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Team:Groningen/OurBiobrick
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<img class="centerimage" src="https://static.igem.org/mediawiki/2012/2/21/Groningen2012_EJ_20120912_psaccmt-RFP-contruct.png" width="200"> | <img class="centerimage" src="https://static.igem.org/mediawiki/2012/2/21/Groningen2012_EJ_20120912_psaccmt-RFP-contruct.png" width="200"> | ||
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| + | This plasmid backbone is designed to integrate a cloned insert into the Bacillus subtilis 168 chromosome at the sacA locus. The user inserts the fragment of interest between the prefix and suffix. The plasmid is transformed into any B. subtilis 168 host with selection for chloramphenicol (cat gene) resistance. This plasmid can be amplified inside E.coli with selection for chloramphenicol or ampicillin and inability to produce red fluorescent protein for inserted plasmid. The red fluorescent protein cannot be produced in B.subtilis. | ||
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{{Template:SponsorsGroningen2012}} | {{Template:SponsorsGroningen2012}} | ||
Revision as of 13:40, 21 September 2012
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 place them in the [http://partsregistry.org 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 [http://partsregistry.org Registry], not on your team wiki. Remember that the goal of proper part documentation is to describe and define a part such that it can be used without a need to refer to the primary literature. The next iGEM team 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.
| <groupparts>iGEM012 Groningen</groupparts> |
This backbone was designed to fulfill the need of a working Bacillus subtilis backbone for our project. This backbone plasmid was derived from pSac-Cm by insertion of biobrick compatible restriction sites (prefixes and suffixes), a terminator (BBa_B0015) after the suffixes sequences, and red fluorescent protein sequence (RFP) in between the prefix and suffix in its multiple cloning sites (MCS). New biobricks can be inserted into this vector by replacement of the RFP biobrick, and selection of the white colonies. This backbone has a multi host replication origin and replicates in E. coli and Bacillus subtilis. The plasmid is designed to integrate a cloned insert into the B. subtilis chromosome via double recombination between plasmid and chromosomal sacA sequences.
The terminator insertion after the suffixes in combination and insertion of the red fluorescent protein (RFP) in between the insertion site were meant to make cloning easier and faster. Transformants with inserts will not produce red color (as the RFP is replaced by the insert).
This plasmid backbone is designed to integrate a cloned insert into the Bacillus subtilis 168 chromosome at the sacA locus. The user inserts the fragment of interest between the prefix and suffix. The plasmid is transformed into any B. subtilis 168 host with selection for chloramphenicol (cat gene) resistance. This plasmid can be amplified inside E.coli with selection for chloramphenicol or ampicillin and inability to produce red fluorescent protein for inserted plasmid. The red fluorescent protein cannot be produced in B.subtilis.
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