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Part:BBa_K2599008

Designed by: YEN-LING CHEN   Group: iGEM18_NCTU_Formosa   (2018-09-16)
Revision as of 10:38, 16 September 2018 by Yen-ling (Talk | contribs)


T7 Promoter+RBS+Subtilosin+intein+CBD

NCTU_Formosa 2018 designed a composite part encoding the Subtilosin A sequence (BBa_K2599000), and then combined with a T7 promoter (BBa_I712074), a ribosome binding site (BBa_B0034), intein and chintin binding domain (CBD).


Figure 1 biobrick picture


Introduction

Subtilosin A is an antimicrobial peptide produced by Bacillus subtilis and belongs to the bacteriocin class V family. It had bacteriocidal activity against some gram-positive bacteria such as Listeria, some species of Bacillus and E.faecium.


Structure Analysis

The amino acid was determined to be as follows:X-Gly-Leu-Gly-Leu-Trp-Gly-Asn-Lys-Gly-Cys-Ala-Thr-Cys-Ser-(sequence; see text) Ile-Gly-Ala-Ala-Cys-Leu-Val-Asp-Gly-Pro-Ile-Pro-Asp-Glx-Ile-Ala-Gly-Ala. The analyses of cross-linking structures revealed that there were linkages between the amino- and carboxyl-termini and between the Cys-19 and the Glx-28 residues through an unknown residue with a residue weight of 163. Consequently, subtilosin A was deduced to be a cyclic peptide antibiotic with a novel cross-linking structure.


Figure 2 structure picture



Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1055
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 778
    Illegal AgeI site found at 868
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 698


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