EPR Plasmid #251: Tb_tethering plasmid library28
Sequence: download sequence
| Purpose of plasmid | Note: This is not a single plasmid, but a pooled library of plasmids. The complete library consists of all the relevant open reading frames from Trypanosoma brucei in a plasmid designed for inducible expression of proteins fused to a lambdaN peptide + 3 myc tags at the N terminus. A description of the composition of the ORFeome can be found in Carter et al 2020. This plasmid library can be used to perform large-scale tethering screens for proteins that modulate mRNA fate. For this purpose, the inducible plasmid library needs to be transfected into trypanosomes that express a reporter mRNA bearing a boxB sequence, which is the recognition element for the lambdaN. Following transfection, the trypanosome library is selected with hygromycin, and the tethering assay performed by inducing the expression of lambdaN-3xmyc fused proteins with tetracycline. Overexpression will result in the binding of the lambdaN-Myc proteins to the boxB element on the reporter via lambdaN. Regulators of mRNA fate can then be determined by evaluating the effect on reporter expression and the ORFs that are responsible identified by high throughput sequencing. The complete library is EPR#224-EPR#253. This pool consists of 370 ORFs ranging in size from 1857-2508 nt. |
| Depositing Lab |
Christine Clayton Heidelberg University |
| Depositing Scientist | Albina Waithaka |
| Antibiotic Resistance | Ampicillin |
| Bacterial Growth Temp | 37 |
| Copy Number | High |
| Selectable Marker | Hygromycin |
| Reference | This plasmid is from an unpublished project. Please acknowledge the depositing scientist. |
| MTA and Terms |
MTA (PDF download) |
How to cite this reagent:
In the Materials & Methods:
Tb_tethering plasmid library28 was a gift from Christine Clayton (European Plasmid Repository plasmid #251).
In the References:
No reference available. (This plasmid is not yet published.)
Tb_tethering plasmid library28 was a gift from Christine Clayton (European Plasmid Repository plasmid #251).
In the References:
No reference available. (This plasmid is not yet published.)
Sequence: download sequence
