A1H Himar1 Transposition Plasmid

Shove any sequence randomly into your microbe via transposon!

This open-source plasmid allows for the direct integration of any payload sequence cloned between the Inverted Repeat flanks into the host microbe's chromosome, in a random location between any TA dinucleotide. It carries a weakly expressing constitutive Himar1 transposase circuit and the necessary inverted repeat flanks to allow for cut-and-paste transposition. Seen below is the result of a chromosomal integration of FuGFP by transforming the cell line, growing in liquid culture, and then selecting for glowy colonies while plating on LB without antibiotics.

It glows faintly due to single(ish) copy insertion so do not expect protein yield similar to that of high copy inducible plasmids. The plasmid is Golden Braid compatible and acts just like an A1 entry vector. Use Bsa1 to assembly multiple Golden Braid parts into a Transcription Unit and then grow the culture up to integrate said circuit directly into your host E. coli's chromosome at a random TA site.

Please refer to my Plasmid Library for the sequence data of this plasmid.

For a protocol on how to use this plasmid, please see my Protocols Library for step-by-step instructions.

For the latest out of my research projects and hints at potential new products, check out my Open Lab Notebook, where all my work lives.

The plasmid is shipped pre-transformed in a safe non-pathogenic laboratory line of E. coli in an agar stab. Please store at 4C in a fridge or at room temp in the dark. DO NOT FREEZE. Properly stored unopened agar stabs have lasted well over 5 years with good recovery in my lab.

To maintain the plasmid in the E. coli it comes in, please use standard LB Agar with 100mg/L Ampicillin and incubate overnight at 37C. Please restreak when you would like to use the cells and only work with freshly grown well isolated colonies. Do not grow cells in liquid media directly from an agar stab as it will carry over stressed cells and lead to uneven colony sizes and poor growth performance.

AIH © 2024 by Sebastian S. Cocioba is licensed under Creative Commons Attribution-ShareAlike 4.0 International

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By purchasing products from New York Botanics, LLC dba "Sebastian's Biotech Bazaar", henceforth referred to as the "Seller", you, the "Purchaser", agree to the following terms:

1. Acknowledgment of Risk: You acknowledge that the purchase and use of the plasmids involve known and unanticipated risks, which could result in physical or chemical damage, and you understand such potential risks.

2. Assumption of Risk: You knowingly, voluntarily, and expressly waive any claim you may have against the Seller for any such damages sustained as a result of purchasing or using the plasmids.

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