version 1.0
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NEBridge SplitSet® Tool 


Request ID

     20240116112829030954


Overhang set

     CTGA,AACC,AATG,AGAA







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






 


CTGA


TCAG


AACC


GGTT


AATG


CATT


AGAA


TTCT







CTGA


0


392


0


0


0


0


0


0







TCAG


392


0


0


0


0


0


0


0







AACC


0


0


0


455


0


0


0


0







GGTT


0


0


455


0


0


0


0


0







AATG


0


0


0


0


0


431


0


0







CATT


0


0


0


0


431


0


0


0







AGAA


0


0


0


0


0


0


0


194







TTCT


0


0


0


0


0


0


194


0










Legend









 

good Watson-Crick pair





 

poor Watson-Crick pair





 

high-count mismatch





 

modest mismatch





 

trace mismatch





Overhang coordinates



Sequence fragments


>1cbs_F1 length=88 coord=1..88 5'-overhang=none 3'-overhang=CTGA
ATGCCCAACTTCTCTGGCAACTGGAAAATCATCCGATCGGAAAACTTCGAGGAATTGCTCAAAGTGCTGGGGGTGAATGTGATGCTGA
>1cbs_F2 length=81 coord=85..165 5'-overhang=CTGA 3'-overhang=AACC
CTGAGGAAGATTGCTGTGGCTGCAGCGTCCAAGCCAGCAGTGGAGATCAAACAGGAGGGAGACACTTTCTACATCAAAACC
>1cbs_F3 length=102 coord=162..263 5'-overhang=AACC 3'-overhang=AATG
AACCTCCACCACCGTGCGCACCACAGAGATTAACTTCAAGGTTGGGGAGGAGTTTGAGGAGCAGACTGTGGATGGGAGGCCCTGTAAGAGCCTGGTGAAATG
>1cbs_F4 length=80 coord=260..339 5'-overhang=AATG 3'-overhang=AGAA
AATGGGAGAGTGAGAATAAAATGGTCTGTGAGCAGAAGCTCCTGAAGGGAGAGGGCCCCAAGACCTCGTGGACCAGAGAA
>1cbs_F5 length=79 coord=336..415 5'-overhang=AGAA 3'-overhang=none
AGAACTGACCAACGATGGGGAACTGATCCTGACCATGACGGCGGATGACGTTGTGTGCACCAGGGTCTCCGTCCGAAAG