Coding

Part:BBa_K4623013

Designed by: Ruixiao Tan   Group: iGEM23_BNU-China   (2023-10-09)
Revision as of 12:49, 10 October 2023 by Tanruixiao (Talk | contribs) (Induction Condition)


Nucleotide Silinker(NS)

Usage and Biology

The Nucleotide Silinker (NS) is a novel engineered protein that covalently links nucleic acid ends to biotinylated adapters. The sequence in FASTA format includes an added HIS tag, enabling purification of the NS protein using a nickel column. Upstream of the mSA sequence, a TrxA (BBa_K3619001) fusion tag is added to aid protein folding and reduce the formation of inclusion bodies in bacterial cells. Following protein expression, cleavage by thrombin exposes the mSA (BBa_K4623001) site, allowing binding to biotinylated functional proteins. The TrwC protein can covalently link specific 5' single-stranded nucleotide sequences, facilitating the conversion between nucleic acids and protein connectors.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 1024
    Illegal PstI site found at 1342
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 1024
    Illegal PstI site found at 1342
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 1024
    Illegal PstI site found at 1342
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 1024
    Illegal PstI site found at 1342
    Illegal AgeI site found at 448
    Illegal AgeI site found at 508
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1147
    Illegal SapI.rc site found at 862

Cultivation, Purification and SDS-PAGE

Induction Condition

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Categories
Parameters
None