3XFlag-pA-Tn5-Fl
(Plasmid #124601)

• Purpose: Expresses protein A and Tn5 Transposase fusion protein in bacterial cells
• Depositing Lab: Steven Henikoff
• Publication: Kaya-Okur et al Nat Commun. 2019 Apr 29;10(1):1930. doi: 10.1038/s41467-019-09982-5.

Backbone

• Vector backbone: pTXB1-Tn5
• Backbone size w/o insert (bp): 8079
• Total vector size (bp): 8679
• Modifications to backbone: Ribosome binding site (RBS) was replaced by a canonical RBS sequence
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): C3013
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: Protein A and hyperactive Tn5 transposase (Tnp) fusion protein
• Alt name: 3XFlag_pA-Tn5
• Species: E. coli
• Promoter: T7
• Tag / Fusion Protein:
  • Protein A and Tn5 transposase fusion protein with N terminal 3XFlag tag (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NcoI (not destroyed)
• 3′ cloning site: NdeI (not destroyed)
• 5′ sequencing primer: CGG TTT AAA CCG GGG ATC TCG
• 3′ sequencing primer: TTG CGC CGC AAC ATT CAC C

Resource Information

• Supplemental Documents:
  • 3XFlag-pA-Tn5-Fl.gbk
• A portion of this plasmid was derived from a plasmid made by: Tn5 transposase: pTXB1-Tn5 plasmid was a gift from Rickard Sandberg (Addgene plasmid # 60240) and for pA, the pK19pA-MN plasmid was a gift from Ulrich Laemmli (available through Addgene plasmid # 86973)
• Articles Citing this Plasmid:
  • 10 References

Terms and Licenses

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

Depositor Comments

Please note that verifying this plasmid by sequencing or digest is challenging due to the presence of the helper plasmid, as multiple bands or mispriming is likely to occur. See Addgene's sequencing results to compare sequences.

This plasmid has been found to be somewhat unstable and prone to concatenation. Concatenation often does not impact plasmid function, but can reduce transformation or transfection efficiencies. If you have trouble isolating the monomeric version of this plasmid, you might consider linearizing, gel extracting, re-ligating, and transforming the plasmid.