NdeI

1ATG CAT CAC CAT CAC CAC CAT ATG GGC GGT AGC GGC GGT TCC GCT AGC ATG GTG AGC AAG GGC GAG GAG CTG ATC

TAC GTA GTG GTA GTG GTG GTA TAC CCG CCA TCG CCG CCA AGG CGA TCG TAC CAC TCG TTC CCG CTC CTC GAC TAG

M H H H H H H M G G S G G S A S M V S K G E E L I

76AAG GAG AAC ATG AGA AGC AAG CTG TAC CTG GAA GGC AGC GTG AAC GGC CAC CAG TTC AAG TGC ACC CAC GAA GGG

TTC CTC TTG TAC TCT TCG TTC GAC ATG GAC CTT CCG TCG CAC TTG CCG GTG GTC AAG TTC ACG TGG GTG CTT CCC

K E N M R S K L Y L E G S V N G H Q F K C T H E G

151GAG GGC AAG CCC TAC GAG GGC AAG CAG ACC AAC AGG ATC AAG GTG GTG GAG GGA GGC CCC CTG CCG TTC GCA TTC

CTC CCG TTC GGG ATG CTC CCG TTC GTC TGG TTG TCC TAG TTC CAC CAC CTC CCT CCG GGG GAC GGC AAG CGT AAG

E G K P Y E G K Q T N R I K V V E G G P L P F A F

226GAC ATC CTG GCC ACC CAC TTT ATG TAC GGG AGC AAG GTG TTC ATC AAG TAC CCC GCC GAC CTC CCC GAT TAT TTT

CTG TAG GAC CGG TGG GTG AAA TAC ATG CCC TCG TTC CAC AAG TAG TTC ATG GGG CGG CTG GAG GGG CTA ATA AAA

D I L A T H F M Y G S K V F I K Y P A D L P D Y F

301AAG CAG TCC TTC CCT GAG GGC TTC ACA TGG GAG AGA GTC ATG GTG TTC GAA GAC GGG GGC GTG CTG ACC GCC ACC

TTC GTC AGG AAG GGA CTC CCG AAG TGT ACC CTC TCT CAG TAC CAC AAG CTT CTG CCC CCG CAC GAC TGG CGG TGG

K Q S F P E G F T W E R V M V F E D G G V L T A T

376CAG GAC ACC AGC CTC CAG GAC GGC GAG CTC ATC TAC AAC GTC AAG GTC AGA GGG GTG AAC TTC CCA GCC AAC GGC

GTC CTG TGG TCG GAG GTC CTG CCG CTC GAG TAG ATG TTG CAG TTC CAG TCT CCC CAC TTG AAG GGT CGG TTG CCG

Q D T S L Q D G E L I Y N V K V R G V N F P A N G

451CCC GTG ATG CAG AAG AAA ACA CTG GGC TGG GAG CCC AGC ACC GAG ACC ATG TAC CCC GCT GAC GGC GGC CTG GAA

GGG CAC TAC GTC TTC TTT TGT GAC CCG ACC CTC GGG TCG TGG CTC TGG TAC ATG GGG CGA CTG CCG CCG GAC CTT

P V M Q K K T L G W E P S T E T M Y P A D G G L E

526GGC AGA TGC GAC AAG GCC CTG AAG CTC GTG GGC GGG GGC CAC CTG CAC GTC AAC TTC AAG ACC ACA TAC AAG TCC

CCG TCT ACG CTG TTC CGG GAC TTC GAG CAC CCG CCC CCG GTG GAC GTG CAG TTG AAG TTC TGG TGT ATG TTC AGG

G R C D K A L K L V G G G H L H V N F K T T Y K S

601AAG AAA CCC GTG AAG ATG CCC GGC GTC CAC TAC GTG GAC CGC AGA CTG GAA AGA ATC AAG GAG GCC GAC AAC GAG

TTC TTT GGG CAC TTC TAC GGG CCG CAG GTG ATG CAC CTG GCG TCT GAC CTT TCT TAG TTC CTC CGG CTG TTG CTC

K K P V K M P G V H Y V D R R L E R I K E A D N E

EcoRI

676ACC TAC GTC GAG CAG TAC GAG CAC GCT GTG GCC AGA TAC TCC AAC CTG GGC GGT TCG AAT TCG GGT GGC GGC TCT

TGG ATG CAG CTC GTC ATG CTC GTG CGA CAC CGG TCT ATG AGG TTG GAC CCG CCA AGC TTA AGC CCA CCG CCG AGA

T Y V E Q Y E H A V A R Y S N L G G S N S G G G S

NotI

751GGT GGT GGC AGC GGC GGT GGC GGC GGT GGT GGC TGC TAA GCG GCC GC

CCA CCA CCG TCG CCG CCA CCG CCG CCA CCA CCG ACG ATT CGC CGG CG

G G G S G G G G G G G C *

Fig. S1. The nucleotide sequence and amino acid sequence of 6His-CyOFP-L-Cys. The amino acid sequence of wild-type CyOFP is shown in red, and its two intrinsic Cys residues are shown in blue. The cleavage sites of restriction enzymes are shaded.

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