pjt126 (Addgene inc)

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Addgene inc pjt126
Pjt126, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/pjt126/pGEM-sema3B-3'-probe+(Plasmid+%2316192)/pm39425622-287-12-13
Average 93 stars, based on 15 article reviews

pjt126 - by Bioz Stars, 2026-10

93/100 stars

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Cloning:

Article Title: High-throughput functional characterization of combinations of transcriptional activators and repressors.
Article Snippet: .. KRAB-Nuclear Pfam screen cloning and cell culture An initial backbone was generated by digesting pJT126 (AddGene #161926) with BsmbI, then ligating in a sequence corresponding to the ZNF10 KRAB domain. .. Then, the nuclear Pfam library was cloned into the downstream GoldenGate cloning site, and used to produce lentivirus identically as in.21 K562 cells were then spinfected in 2 separate replicates with this KRAB+nuclear Pfam library lentivirus at an average MOI of 0.015, corresponding to an infection coverage of 85x per replicate.

Cell Culture:

Generated:

Article Title: High-throughput functional characterization of combinations of transcriptional activators and repressors
Article Snippet: .. An initial backbone was generated by digesting pJT126 (AddGene #161926) with BsmbI, then ligating in a sequence corresponding to the ZNF10 KRAB domain. .. Then, the nuclear Pfam library was cloned into the downstream GoldenGate cloning site, and used to produce lentivirus identically as in .

Sequencing:

Clone Assay:

Article Title: Prediction and design of transcriptional repressor domains with large-scale mutational scans and deep learning
Article Snippet: .. RDs were ordered as 300bp gBlock gene fragments (Integrated DNA Technologies (IDT)) with Esp3I/BsmBI restriction sites and cloned as fusions with rTetR(SE-G72P) in pJT126 (Addgene #161926). gBlocks were resuspended to 10-20 ng/μL in EB. pJT126 (Addgene no. 161926) was predigested with Esp3I (NEB, catalog no. R0734L) for 15 mins at 37°C and then purified by gel extraction. .. 10μL Golden Gate reactions were performed using a 2:1 molar ratio of insert to digested backbone with 75 ng pJT126 and 5.3 ng of each 300bp insert, with 1 μL T4 DNA Ligase Reaction Buffer (NEB, catalog no. B0202S), 0.125 μL T4 DNA Ligase (NEB, catalog no. M0202T), 0.75 μL Esp3I (NEB, catalog no. R0734L), and nuclease free dH 2 O up to the final volume.

Purification:

Gel Extraction:

Plasmid Preparation:

Article Title: A High-Throughput Screen for Antiproliferative Peptides in Mammalian Cells Identifies Key Transcription Factor Families.
Article Snippet: HeLa and HEK293T cells were grown in DMEM (Gibco) with 10% FBS, 2 mM L-glutamine, and 1% penicillin/streptomycin. .. All plasmid backbones and peptide sequences are listed in Supporting Table 2. pJT126 (Addgene #161926) was modified to produce pGH1173. .. Specifically, an HA tag-PYL1-NLS gene block was constructed (IDT) and inserted in the place of the rTetR protein via Gibson Assembly.

Review