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URL: http://www.addgene.org/11572
Proper Citation: RRID:Addgene_11572
Insert Name: CaMV 35S enhancers
Bacterial Resistance: Spectinomycin
Defining Citation: PMID:10759496
Vector Backbone Description: Backbone Marker:Hajdukiewicz et al 1994; Backbone Size:11073; Vector Backbone:pPZP212; Vector Types:Plant Expression; Bacterial Resistance:Spectinomycin
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.
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Source: Addgene
