Difference between revisions of "Part:BBa K3352000"
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We optimized the DNA sequence for expression in E. coli and removed the PstI cutting site. We attached a 6x histidine tag (6x His-Tag) upstream of the SplintR ligase sequence for purification purposes followed by a glycine-serine linker (GS linker) to form our ORF (BBa_K3352001). This served as our open reading frame (ORF; BBa_K3352000). | We optimized the DNA sequence for expression in E. coli and removed the PstI cutting site. We attached a 6x histidine tag (6x His-Tag) upstream of the SplintR ligase sequence for purification purposes followed by a glycine-serine linker (GS linker) to form our ORF (BBa_K3352001). This served as our open reading frame (ORF; BBa_K3352000). | ||
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Revision as of 04:07, 23 October 2020
SplintR Ligase with His-Tag and GS Linker Sequence
SplintR Ligase catalyzes the ligation of adjacent single-stranded DNA splinted by complementary RNA strands [4]. SplintR ligase has been previously shown to be capable of differentiating these ligation junctions to SNP levels and ligate padlock probes [4].
Construct Design
We optimized the DNA sequence for expression in E. coli and removed the PstI cutting site. We attached a 6x histidine tag (6x His-Tag) upstream of the SplintR ligase sequence for purification purposes followed by a glycine-serine linker (GS linker) to form our ORF (BBa_K3352001). This served as our open reading frame (ORF; BBa_K3352000).
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 790
- 1000COMPATIBLE WITH RFC[1000]