Coding

Part:BBa_K4449001:Design

Designed by: SRIJANI SAHA , RASHMI RANJAN BEHERA , SATYAM KUMAR SINGH   Group: iGEM22_IISER_Berhampur   (2022-09-28)


FimH2


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 232
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 232
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 232
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 232
    Illegal AgeI site found at 390
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

A single G base was inserted at the 13th base of the forward primer sequence while designing primers to maintain the reading frame of the sequence during cloning in expression vector Pet15b. Deleted the ATG start codon from the coding sequence since the ATG start codon is already present in the restriction enzyme recognition site (Nco 1)which will be added in the primers.


Source

The part was retrieved from pBAD-FimH -9X His plasmid (Addgene - https://www.addgene.org/97305/) which ranged from 346 bp to 882bp of the plasmid backbone.

References


1. pBAD-FimH-9XHis https://www.addgene.org/97305/
2. Schembri, M.A., Hasman, H. and Klemm, P. (2000). Expression and purification of the mannose recognition domain of the FimH adhesin. FEMS Microbiology Letters, 188(2), pp.147–151.DOI: 10.1111/j.1574-6968.2000.tb09186.x