MIGS5.2 (FF576)
(Plasmid
#35254)
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Purpose(Empty Backbone)
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 35254 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepGreenII
- Backbone size (bp) 8002
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Modifications to backbone1) Addition of an expression cassette with the miR173, however with no promoter, this should be added by multi-site gateway; 2) No promoter for expression of the MIGS construct. Instead, a promoter of choice can be cloned using multi-site gateway. See original article: de felippes et all, 2012, the Plant Journal.
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Vector typePlant Expression
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Selectable markerskanamycin
Growth in Bacteria
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Bacterial Resistance(s)Chloramphenicol and Spectinomycin, 25 & 50 μg/mL
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Growth Temperature37°C
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Growth Strain(s)ccdB Survival
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Growth instructions1) Before recombination Chloramphenicol might be used for bacteria selection (resistance lost after recombination). 2) E. coli DB3.1 strain (or some other ccdB survival strain) should be used for the amplification of the plasmid due to the presence of the lethal ccdB gene. After recombination, any common strain, such as DH5alpha is recommended. 3) Due to pGreen backbone characteristics, the helper plasmid pSOUP is required for replication of MIGS vector in Agrobacterium. PSOUP can be requested directly at http://www.pgreen.ac.uk/.
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Copy numberHigh Copy
Terms and Licenses
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Academic/Nonprofit Terms
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Industry Terms
- Not Available to Industry
Trademarks:
- Zeocin® is an InvivoGen trademark.
Depositor Comments
MIGS vectors are based on pGREEN series:
Hellens, R.P., Edwards, E.A., Leyland, N.R., Bean, S. and Mullineaux, P.M. (2000)
pGreen: a versatile and flexible binary Ti vector for Agrobacterium-mediated plant
transformation. Plant Mol. Biol., 42, 819-832.
obtained from John Innes Centre, UK
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.
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For your Materials & Methods section:
MIGS5.2 (FF576) was a gift from Detlef Weigel (Addgene plasmid # 35254 ; http://n2t.net/addgene:35254 ; RRID:Addgene_35254) -
For your References section:
MIGS: MiRNA Induced Gene Silencing. de Felippes FF, Wang JW, Weigel D. Plant J. 2011 Dec 28. doi: 10.1111/j.1365-313X.2011.04896.x. 10.1111/j.1365-313X.2011.04896.x PubMed 22211571