Difference between revisions of "Part:BBa K1763436"
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<partinfo>BBa_K1763436 short</partinfo> | <partinfo>BBa_K1763436 short</partinfo> | ||
| − | + | This is a 12-mer construct of MaSp1 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. | |
| − | <!-- Add more about the biology of this part here | + | <!-- Add more about the biology of this part here --> |
| − | === | + | ===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. | ||
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<partinfo>BBa_K1763436 SequenceAndFeatures</partinfo> | <partinfo>BBa_K1763436 SequenceAndFeatures</partinfo> | ||
| + | ===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:BBa_K1763437 BBa_K1763437], 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:8 28 2015 UCLA ICA M1 12.jpg|none|thumb|500px|'''Fig. 1''' PCR amplification of MaSp1-12 using post-elution primers. The expected band size is 1324 bp.]] | ||
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===Functional Parameters=== | ===Functional Parameters=== | ||
<partinfo>BBa_K1763436 parameters</partinfo> | <partinfo>BBa_K1763436 parameters</partinfo> | ||
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Latest revision as of 21:19, 18 September 2015
MaSp1-12(1C3)
This is a 12-mer construct of MaSp1 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 1264
- 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_K1763437, 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.
