Yellow Cameleon-Nano15/pBig
(Plasmid #51962)

• Purpose: Expression of Yellow Cameleon-Nano15, an ultrasensitive Ca2+ indicator with Kd = 15 nM, in Dictyostelium
• Depositing Lab: Takeharu Nagai
• Publication: Horikawa et al Nat Methods. 2010 Sep;7(9):729-32. doi: 10.1038/nmeth.1488. Epub 2010 Aug 8.

Backbone

• Vector backbone: pBig delta BamHI
• Vector type: Bacterial Expression
• Selectable markers: Gentamicin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Yellow Cameleon-Nano15
• Species: Synthetic
• Insert Size (bp): 2000

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (not destroyed)
• 3′ cloning site: SacI (not destroyed)
• 5′ sequencing primer: Neo-R
• 3′ sequencing primer: M13-F20

Terms and Licenses

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

Depositor Comments

The vector pBIG is derived from the published construct pATANB43 (Dynes and Firtel, 1989). pBIG contains a 5.9 kb fragment of the native Dictyostelium extrachromosomal plasmid Ddpl that confers autonomous replication in Dictyostelium, a 2.2 kb actm 6 promoter-G418-resistant gene cartridge, and vector sequences from pBluescript (11) SK (Stratagene). (reference: Cell, Vol 75, Issue 2, 1993, 361-371).