On VectorBuilder’s homepage, click Design My Vector, and you will be taken to the Choose Vector System page. Here, you will see a comprehensive list of available vector systems. You can then select your desired vector system by clicking the Go to Design button next to it. For example, if you want to design a lentiviral vector for expressing your gene of interest in mammalian cells, you would click the Go to Design button next to Lentivirus under the Mammalian Gene Expression Vectors section.
Detailed technical information for all our vector systems is available in the Guide to Vector Systems page.
Upon selecting a vector system, you will enter the Vector Design Studio page. Here, you will be presented with a map for the vector system of your choice. Two types of components are marked on the map. One is fixed components that you cannot change, which serve as the backbone of the vector. The other is custom components that you can add onto the backbone. You can enter your custom components one by one, such as the promoter for driving gene expression, the ORF to be placed behind the promoter, and accessory features such as the drug-selection marker or fluorescent reporter. When adding a component, you will have the option of selecting it from VectorBuilder’s component database or pasting your own sequence. When adding an ORF, you will also have the option to insert epitope tags or introduce mutations to the ORF. Below are some useful tips:
Selecting a component from database: Our extensive component databases contain two categories of sequences. One is popular components presented to you as a list for you to easily choose from (e.g. popular promoters such as CMV, EF1A and CAG). The other is gene-based databases for ORFs, shRNAs and gRNAs, which you can search by gene name and species to retrieve the component corresponding to your gene of interest. For example, if you wish to design a lentiviral vector that expresses the human BMP2 gene, then in the drop-down menu of the Add ORF step, you will need to choose Select from ORF database and search for human BMP2. The search should return a list of all the RefSeq transcripts for this gene, one of which is recommended by VectorBuilder. You can then select the ORF of one of these transcripts to add to your vector. You can also edit the ORF sequence (e.g. inserting epitope tags or introducing mutations into the ORF) before adding it to the vector.
Detailed technical information is available for all our popular components in the Guide to Vector Components page.
ORF editing capability: You can insert epitope tags or introduce point mutations into your ORF. You can do so after you have already added an ORF to your vector. Just click on the name of the ORF on the vector map and choose Edit ORF from the drop-down menu. This should open up a sequence editor window where you can make changes. Click the Add tag button below the sequence to reveal a popular set of tags that you can choose from to append to either the N or C terminus of your ORF. You can also directly edit your sequence by entering or deleting nucleotides. If you are editing a component retrieved from VectorBuilder’s database, then all edits will be tracked to record the changes you made relative to the original reference sequence. In the sequence editor window, there are other handy functions for you to use, such as translating nucleotide sequence into amino acid sequence, finding ORFs, and showing the sequence of the reverse-complementary strand.
Placing multiple ORFs behind a single promoter: When you design a gene expression vector, you can place up to 4 ORFs behind a promoter as a polycistron. The multiple ORFs can be expressed as a single fusion protein, or as distinct proteins separated by 2A or IRES linkers. The Select number of ORFs option on the upper-left corner of the Vector Design Studio window allows you to set the number of ORFs that you wish to place behind the promoter.