Part:BBa_K2100055
pENTR L4_EGSH 4x k-turn_R1
This basic part entry vector is flanked by L4 and R1 sites, which are used to denote a promoter. This can be cascade with a gene (flanked by L1, L2 sites) using an LR reaction and cloned into a backbone that has a negative selection marker between R4 and R2 sites. This part adheres to RFC 65 for recombination based cloning of mammalian parts. k-turn stands for kink turn motif in the messenger RNA. The k-turn motif is a part of the L7Ae/k-turn RNA-based repressing system. By putting the k-turn motifs in pENTR promoter vector, the k-turns will be added into the 5'UTR of the gene after LR reaction. The k-turn adds in a higher gene regulation level to reduce the basal expression of the gene downstream due to leaky expression of the EGSH promoter. Theoretically, the number of k-turn motifs would affect the efficiency of L7Ae/k-turn repressing system. Thus, promoter with 4x k-turn would have higher efficiency in suppressing leaky expression than promoter with just 2x k-turn pENTR EGSH 2x k-turn
Sequence and Features
- 10INCOMPATIBLE 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 - 12INCOMPATIBLE 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 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 1265
Illegal BglII site found at 1388 - 23INCOMPATIBLE 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 - 25INCOMPATIBLE 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 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 687
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