Coding

Part:BBa_K4449000:Design

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


FimH1


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 238
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 238
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 238
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 238
    Illegal AgeI site found at 396
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

A single T base was inserted at the 13 th base of the forward primer sequence while designing primers so as to maintain the reading frame of the sequence during cloning in expression vector Pet28b+. 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

This 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.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


2.Tao Wang , Wang Yin,Hadi AlShamaileh , Yumei Zhang , Phuong Ha-Lien Tran , Tuong Ngoc-Gia Nguyen , Yong Li , Kuisheng Chen , Miaomiao Sun, Yingchun Hou , Weihong Zhang, Qingxia Zhao, Changying Chen , Pei-Zhuo Zhang, and Wei Duan, 2019 Feb;30(1),A detailed protein-SELEX protocol allowing visual assessments of individual steps for high success rate, DOI: 10.1089/hgtb.2018.237