TUNEYALI Kit
(Kit #
1000000255
)
Depositing Lab: Irina Borodina
The TUNEYALI collection is a modular toolkit for the precise regulation of transcription factor expression in Yarrowia lipolytica. This kit is comprised of 64 plasmids: 56 base plasmids targeting the promoter region of a specific transctiption factor, and 8 plasmids encoding synthetic promoters with varying expression strengths.
This kit will be sent as bacterial glycerol stocks in 96-well plate format.
Description
The TUNEYALI kit consists of 64 plasmids: 56 base plasmids and 8 plasmids encoding synthetic promoters. Each base plasmid is designed to delete the native promoter of one specific transcription factor among a set of 56, allowing targeted modulation of gene regulation. The synthetic promoters span a range of strengths and can be inserted into the base plasmids using a SapI-based Golden Gate reaction to replace the native promoter.
This modular design enables precise tuning of transcription factor expression by selecting appropriate promoter-plasmid combinations. Moreover, multiple promoters and base plasmids can be assembled simultaneously in a single one-pot SapI-based Golden Gate reaction, enabling efficient construction of a TUNEYALI transcription factor (TF) plasmid library. This high-throughput approach supports systematic studies of TF function, regulatory networks, and synthetic biology applications in Y. lipolytica.
- Figure 1: Schematic overview of the TUNEYALI toolkit and workflow for tuning transcription factor expression in Yarrowia lipolytica. (A) A synthetic promoter is inserted into a base plasmid via SapI-based Golden Gate assembly to generate a TF expression construct. The engineered construct replaces the native promoter of the target transcription factor, enabling tunable expression. (B) Multiple promoters and base plasmids can be combined in a one-pot Golden Gate reaction to build a diverse TF plasmid library. The resulting constructs are transformed into Y. lipolytica, followed by automated screening and phenotypic analysis to evaluate the effects of transcription factor modulation.
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 TUNEYALI Kit was a gift from Irina Borodina (Addgene kit #1000000255).”
TUNEYALI Kit - #1000000255
- 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 | |
| Kanamycin |
Inventory
| Well | Plasmid | Resistance |
|---|---|---|
| A / 1 | pCfb13191 | Ampicillin |
| A / 2 | pCfb13192 | Ampicillin |
| A / 3 | pCfb13193 | Ampicillin |
| A / 4 | pCfb13194 | Ampicillin |
| A / 5 | pCfb13195 | Ampicillin |
| A / 6 | pCfb13196 | Ampicillin |
| A / 7 | pCfb13197 | Ampicillin |
| A / 8 | pCfb13198 | Ampicillin |
| A / 9 | pCfb13199 | Ampicillin |
| A / 10 | pCfb13200 | Ampicillin |
| A / 11 | pCfb13201 | Ampicillin |
| A / 12 | pCfb13202 | Ampicillin |
| B / 1 | pCfb13203 | Ampicillin |
| B / 2 | pCfb13204 | Ampicillin |
| B / 3 | pCfb13205 | Ampicillin |
| B / 4 | pCfb13206 | Ampicillin |
| B / 5 | pCfb13207 | Ampicillin |
| B / 6 | pCfb13208 | Ampicillin |
| B / 7 | pCfb13209 | Ampicillin |
| B / 8 | pCfb13210 | Ampicillin |
| B / 9 | pCfb13211 | Ampicillin |
| B / 10 | pCfb13212 | Ampicillin |
| B / 11 | pCfb13213 | Ampicillin |
| B / 12 | pCfb13214 | Ampicillin |
| C / 1 | pCfb13215 | Ampicillin |
| C / 2 | pCfb13216 | Ampicillin |
| C / 3 | pCfb13217 | Ampicillin |
| C / 4 | pCfb13218 | Ampicillin |
| C / 5 | pCfb13219 | Ampicillin |
| C / 6 | pCfb13220 | Ampicillin |
| C / 7 | pCfb13221 | Ampicillin |
| C / 8 | pCfb13222 | Ampicillin |
| C / 9 | pCfb13223 | Ampicillin |
| C / 10 | pCfb13224 | Ampicillin |
| C / 11 | pCfb13225 | Ampicillin |
| C / 12 | pCfb13226 | Ampicillin |
| D / 1 | pCfb13227 | Ampicillin |
| D / 2 | pCfb13228 | Ampicillin |
| D / 3 | pCfb13229 | Ampicillin |
| D / 4 | pCfb13231 | Ampicillin |
| D / 5 | pCfb13233 | Ampicillin |
| D / 6 | pCfb13234 | Ampicillin |
| D / 7 | pCfb13235 | Ampicillin |
| D / 8 | pCfb13236 | Ampicillin |
| D / 9 | pCfb13237 | Ampicillin |
| D / 10 | pCfb13238 | Ampicillin |
| D / 11 | pCfb13239 | Ampicillin |
| D / 12 | pCfb13240 | Ampicillin |
| E / 1 | pCfb13242 | Ampicillin |
| E / 2 | pCfb13244 | Ampicillin |
| E / 3 | pCfb13245 | Ampicillin |
| E / 4 | pCfb13246 | Ampicillin |
| E / 5 | pCfb13247 | Ampicillin |
| E / 6 | pCfb13248 | Ampicillin |
| E / 7 | pCfb13249 | Ampicillin |
| E / 8 | pCfb13250 | Ampicillin |
| E / 9 | pCfb12561 | Kanamycin |
| E / 10 | pCfb12562 | Kanamycin |
| E / 11 | pCfb13186 | Kanamycin |
| E / 12 | pCfb12565 | Kanamycin |
| F / 1 | pCfb12566 | Kanamycin |
| F / 2 | pCfb12567 | Kanamycin |
| F / 3 | pCfb12620 | Kanamycin |
| F / 4 | pCfb12622 | Kanamycin |