Difference between revisions of "Part:BBa K1351009"
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|suffix with AgeI, SpeI, NotI and PstI: | |suffix with AgeI, SpeI, NotI and PstI: | ||
| − | |<span style="color:orange">ACCGGT</span><u>TAA</u>T<span style="color:red">AC<u>TAG</u><u>T</u></span><u>A</u><span style="color:green"><u>G</u> | + | |<span style="color:orange">ACCGGT</span><u>TAA</u>T<span style="color:red">AC<u>TAG</u><u>T</u></span><u>A</u><span style="color:green"><u>G</u>CGGCCG</span><span style="color:blue">CTGCAG</span> |
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Sites of restriction enzymes generating compatible overhangs have the same color: | Sites of restriction enzymes generating compatible overhangs have the same color: | ||
Revision as of 15:50, 5 October 2014
57 aa flexible protein linker
Flexible linker of 56 amino acids for translational fusion of protein domains. Used in the [http://2014.igem.org/Team:LMU-Munich/Project/Bakillus BaKillus project] to display pathogen-specific peptides on the surface of B. subtilis with the YhcR/YhcS system to mediate adhesion of B. subtlis to pathogens.
The YhcR/YhcS systems consists of four components: the sortase substrate with a signal peptide (BBa_K1351008), a linker (this part) and a cell wall-anchoring domain (BBa_K1351010) as well as the sortase YhcS itself (BBa_K1351011), whose overexpression increases the efficiency of the surface display.
This part was generated in a modified version of RFC25, where a strong Shine Dalgarno Sequence (SD) is included, and has the following prefix and suffix:
| prefix with EcoRI, NotI, XbaI, SD and NgoMIV: | GAATTCGCGGCCGCTTCTAGAGTAAGGAGGAGCCGGC |
| suffix with AgeI, SpeI, NotI and PstI: | ACCGGTTAATACTAGTAGCGGCCGCTGCAG |
Sites of restriction enzymes generating compatible overhangs have the same color:
EcoRI and PstI in blue, NotI in green, XbaI and SpeI in red, NgoMIV and AgeI in orange. Shine-Dalgarno sequence and stop codons are underlined.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 95