MIGS5.2 (FF576)
(Plasmid #35254)

• Purpose: (Empty Backbone)
• Depositing Lab: Detlef Weigel
• Publication: de Felippes et al Plant J. 2011 Dec 28. doi: 10.1111/j.1365-313X.2011.04896.x.

Backbone

• Vector backbone: pGreenII
• Backbone size (bp): 8002
• Modifications to backbone: 1) 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.
• Vector type: Plant Expression
• Selectable markers: kanamycin

Growth in Bacteria

• Bacterial Resistance(s): Chloramphenicol and Spectinomycin, 25 & 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): ccdB Survival
• Growth instructions: 1) 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/.
• Copy number: High Copy

Terms and Licenses

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

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.

http://www.pgreen.ac.uk/

obtained from John Innes Centre, UK