aa2ar_human
NAME
3VGA
ORGANISM
Human (Homo sapiens)
PROTEIN NAME
aa2ar_human
SOURCE
SWISSPROT
SEQUENCE
Perspective:
Wild-type
Construct
TM1 | |||||||||
M
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P
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I
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M
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G
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S
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S
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V
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Y
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I
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10 |
T
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V
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E
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L
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A
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I
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A
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V
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L
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A
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20 |
I
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L
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G
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N
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V
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L
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V
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C
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W
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A
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30 |
ICL1 | |||||||||
V
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W
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L
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N
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S
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N
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L
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Q
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N
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V
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40 |
TM2 | |||||||||
T
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N
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Y
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F
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V
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V
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S
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L
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A
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A
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50 |
A
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D
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I
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A
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V
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G
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V
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L
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A
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I
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60 |
P
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F
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A
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I
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T
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I
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S
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T
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G
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F
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70 |
ECL1 | TM3 | ||||||||
C
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A
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A
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C
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H
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G
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C
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L
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F
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I
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80 |
A
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C
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F
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V
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L
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V
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L
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T
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Q
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S
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90 |
S
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I
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F
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S
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L
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L
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A
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I
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A
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I
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100 |
D
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R
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Y
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I
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A
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I
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R
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I
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P
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L
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110 |
ICL2 | TM4 | ||||||||
R
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Y
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N
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G
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L
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V
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T
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G
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T
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R
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120 |
A
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K
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G
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I
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I
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A
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I
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C
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W
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V
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130 |
L
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S
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F
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A
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I
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G
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L
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T
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P
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M
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140 |
ECL2 | |||||||||
L
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G
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W
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N
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N
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C
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G
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Q
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P
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K
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150 |
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|||||||||
E
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G
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K
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N
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H
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S
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Q
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G
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C
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G
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160 |
E
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G
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Q
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V
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A
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C
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L
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F
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E
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D
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170 |
TM5 | |||||||||
V
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V
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P
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M
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N
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Y
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M
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V
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Y
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F
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180 |
N
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F
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F
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A
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C
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V
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L
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V
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P
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L
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190 |
L
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L
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M
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L
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G
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V
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Y
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L
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R
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I
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200 |
F
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L
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A
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A
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R
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R
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Q
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L
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K
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Q
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210 |
ICL3 | |||||||||
M
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E
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S
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Q
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P
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L
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P
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G
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E
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R
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220 |
TM6 | |||||||||
A
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R
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S
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T
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L
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Q
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K
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E
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V
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H
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230 |
A
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A
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K
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S
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L
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A
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I
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I
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V
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G
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240 |
L
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F
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A
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L
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C
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W
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L
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P
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L
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H
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250 |
I
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I
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N
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C
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F
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T
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F
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F
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C
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P
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260 |
ECL3 | TM7 | ||||||||
D
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C
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S
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H
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A
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P
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L
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W
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L
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M
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270 |
Y
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L
