Difference between revisions of "Part:BBa K2996003"

Latest revision as of 04:17, 20 October 2019

Tac promoter and operator plus RBS

Tac-Promoter (abbreviated as Ptac) is a synthetically produced DNA promoter, produced from the combination of promoters from the trp and lac operons. It is commonly used for protein production in Escherichia coli

We constructed the pRead plasmid for information storage, where RSRL array and EGFP are fused downstream of this promotor. Spacer acquisition carried out by cas1/2 would cause an addition of 61 base pairs, a protospacer and another repeat sequence, thus resulting in frame shifting. In that case, the stop codon would disappear, and EGFP enter the correct reading frame.

Whenever you want to read the information, add IPTG to a final concentration of 0.5M and incubate at 20℃, leading to the expression of fluorescent protein.

For more detailed results, click BBa_K2996002.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K2996003 short</partinfo>
+Tac-Promoter (abbreviated as Ptac) is a synthetically produced DNA promoter, produced from the combination of promoters from the trp and lac operons. It is commonly used for protein production in Escherichia coli
-The cassette consists of 69 bp of the respective leader, including the sequence elements essential for adaptation, and one spacer flanked by two repeats. Wildtype E.coli has two types of CRISPRER array, with their repeat sequence differ in one base pair. Thus, we name them 1T and 2A.
+We constructed the pRead plasmid for information storage, where RSRL array and EGFP are fused downstream of this promotor.
+Spacer acquisition carried out by cas1/2 would cause an addition of 61 base pairs, a protospacer and another repeat sequence, thus resulting in frame shifting. In that case, the stop codon would disappear, and EGFP enter the correct reading frame.
+Whenever you want to read the information, add IPTG to a final concentration of 0.5M and incubate at 20℃, leading to the expression of fluorescent protein.
-#Usage and Biology
+For more detailed results, click BBa_K2996002.
-In our design, this integration cassette RSRL in pRead consists of a leader and two repeats interspaced by a spacer fused downstream, to the complete coding sequence of an out-of-frame EGFP gene (EGFP +2).
-Originally, there is a stop codon within the RSRL sequence and the fluorescent protein gene was not in the correct reading frame, so there would be no fluorescent protein expressed. After illuminating, Cas1/2 would insert 61bp protospacer and another repeat sequence, which would result in frame shifting. In that case, the stop codon would disappear, and EGFP enter the correct reading frame.
-When you want to read the information, inducer should be added and expression of fluorescent protein would be activated.