Coding

Part:BBa_K4317099

Designed by: Chris Song   Group: iGEM22_Seoul_Korea   (2022-10-12)
Revision as of 13:29, 12 October 2022 by Envien mc (Talk | contribs)


Secretion signal library

This part contains 12 signal peptides that can be used in E. coli and Bacillus subtilis. This part is a composite part composed of the previously registered basic part and the basic part newly registered by our team. Therefore, it is different from the nucleotide sequence we actually used. Therefore, we registered the actual base sequence of the library we used in the basic part. (BBa_K4317099)

800px-Secretion_signal_library.png

Usage and Biology

We did not digest the plasmid DNA directly with NdeI or NcoI, but cut the library part after amplification using the primers (AIO F- GGATAACCGTATTACCGCCT TTGAG, AIO R- CACAAACAGACGATAACGGCTCTC) shown in Fig. 1 (Fig. 2, 3). Secretion signal fragments were ligated with the DNA fragments coding for each enzyme with Bacillus and E. coli expression vectors and transformed (Fig. 4).


Figure 1. Plasmid map of signal peptide library

529px-Library_PCR.png

Figure 2. PCR product containing the signal peptide library

452px-Re.png

Figure 3. secretion signal fragments digested by NdeI, NcoI

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 NgoMIV site found at 820
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None