EENdb - Utilities  >  TALEN framework & linker peptides
Summary
The framework/scaffold of TALEN contains the sequences beside the RVDs of each repeat units. Most natural TALEs have similar frameworks, except for the numbers of repeat units. N- and C-terminal regions of the terdem repeat region of TALE are often trancated in artificial TALENs / TAL effectors. Different lengths of the C-terminal region have been reported to prefer different lengths of TALEN spacer.

The most frequently used framework in artificial TALENs is "+63", which was first reported by 21179091, with 63 aa remained in the C-terminus. Another framework also has a 63-aa length C-terminus called "+63-Goldy" was reported and compared to the previous one by 23000899.

The linker peptide of TALEN is defined as the amino acid sequence between the C-terminus region of TALE DNA-binding domain and the beginning of FokI cleavage domain (QLVKS...), which are often very short when compared to the C-terminus region.

Sequence of a typical TALEN with framework "+63"
Sequence (posterior region of FokI cleavage domain omitted & only 4.5 repeat units remained), based on 21179091

M A P K K K R K V D Y K D H D G D Y K D H D I D Y K D D D D K G T V D L R T L G Y S Q Q Q Q E K I K P K V R S T V A Q H H E A L V G H G F T H A H I V A L S Q H P A A L G T V A V K Y Q D M I A A L P E A T H E A I V G V G K Q W S G A R A L E A L L T V A G E L R G P P L Q L D T G Q L L K I A K R G G V T A V E A V H A W R N A L T G A P L N L T P D Q V V A I A S N I G G K Q A L E T V Q R L L P V L C Q D H G L T P E Q V V A I A S H D G G K Q A L E T V Q R L L P V L C Q A H G L T P D Q V V A I A S N N G G K Q A L E T V Q R L L P V L C Q A H G L T P A Q V V A I A S N G G G K Q A L E T V Q R L L P V L C Q D H G L T P D Q V V A I A S N G G K Q A L E T V Q R L L P V L C Q D H G . . . . . . . . . . . . L T P E Q V V A I A S N G G G R P A L E S I V A Q L S R P D P A L A A L T N D H L V A L A C L G G R P A L D A V K K G L P H A P A L I K R T N R R I P E R T S H R V A G S . . .


Sequence with annotation:

|< SV40 NLS>| |<---------- Tag (e.g. 3xFlag) ---------->| GT: Tag-TAL linker peptide (L.) M A P K K K R K V D Y K D H D G D Y K D H D I D Y K D D D D K G T |<---------------- N-terminal region (Δ152; 136 aa remained) V D L R T L G Y S Q Q Q Q E K I K P K V R S T V A Q H H E A L V G H G F T H A H I V A L S Q H P A A L G T V A V K Y Q D M I A A L P E A T H E A I V G V G K Q W S G A R A L E A L L T V A G E L R G P P L Q N-terminal region ----------------->| L D T G Q L L K I A K R G G V T A V E A V H A W R N A L T G A P L N |<---------------- repeat units (33-34 aa each) ----------------->| * A/D/E |-| RVD * A/D L T P D Q V V A I A S N I G G K Q A L E T V Q R L L P V L C Q D H G L T P E Q V V A I A S H D G G K Q A L E T V Q R L L P V L C Q A H G L T P D Q V V A I A S N N G G K Q A L E T V Q R L L P V L C Q A H G L T P A Q V V A I A S N G G G K Q A L E T V Q R L L P V L C Q D H G |-| RVD "N*" of a 33-aa repeat unit L T P D Q V V A I A S N G G K Q A L E T V Q R L L P V L C Q D H G . . . . . . . . . . . . last 0.5 unit only have 20 aa L T P E Q V V A I A S N G G G R P A L E |<-------------- C-terminal region (+63; 63 aa remained) S I V A Q L S R P D P A L A A L T N D H L V A L A C L G G R P A L D C-terminal region ----------------->| L. |<- FokI A V K K G L P H A P A L I K R T N R R I P E R T S H R V A G S . . .



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