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A
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I
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V
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L
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S
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H
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T
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N
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280 |
S
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V
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V
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N
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P
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F
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I
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Y
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A
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Y
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290 |
H8 | |||||||||
R
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I
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R
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E
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F
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R
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Q
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T
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F
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R
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300 |
K
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I
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I
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R
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S
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H
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V
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L
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R
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Q
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310 |
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C-term | |||||||||
Q
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E
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P
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F
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K
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A
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A
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G
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T
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S
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320 |
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A
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R
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V
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L
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A
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A
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H
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G
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S
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D
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330 |
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|
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G
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E
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Q
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V
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S
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L
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R
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L
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N
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G
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340 |
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|
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H
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P
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P
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G
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V
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W
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A
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N
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G
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S
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350 |
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A
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P
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H
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P
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E
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R
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R
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P
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N
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G
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360 |
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|
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Y
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A
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L
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G
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L
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V
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S
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G
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G
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S
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370 |
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|
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A
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Q
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E
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S
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Q
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G
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N
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T
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G
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L
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380 |
|
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P
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D
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V
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E
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L
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L
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S
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H
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E
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L
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390 |
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K
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G
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V
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C
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P
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E
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P
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P
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G
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L
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400 |
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D
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D
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P
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L
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A
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Q
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D
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G
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A
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G
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410 |
|
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V
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S
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α-mating fa | ||||||||||
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FLAG | |||||||||
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TM1 | |||||||||
M
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P
|
I
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M
|
G
|
S
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S
|
V
|
Y
|
I
|
10 |
T
|
V
|
E
|
L
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A
|
I
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A
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V
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L
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A
|
20 |
I
|
L
|
G
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N
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V
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L
|
V
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C
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W
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A
|
30 |
ICL1 | |||||||||
V
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W
|
L
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N
|
S
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N
|
L
|
Q
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N
|
V
|
40 |
TM2 | |||||||||
T
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N
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Y
|
F
|
V
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V
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S
|
L
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A
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A
|
50 |
A
|
D
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I
|
A
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V
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G
|
V
|
L
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A
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I
|
60 |
P
|
F
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A
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I
|
T
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I
|
S
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T
|
G
|
F
|
70 |
ECL1 | TM3 | ||||||||
C
|
A
|
A
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C
|
H
|
G
|
C
|
L
|
F
|
I
|
80 |
A
|
C
|
F
|
V
|
L
|
V
|
L
|
T
|
Q
|
S
|
90 |
S
|
I
|
F
|
S
|
L
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L
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A
|
I
|
A
|
I
|
100 |
D
|
R
|
Y
|
I
|
A
|
I
|
R
|
I
|
P
|
L
|
110 |
ICL2 | TM4 | ||||||||
R
|
Y
|
N
|
G
|
L
|
V
|
T
|
G
|
T
|
R
|
120 |
A
|
K
|
G
|
I
|
I
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A
|
I
|
C
|
W
|
V
|
130 |
L
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S
|
F
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A
|
I
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G
|
L
|
T
|
P
|
M
|
140 |
ECL2 | |||||||||
L
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G
|
W
|
N
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N
|
C
|
G
|
Q
|
P
|
K
|
150 |
|
|||||||||
E
|
G
|
K
|
Q
|
H
|
S
|
Q
|
G
|
C
|
G
|
160 |
E
|
G
|
Q
|
V
|
A
|
C
|
L
|
F
|
E
|
D
|
170 |
TM5 | |||||||||
V
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V
|
P
|
M
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N
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Y
|
M
|
V
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Y
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F
|
180 |
N
|
F
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F
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A
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C
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V
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L
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V
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P
|
L
|
190 |
L
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L
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M
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L
|
G
|
V
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Y
|
L
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R
|
I
|
200 |
F
|
L
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A
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A
|
R
|
R
|
Q
|
L
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K
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Q
|
210 |
ICL3 | |||||||||
M
|
E
|
S
|
Q
|
P
|
L
|
P
|
G
|
E
|
R
|
220 |
TM6 | |||||||||
A
|
R
|
S
|
T
|
L
|
Q
|
K
|
E
|
V
|
H
|
230 |
A
|
A
|
K
|
S
|
L
|
A
|
I
|
I
|
V
|
G
|
240 |
L
|
F
|
A
|
L
|
C
|
W
|
L
|
P
|
L
|
H
|
250 |
I
|
I
|
N
|
C
|
F
|
T
|
F
|
F
|
C
|
P
|
260 |
ECL3 | TM7 | ||||||||
D
|
C
|
S
|
H
|
A
|
P
|
L
|
W
|
L
|
M
|
270 |
Y
|
L
|
A
|
I
|
V
|
L
|
S
|
H
|
T
|
N
|
280 |
S
|
V
|
V
|
N
|
P
|
F
|
I
|
Y
|
A
|
Y
|
290 |
H8 | |||||||||
R
|
I
|
R
|
E
|
F
|
R
|
Q
|
T
|
F
|
R
|
300 |
K
|
I
|
I
|
R
|
S
|
H
|
V
|
L
|
R
|
Q
|
310 |
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|||||||||
C-term | |||||||||
Q
|
E
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P
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F
|
K
|
A
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|||
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His(10) | |||||||||
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SCHEMATIC
Perspective:
Wild-type
Construct
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DIAGRAMS
MUTATIONS
Position | Wild-type | Mutated |
---|---|---|
154 | | N | Q |
DELETIONS
MODIFICATIONS
AUXILIARY PROTEINS
Protein_type: signal
Name: α-mating factor secretion signal
Sequence: None
Deletions:
Positions: N-term_0 None:None
Presence in Crystal:NO
Remarks: None
Protein_type: tag
Name: FLAG
Sequence: None
Deletions:
Positions: N-term_1 None:None
Presence in Crystal:NO
Remarks: None
Protein_type: tag
Name: His(10)
Sequence: None
Deletions:
Positions: C-term_2 None:None
Presence in Crystal:YES
Remarks: None
EXPRESSION
Expression method | Host cell type | Host cell | Remarks |
---|---|---|---|
P.pastoris | P. pastoris SMD1163 | Yeast |
SOLUBILIZATION
Solvent type | Item | Concentration |
---|---|---|
additive | CHS | 0.2 %w/v |
detergent | DDM | 1 %w/v |
Remarks:
PURIFICATION
Purification type | Description |
---|---|
Deglycosylation by ConA | None |
Remarks:None
CRYSTALLIZATION
METHOD: vapour diffusion
TYPE: hanging drop ()
PROTEIN CONCENTRATION: 20 mg/ml
Temperature: 20°C
ph : 6.5-6.5
Remarks: Purified A2AAR and the Fab fragments were mixed in a 1:1.2 molar ratio
Chemical lists
Type | Item | Concentration |
---|---|---|
Additional | ||
unknown | PEG 400 | 30 % v/v |
unknown | MES, pH 6.5 | 0.1 M |
unknown | Magnesium Chloride | 0.2 M |
Detergent | ||
detergent | OTG(octylthioglucoside) | 0.3-0.6 % w/v |