Viraless

Overview 

Our team planned to use 12 basic parts. We inserted the B5R gene (with 1st sushi and all 4 sushi ectodomains) into pET23a plasmid transformed into E. coli BL21(DE3) (no. 1 and 2). We used 3 primers for B5R gene cloning from pVAX plasmid (one forward - no. 8 - and two reverse - no. 9 and 10). Other possible genetic models for B5R protein synthesis can be Codon optimized B5R (all 4 sushi domains) sequence (no. 11) and B5R sequence (all 4 sushi domains) with altered Cys140 to Ser (no. 12). 

    Moreover, since we used the L1, A33, and A27 proteins of the Vaccinia virus for optic fiber functionalization, we included them as other basic parts (no. 5-7). We also included aptamer sequences for L1 protein as the example reference for aptamer modeling in MAWS software made by the iGEM Heidelberg 2021 team. 

Parts List

 

Code

Description 

1

BBa_K4389000

B5R (all 4 sushi domains)

2

BBa_K4389001

B5R 1st sushi

3

BBa_K4389003

DNA aptamer for L1 protein

4

BBa_K4389004

DNA aptamer for L1 protein (2)

5

BBa_K4389005

L1 protein encoding sequence

6

BBa_K4389006

A33 protein encoding sequence

7

BBa_K4389007

A27 protein encoding sequence

8

BBa_K4389008

Forward B5R sequence primer

9

BBa_K4389009

Reverse primer for B5 protein (all 4 sushi domains)

10

BBa_K4389010

Reverse primer B5R sequence (only 1st sushi)

11

BBa_K4389011

Codon-optimized B5R (all 4 sushi domains)

12

BBa_K4389012

B5R sequence with altered Cys140 to Ser