Difference between revisions of "Part:BBa K3868097"

Revision as of 08:59, 18 October 2021

pCBE

The pCBE plasmid contains the lambda operator, cytidine deaminase, Uracil DNA glycosylase inhibitor and LVA degradation labels. The CBE / sgRNA complex can bind to the double-stranded DNA to form an R-loop in a sgRNA and PAM-dependent manner. CDA catalyzes the deamination of cytosines located at the top (non-complementary) strand within 15–20 bases upstream from PAM, which results in C-to-T mutagenesis. Sequence and Features

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Illegal EcoRI site found at 1572
12
INCOMPATIBLE WITH RFC[12]
Illegal EcoRI site found at 1572
Illegal NheI site found at 1331
21
INCOMPATIBLE WITH RFC[21]
Illegal EcoRI site found at 1572
Illegal BglII site found at 5013
Illegal BamHI site found at 3610
Illegal XhoI site found at 4616
23
INCOMPATIBLE WITH RFC[23]
Illegal EcoRI site found at 1572
25
INCOMPATIBLE WITH RFC[25]
Illegal EcoRI site found at 1572
1000
COMPATIBLE WITH RFC[1000]

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 <partinfo>BBa_K3868097 short</partinfo>
-pET--MME-eGFP plasmid contains pT7, lacO, MME, thrombin, eGFP-His-tag etc.Based on the plasmid of pET-24a, the C-terminal of MME was fused with eGFP in order to characterize the yield of MME. Moreover, in order to purify the MME , the enzyme loci of thrombin was inserted between MME and eGFP, and the label of 6*His was inserted at the end of eGFP, yielding the plasmid of pET-MME-eGFP. The fluorescence intensity of eGFP could be used to indicate the expression level of MME.
+The pCBE plasmid contains the lambda operator, cytidine deaminase, Uracil DNA glycosylase inhibitor and LVA degradation labels. The CBE / sgRNA complex can bind to the double-stranded DNA to form an R-loop in a sgRNA and PAM-dependent manner. CDA catalyzes the deamination of cytosines located at the top (non-complementary) strand within 15–20 bases upstream from PAM, which results in C-to-T mutagenesis.
 <!-- Add more about the biology of this part here
 ===Usage and Biology===