pQE30::CLOMELEON:Strep
(Plasmid #247989)

• Purpose: Expression of the indicator protein CLOMELEON in E. coli.
• 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: pQE30
• Backbone manufacturer: Qiagene
• Backbone size w/o insert (bp): 3461
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): NEB Stable
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: CLOMELEON (CFP-YFP fusion with ENLYFQG-linker)
• Species: Synthetic
• Insert Size (bp): 1529
• Mutation: CFP-variant: K26R, F64L, S65T, Y66W, N146I, M153T, V163A, N164H, H231L; YFP-variant: S65G, S72A, K79R, T203Y, H231L
• Promoter: T5-phage promoter
• Tags / Fusion Proteins:
  • 6xHis (N terminal on insert)
  • Strep (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: KpnI (unknown if destroyed)
• 3′ cloning site: HindIII (unknown if destroyed)
• 5′ sequencing primer: GTATCACGAGGCCCTTTCG
• 3′ sequencing primer: GTTCTGAGGTCATTACTGG

Resource Information

• Supplemental Documents:
  • Description of plasmids

Terms and Licenses

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

Depositor Comments

pQE30::CLOMELEON:Strep is a vector for bacterial expression of the CLOMELEON protein (Kuner, T. & Augustine, G.J.; PubMed: 11055428; doi: 10.1016/S0896-6273(00)00056-8) with N-terminal 6xHis-tag and C-terminal Strep-tag (WSHPQFEK). The gene coding for CLOMELEON with Strep-tag was introduced into the commercial expression vector pQE30 (Qiagen) between KpnI and HindIII sites. See the Supplemental Document under Resource Information for more details, including annotated maps.