version 1.0

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NEBridge SplitSet® Tool

Request ID

     20240116113747031884

Overhang set

     TTGT,GGTA,TAAG,CTGA



Estimated ligation fidelity: 100%


Using the given set of overhangs, Golden Gate Assembly is predicted to yield 100% of correctly-ligated products.


Ligation frequency matrix
 
TTGT
ACAA
GGTA
TACC
TAAG
CTTA
CTGA
TCAG
TTGT
0
155
0
0
0
0
0
0
ACAA
155
0
0
0
0
0
0
0
GGTA
0
0
0
414
0
0
0
0
TACC
0
0
414
0
0
0
0
0
TAAG
0
0
0
0
0
288
0
0
CTTA
0
0
0
0
288
0
0
0
CTGA
0
0
0
0
0
0
0
392
TCAG
0
0
0
0
0
0
392
0
Legend

 
good Watson-Crick pair
 
poor Watson-Crick pair
 
high-count mismatch
 
modest mismatch
 
trace mismatch

Overhang coordinates
Sequence fragments
>3k2y_F1 length=71 coord=1..71 5'-overhang=none 3'-overhang=TTGT
ATGACAGGCGATGCAGCCGTCGCATTGGATACAGTCACGGTCGTTGGCGAACGGTATGTCGATGATATTGT
>3k2y_F2 length=33 coord=68..100 5'-overhang=TTGT 3'-overhang=GGTA
TTGTGGCAACGTTAACGACGCTCAGAGTGGGTA
>3k2y_F3 length=84 coord=97..180 5'-overhang=GGTA 3'-overhang=TAAG
GGTATGGCGGTGTTGCTCCAGCGAGAAAGTGGCAATCAATACGATGATAATGCCATCTCAGTGTGGACGTTGCAACATGCTAAG
>3k2y_F4 length=50 coord=177..226 5'-overhang=TAAG 3'-overhang=CTGA
TAAGTTAGGCTACATTGCCCGGTATCAGAACCAGCCGTACGCGACACTGA
>3k2y_F5 length=84 coord=223..307 5'-overhang=CTGA 3'-overhang=none
CTGATGGATCAGGGGCAACGATTGTATGGCATCGTGACGGTATTGGATCAGCAAAAACAGCATCTTGAATTGATGCTATGGCGG