Difference between revisions of "Part:BBa K1351009"
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__NOTOC__ | __NOTOC__ | ||
<partinfo>BBa_K1351009 short</partinfo> | <partinfo>BBa_K1351009 short</partinfo> | ||
− | Flexible linker of | + | Flexible linker of 57 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 ([https://parts.igem.org/Part:BBa_K1351008 BBa_K1351008]), a linker (this part) and a cell wall-anchoring domain ([https://parts.igem.org/Part:BBa_K1351010 BBa_K1351010]) as well as the sortase YhcS itself ([https://parts.igem.org/Part:BBa_K1351011 BBa_K1351011]), whose overexpression increases the efficiency of the surface display. | The YhcR/YhcS systems consists of four components: the sortase substrate with a signal peptide ([https://parts.igem.org/Part:BBa_K1351008 BBa_K1351008]), a linker (this part) and a cell wall-anchoring domain ([https://parts.igem.org/Part:BBa_K1351010 BBa_K1351010]) as well as the sortase YhcS itself ([https://parts.igem.org/Part:BBa_K1351011 BBa_K1351011]), whose overexpression increases the efficiency of the surface display. | ||
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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> |
|} | |} | ||
Sites of restriction enzymes generating compatible overhangs have the same color: | Sites of restriction enzymes generating compatible overhangs have the same color: | ||
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<!-- --> | <!-- --> | ||
− | + | ===Sequence and Features=== | |
<partinfo>BBa_K1351009 SequenceAndFeatures</partinfo> | <partinfo>BBa_K1351009 SequenceAndFeatures</partinfo> | ||
Latest revision as of 00:41, 18 October 2014
57 aa flexible protein linker
Flexible linker of 57 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