Difference between revisions of "Part:BBa K1763430"

 
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<partinfo>BBa_K1763430 short</partinfo>
 
<partinfo>BBa_K1763430 short</partinfo>
  
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MaSp2 12 mer in pSB1C3. Does not contain regulatory elements
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This is a 12-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. This construct includes an N-terminal 8-his tag which can be used purify this protein using nickel based affinity purification techniques.
  
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===Usage and Biology===
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===Biology===
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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.
  
 
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<partinfo>BBa_K1763430 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K1763430 SequenceAndFeatures</partinfo>
  
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===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_K1763431 BBa_K1763431], 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.
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[File:7 16 2015 UCLA ICA.jpg|none|thumb|500px|'''Fig. 1''' Post-elution primer amplification of MaSp2-12. The expected band product is 1324 bp.]]
  
 
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Revision as of 20:57, 18 September 2015

MaSp2-12(1C3)

This is a 12-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. This construct includes an N-terminal 8-his tag which can be used purify this protein using nickel based affinity purification techniques.

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 1264
  • 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_K1763431, 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. [File:7 16 2015 UCLA ICA.jpg|none|thumb|500px|Fig. 1 Post-elution primer amplification of MaSp2-12. The expected band product is 1324 bp.]]