Team:TCU Taiwan/Parts

From 2014.igem.org

 
Parts
 
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 change width 586 to 520 & 745 Fig.1
BBa_K1473005  
   
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 phagemid-carrying phages. 

pBluescript contains a f1 ori while M13KO7 helper phage’s f1 ori has been mutated. So after the infection of M13KO7, its genome can produce protein and replicate but these proteins will package pBluescript as "genome". As a result, the pahemid-carrying phaes will only contain pBluescript, they cannot make next generation.
BBa_K1473005
 
 
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 it on or off.
SourceĀ :Science

BBa_K1473009
 
 
 
Parts Sandbox
 
Click on the name of the parts for detailed information.
 
NameTypeDescriptionDesignerLength
  BBa_K1473001DNAgRNA 1 Shan, Zhe22
  BBa_K1473002DNAgRNA 2Shan, Zhe22
  BBa_K1473003DNAgRNA 3Shan, Zhe22
  BBa_K1473004DNAgRNA 4Shan, Zhe22
  BBa_K1473005Plasmid_Backbonephagemid pBluescript Shan, Zhe2929
  BBa_K1473006DNAgRNA for ampR 1Shan, Zhe22
  BBa_K1473007DNAgRNA for ampR 2Shan, Zhe22
  BBa_K1473008DNAgRNA for ampR 3Shan, Zhe22
 BBa_K1473009CodingpRha induced CRISPRShan, Zhe5011
  BBa_K1473010DNAgRNA for ampR 4Shan, Zhe22
  BBa_K1473011DNAgRNA for ampR 5Shan, Zhe22
  BBa_K1473012DNAgRNA for NeoR/kanR 1Shan, Zhe22
  BBa_K1473013DNAgRNA for NeoR/kanR 2Shan, Zhe22
  BBa_K1473014DNAgRNA for NeoR/kanR 3Shan, Zhe22
  BBa_K1473015DNAgRNA for NeoR/kanR 4Shan, Zhe22
  BBa_K1473016DNAgRNA for NeoR/kanR 5Shan, Zhe22
  BBa_K1473017DNAgRNA for tetR 1Shan, Zhe22
  BBa_K1473018DNAgRNA for tetR 2Shan, Zhe22
  BBa_K1473019DNAgRNA for tetR 3Shan, Zhe22
  BBa_K1473020DNAgRNA for tetR 4Shan, Zhe22
  BBa_K1473021DNAgRNA for tetR 5Shan, Zhe22
TCU_Taiwan 2014 iGEM Team Parts
gRNA
 
Fig.2
 
Fig.3

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 surffix. Then we cut this recombinant plasmid with sacI and run gel electrophoresis for checking. 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 Table

This table is from the Database of iGEM.

Any parts our team has created will appear in this table below:
<groupparts>iGEM014 TCU_Taiwan</groupparts>
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Team Members Project Parts Human Pratics Modeling Safety Notebook Attributions

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