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Standardized open source DNA vectors

Creation of modular, well characterized, and open source DNA vectors.

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Biohacking currently has a serious lack of modular, well characterized, and open source DNA vectors, or DNAs that act as 'backbones' for more interesting DNA and gene related projects. Normally, in order to modify an organism, you simply take a DNA of interest, which can be synthesized or taken form other organisms, and put it into a DNA vector. Once in the vector, it is transferred to an organism of interest.

Before any interesting work can be done in bacteria, plants, yeast, or any other organism, it must be cloned into a DNA vector. This can be cumbersome and it takes away time from the actual hacking of the target organism. In order to solve this problem, I want to create modular, well characterized, and open source DNA vectors.

To modularize vectors, I have made my 'base' DNA vector as compatible as possible. It allows simple transfer of DNA from one context to another, meaning you can use your DNA is as many organisms as possible. I am creating modularized vectors so you can transfer your DNA of interest to as many organisms as possible, including baker's yeast, B subtilis natto, and possibly in the future, plants.

To well characterize my vectors, I am carefully analyzing and testing all conditions. You can see some of my current characterization here. I plan on testing and optimizing cloning conditions using gibson, SLiCE, GoldenGate, classic cloning, and recombination in order to facilitate anyway you plan to clone your DNA of interest.

To open my DNA vectors to everyone, I will not require any MTAs or any other restrictive licenses, but instead open up my DNAs under the Creative Commons Attribution 4.0 International license. I am also characterizing a method to share DNA more efficiently using a virus to safely package the DNA.

Further information can be found on koengb.io

pKGs2-sacB7_RFP.dna

First vector-part plasmid. Used to store parts to create new vectors with.

x-snapgene - 1.18 MB - 04/29/2016 at 16:31

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pKGs1-GFP.dna

open source base vector.

x-snapgene - 466.18 kB - 04/29/2016 at 16:31

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  • Updated website

    koeng10105/04/2016 at 02:01 0 comments

    Updated my website with additional information. Will be posting soon on planned tests and updates on pKG basic vectors.

  • Plants and yeast

    koeng10105/01/2016 at 03:08 0 comments

    Did an overview of work it would take for a 'hello world' plant experiment. Going to begin learning more about specific plan tissue culture to perhaps make a compatible series of plasmids.

    I have several yeast plasmids I would like to get working with soon. Will make compatible plasmids and reclone them.

    Good progress, pKGs3 synthesis has finished and I am going to clone it next week. Hopefully I will be able to start adding parts by the end of May.

  • pKGs2-sacB7_RFP stabilized and pKGs1 synthesized

    koeng10104/29/2016 at 16:29 0 comments

    Interesting few weeks. As a preface-

    pKGs2-sacB7_RFP is a plasmid for cloning vector parts. These vector parts can be used in a modular fashion to create new vectors. More information on this vector can be found on my website. Soon it will be updated to have viral(M13) packaging compatibility.

    pKGs1 is the base vector. Parts can be put in this vector for storage before use in other contexts. It has a few advantages over pKGs2-sacB7_RFP in that it has superior transcriptional isolation, better primer binding sites, and a form of the T7 terminator at the 3' end.

    I had some issues with pKGs2-sacB7_RFP stability, but it appears this was just because of old cam plates. Plasmid loss was rather slow, but still occurred. Perhaps it was because of the media, which was autoclaved with glucose. Anyway, it appears to be a stable plasmid. Now to just modify it with viral packaging! Updated log with files. Open with SnapGene viewer ( http://www.snapgene.com/ ).

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Julian Hofmann wrote 09/20/2016 at 18:59 point

But you are aware that there is iGEM and with its whole library of standardized, well tested and documented DNA bricks/parts/fragments (including vectors and everything someone needs too built one?

If not you should check out http://parts.igem.org/Main_Page (and maybe this for some explenation http://parts.igem.org/Help:Philosophy)

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