The Cloning Vector Catalog, CloningVectors.sbp, was installed with Lasergene. It can be opened and edited like any other SeqBuilder Project. When you add a vector to this project and save the project, the vector is added to the Catalog and can be accessed for use in future SeqBuilder Pro projects. The next time you perform a cloning project, the custom vector will be available in the Vector drop-down list in the Clone tab of the Project window.
- Use File > Open and open the Vector Catalog project CloningVectors.sbp.
- Win: C:\Users\Public\Documents\DNASTAR\Lasergene ‘x’ Data\CloningVectors.sbp
- Mac: Hard Drive:Applications:DNASTAR:Lasergene ‘x’ Data:CloningVectors.sbp
… where ‘x’ represents the version number.
The Project window opens to display folders containing different categories of vectors.
- Click on the plus signs to the left of folders to expand and view their contents.
- Decide which folder is best for your custom vector, or add a new folder by right-clicking and choosing New Folder. Click on the desired folder to select it. Then right-click and choose Import.
- Select your custom vector sequence and press Open to add a copy of it to the Cloning Vector Catalog. Gateway cloning vectors must be formatted as specified below.
- Choose File > Save Project.
- (optional) If you would like to remove the vector you just added, click on it to select it. Then right-click on it and choose Delete. Then use File > Save Project again.
In subsequent Lasergene updates, DNASTAR will update the Cloning Vector Catalog, but will not overwrite any vectors that you have imported or customized.
If you plan to import a Gateway cloning vector in Step 4, above, you should open it in SeqBuilder Pro in advance, and prepare it as follows:
- Set the type to circular by selecting Sequence > Circular.
- Add misc_recomb features to your sequence on the appropriate sub-sequences where recombination occurs. Note that while it is preferred for the entire recombination site to exist within your sequence, SeqBuilder Pro at minimum requires the appropriate att core sequence for each of the five types of recombination sites. SeqBuilder Pro uses these core sequences along with the feature labels to correctly identify each Gateway plasmid:
- Type 1: GTACAAA (forward strand)
- Type 2: CTTGTAC (reverse strand)
- Type 3: ATTATAC (reverse strand)
- Type 4: GTATAGA (forward strand)
- Type 5: GTATACA (forward strand)
- Label the features as follows:
- For Expression Clones: attB1, attB2, attB3, attB4, and attB5
- For Donor Vectors: attP1, attP2, attP3, attP4, and attP5
- For Entry Clones: attL1, attL2, attL3, attL4, and attL5
- For Destination Vectors: attR1, attR2, attR3, attR4, and attR5