Difference between revisions of "Part:BBa K2381012"

Revision as of 06:27, 28 October 2017

3'-OriC gRNA

 It’s a gRNA plasmid which helps our CRISPR-System target to the MG1655 OriC sequence in order to achieve our expectation this year. Its backbone is taken from iGEM Kit Plate named BBa_K165005, but the BsaI restriction site which was in Amp. coding sequence has been deleted and a synonymous mutation has occurred by overlapping PCR. We used J23119 promoter, a high-intensity promoter, to obtain a higher expression of gRNA than dCas9. The 21bp-sequence comes after is a kind of sequence types taken from the article that is complementary to the Oric sequence and has a nice effect on our project. The full length gRNA scaffold is also designed after consulting literatures.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BamHI site found at 4
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K2381012 short</partinfo>
-It's a gRNA targeting towards OriC
+  It’s a gRNA plasmid which helps our CRISPR-System target to the MG1655 OriC sequence in order to achieve our expectation this year. Its backbone is taken from iGEM Kit Plate named BBa_K165005, but the BsaI restriction site which was in Amp. coding sequence has been deleted and a synonymous mutation has occurred by overlapping PCR. We used J23119 promoter, a high-intensity promoter, to obtain a higher expression of gRNA than dCas9. The 21bp-sequence comes after is a kind of sequence types taken from the article that is complementary to the Oric sequence and has a nice effect on our project. The full length gRNA scaffold is also designed after consulting literatures.
 <!-- Add more about the biology of this part here