pTRE Tight ArchT-GFP
PurposeTetracycline-dependent expression of ArchT-EGFP in mammalian cells
Full plasmid sequence is not available for this item.
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||79627||Plasmid sent as bacteria in agar stab||1||$65|
This material is available to academics and nonprofits only.
- Backbone size w/o insert (bp) 2605
Vector typeMammalian Expression
Growth in Bacteria
Insert Size (bp)1668
MutationArchT is a more sensitive version of Arch (archaerhodpsin; light-driven outward proton pump)
- Promoter Tet responsive Ptight promoter
/ Fusion Protein
- EGFP (C terminal on backbone)
- Cloning method Restriction Enzyme
- 5′ cloning site BamHI (not destroyed)
- 3′ cloning site ClaI (not destroyed)
- 5′ sequencing primer AGGCGTATCACGAGGCCCTTTCGT
- 3′ sequencing primer tattaccgcctttgagtgagctga (Common Sequencing Primers)
A portion of this plasmid was derived from a plasmid made byDr. Ed Boyden at MIT
Terms and Licenses
SV40 polyA in pTRE-Tight does not work in TG animal for some reason, additional intron + polyA from pMSG vector was inserted (please see vector map).
To make plasmid, intron+polyA part from pMSG was PCR amplified and ligated to the HindIII/XbaI sites of pTRE-Tight (3’ primer has AatII site also for linearization later). ArchT-GFP was subcloned into pTRE-Tight with BamHI/ClaI.
Linearize the vector with AatII digestion to make transgenic mouse
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
For your Materials & Methods section:pTRE Tight ArchT-GFP was a gift from Yasunori Hayashi (Addgene plasmid # 79627)
For your References section:Inhibiting the Activity of CA1 Hippocampal Neurons Prevents the Recall of Contextual Fear Memory in Inducible ArchT Transgenic Mice. Sakaguchi M, Kim K, Yu LM, Hashikawa Y, Sekine Y, Okumura Y, Kawano M, Hayashi M, Kumar D, Boyden ES, McHugh TJ, Hayashi Y. PLoS One. 2015 Jun 15;10(6):e0130163. doi: 10.1371/journal.pone.0130163. eCollection 2015. PONE-D-15-13315 [pii] PubMed 26075894
Map uploaded by the depositor.