GAATTCGAGCTCATATAGTTGGTATGTATTCTAATTCCAGACTACTGGCGTGGATAAACATGTCCCCTGGAATTCGAGCTCATATAGTTGGTATGTATTCTAATTCCAGACTACTGGCGTGGATAAACATGTCCCCTGATTAAAAGGAAAGATTCCATAGCCCATTAGTGCAAAGATAATTGGTACACCGTCAAAAACACATGGGAGACCACACGATACGACACTGTGACCGTAGCATCAATTCCGCACTCGATATAACTCTATCGAAGAACTTCCATGGTACAAACTGGTCGCCACGGTCTGCTCCACACGGAGATCATCGTATCATTTTTATCGATAGCGGCCGCTATCGATGTATGCCTTGGCCCTGATGGCATGCCAATTTCACTACAACGGACTATGTGGACCCCGTTATCATGGAAATGCGCTAGTTGGAGGAAGTTAGACCGCGCCGGAAAAGAGAGGGGTAGAGAAAATGACAACTTGGAAGATATCCACACACGCACGGTGAAACGTTAGCAACAATTATTAGGGAAGCACGCTTGCCCTAGTCCCACGAGTAACCAAGACTCCAAAAGCCTTTCTGGCACAGAGAGCGAGCCGAAATGGAAAAGAGAAACAATGCCTGCACTAACACTACTGAGCGATTCGCCTCGCCGCGGAGGACCGAATACTAATAACGACACTTGCGGTCAAAAAGTAGAAGAACAAATGCTCAACGATGAGTGAATCAGGTTAGGGTAGTTGGAAAATTATACAGAATGTCTTTGGCAACACACCGGTACCGTCATTGGGGTTAAGCGGAAAGGTGTGTGTCAGTAGGTTGTGAGGTGAAAGCGTTTTCAGATGCATAGTGAGCTTAATGTCCTTTTCACAGTATATCGTGTCTGATAGGTATCTCTTATTAGTATAGTCGAATACTAGTCAATAGTGCGTTTTGTGCAAAATGTCCATTTTGTGGCAGTGATGGGGTTGTTTTATGCTATTCCGTGTCTCTGGGTGGGCGTGCATTGAAAATAGGGGTTATCGGGTATTAAAAGGAAAGATTCCATAGCCCATTAGTGCAAAGATAATTGGTACACCGTCAAAAACACATGGGAGACCACACGATACGACACTGTGACCGTAGCATCAATTCCGCACTCGATATAACTCTATCGAAGAACTTCCATGGTACAAACTGGTCGCCACGGTCTGCTCCACACGGAGATCATCGTATCATTTTTATCGATAGCGGCCGCTATCGATGTATGCCTTGGCCCTGATGGCATGCCAATTTCACTACAACGGACTATGTGGACCCCGTTATCATGGAAATGCGCTAGTTGGAGGAAGTTAGACCGCGCCGGAAAAGAGAGGGGTAGAGAAAATGACAACTTGGAAGATATCCACACACGCACGGTGAAACGTTAGCAACAATTATTAGGGAAGCACGCTTGCCCTAGTCCCACGAGTAACCAAGACTCCAAAAGCCTTTCTGGCACAGAGAGCGAGCCGAAATGGAAAAGAGAAACAATGCCTGCACTAACACTACTGAGCGATTCGCCTCGCCGCGGAGGACCGAATACTAATAACGACACTTGCGGTCAAAAAGTAGAAGAACAAATGCTCAACGATGAGTGAATCAGGTTAGGGTAGTTGGAAAATTATACAGAATGTCTTTGGCAACACACCGGTACCGTCATTGGGGTTAAGCGGAAAGGTGTGTGTCAGTAGGTTGTGAGGTGAAAGCGTTTTCAGATGCATAGTGAGCTTAATGTCCTTTTCACAGTATATCGTGTCTGATAGGTATCTCTTATTAGTATAGTCGAATACTAGTCAATAGTGCGTTTTGTGCAAAATGTCCATTTTGTGGCAGTGATGGGGTTGTTTTATGCTATTCCGTGTCTCTGGGTGGGCGTGCATTGAAAATAGGGGTTATCGGGTAGGGATCTCCCTATCAGTGATAGAGATCTCCCTATCAGTGATAGAGATCCCTGAGTACTGAGTTTAACATGTTCTCGTCCCGGGCTGCACGCGCCTTCGAGTTTTTTTTCCTTTTCCCCATTTTTTTCAACTTGAAGACTTCAATTACACCAAAAAGTAAAATTCACAAGCTACGCGTATGGACGCACAAACACGACGACGTGAGCGTCGCGCTGAGAAACAAGCTCAATGGAAAGCTGCAAACGGCGGGCTAGCCACCGTCGAGGGCGGCCGATCGAGATCCGAGGAGCAGAAGCTGATCTCGGAGGAGGATCTGCTGAGATCCGAGGAGCAgAAGCTGATCTCGGAGGAGGATCTGCTGAGATCCGAGGAGCAGAAGCTGATCTCGGAGGAGGATCTGCTGTCAAGCTTACAAGTTTGTACAAAAAAGCAGGCT[Gene of interest