Tol2-elavl3-H2B-GCaMP6s
(Plasmid #59530)

• Purpose: Expressed nuclear-localized GCaMP6-slow, a calcium indicator, in neurons in zebrafish larva. The plasmid is in Tol2 transposon backbone
• Depositing Lab: Misha Ahrens
• Publication: Freeman et al Nat Methods. 2014 Jul 27. doi: 10.1038/nmeth.3041.

Backbone

• Vector backbone: MiniTol2
• Vector type: Transposable element

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert 1

• Gene/Insert name: elavl3 promoter
• Alt name: HuC
• Species: D. rerio (zebrafish)
• Insert Size (bp): 8700
• Entrez Gene: elavl3 (a.k.a. HuC, elrc, id:ibd1248, wu:fb77b03, zHuC)

Cloning Information for Gene/Insert 1

• Cloning method: Restriction Enzyme
• 5′ cloning site: Xho1 (not destroyed)
• 3′ cloning site: Age1 (not destroyed)
• 5′ sequencing primer: TGATCTGCAAAAGACGTGAATATC
• 3′ sequencing primer: tttaacctctcagcgagaatgc

Gene/Insert 2

• Gene/Insert name: H2B
• Alt name: Histone
• Species: H. sapiens (human)
• Insert Size (bp): 378

Cloning Information for Gene/Insert 2

• Cloning method: Gibson Cloning
• 5′ sequencing primer: tttaacctctcagcgagaatgc

Gene/Insert 3

• Gene/Insert name: GCaMP6s
• Species: Synthetic
• Insert Size (bp): 1353

Cloning Information for Gene/Insert 3

• Cloning method: Gibson Cloning
• 5′ sequencing primer: ACTCCAGCAGGACCATGTGAT
• 3′ sequencing primer: CATCGATGGTACAGTAAAACGACGG

Resource Information

• A portion of this plasmid was derived from a plasmid made by: GCaMP6s is from HHMI-Janelia Farm. There might be a patent. MiniTol2 backbone is from National Institute of Genetics Japan. Zebrafish elavl3 promoter is originally came from Brain Science Institute, RIKEN, Japan
• Articles Citing this Plasmid:
  • 4 References

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • National Institute of Genetics Limited Use Label License for Tol2 System
• Industry Terms:
  • Not Available to Industry

Depositor Comments

Addgene NGS QC analysis identified multiple variants in the HuC promoter sequence.