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SNAPGENE VIEWER RESTRICTION ENZYME DIGEST HOW TO
SNAPGENE VIEWER RESTRICTION ENZYME DIGEST FULL

SNAPGENE VIEWER RESTRICTION ENZYME DIGEST HOW TO

See the introduction tutorial video and the working with features tutorial video for further details on how to annotate sequences in SnapGene.

Change name of the feature in mRFP1-Rab5įill in the names, start and stop positions of the two genes and click OK:.

Right click ORF frame 2 in the map and select Edit Feature:Ĭhange name of the feature in NeoR/KanR and click OK:Ĭhange the name of ORF frame 1 and annotate it as a fusion. To switch back to mRFPrab5 click Window in the top menu and select mRFPrab5.

Rab5 from position 1330 to position 1977.

mRFP1 from position 613 to position 1329.

We are going to correct the annotation: change the names of the ORFs and split ORF frame 1 into two parts:

what is called ORF frame 1 in the map is a fusion between mRFP1 (a red fluorescent protein) and human Rab5 (a small GTPase that is a key regulator of intracellular membrane trafficking) gene.

what is called ORF frame 2 in the map is a neomycin/kanamycin resistance gene.

Since it's a vector sequence, select circular:Ĭlick OK to open the sequence in SnapGene.Ĭhanging annotation of sequences in SnapGeneĪlthough we imported mRFP1-Rab5 in Genbank format, the annotation that was included in the file was not complete: SnapGene will ask you wether the sequence is linear or circular. Open the annotated sequence of mRFP-Rab5 in SnapGene.Ĭlick the Open button in the top menu, browse to the mRFPrab5.txt file and open it. To include the annotation, you need the sequence in Genbank format:Ĭopy the text and paste it in Notepad. On the next page select the Sequence tab:

SNAPGENE VIEWER RESTRICTION ENZYME DIGEST FULL

On the next page containing the full sequence of the vector, click the Analyze Sequence button on the right: On the webpage of mRFP-Rab5 go to the SEQUENCE INFORMATION section and click Full (1) next to Depositor Sequences: Obtain the annotated sequence of mRFP-Rab5 from Addgenes. If you load the annotations (in Genbank format) SnapGene will automatically show them. Genbank format not only contains a sequence but also all annotations. Even if you don't load them from Genbank it is still a good idea to import them in Genbank format (and not in Fasta format). In SnapGene you can load sequences directly from Genbank or you can load them by importing a file. Fortunately, many vectors are available from Addgene, an organization that improves sharing of plasmids. However, not all vector sequences are stored in the SnapGene database.

Loading vector sequences from other resources Go back to SnapGene and click the Open button in the top menu:īrowse to the pAcGFP1-C1.dna file and open it. On the webpage of pAcGFP1-C1 click pAcGFP1-C1.dna: Try to load it without peeking at the solution. This vector is available in SnapGene's list of fluorescent protein genes and plasmids. You now see the circular view of the vector with restriction sites automatically visualized:Įxercise: load the sequence of vector pAcGFP1-C1 Open SnapGene and select Open in the startup menu:īrowse to the pcDNA4 TO.dna file and open it. Open the annotated sequence of pcDNA4/TO in SnapGene. On the webpage of pcDNA4/TO click the pcDNA4 TO.dna link to save the file: Annotation: every possible piece of information about a sequenceĭownload the annotated sequence of pcDNA4/TO from SnapGene.