ORF]ACCCAGCTTTcTtGTACAAAGTGGTAGATCCTAACACCGGGTTGTGTGGCCAAAATTGTTCTGTAGTCGCTGTGAGTTGACACGGCTAGTGCTTATGATTTTCCTCGCGTGTGGTGCCTGTACTCAGCCCTATGCCTTATTTGCAACACATTTACGTACAGCGCACAAGAGAAGAGAAGATCACTTGAAGATAATAAATATAGGGTTGTAGGCATCTTGTTTAACTCAAATTTTCTCGTCTTGGTGTGTCGACATGATTGAAATAGTGCCACCAGTTGTGTTTGATGCGTTTGTTATCTATGCAGTATTCTGCAGTTTAATTTGTTGGATGAGCTATTTCATTAATTTTTTTGCCTTCTCTTCTTTTGGGTTCGCTCGAGTTTCCACCCAGCGCGGGTGCATTCTGGCTCTTATATATACTTATTGTCATGACAGAGTATATTGTACTGTGTTGATAAGGGACGGGTAACTGTATTGAAGAGCCGATGCTTTTGACATGTTAGATATAATATGTTTTATTGTAAAGTCAATACAACACACAATAGGATAATAATGATAAAGTTAAAAAAGTATATATAGTAATAGAAATATATCTTATATAGGAAAGATTAAGCAGTAAAAGTAGCGCTTACGGCGTACGGAGCAGGAGAGCAACTGACCGCTCTCAGAgcccGGGCACAGCAAGGTCTTCTGAAATTCATGTTTTTTTTTTTTTTACTCTGCATTGCAGTCTCCGCTCTTATTTAGTTTTGCTTTACGTAAGGTCTCGTTGCTGCCATAAAATAAGCTACTAGTGATGAAAAAGCCTGAACTCACCGCGACGTCTGTCGAGAAGTTTCTGATCGAAAAGTTCGACAGCGTCTCCGACCTGATGCAGCTCTCGGAGGGCGAAGAATCTCGTGCTTTCAGCTTCGATGTAGGAGGGCGTGGATATGTCCTGCGGGTAAATAGCTGCGCCGATGGTTTCTACAAAGATCGTTATGTTTATCGGCACTTTGCATCGGCCGCGCTCCCGATTCCGGAAGTGCTTGACATTGGGGAATTCAGCGAGAGCCTGACCTATTGCATCTCCCGCCGTGCACAGGGTGTCACGTTGCAAGACCTGCCTGAAACCGAACTGCCCGCTGTTCTGCAGCCGGTCGCGGAGGCCATGGATGCGATCGCTGCGGCCGATCTTAGCCAGACGAGCGGGTTCGGCCCATTCGGACCGCAAGGAATCGGTCAATACACTACATGGCGTGATTTCATATGCGCGATTGCTGATCCCCATGTGTATCACTGGCAAACTGTGATGGACGACACCGTCAGTGCGTCCGTCGCGCAGGCTCTCGATGAGCTGATGCTTTGGGCCGAGGACTGCCCCGAAGTCCGGCACCTCGTGCACGCGGATTTCGGCTCCAACAATGTCCTGACGGACAATGGCCGCATAACAGCGGTCATTGACTGGAGCGAGGCGATGTTCGGGGATTCCCAATACGAGGTCGCCAACATCTTCTTCTGGAGGCCGTGGTTGGCTTGTATGGAGCAGCAGACGCGCTACTTCGAGCGGAGGCATCCGGAGCTTGCAGGATCGCCGCGGCTCCGGGCGTATATGCTCCGCATTGGTCTTGACCAACTCTATCAGAGCTTGGTTGACGGCAATTTCGATGATGCAGCTTGGGCGCAGGGTCGATGCGACGCAATCGTCCGATCCGGAGCCGGGACTGTCGGGCGTACACAAATCGCCCGCAGAAGCGCGGCCGTCTGGACCGATGGCTGTGTAGAAGTACTCGCCGATAGTGGAAACCGACGCCCCAGCACTCGTCCGAGGGCAAAGGAATAGAGTAGATGCCGACCGGGATCGATCCAAGCTAGCTTGTATTCTATAGTGTCACCTAAATCGTATGTGTATGATACATAAGGTTATGTATTAATTGTAGCCGCGTTCTAACGACAATATGTACAAGCCTAATTGTGTAGCATCTGGCTTACTGAAGCAGACCCTATCATCTCTCTCGTAAACTGCCGTCAGAGTCGGTTTGGTTGGACGAACCTTCTGAGTTTCTGGTAACGCCGTTCCGCACCCCGGAAATGGTCAGCGAACCAATCAGCAGGGTCATCGCTAGCCAGATCCTCTACGCCGGACGCATCGTGGCCGGCATCACCGGCGCCACAGGTGCGGTTGCTGGCGCCTATATCGCCGACATCACCGATGGGGAAGATCGGGCTCGCCACTTCGGGCTCATGAGCGCTTGTTTCGGCGTGGGTATGGTGGCAGGCCCCGTGGCCGGGGGACTGTTGGGCGCCATCTCCTTGCACCATTCCTTGCGGCGGCGGTGCTCAACGGCCTCAACCTACTACTGGGCTGCTTCCTAATGCAGGAGTCGCATAAGGGAGAGCGTCGATATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCTTATCGAAATTAATACGACTCACTATAGGGAGACCG
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