C. Athale's page // IISER Pune
Bio122: Introduction to synthetic biology
Here we aim to introduce concepts in synthetic biology and run you through a
hands on exercise in virtual cloning of a gene for expression. The target gene
- APE: a plasmid editor program
- pSB1A3 (Amp selection) http://parts.igem.org/Part:pSB1A3
- lacP promoter sequence (BBa_R0010) inside a pSB1C3 backbone
- rbs + GFPmut3 + ter1 + ter2 sequence allows us to include a
‘functional’ version of the sequence for translation (BBa_I13504)
All plasmids* can be downloaded from here.
Lab notebook: Questions to address
In your lab-notebooks note the following:
- The size of the 3 plasmids
- The schematic of the cloning strategy (i.e. draw the 3 plasmids you
started with, the enzymes you digested them with virtually, the ligation
reaction, planned transformation and selection.
- Sketch a representative figure of the DNA digest (in APE, choose "Enzyme
Selection" ---> "Digest")
- Note the sequence of the transcriptional terminator in your
- How the transcriptional terminator function?
Download the file: [pdf]
3A Assembly explained graphically
Some notes from the presentation in lab (LHC303)
Last updated: 25-Feb-2019