Table 1.
Plasmids used and constructed within this study with properties and multiple cloning site characteristics
| Plasmid name | Resistance | Affinity tags | Leader sequence | Unique sticky end restriction sites in MCS |
|---|---|---|---|---|
| pTD-NStrepHis |
Sm |
Strep-tagII/His6 |
- |
MCSI: NheI, BstBI, EcoRI, Acc65I, XmaI, BamHI, XhoI, SalI, NcoI, HindIII1, KpnI, PstI, SacI |
| pTD-NTwinStrep_Sm |
Sm |
TwinStrep/His6 |
- |
MCSII: EcoRI, Acc65I, XmaI, BamHI, XhoI, SalI, NcoI, HindIII1, SacI PstI, KpnI, |
| pTD-NTwinStrep_Km |
Km |
TwinStrep/His6 |
- |
MCSII: EcoRI, Acc65I, XmaI, BamHI, XhoI, SalI, NcoI, HindIII1, PstI, KpnI, SacI |
| pTD-CTwinStrep |
Sm |
TwinStrep |
- |
MCSIII: BamHI, EcoRI, SacI, SalI, XbaI, XhoI, HindIII2 |
| pTDpelB-NTwinStrep |
Sm |
TwinStrep/His6 |
PelB |
MCSIV: EcoRI, Acc65I, XmaI, BamHI, XhoI, SalI, SacI, KpnI, PstI, HindIII1 |
| pTDpelB-CTwinStrep |
Sm |
TwinStrep |
PelB |
MCSV: NcoI, EcoRI, SacI, XbaI, XhoI, SacI, KpnI, PstI, HindIII2 |
| pTD-C_eYFPTwinStrep |
Sm |
eYFP-TwinStrep |
- |
MCSVI: BamHI, EcoRI, XbaI, XhoI, HindIII2 |
| pTD-C_sfYFPTwinStrep |
Sm |
sfYFP-TwinStrep |
- |
MCSVI: BamHI, EcoRI, XbaI, XhoI, HindIII2 |
| pTDpelB-C_eYFPTwinStrep |
Sm |
eYFP-TwinStrep |
PelB |
MCSVII: NcoI, EcoRI, XbaI, XhoI, HindIII2 |
| pTDpelB-C_sfYFPTwinStrep | Sm | sfYFP-TwinStrep | PelB | MCSVII: NcoI, EcoRI, XbaI, XhoI, HindIII2 |
1 use of this restriction site results in a construct without the C-terminal HIS6-tag, addition of a stop codon is recommended; 2 use of this restriction site results in a construct without the C-terminal Twin-Strep-tag® addition of a stop codon is recommended. Note MCSI, II, IV, contain PshAI sites restricting the SEVA selection marker exchangeability.