Part:BBa_K1351009:Design

57 aa flexible protein linker

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
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI site found at 95
  

Design Notes

The linker sequence corresponds to the linker used in Liew et. al., 2012, and Chen et. al., 2007, except for a change from C to G at position 20 to remove a SpeI site, changing amino acid 7 from Thr to Ser.

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

References

Chen, C. L., S. C. Wu, W. M. Tjia, C. L. Wang, M. J. Lohka and S. L. Wong. "Development of a Lyte-Based High-Density Surface Display System in Bacillus Subtilis." Microb Biotechnol 1, no. 2 (2008): 177-90. [http://www.ncbi.nlm.nih.gov/pubmed/21261835 PubMed]

Liew, P. X., C. L. Wang and S. L. Wong. "Functional Characterization and Localization of a Bacillus Subtilis Sortase and Its Substrate and Use of This Sortase System to Covalently Anchor a Heterologous Protein to the B. Subtilis Cell Wall for Surface Display." J Bacteriol 194, no. 1 (2012): 161-75. [http://www.ncbi.nlm.nih.gov/pubmed/22020651 PubMed]