Plasmid Guide

GenBank files of each plasmid’s sequence are available for download below.

Each plasmid has one distinct sequence but may have several cloning options based on a combination of different restriction enzymes. Reference the tables for detailed breakdowns of the elements that are included or excluded in each cloning option.

Multi-Subunit Vector

Plasmid 1: Contains the ttRNAP protein – a model DNA/RNA-binding multi-subunit protein – with a C-terminal CL7 tag.

  • Plasmid 1, Options 1-3 - Bsu36I/SwaI, HindIII/SfiI, and AhdI/SpeI - excise subunits 1, 2, and 3, respectively

multi-subunit vector table

Single Subunit Vectors

Plasmid 1: Contains the ttRNAP protein – a model DNA/RNA-binding multi-subunit protein – with a C-terminal CL7 tag.

Plasmid 3: Serves as a template for tagging target proteins with an N-terminal CL7 tag.

Plasmid 4: Serves as a control; it encodes the CL7 model (CL7M) protein, which directly binds the Im7 protein immobilized on the Im7 Resin.

Group 1: Vectors for Membrane Proteins

Plasmid 8: Plasmid #8 is used for the expression of membrane proteins using an engineered signal peptide. It includes a C-terminal His8 tag and CL7 tags with cleavage sites before the His8 tag and in between the His8 tag and the CL7 tag.

    Plasmid 9: Optimized for the expression of membrane proteins with C-terminal His8 and CL7
      Plasmid 10: Allows for the purification of membrane proteins. It includes an N-terminal His8 tag, a C-terminal CL7 tag, and an engineered signal peptide from the Bacillus halodurans YidC gene.

        Group 20: N-terminal CL7 and Expression Tags

        Plasmid 21: Enables target proteins to be tagged with N-terminal CL7 and His8.  There are three options for the 5’ placement of target genes, and XhoI is the 3’ cloning site.

        • Plasmid 21, Option 1 - HindIII/XhoI Insertion Site – Trx | CL7 | PSC | 44-Amino Acid Linker | PSC | His8 Tag | SUMO | 13-Amino Acid Linker | SRT | Gene of Interest
        • Plasmid 21, Option 2 - KpnI/XhoI Insertion Site – Trx | CL7 | PSC | 44-Amino Acid Linker | PSC | His8 Tag | SUMO | 3-Amino Acid Linker | Gene of Interest
        • Plasmid 21, Option 3 - Bsu36I/XhoI Insertion Site – Trx | CL7 | PSC | 44-Amino Acid Linker | PSC | His8 Tag | SUMO | Gene of Interest

        Plasmid 22: Enables the tagging of target proteins with a cleavable CL7 at the N-terminus and an uncleavable His8 at the C-terminus. There are three options for the 5’ placement of target genes, and SpeI is the 3’ cloning site. 

        • Plasmid 22, Option 1 - HindIII/SpeI Insertion Site – Trx | CL7 | 20-Amino Acid Linker | SUMO | SRT | 13-Amino Acid Linker | Gene of Interest | His8
        • Plasmid 22, Option 2 - KpnI/SpeI Insertion Site – Trx | CL7 | 20-Amino Acid Linker | SUMO | 3 aa | Gene of Interest | His8
        • Plasmid 22, Option 3 - Bsu36I/SpeI Insertion Site – Trx | CL7 | 20-Amino Acid Linker | SUMO | Gene of Interest | His8
        • Plasmid 22, Option 4 - HindIII/XhoI Insertion Site – Trx | CL7 | 20-Amino Acid Linker | SUMO | SRT | 13-Amino Acid Linker | Gene of Interest 
        • Plasmid 22, Option 5 - KpnI/XhoI Insertion Site – Trx | CL7 | 20-Amino Acid Linker | SUMO | 3 aa | Gene of Interest 
        • Plasmid 22, Option 6 - Bsu36I/XhoI Insertion Site – Trx | CL7 | 20-Amino Acid Linker | SUMO | Gene of Interest 
        group 20 vector table

        Group 30: C-terminal CL7 and N-terminal Expression Tags

        Plasmid 31: Enables the labeling of target proteins with both cleavable N-terminal His8 and cleavable C-terminal CL7 tags.

        • Plasmid 31, Option 1 - HindIII/SpeI Insertion Site – Trx | SUMO | His8 | TEV | Gene of Interest | SRT | PSC | PSC | PSC | CL7
        • Plasmid 31, Option 2 - KpnI/SpeI Insertion Site – Trx | SUMO | His8 | Gene of Interest | SRT | PSC | PSC | PSC | CL7
        • Plasmid 31, Option 3 - Bsu36I/SpeI Insertion Site – Trx | SUMO | Gene of Interest | SRT | PSC | PSC | PSC | CL7
        Plasmid 32: Enables target proteins to be labeled with an uncleavable N-terminal His8 and a cleavable C-terminal CL7 tag.

            Plasmid 33: Enables target proteins to be labeled with C-terminal His8 and CL7, retaining the His8 tag depending on the protease utilized.

            • Plasmid 33, Option 1 - HindIII/SpeI Insertion Site – Trx | SUMO | Gene of Interest | SRT | His8 | PSC | PSC | PSC | CL7
            • Plasmid 33, Option 2 - KpnI/SpeI Insertion Site – Trx | SUMO | Gene of Interest | SRT | His8 | PSC | PSC | PSC | CL7
            • Plasmid 33, Option 3 - Bsu36I/SpeI Insertion Site – Trx | SUMO | Gene of Interest | SRT | His8 | PSC | PSC | PSC | CL7

            Plasmid 34: Enables target proteins to be labeled with C-terminal His8 and CL7.  Protease cleavage removes both tags.

            • Plasmid 34, Option 1 - HindIII/SpeI Insertion Site – Trx | SUMO | Gene of Interest | SRT | PSC | PSC | PSC | His8 | CL7
            • Plasmid 34, Option 2 - KpnI/SpeI Insertion Site – Trx | SUMO | Gene of Interest | SRT | PSC | PSC | PSC | His8 | CL7
            • Plasmid 34, Option 3 - Bsu36I/SpeI Insertion Site – Trx | SUMO | Gene of Interest | SRT | PSC | PSC | PSC | His8 | CL7