Coding

Part:BBa_K3352000

Designed by: Joyce Ting   Group: iGEM20_TAS_Taipei   (2020-10-17)
Revision as of 04:07, 23 October 2020 by Joyceting (Talk | contribs)


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).

T--TAS_Taipei--Registry_1.png


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
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 790
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None