pART27::CLOMELEON
(Plasmid #247988)

• Purpose: Stable expression of the indicator protein CLOMELEON for monitoring chloride concentrations in the cytoplasm of plant cells.
• Depositing Lab: Christoph Plieth
• Publication: Lorenzen et al Plant J. 2004 May;38(3):539-44. doi: 10.1111/j.0960-7412.2004.02053.x.

Backbone

• Vector backbone: pART27
• Backbone manufacturer: Andrew Gleave
• Backbone size w/o insert (bp): 11612
• Vector type: Plant Expression
• Selectable markers: lacZ (blue-white screening) and Kanamycin (in plants)

Growth in Bacteria

• Bacterial Resistance(s): Spectinomycin and Streptomycin, 50 & 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): EPI300
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: CLOMELEON (CFP-YFP fusion with ENLYFQG-linker)
• Species: Synthetic
• Insert Size (bp): 1508
• Mutation: CFP-variant: K26R, F64L, S65T, Y66W, N146I, M153T, V163A, N164H, H231L; YFP-variant: S65G, S72A, K79R, T203Y, H231L
• Promoter: Cauliflower Mosaic Virus 35S promoter (GenBank AJ007625), without artificial 40bp insert
• Tag / Fusion Protein:
  • 6xHis (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NotI (unknown if destroyed)
• 3′ cloning site: NotI (unknown if destroyed)
• 5′ sequencing primer: (1) CGCCAAGCTATTTAGGTGAC, (2) CAACCACGTCTTCAAAGC
• 3′ sequencing primer: (1) CGGCCAGTGAATTGTAATACG, (2) GGATCTGAGCTACACATGC

Resource Information

• Supplemental Documents:
  • Description of plasmids

Terms and Licenses

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

Depositor Comments

pART27::CLOMELEON is a binary pART27 vector harbouring a NotI-fragment with a gene cassette subcloned from pART7::CLOMELEON to stably insert the CLOMELEON gene (Kuner, T. & Augustine, G.J.; PubMed: 11055428; doi: 10.1016/S0896-6273(00)00056-8) into the genome of plants by Agrobacterium-mediated gene transfer, e.g. by floral dip (Clough & Bent 1998; PubMed: 10069079; doi: 10.1007/BF00028910). See the Supplemental Document under Resource Information for more details, including annotated maps.