DNA

Part:BBa_K3683002:Design

Designed by: Leyi Zheng, Xu Lin Zi Cai, JieYing Gan, Guanghui Sun, Ouqiao Li, Charlotte Zhang, Taylor Khouw Shimizu, Emily Roshan   Group: iGEM20_PYMS_GZ_China   (2020-10-19)


S(1-1254aa) DNA D614G


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 3094
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1435
    Illegal NgoMIV site found at 1951
    Illegal NgoMIV site found at 2014
    Illegal NgoMIV site found at 2509
    Illegal NgoMIV site found at 2653
    Illegal NgoMIV site found at 2686
    Illegal NgoMIV site found at 2963
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 647
    Illegal BsaI.rc site found at 898
    Illegal BsaI.rc site found at 2558
    Illegal SapI.rc site found at 915
    Illegal SapI.rc site found at 1596


Design Notes

Cell culture

293T cells were maintained in Dulbecco’s Modified Eagle Medium (Invitrogen) supplemented with 10% fetal bovine serum (FBS; Gibco, Rockville, MD, USA), 100 mg/mL of streptomycin, and 100 unit/mL of penicillin at 37 °C in 5% CO2. HEK293T cells transfected with human ACE2 (293T-ACE2) were cultured under the same conditions with the addition of G418 (0.5 mg/mL) to the medium.


Source

HEK293T cells and ACE2-293T cells (cells transfected with human ACE2) were purchased from ProCell (Wuhan, China).

Serum sample (harvested at Day 35 post vaccination by an RBD vaccine was used for infectivity inhibition experiment. Detailed information is provided by Yang et al, Nature 2020 and was a kindly gift from Dr. Jingyun Yang (West China Hospital, Sichuan University).

References