Phagemid pBluescript is a special plasmid, it functions as a normal plasmid when transformed into bacteria. But when helper phage M13KO7 infect the bacteria, it will produce progeny M13 phage as vector for pBluescript.
pBluescript contains a f1 ori while M13KO7 helper phage’s f1 ori has been knock-down. So after the infection of M13KO7, its genome can produce protein and replicate but these proteins will package pBluescript as their true “genome”. As a result, the progeny phage’s genome will only contains pBluescript’s MCS, they cannot make next generation.
Source : XVIVO/Barcroft USA
BBa_K1473009
Here we combined a CRISPR system(BBa_K1218011) with an inducible promoter(BBa_K914003). The CRISPR system contains a tracrRNA , a Cas9 protein and a minimal
CRISPR array, it functions to knockout target gene. We give this system an inducible promoter so we can decided when to switch if on or off.
We put a sacI restriction cutting site downstream to gRNA (not shown in sequence information) and ligated it back to pSB1C3 with correct prefix and surfix. Then we cut this recombinant plasmid with sacI and run gel electrophoresis for testing. As we can see in this image, because pSB1C3 only contains 1 sacI cutting site itself, so it would not be cut into 2 parts when no gRNA is ligated inside. But if there is, then sacI will cut this recombinant plasmid into 2 parts, whose lengths are 939 bp and 1159 bp. Obviously, we have successfully synthesized this gRNA.
Parts Submitted to the Registry
What information do I need to start putting my parts on the Registry?
An important aspect of the iGEM competition is the use and creation of standard biological parts. Each team will make new parts during iGEM and will submit them to the Registry of Standard Biological Parts. The iGEM software provides an easy way to present the parts your team has created. The "groupparts" tag will generate a table with all of the parts that your team adds to your team sandbox.
Note that if you want to document a part you need to document it on the Registry, not on your team wiki. Future teams and other users and are much more likely to find parts on the Registry than on your team wiki.
Remember that the goal of proper part documentation is to describe and define a part, so that it can be used without a need to refer to the primary literature. Registry users in future years should be able to read your documentation and be able to use the part successfully. Also, you should provide proper references to acknowledge previous authors and to provide for users who wish to know more.
When should you put parts into the Registry?
As soon as possible! We encourage teams to start completing documentation for their parts on the Registry as soon as you have it available. The sooner you put up your parts, the better recall you will have of all details surrounding your parts. Remember you don't need to send us the DNA to create an entry for a part on the Registry. However, you must send us the sample/DNA before the Jamboree. Only parts for which you have sent us samples/DNA are eligible for awards and medal requirements.
The information needed to initially create a part on the Registry is:
Part Name
Part type
Creator
Sequence
Short Description (60 characters on what the DNA does)
Long Description (Longer description of what the DNA does)
Design considerations
We encourage you to put up much more information as you gather it over the summer. If you have images, plots, characterization data and other information, please also put it up on the part page. Check out part BBa_K404003 for an excellent example of a highly characterized part.
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BBa_K1473005
BBa_K1473009
