Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Mona Dotzler
Date created: 2014-10-04 11:00:00
Date modified: 2015-05-08 01:10:01
57 aa flexible protein linker
| Types | DnaRegion |
| Roles | CDS Coding |
| Sequences | BBa_K1351009_sequence (Version 1) |
Description
Flexible linker of 56 amino acids for translational fusion of protein domains. Used in the 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:
|ACCGGTTAATACTAGTAGCGGCCGCCTGCAG
|}
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.
Notes
The linker sequence corresponds to the linker used in Liew et. al., 2012.Oligonucleotides were designed as precut by EcoRI and PstI to shorten the synthesized fragments and to simplify the cloning into pSB1C3
Source
This part was generated by annealing synthesized oligonucleotides followed by ligation in pBS1C3:L56_ENX_SD_Ngo_precutE_fwd1: AATTCgcggccgctTCTAGAgtaaggaggaaGCCGGCATG ATCGATCCGGCAGGAAGTAGTCCGTCGACA
L56_fwd2: GAGCTTGCATTTGACGAAGAAGACGACGATGATGAAGAGCCTGAAGATGCAGGCCCGGGAGATGGTGTGCTCGGATTCGAA
L56_SNP_Age_precutP_fwd3: TTCGTCAAATGCAAGCTCCGGCACGACGACTACACCTGATCTTCCTGGGTCACCG ACCGGTtaatACTAGTagcggccgCTGCA
L56_SNP_Age_precutP_rev1: GcggccgctACTAGTattaACCGGT CGGTGACCCAGGAAGATCAGGTGTAGTCGTCGTGCCG
L56_rev2: GAGCTTGCATTTGACGAAGAAGACGACGATGATGAAGAGCCTGAAGATGCAGGCCCGGGAGATGGTGTGCTCGGATTCGAA
L56_ENX_SD_Ngo_precutE_rev3: GGAGTACTAGGTCCCTCAGGTGTCGACGGACTACTTCCTGCCGGATCGAT CATGCCGGCttcctccttacTCTAGAagcggccgcG