Composite

Part:BBa_K3490021:Design

Designed by: Yao Tien Cheng   Group: iGEM20_NCKU_Tainan   (2020-10-23)
Revision as of 05:41, 25 October 2020 by Jason Tsai (Talk | contribs)


Verify the function of EL222-pBlind and FLP-FRT systems.


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1573
    Illegal SpeI site found at 1891
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1573
    Illegal NheI site found at 7
    Illegal NheI site found at 30
    Illegal NheI site found at 3035
    Illegal NheI site found at 3058
    Illegal SpeI site found at 1891
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1573
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1573
    Illegal SpeI site found at 1891
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1573
    Illegal SpeI site found at 1891
    Illegal NgoMIV site found at 142
    Illegal AgeI site found at 367
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 2882
    Illegal BsaI.rc site found at 3115


Design Notes

during the experiment process, we had difficulty constructing both plasmids. With the doubt that the EL222-FLP system might be harmful to the growth of bacteria, we changed the design of blue-sensitive protein EL222 into thermo-sensitive protein CI857 on the plasmid pCP20 provided by our PI, Prof. Ng


Source

Derived from IDT synthesis.

References


[1]Guglielmini J, Van Melderen L. Bacterial toxin-antitoxin systems. Mobile Genetic Elements. 2011;1(4):283-306.
[2]Jørgensen MG, Pandey DP, Jaskolska M, Gerdes K. HicA of Escherichia coli Defines a Novel Family of Translation-Independent mRNA Interferases in Bacteria and Archaea. Journal of Bacteriology. 2008;191(4):1191-1199.‌
[3]Motta-Mena LB, Reade A, Mallory MJ, et al. An optogenetic gene expression system with rapid activation and deactivation kinetics. Nature Chemical Biology. 2014;10(3):196-202.
[4]Branda CS, Dymecki SM. Talking about a Revolution. Developmental Cell. 2004;6(1):7-28.
[5]Jechlinger W, Szostak MP, Witte A, Lubitz W. Altered temperature induction sensitivity of the lambda pR/cI857 system for controlled gene expression in Escherichia coli. FEMS Microbiology Letters. 1999;173(2):347-352.
[6]Ringrose L. Quantitative comparison of DNA looping in vitro and in vivo: chromatin increases effective DNA flexibility at short distances. The EMBO Journal. 1999;18(23):6630-6641.
[7]Park Y-N, Masison D, Eisenberg E, Greene LE. Application of the FLP/FRT system for conditional gene deletion in yeast Saccharomyces cerevisiae. Yeast. 2011;28(9):673-681.