Human Kinase Domain Constructs Kit
(Kit #
1000000094
)
Depositing Lab: John Chodera, Nicholas Levinson, and Markus Seeliger
These kinase plasmids were developed through a large-scale expression screen with the goal of generating a library of His-tagged human kinase constructs that express well in a simple automated bacterial expression system.
The kit contains 68 human kinase catalytic domain constructs and the respective phosphatases that enhance their bacterial expression. Lambda phosphatase enhances bacterial expression of Ser/Thr kinases while YopH (residues 164-468) enhances expression of Tyr kinases.
Original Publication
The construction and expression screening of these plasmids were initially described in the following preprint:
An open library of human kinase domain constructs for automated bacterial expression. Parton DL, Hanson SM, RodrÃguez-Laureano L, Albanese SK, Gradia S, Jeans C, Seeliger M, Levinson NM, Chodera JD. bioRxiv Preprint.
The preprint article has subsequently been published as:
An open library of human kinase domain constructs for automated bacterial expression. Albanese SK, Parton DL, Isik M, RodrÃguez-Laureano L, Hanson SM, Behr JM, Gradia S, Jeans C, Levinson NM, Seeliger MA, Chodera JD. Biochemistry. 2018 Jul 13. doi: 10.1021/acs.biochem.7b01081. PMID: 30004690.
Description
This kit consists of constructs containing His-tagged human kinases that have been optimized for automated bacterial expression. Constructs were generated to be co-expressed with a phosphatase (YopH or lambda) in order to enhance yield.
Phylogenetic tree representing kinase domain expression. Authors note that the image was created using KinMap.
Kit Documentation
Plasmid Sequences
Sequences for kit plasmids (CSV format)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 Human Kinase Domain Constructs Kit for Automated Bacterial Expression was a gift from John Chodera (Addgene kit #1000000094).”
For your Reference section:
An open library of human kinase domain constructs for automated bacterial expression. Albanese SK, Parton DL, Isik M, RodrÃguez-Laureano L, Hanson SM, Behr JM, Gradia S, Jeans C, Levinson NM, Seeliger MA, Chodera JD. Biochemistry. 2018 Jul 13. doi: 10.1021/acs.biochem.7b01081. PMID: 30004690.
Human Kinase Domain Constructs for Automated Bacterial Expression - #1000000094
- 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 | |
| Spectinomycin |
Inventory
| Well | Plasmid | Resistance |
|---|---|---|
| A / 1 | CDK16_HUMAN_D0 | Ampicillin |
| A / 2 | KCC1G_HUMAN_D0 | Ampicillin |
| A / 3 | KAPCA_HUMAN_D0 | Ampicillin |
| A / 4 | STK3_HUMAN_D0 | Ampicillin |
| A / 5 | STK24_HUMAN_D0 | Ampicillin |
| A / 6 | KCC2D_HUMAN_D0 | Ampicillin |
| A / 7 | GAK_HUMAN_D0 | Ampicillin |
| A / 8 | PMYT1_HUMAN_D0 | Ampicillin |
| A / 9 | HASP_HUMAN_D0 | Ampicillin |
| A / 10 | CDKL1_HUMAN_D0 | Ampicillin |
| A / 11 | BMPR2_HUMAN_D0 | Ampicillin |
| A / 12 | KC1G1_HUMAN_D0 | Ampicillin |
| B / 1 | DYR1A_HUMAN_D0 | Ampicillin |
| B / 2 | MK01_HUMAN_D0 | Ampicillin |
| B / 3 | VRK3_HUMAN_D0 | Ampicillin |
| B / 4 | VRK2_HUMAN_D0 | Ampicillin |
| B / 5 | KCC2A_HUMAN_D0 | Ampicillin |
| B / 6 | CSKP_HUMAN_D0 | Ampicillin |
| B / 7 | STK16_HUMAN_D0 | Ampicillin |
| B / 8 | STK10_HUMAN_D0 | Ampicillin |
| B / 9 | FGFR1_HUMAN_D0 | Ampicillin |
| B / 10 | VGFR1_HUMAN_D0 | Ampicillin |
| B / 11 | KC1E_HUMAN_D0 | Ampicillin |
| B / 12 | SRC_HUMAN_D0 | Ampicillin |
| C / 1 | KC1D_HUMAN_D0 | Ampicillin |
| C / 2 | MERTK_HUMAN_D0 | Ampicillin |
| C / 3 | FGFR3_HUMAN_D0 | Ampicillin |
| C / 4 | EPHB1_HUMAN_D0 | Ampicillin |
| C / 5 | ABL1_HUMAN_D0 | Ampicillin |
| C / 6 | EPHB2_HUMAN_D0 | Ampicillin |
| C / 7 | MK08_HUMAN_D0 | Ampicillin |
| C / 8 | EPHB3_HUMAN_D0 | Ampicillin |
| C / 9 | M3K5_HUMAN_D0 | Ampicillin |
| C / 10 | E2AK2_HUMAN_D0 | Ampicillin |
| C / 11 | CSK22_HUMAN_D0 | Ampicillin |
| C / 12 | DAPK1_HUMAN_D0 | Ampicillin |
| D / 1 | PAK7_HUMAN_D0 | Ampicillin |
| D / 2 | SLK_HUMAN_D0 | Ampicillin |
| D / 3 | KCC4_HUMAN_D0 | Ampicillin |
| D / 4 | KCC2G_HUMAN_D0 | Ampicillin |
| D / 5 | KC1G3_HUMAN_D0 | Ampicillin |
| D / 6 | CSK21_HUMAN_D0 | Ampicillin |
| D / 7 | NEK7_HUMAN_D0 | Ampicillin |
| D / 8 | CHK2_HUMAN_D0 | Ampicillin |
| D / 9 | PAK4_HUMAN_D0 | Ampicillin |
| D / 10 | MK11_HUMAN_D0 | Ampicillin |
| D / 11 | NEK1_HUMAN_D0 | Ampicillin |
| D / 12 | KS6A1_HUMAN_D1 | Ampicillin |
| E / 1 | MARK3_HUMAN_D0 | Ampicillin |
| E / 2 | MK14_HUMAN_D0 | Ampicillin |
| E / 3 | MAPK3_HUMAN_D0 | Ampicillin |
| E / 4 | VRK1_HUMAN_D0 | Ampicillin |
| E / 5 | PAK6_HUMAN_D0 | Ampicillin |
| E / 6 | MK03_HUMAN_D0 | Ampicillin |
| E / 7 | EPHA3_HUMAN_D0 | Ampicillin |
| E / 8 | FES_HUMAN_D0 | Ampicillin |
| E / 9 | MK13_HUMAN_D0 | Ampicillin |
| E / 10 | DMPK_HUMAN_D0 | Ampicillin |
| E / 11 | PHKG2_HUMAN_D0 | Ampicillin |
| E / 12 | CSK_HUMAN_D0 | Ampicillin |
| F / 1 | CDK2_HUMAN_D0 | Ampicillin |
| F / 2 | DAPK3_HUMAN_D0 | Ampicillin |
| F / 3 | DYRK2_HUMAN_D0 | Ampicillin |
| F / 4 | KCC2B_HUMAN_D0 | Ampicillin |
| F / 5 | AURKA_HUMAN_D0 | Ampicillin |
| F / 6 | FAK2_HUMAN_D0 | Ampicillin |
| F / 7 | KC1G2_HUMAN_D0 | Ampicillin |
| F / 8 | AAPK2_HUMAN_D0 | Ampicillin |
| F / 9 | YopH (residues 164-468) | Spectinomycin |
| F / 10 | Lambda Phosphatase | Spectinomycin |