Difference between revisions of "Part:BBa K3081056"
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SgRNA Generator was designed base on the type II CRISPR/Cas9 system including T7 promotor, lacZa', crRNA(tracrRNA) and T7 terminator, with two BsaI cutting sites flanking lacZa' coding sequence. To facilitate the construction of sgRNA expression arrays for multiplex DnaA boxes, the Golden Gate Assembly method was adopted to insert the target-specific sequence between two BsaI cutting sites. For constant expression of sgRNA, they were placed under the control of J23119 promoter. | SgRNA Generator was designed base on the type II CRISPR/Cas9 system including T7 promotor, lacZa', crRNA(tracrRNA) and T7 terminator, with two BsaI cutting sites flanking lacZa' coding sequence. To facilitate the construction of sgRNA expression arrays for multiplex DnaA boxes, the Golden Gate Assembly method was adopted to insert the target-specific sequence between two BsaI cutting sites. For constant expression of sgRNA, they were placed under the control of J23119 promoter. | ||
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| + | https://2019.igem.org/wiki/images/2/27/T--Peking--sgRNA_generator.png | ||
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| + | Figure1 Schematic cartoon of the DNA construct of <partinfo>BBa_K3081056</partinfo> | ||
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| + | This part is used in our CRISPRri system<partinfo>BBa_K3081058</partinfo> to generate specific 20bp sgRNA binding to different DNA boxes on the OriC,the genome replication origin of <i>E.coli</i>. For more information, see <partinfo>BBa_K3081058</partinfo>. | ||
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Latest revision as of 09:44, 21 October 2019
sgRNA generator with promoter J23119
SgRNA Generator was designed base on the type II CRISPR/Cas9 system including T7 promotor, lacZa', crRNA(tracrRNA) and T7 terminator, with two BsaI cutting sites flanking lacZa' coding sequence. To facilitate the construction of sgRNA expression arrays for multiplex DnaA boxes, the Golden Gate Assembly method was adopted to insert the target-specific sequence between two BsaI cutting sites. For constant expression of sgRNA, they were placed under the control of J23119 promoter.
Figure1 Schematic cartoon of the DNA construct of BBa_K3081056
This part is used in our CRISPRri systemBBa_K3081058 to generate specific 20bp sgRNA binding to different DNA boxes on the OriC,the genome replication origin of E.coli. For more information, see BBa_K3081058.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 251
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 441
Illegal BsaI.rc site found at 37