Coding

Part:BBa_K1351011:Design

Designed by: Mona Dotzler   Group: iGEM14_LMU-Munich   (2014-10-05)


Bacillus subtilis sortase YhcS


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 34
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 199
    Illegal SapI.rc site found at 508


Design Notes

none


Source

This part was generated by amplification from B. subtilis W168 gDNA with the primers listed below, followed by digestion with EcoRI and PstI and ligation into pSB1C3.

YhcS_ENX_SD_Ngo_fwd: gatcGAATTCgcggccgctTCTAGAgtaaggaggaaGCCGGC GTGAAAAAAGTTATTCCACTATTCATCATTGC YhcS_SNP_rev: gatcCTGCAGcggccgctACTAGTa TTAAGTCACTCGTTTTCCATATATAATATAGCG

An EcoRI site was removed by changing T to C at position 399 without altering the protein sequence by fusion PCR with the following mutagenesis primers:

YhcS_EcoRI435mut_fwd: CGTCTTCTCCTGTCATATGGAGAATTtACATATGAAATTGTAAAAACAAAAATTGTCG YhcS_EcoRI435mut_rev: CGACAATTTTTGTTTTTACAATTTCATATGTaAATTCTCCATATGACAGGAGAAGACG

References