Difference between revisions of "Part:BBa K1351006"

Latest revision as of 15:49, 5 October 2014

CWB domain of B. subtilis major autolysin LytC

Cell wall binding domain and signal peptide (amino acids 1 to 333) of Bacillus subtilis major autolysin LytC (N-acetylmuramoyl-L-alanine amidase, Uniprot [http://www.uniprot.org/uniprot/Q02114 Q02114]). This domain has been used for displaying heterologous proteins on the cell surface of B. subtilis (Tsuchiya et. al., 1999). Used in the [http://2014.igem.org/Team:LMU-Munich/Project/Bakillus BaKillus project] to display pathogen-specific peptides by C-terminal fusions to mediate adhesion of B. subtlis to pathogens.

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

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

@@ Line 13: / Line 13: @@
 |-
 |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>CGGCCGC</span><span style="color:blue">CTGCAG</span>
+|<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: