified Shipping Schedule: Addgene will be closed November 23rd & 24th for the Thanksgiving holiday. Order processing and shipping may be delayed the week of Nov 20 - 24. If you have any questions, please contact us at [email protected].
This website uses cookies to ensure you get the best experience. By continuing to use this site, you agree to the use of cookies.

Please note: Your browser does not support the features used on Addgene's website. You may not be able to create an account or request plasmids through this website until you upgrade your browser. Learn more

Please note: Your browser does not fully support some of the features used on Addgene's website. If you run into any problems registering, depositing, or ordering please contact us at [email protected] Learn more

pMN20
(Plasmid #11573)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 11573 Plasmid sent as bacteria in agar stab 1 $65

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pPZP212
  • Backbone manufacturer
    Hajdukiewicz et al 1994
  • Backbone size w/o insert (bp) 11073
  • Vector type
    Plant Expression
  • Selectable markers
    kanamycin

Growth in Bacteria

  • Bacterial Resistance(s)
    Spectinomycin
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Growth instructions
    Plasmid 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 number
    High Copy

Gene/Insert

  • Gene/Insert name
    CaMV 35S enhancers

Cloning Information

  • 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)

Resource Information

  • A portion of this plasmid was derived from a plasmid made by
    Max Planck Institute
  • Terms and Licenses

Depositor Comments

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.

How to cite this plasmid ( Back to top)

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)
  • 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