Development and comparison of cell-free protein synthesis systems derived from typical bacterial chassis

Bioresources and Bioprocessing

Table 1 RBS sequences designed by RBS Library Calculator

Bacterial specie	RBS number	RBS sequence	Translation initiation rate
E. coli	Original	GAAGGAG	6748.38
	1	CCCGAUGGGAUCACGCAUCUAAGG	0.13
	2	UCCGAUGCGACAAGGCAGGUCCUA	9.43
	3	UCCGAUGGGACAAGGCAGAUCCUA	38.37
	4	CCCGAUGCGAUCAAGGAUCUAAGG	158.33
	5	CCCGAUGGGAUCAAGAAUGUAAGG	642.63
	6	UCCGAUGGGACAAAGGAGGUUGUA	67,421.00
B. subtilis	Original	GAAGGAG	9491.88
	1	UUAUAGACAUUGGACAGUUCCUCGUUA	0.35
	2	UUAUAGACAUUGGGAAGUUCCUCCUUA	32.55
	3	UUAUAGACAUUGGAAAGUUACUCCUUA	130.15
	4	ACAUAGACAUUGGAAAGUUACUUCUA	464.48
	5	ACGAUGUAGUGAAGGGGUUACUUAUA	2053.54
	6	ACAAUGUAGUGAAGGGGGUACUUAUU	189,477.64
C. glutamicum	Original	GAAGGAG	8338.00
	1	UUCGGUUUACUAAACGCGCCCUUAU	0.12
	2	UGCGGUUUACUAAACGGACCCUUAU	8.12
	3	UGCGGUUUACUAAAAGCACCCAUAU	32.02
	4	UUCAGUUGGUAAAGGGAACCCAUAU	95.11
	5	GUCAGAUGGUAAAAAGGAGUUAAUCC	416.15
	6	UAAUAGUCACUUUAAGGAGGUUUAU	50,737.39
V. natriegens	Original	GAAGGAG	6748.83
	1	CCGUAUUUUUUCGACGCCGGUAUCCU	0.15
	2	CAGUCUUUUUUCAGCGCAGGUAAACA	12.18
	3	CCGUAUUUUUUCAAGGCCGGUAACAU	50.95
	4	CCGUAUUUUUUCAAUGGCGGUAAUAU	201.77
	5	CCGUAUUUUUUCUACGGAGGUAUUCU	1184.25
	6	CCGUAUUUUUUCAAGGGAGGUUAUAU	31,367.00