DNA

Part:BBa_K2100055:Design

Designed by: Trinh Nguyen   Group: iGEM16_MIT   (2016-10-19)


pENTR L4_EGSH 4x k-turn_R1


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1265
    Illegal XbaI site found at 2225
    Illegal PstI site found at 1758
    Illegal PstI site found at 2232
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1265
    Illegal NheI site found at 810
    Illegal NheI site found at 1076
    Illegal PstI site found at 1758
    Illegal PstI site found at 2232
    Illegal NotI site found at 2239
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1265
    Illegal BglII site found at 1388
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1265
    Illegal XbaI site found at 2225
    Illegal PstI site found at 1758
    Illegal PstI site found at 2232
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1265
    Illegal XbaI site found at 2225
    Illegal PstI site found at 1758
    Illegal PstI site found at 2232
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 687


Design Notes

The distance from the k-turn repeats to the open reading frame of the downstream gene determines the efficiency of L7Ae/k-turn repressing system. Thus, we were considering adding k-turns right after the promoter (in the pENTR promoter vector) or right in front of the gene coding sequence (in the pENTR gene vector).



Source

EGSH was obtained using PCR extension from pENTR pEGSH (BBa_K2100021). The k-turn sequence was obtained from addgene (plasmids deposite by Dr. Ron Weiss' lab). We ordered the k-turns as single-stranded oligos with different Q sites for Golden Gate reaction.


References