Coding

Part:BBa_K844005

Designed by: Ryan Putman   Group: iGEM12_Utah_State   (2012-10-02)

Spider Silk 1x 1E Subunit "U" with Met (ATG) start codon

Spider silk subunit native sequence; contains one elasticity domain and one strength domain, and Met (ATG) start codon on 5’ end.

IMPORTANT NOTE: This part uses Assembly Standard #23 (Silver Fusion) scar sites, which the sequence data below and on composite parts using this part will not reflect this scar sequence (it shows Assembly Standard #10 scars).


Usage and Biology

The sequence for this subunit was derived from the 1E silk subunit from Brooks et al. (2008), which is based on the native sequence of the MaSp2 silk gene in Argiope aurantia. See part BBa_K844001 for additional sequence details. This part contains an added Met (ATG) codon at the 5’ end and is designed for use as the first subunit in a longer spider silk construct. This part should be followed by numerous repeats of the BBa_K844001 silk subunit, and the silk coding region should end with a BBa_K844000 part, which contains a 10-Histidine tag and a double stop codon (TAATAA)

The silk subunit contains three main domains: an elasticity domain, a linker domain, and a strength domain. The elasticity domain contains sequences coding for beta-helices and beta-spiral in the protein; these structures increase the elasticity of the spider silk. The linker domain joins the strength and elasticity domains together. The strength domain will form beta-sheets and strengthen the silk fibers by cross-linking silk strands.


Related parts:

BBa_K844001 – The “U” silk subunit with 1 elasticity domain and one strength domain without any added Met (ATG) codons.


References:

Brooks AE, Stricker SM, Joshi SB, Kamerzell TJ, Middaugh CR, and Lewis RV. 2008. Properties of synthetic spider silk fibers based on Argiope aurantia MaSp2. Biomacromolecules 9:1506–10.


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 22
  • 1000
    COMPATIBLE WITH RFC[1000]


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