SuperFolder GFP (sfGFP) has
1, Cycle3 mutations: F99S, M153T, and V163A;
2, Enhanced GFP mutations: F64L and S65T;
3, Six additional mutations: S30R, Y39N, N105T, Y145F, I171V, A206V;
4, Q80R mutation
5, N39I, T105K, E111V, I128T, K166T, I167V and S205T

Characteristics of SuperFolder GFP:

1, 100% fluorescence recovery after denaturing by urea
2, Very bright
3, Faster chromophore formation
3, 50%ssfGFP 1-10 OPT expressed in E. coli is soluble
Availability of SuperFolder GFP

Gene synthesis from http://www.Excellgen.com

GFP 1-10 OPT nucleotide sequence:
ATGAGCAAAGGAGAAGAACTTTTCACTGGAGTTGTCCCAATTCTTGTTGAATTAGATGGT 60
GATGTTAATGGGCACAAATTTTCTGTCAGAGGAGAGGGTGAAGGTGATGCTACAATCGGA 120
AAACTCACCCTTAAATTTATTTGCACTACTGGAAAACTACCTGTTCCATGGCCAACACTT 180
GTCACTACTCTGACCTATGGTGTTCAATGCTTTTCCCGTTATCCGGATCACATGAAAAGG 240
CATGACTTTTTCAAGAGTGCCATGCCCGAAGGTTATGTACAGGAACGCACTATATCTTTC 300
AAAGATGACGGGAAATACAAGACGCGTGCTGTAGTCAAGTTTGAAGGTGATACCCTTGTT 360
AATCGTATCGAGTTAAAGGGTACTGATTTTAAAGAAGATGGAAACATTCTCGGACACAAA 420
CTCGAGTACAACTTTAACTCACACAATGTATACATCACGGCAGACAAACAAAAGAATGGA 480
ATCAAAGCTAACTTCACAGTTCGCCACAACGTTGAAGATGGTTCCGTTCAACTAGCAGAC 540
CATTATCAACAAAATACTCCAATTGGCGATGGCCCTGTCCTTTTACCAGACAACCATTAC 600
CTGTCGACACAAACTGTCCTTTCGAAAGATCCCAACGAAAAGGGTACCTAA

GFP 11 WT nucleotide sequence:
AAGCGTGACCACATGGTCCTTCTTGAGTTTGTAACTGCTGCTGGGATTACAGGTAC CTAA

Soluble SuperFolder GFP nucleotide sequence
ATGAGCAAAGGAGAAGAACTTTTCACTGGAGTTGTCCCAATTCTTGTTGAATTAGATGGT 60
GATGTTAATGGGCACAAATTTTCTGTCAGAGGAGAGGGTGAAGGTGATGCTACAATCGGA 120
AAACTCACCCTTAAATTTATTTGCACTACTGGAAAACTACCTGTTCCATGGCCAACACTT 180
GTCACTACTCTGACCTATGGTGTTCAATGCTTTTCCCGTTATCCGGATCACATGAAAAGG 240
CATGACTTTTTCAAGAGTGCCATGCCCGAAGGTTATGTACAGGAACGCACTATATCTTTC 300
AAAGATGACGGGAAATACAAGACGCGTGCTGTAGTCAAGTTTGAAGGTGATACCCTTGTT 360
AATCGTATCGAGTTAAAGGGTACTGATTTTAAAGAAGATGGAAACATTCTCGGACACAAA 420
CTCGAGTACAACTTTAACTCACACAATGTATACATCACGGCAGACAAACAAAAGAATGGA 480
ATCAAAGCTAACTTCACAGTTCGCCACAACGTTGAAGATGGTTCCGTTCAACTAGCAGAC 540
CATTATCAACAAAATACTCCAATTGGCGATGGCCCTGTCCTTTTACCAGACAACCATTAC 600
CTGTCGACACAAACTGTCCTTTCGAAAGATCCCAACGAAAAGGGTACCAAGCGTGACCAC 660
ATGGTCCTTCTTGAGTTTGTAACTGCTGCTGGGATTACAGGTACC

Amino acid sequence
GFP 11 WT KRDHMVLLEFVTAAGITGT
GFP 11 M1 (L221H) KRDHMVLHEFVTAAGITGT
GFP 11 M2 (L221H, F223S, T225N) KRDHMVLHESVNAAGGT
GFP 11 M3 (L221H, F223Y, T225N) RDHMVLHEYVNAAGIT

Blast vs superfolderGFP

Score = 459 bits (1180), Expect = 3e-134, Method: Compositional matrix adjust.
Identities = 223/233 (95%), Positives = 225/233 (96%), Gaps = 3/233 (1%)

sfGFP 1 MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKFICTTGKLPVPWPTL 60
MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDAT GKLTLKFICTTGKLPVPWPTL
ssfGFP 1 MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATIGKLTLKFICTTGKLPVPWPTL 60

Query 61 VTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGTYKTRAEVKFEGDTLV 120
VTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG YKTRA VKFEGDTLV
Sbjct 61 VTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGKYKTRAVVKFEGDTLV 120

Query 121 NRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKANFKIRHNVEDGSVQLAD 180
NRIELKG DFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKANF +RHNVEDGSVQLAD
Sbjct 121 NRIELKGTDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKANFTVRHNVEDGSVQLAD 180

Query 181 HYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNE—KRDHMVLLEFVTAAGIT 230
HYQQNTPIGDGPVLLPDNHYLSTQ+VLSKDPNE KRDHMVLLEFVTAAGIT
Sbjct 181 HYQQNTPIGDGPVLLPDNHYLSTQTVLSKDPNEKGTKRDHMVLLEFVTAAGIT 233