pMN20
(Plasmid #11573)

• Depositing Lab: Detlef Weigel
• Publication: Weigel et al Plant Physiol. 2000 Apr . 122(4):1003-13.

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, 50 μg/mL
• 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

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Max Planck Institute
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

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.