Difference between revisions of "Part:BBa K1763424"

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===Usage===
 
===Usage===
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In order to express this construct as a protein, it is necessary to subclone it after a promoter and RBS. The 2015 UCLA iGEM team has made a composite part, [https://parts.igem.org/Part:BBa_K1763424 BBa_K1764424], has placed this part under control of the T7-RBS regulatory elements.
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This construct was successfully amplified using the [http://2015.igem.org/Team:UCLA/Notebook/Spider_Silk_Genetics/ICA_Oligo_Sequences post-elution primers] after ICA. The results of the amplification are shown below.
 
This construct was successfully amplified using the [http://2015.igem.org/Team:UCLA/Notebook/Spider_Silk_Genetics/ICA_Oligo_Sequences post-elution primers] after ICA. The results of the amplification are shown below.
 
[[File:7 7 2015.jpg|none|thumb|500px|'''Fig. 1''' E-gel of post-elution primer amplification of ICA on M-270 beads. The expected size of the desired product is ~406 bp. In addition to the expected bands, there are bands present at ~100 bp and at ~750 bp.]]
 
[[File:7 7 2015.jpg|none|thumb|500px|'''Fig. 1''' E-gel of post-elution primer amplification of ICA on M-270 beads. The expected size of the desired product is ~406 bp. In addition to the expected bands, there are bands present at ~100 bp and at ~750 bp.]]

Revision as of 18:58, 18 September 2015

MaSp2-3(1C3)

This is a 3-mer construct of MaSp2 constructed using Iterative Capped Assembly (ICA) protocols as outlined on the 2015 UCLA iGEM page [http://2015.igem.org/Team:UCLA/Notebook/Spider_Silk_Genetics/Protocols here]. This part is a coding sequence, with no regulatory elements present.

Biology

Native spider silk genes consist of many repeats of the MaSp genes, up to 100 repeats. The extensive repetition confers the properties of strength and elasticity that are commonly associated with spider silk threads. In order to examine the role of protein length and composition on the final properties of spider silk, we created this construct using Iterative Capped Assembly.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 346
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

Usage

In order to express this construct as a protein, it is necessary to subclone it after a promoter and RBS. The 2015 UCLA iGEM team has made a composite part, BBa_K1764424, has placed this part under control of the T7-RBS regulatory elements.

This construct was successfully amplified using the [http://2015.igem.org/Team:UCLA/Notebook/Spider_Silk_Genetics/ICA_Oligo_Sequences post-elution primers] after ICA. The results of the amplification are shown below.

Fig. 1 E-gel of post-elution primer amplification of ICA on M-270 beads. The expected size of the desired product is ~406 bp. In addition to the expected bands, there are bands present at ~100 bp and at ~750 bp.