Difference between revisions of "Part:BBa K1638018:Design"
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===Source=== | ===Source=== | ||
| − | + | pUT18C | |
| − | + | <br> | |
| + | Human thioredoxin sequence: UniProt (AC: P10599) | ||
===References=== | ===References=== | ||
| + | [1]: Karimova G, Pidoux J, Ullmann A, Ladant D. A bacterial two-hybrid system based on a reconstituted signal transduction pathway. Proceedings of the National Academy of Sciences of the United States of America. 1998;95(10):5752-6. | ||
| + | <br> | ||
| + | [2]: Borghouts C, Kunz C, Delis N, Groner B. Monomeric Recombinant Peptide Aptamers Are Required for Efficient Intracellular Uptake and Target Inhibition. Molecular Cancer Research. 2008;6(2):267-81. | ||
Latest revision as of 10:05, 18 September 2015
hTrx-based scaffold fused to T18 through a flexible linker
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 624
Illegal XhoI site found at 753 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 165
Illegal NgoMIV site found at 575
Illegal AgeI site found at 381 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
This part was synthesized by IDT as a gBlock gene fragment.
Library
- Choose the size of your library. Typically the length of the variable peptide loop is between 10 and 20 amino acids.
- Dependent on the size your variable peptide loop, you can for example contruct the library as following: ATATGCGCTCGAG(NNK)xGAGCTCATTACCG (where N represent A, T, G or C and K represent G or C). The NNK codon minimizes the number of stop codons. The XhoI restriction included allows insertion of library into the scaffold.
- Order the library as duplex-DNA
- When delivered, cut the library and scaffold with XhoI restriction enzyme and ligate.
Important note: This part cannot be used in the standard backbones pSB1C3 or pSB1K3, as they include a xhoI restriction site. Instead we recommend using the pSB1A2 backbone
Source
pUT18C
Human thioredoxin sequence: UniProt (AC: P10599)
References
[1]: Karimova G, Pidoux J, Ullmann A, Ladant D. A bacterial two-hybrid system based on a reconstituted signal transduction pathway. Proceedings of the National Academy of Sciences of the United States of America. 1998;95(10):5752-6.
[2]: Borghouts C, Kunz C, Delis N, Groner B. Monomeric Recombinant Peptide Aptamers Are Required for Efficient Intracellular Uptake and Target Inhibition. Molecular Cancer Research. 2008;6(2):267-81.