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Team:WHU-China/Parts/Data
From 2011.igem.org
Data for our favorite part
BBa_K560000 Main page - Blue Light Sensing and Reporting System: This part shows the complete system of light sensing and reporting regulation in E.coli. Although it is not very stable, it can truly work in TrpR(-) E.coli.
Data for our plasmid backbone parts
1. BBa_K560012, BBa_K560013, BBa_K560014, BBa_K560015 - BsaI Golden Gate Assembly No. 1~4: Vectors of the Golden Gate Series (adapted from pSB1C3) .To make this set of vectors compatible with the standard biobricks, we have maintained PstI and XbaI recognition sites. Flanking these two sites are the cleavage and recognition sites of BsaI, a kind of Type IIS restriction endonuclease. This kind of restriction endonucleases cleaves outside their recognition sites thus creating different sticky ends. If you want to assemble four biobricks at a time, you can put the biobricks in order into corresponding vector by the digestion of PstI and Xbal. Then mix these assembled vectors and a backbone (PFUS4 from TALEN Golden Gate kit). Under the effect of ligase and Bsal, these biobricks will have sticky ends nose to tails and ligate in order in a single reaction. 2. BBa_K560016 Main page - The backbone for golden gate assembly: This backbone is taken from TALEN Golden gate Kit, and it can expedite golden gate cloning together with a set of vectors for golden gate submitted by WHU-igem team. This plasmid contains the coding sequence for α-strand of LacZ. Flanking LacZ are two recognition sites of Bsal, a kind of type IIS restriction endonuclease. If you want to assembly four biobricks at a time, you can mix this backbone with four assembled vectors(BBa_K560012, BBa_K560013, BBa_K560014, BBa_K560015), and add adequate amount of ligase and Bsal.
Data for pre-existing parts
1. Experience – BBa_I15010 Cph8 (Cph1/EnvZ fusion) (UTAustin, iGEM2004): This part was used by 2011 WHU_China team as a part of light sensing system. However, the result of sequencing proves that this part in the 2011 kit turns out to be a vector without the fragment of cph8. 2011 WHU_China requested the same part from the 2011 kit of HUST_China, but the result of pcr and restriction enzymes cleavage shows that their plasmids have the same problem. 2. Experience – BBa_E0040 Green fluorescent protein derived from jellyfish Aequeora victoria wild-type GFP (Endy Lab, iGEM2004): This part was used by WHU_China team as a reporter of long-term oscillator. Information from the website of this part shows that its length is about 720bp; however, the result of restriction enzymes cleavage experiment showed that it is actually about 1kb. And after several attempts, we failed to assemble this part with other biobricks.
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