pAAV_hSyn_HA-TALE(NG)-Tet1FL-NLS-SID4X_2A_EGFP_W3SL
(Plasmid #172205)

• Purpose: AAV vector expressing an HA-tagged TALE-SID4X transcriptional repressor protein targeting the mouse Tet1FL promoter. Synapsin promoter for neuronal expression. C-terminal, self-cleaving EGFP marker.
• Depositing Lab: Garrett Kaas
• Publication: Greer et al J Neurosci. 2021 Jan 27;41(4):578-593. doi: 10.1523/JNEUROSCI.1821-20.2020. Epub 2020 Dec 1.

Backbone

• Vector backbone: pAAV_hSyn_WPRE_bGHpA
• Backbone manufacturer: Zhang Lab
• Backbone size w/o insert (bp): 2884
• Total vector size (bp): 7335
• Modifications to backbone: Replaced the WPRE_bGHpA cassette with the modified W3SL cassette (Choi et al Mol Brain. 2014) to maximize insert size and remain within AAV packaging limits
• Vector type: Mammalian Expression, Mouse Targeting, AAV ; TALE

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: HA-TALE(NG)-Tet1FL-SID4X-NLS_2A_EGFP_W3SL
• Alt name: Tet1FL-SID4X
• Species: Synthetic
• Insert Size (bp): 4451
• Promoter: hSyn human synapsin (mammalian neuronal expression)
• Tags / Fusion Proteins:
  • HA (N terminal on insert)
  • T2A-EGFP (C terminal on insert)
  • NLS

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: hSyn_promoter_seqF 5'-CCTCAGCACTGAAGGC-3'
• 3′ sequencing primer: W3SL_seqR 5'-CGTATGCGGTGTGAAATA-3'

Resource Information

• A portion of this plasmid was derived from a plasmid made by: The pAAV_hSyn_HA-TALE(NG)-Tet1FL-NLS-SID4X_2A_EGFP_W3SL backbone vector was generated via Gibson Assembly using PCR fragments amplified from pAAV_CW3SL-EGFP, a gift from Bong-Kiun Kaang (Addgene plasmid # 61463) as well as pAAV_hSyn_TALEBB(NG)-NLS-VP64_2A_GFP_WPRE_bGHpA and pAAV_hSyn_TALEBB(NG)-NLS-SID4X_2A_phiLOV2.1_WPRE_bGHpA, gifts from Feng Zhang (Addgene plasmids # 47447, # 47451, respectively).

Terms and Licenses

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