pTol2-ziUAS:GPHN.FingR-mCherry-ZFI (IL2RGTC)-KRAB(A)
(Plasmid #72639)

• Purpose: Tol2 based UAS modified with a IL2RGTC binding site upstream of it, driving GPHN-FingR fused to mCherry, IL2RGTC zinc finger, and KRAB(A) repressor domain
• Depositing Lab: Joshua Bonkowsky
• Publication: Bonkowsky lab FingR plasmids (unpublished)

Backbone

• Vector backbone: pDONR 221
• Backbone manufacturer: Invitrogen
• Total vector size (bp): 7182
• Vector type: Gateway

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: UAS upstream of GPHN-FingR with ZFI(IL2RFTC) and KRAB(A)
• Species: M. musculus (mouse)
• Insert Size (bp): 2694
• Mutation: Stop codon after mCherry- SEE DEPOSITOR COMMENTS BELOW

Cloning Information

• Cloning method: Gateway Cloning
• 5′ sequencing primer: T7

Resource Information

• A portion of this plasmid was derived from a plasmid made by: from Don Arnold lab (Gross et al., Neuron, 2013): Plasmids pCAG_PSD95.FingR-eGFP-CCR5TC-KRAB(A) and pCAG_GPHN.FingR-mKate2-IL2RGTCKRAB(A) were obtained from Addgene (plasmids #46295 and #46297).

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • National Institute of Genetics Limited Use Label License for Tol2 System
  • Takara Bio Limited Use Label License (formerly Clontech)
• Industry Terms:
  • Not Available to Industry

Depositor Comments

PLEASE NOTE- this plasmid has a Stop codon after mCherry which is an error and will cause the construct to not be specifically targeted to the synapse.

The Bonkowsky Lab has a corrected plasmid available which will soon be available at Addgene: pTol2-ziUAS:GPHN.FingR-mKate2-ZFI (IL2RGTC)-KRAB(A).

Details of cloning are in:
Son et al., Scientific Reports, 2016. Transgenic FingRs for Live Mapping of Synaptic Dynamics in Genetically-Defined Neurons.