Project

Part:BBa_K3646010:Design

Designed by: Sukanya Chakraborty   Group: iGEM20_IISER_Berhampur   (2020-10-25)


FRaPPeT2:NS-5_CID_FRET


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1450
    Illegal EcoRI site found at 2880
    Illegal XbaI site found at 2533
    Illegal XbaI site found at 2826
    Illegal XbaI site found at 4014
    Illegal PstI site found at 859
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1450
    Illegal EcoRI site found at 2880
    Illegal PstI site found at 859
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1450
    Illegal EcoRI site found at 2880
    Illegal BamHI site found at 3919
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1450
    Illegal EcoRI site found at 2880
    Illegal XbaI site found at 2533
    Illegal XbaI site found at 2826
    Illegal XbaI site found at 4014
    Illegal PstI site found at 859
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1450
    Illegal EcoRI site found at 2880
    Illegal XbaI site found at 2533
    Illegal XbaI site found at 2826
    Illegal XbaI site found at 4014
    Illegal PstI site found at 859
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 3120
    Illegal BsaI.rc site found at 3280
    Illegal BsaI.rc site found at 3686


Design Notes

Gene synthesized using Splicing by Overlap Extension. Used as a part of FRaPPe combined CID-FRET reporter to study host-viral PPIs in mammalian HEK293 cells. Other components sourced from Addgene Plasmid 20148 https://www.addgene.org/20148/



Source

Gene sequence of NS-5 was sourced from https://www.ncbi.nlm.nih.gov.

TSV08 Strain, Serotype 1, GenBank Accession : KR919821.1

eYFP and FRB CDS is a part of the YR plasmid from Addgene used for designing the FRaPPe test construct T2.

References