Difference between revisions of "Part:BBa K1763424"
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<partinfo>BBa_K1763424 short</partinfo> | <partinfo>BBa_K1763424 short</partinfo> | ||
| − | 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. | + | 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. 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=== | ===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, [https://parts.igem.org/Part: | + | 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_K1763425 BBa_K1763425], 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. | 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. | ||
Latest revision as of 20:55, 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. 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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 346
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE 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_K1763425, 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.
