Komagataeibacter Tool Kit (KTK)
(Kit #
1000000216
)
Depositing Lab: Tom Ellis
The Komagataeibacter Tool Kit (KTK) is a standardized modular cloning system based on Golden Gate DNA assembly that allows DNA parts to be combined to build complex multigene constructs expressed from plasmids in cellulose-producing Komagataeibacter species.
This kit will be sent as bacterial glycerol stocks in 96-well plate format.
Original Publication
Komagataeibacter Tool Kit (KTK): A Modular Cloning System for Multigene Constructs and Programmed Protein Secretion from Cellulose Producing Bacteria. Goosens VJ, Walker KT, Aragon SM, Singh A, Senthivel VR, Dekker L, Caro-Astorga J, Buat MLA, Song W, Lee KY, Ellis T. ACS Synth Biol. 2021 Dec 17;10(12):3422-3434. doi: 10.1021/acssynbio.1c00358. PubMed (Link opens in a new window) Article (Link opens in a new window).
Description
The Komagataeibacter Tool Kit (KTK) consists of standardized plasmid vectors to facilitate modular cloning of DNA constructs of increasing numbers of parts, and plasmids containing basic parts that can be used in this system including promoters, fusion tags, and reporter proteins. The plasmid vectors are designed to contain replication and selection machinery that enable cloning steps in E. coli and then propagation and function in Komagataeibacter. Additional plasmids are also provided that enable genome integration of DNA to be attempted in Komagataeibacter.
A Golden Gate cloning toolkit for standardised assembly of modular parts from part libraries into plasmids for expression and genome integration in Komagataeibacter. (A) Different levels of plasmids that can be constructed by KTK cloning. The restriction enzymes (BsmBI, BsaI, and BpiI) used for cloning at each level are depicted along with the bacterial selection marker (AmpR, CmR, and SpecR) and phenotypic marker (X-Gal, GFP). (B) Entry level parts can be combined with a level 1 backbone to form a basic transcriptional unit (TU) or fused CDS TU. Two backbone vectors are available at each level, allowing for 4 TUs in level 3 cloning. (Image from bioRxiv, reprinted under the terms of the CC-BY-NC (Link opens in a new window)).
Please visit https://www.biorxiv.org/content/10.1101/2021.06.09.447691v1 (Link opens in a new window) for bioRxiv preprint.
Kit Documentation
KTK Guide (PDF, 932 KB) (Link opens in a new window)How to Cite this Kit
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which they were created, and include Addgene in the Materials and Methods of your future publications.
For your Materials and Methods section:
“The Komagataeibacter Tool Kit (KTK) was a gift from Tom Ellis (Addgene kit #1000000216).”
For your Reference section:
Komagataeibacter Tool Kit (KTK): A Modular Cloning System for Multigene Constructs and Programmed Protein Secretion from Cellulose Producing Bacteria. Goosens VJ, Walker KT, Aragon SM, Singh A, Senthivel VR, Dekker L, Caro-Astorga J, Buat MLA, Song W, Lee KY, Ellis T. ACS Synth Biol. 2021 Dec 17;10(12):3422-3434. doi: 10.1021/acssynbio.1c00358. PubMed (Link opens in a new window) Article (Link opens in a new window).
Komagataeibacter Tool Kit (KTK) - #1000000216
- Resistance Color Key
Each circle corresponds to a specific antibiotic resistance in the kit plate map wells.
- Inventory
Searchable and sortable table of all plasmids in kit. The Well column lists the plasmid well location in its plate. The Plasmid column links to a plasmid's individual web page.
- Kit Plate Map
96-well plate map for plasmid layout. Hovering over a well reveals the plasmid name, while clicking on a well opens the plasmid page.
Resistance Color Key
| Ampicillin | |
| Chloramphenicol | |
| Spectinomycin |
Inventory
| Well | Plasmid | Resistance |
|---|---|---|
| A / 1 | KTK_001 | Ampicillin |
| A / 2 | KTK_354 | Ampicillin |
| A / 3 | KTK_034 | Ampicillin |
| A / 4 | KTK_002 | Ampicillin |
| A / 5 | KTK_032 | Ampicillin |
| A / 6 | KTK_247 | Ampicillin |
| A / 7 | KTK_042 | Ampicillin |
| A / 8 | KTK_043 | Ampicillin |
| A / 9 | KTK_044 | Ampicillin |
| A / 10 | KTK_218 | Ampicillin |
| A / 11 | KTK_219 | Ampicillin |
| A / 12 | KTK_220 | Ampicillin |
| B / 1 | KTK_221 | Ampicillin |
| B / 2 | KTK_222 | Ampicillin |
| B / 3 | KTK_223 | Ampicillin |
| B / 4 | KTK_225 | Ampicillin |
| B / 5 | KTK_113 | Ampicillin |
| B / 6 | KTK_003 | Ampicillin |
| B / 7 | KTK_029 | Ampicillin |
| B / 8 | KTK_045 | Ampicillin |
| B / 9 | KTK_046 | Ampicillin |
| B / 10 | KTK_033 | Ampicillin |
| B / 11 | KTK_251 | Ampicillin |
| B / 12 | KTK_316 | Ampicillin |
| C / 1 | KTK_344 | Ampicillin |
| C / 2 | KTK_346 | Ampicillin |
| C / 3 | KTK_345 | Ampicillin |
| C / 4 | KTK_047 | Ampicillin |
| C / 5 | KTK_226 | Ampicillin |
| C / 6 | KTK_252 | Ampicillin |
| C / 7 | KTK_110 | Ampicillin |
| C / 8 | KTK_005 | Ampicillin |
| C / 9 | KTK_031 | Ampicillin |
| C / 10 | KTK_035 | Ampicillin |
| C / 11 | KTK_036 | Ampicillin |
| C / 12 | KTK_037 | Ampicillin |
| D / 1 | KTK_038 | Ampicillin |
| D / 2 | KTK_039 | Ampicillin |
| D / 3 | KTK_040 | Ampicillin |
| D / 4 | KTK_007 | Ampicillin |
| D / 5 | KTK_048 | Ampicillin |
| D / 6 | KTK_139 | Chloramphenicol |
| D / 7 | KTK_009 | Chloramphenicol |
| D / 8 | KTK_010 | Chloramphenicol |
| D / 9 | KTK_136 | Chloramphenicol |
| D / 10 | KTK_137 | Chloramphenicol |
| D / 11 | KTK_095 | Chloramphenicol |
| D / 12 | KTK_324 | Spectinomycin |
| E / 1 | KTK_015 | Spectinomycin |
| E / 2 | KTK_016 | Spectinomycin |