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Addgene

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.

Add to Cart
$400 USD + shipping
Available to academics and nonprofits only.

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 two paneled (A and B) figure depicting the Komagataeibacter Tool Kit. Panel A displays the process for moving from entry level constructs to level 1, 2, and 3+ using varying restriction enzymes and selection methods. Panel B depicts entry level components (promoter, RBS, CDS, terminator) that can be used to create level 1 expression constructs, or higher-level constructs with multiple components.

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
Data calculated @ 2024-06-13

Kit Plate Map - #1000000216

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