Part:BBa_K2246000
BSLA: siRNA production and reporter cassette
This part is for producing any small interfering RNA (siRNA), it is produced attached to a mRNA for expressing blue chromoprotein as a reporter. Cassette designed for E.coli siRNA production. This part may express any siRNA sequence desired, as it includes a BamHI and HincII cut sites limiting the region where the siRNA sequence is to be inserted. Also include a blue chromoprotein coding region as a reporter of production of the mRNA. The region designed for siRNA insertion contains the AWD siRNA coding region in the format (sense, 9 bp loop, anti-sense). This part is a universal expression cassette for small interfering RNA (siRNA) in E.coli. This cassette contains the coding sequence of amilCP Blue Chromoprotein (BBa_K592009) as a reporter of expression. This method, designed by team TecCEM 2017, is named BSLA, which stands for Blue chromoprotein, Sense, Loop, Antisense; as it is its structure. The desired siRNA must be produced with the format (BamHI cut-site, sense, loop, antisense ending with blunt end). It is recommended to synthetisize two DNA oligos with the previously metioned format, then align them. The cassette must be digested with BamHI and HincII, then ligated with the siRNA oligos. Finally the ligation is transformed into E.coli HT115 to maximize the production. This part comes with the insert for producing AWD siRNA (BBa_K2246002), and it must be removed for the production of other siRNA.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 822
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 750
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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