aa1r_human
NAME
5UEN
ORGANISM
Human (Homo sapiens)
PROTEIN NAME
aa1r_human
SOURCE
SWISSPROT
SEQUENCE
Perspective:
Wild-type
Construct
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N-term | TM1 | ||||||||
M
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P
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P
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S
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I
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S
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A
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F
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Q
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A
|
10 |
A
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Y
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I
|
G
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I
|
E
|
V
|
L
|
I
|
A
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20 |
L
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V
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S
|
V
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P
|
G
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N
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V
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L
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V
|
30 |
I
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W
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A
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V
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K
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V
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N
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Q
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A
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L
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40 |
ICL1 | TM2 | ||||||||
R
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D
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A
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T
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F
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C
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F
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I
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V
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S
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50 |
L
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A
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V
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A
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D
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V
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A
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V
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G
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A
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60 |
L
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V
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I
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P
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L
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A
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I
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L
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I
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N
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70 |
ECL1 | TM3 | ||||||||
I
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G
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P
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Q
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T
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Y
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F
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H
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T
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C
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80 |
L
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M
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V
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A
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C
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P
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V
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L
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I
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L
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90 |
T
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Q
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S
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S
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I
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L
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A
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L
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L
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A
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100 |
I
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A
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V
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D
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R
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Y
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L
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R
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V
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K
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110 |
ICL2 | |||||||||
I
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P
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L
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R
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Y
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K
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M
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V
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V
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T
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120 |
TM4 | |||||||||
P
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R
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R
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A
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A
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V
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A
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I
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A
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G
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130 |
C
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W
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I
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L
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S
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F
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V
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V
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G
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L
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140 |
ECL2 | |||||||||
T
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P
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M
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F
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G
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W
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N
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N
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L
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S
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150 |
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|||||||||
A
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V
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E
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R
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A
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W
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A
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A
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N
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G
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160 |
S
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M
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G
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E
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P
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V
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I
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K
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C
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E
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170 |
TM5 | |||||||||
F
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E
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K
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V
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I
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S
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M
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E
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Y
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M
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180 |
V
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Y
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F
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N
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F
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F
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V
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W
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V
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L
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190 |
P
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P
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L
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L
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L
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M
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V
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L
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I
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Y
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200 |
L
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E
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V
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F
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Y
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L
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I
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R
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K
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Q
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210 |
|
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ICL3 | |||||||||
L
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N
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K
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K
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V
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S
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A
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S
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S
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G
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220 |
|
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TM6 | |||||||||
D
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P
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Q
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K
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Y
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Y
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G
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K
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E
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L
|
230 |
K
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I
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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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L
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I
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L
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240 |
F
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L
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F
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A
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L
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S
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W
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L
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P
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L
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250 |
H
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I
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L
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N
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C
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I
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T
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L
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F
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C
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260 |
ECL3 | TM7 | ||||||||
P
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S
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C
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H
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K
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P
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S
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I
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L
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T
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270 |
Y
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I
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A
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I
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F
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L
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T
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H
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G
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N
|
280 |
S
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A
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M
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N
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P
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I
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V
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Y
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A
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F
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290 |
H8 | |||||||||
R
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I
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Q
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K
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F
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R
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V
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T
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F
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L
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300 |
K
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I
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W
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N
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D
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H
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F
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R
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C
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Q
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310 |
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C-term | |||||||||
P
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A
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P
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P
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I
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D
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E
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D
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L
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P
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320 |
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|
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E
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E
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R
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P
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D
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D
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hemagglutin | ||||||||||
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FLAG | |||||||||
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M4 muscarin | ||||||||||
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PreScission | ||||||||||
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N-term | TM1 | ||||||||
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P
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P
|
S
|
I
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S
|
A
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F
|
Q
|
A
|
10 |
A
|
Y
|
I
|
G
|
I
|
E
|
V
|
L
|
I
|
A
|
20 |
L
|
V
|
S
|
V
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P
|
G
|
N
|
V
|
L
|
V
|
30 |
I
|
W
|
A
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V
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K
|
V
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N
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Q
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A
|
L
|
40 |
ICL1 | TM2 | ||||||||
R
|
D
|
A
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T
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F
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C
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F
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I
|
V
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S
|
50 |
L
|
A
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V
|
A
|
D
|
V
|
A
|
V
|
G
|
A
|
60 |
L
|
V
|
I
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P
|
L
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A
|
I
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L
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I
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N
|
70 |
ECL1 | TM3 | ||||||||
I
|
G
|
P
|
Q
|
T
|
Y
|
F
|
H
|
T
|
C
|
80 |
L
|
M
|
V
|
A
|
C
|
P
|
V
|
L
|
I
|
L
|
90 |
T
|
Q
|
S
|
S
|
I
|
L
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A
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L
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L
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A
|
100 |
I
|
A
|
V
|
D
|
R
|
Y
|
L
|
R
|
V
|
K
|
110 |
ICL2 | |||||||||
I
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P
|
L
|
R
|
Y
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K
|
M
|
V
|
V
|
T
|
120 |
TM4 | |||||||||
P
|
R
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R
|
A
|
A
|
V
|
A
|
I
|
A
|
G
|
130 |
C
|
W
|
I
|
L
|
S
|
F
|
V
|
V
|
G
|
L
|
140 |
ECL2 | |||||||||
T
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P
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M
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F
|
G
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W
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N
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N
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L
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S
|
150 |
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|||||||||
A
|
V
|
E
|
R
|
A
|
W
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A
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A
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A
|
G
|
160 |
S
|
M
|
G
|
E
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P
|
V
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I
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K
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C
|
E
|
170 |
TM5 | |||||||||
F
|
E
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K
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V
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I
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S
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M
|
E
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Y
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M
|
180 |
V
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Y
|
F
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N
|
F
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F
|
V
|
W
|
V
|
L
|
190 |
P
|
P
|
L
|
L
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L
|
M
|
V
|
L
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I
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Y
|
200 |
L
|
E
|
V
|
F
|
Y
|
L
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I
|
R
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K
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Q
|
210 |
|
|
L
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|
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BRIL (b562R | ||||||||||
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|||||||
TM6 | |||||||
K
|
E
|
L
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K
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I
|
A
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K
|
S
|
235 |
L
|
A
|
L
|
I
|
L
|
F
|
L
|
F
|
A
|
L
|
245 |
S
|
W
|
L
|
P
|
L
|
H
|
I
|
L
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N
|
C
|
255 |
ECL3 | |||||||||
I
|
T
|
L
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F
|
C
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P
|
S
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C
|
H
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K
|
265 |
TM7 | |||||||||
P
|
S
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I
|
L
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T
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Y
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I
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A
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I
|
F
|
275 |
L
|
T
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H
|
G
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N
|
S
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A
|
M
|
N
|
P
|
285 |
H8 | |||||||||
I
|
V
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Y
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A
|
F
|
R
|
I
|
Q
|
K
|
F
|
295 |
R
|
V
|
T
|
F
|
L
|
K
|
I
|
W
|
N
|
D
|
305 |
H
|
F
|
R
|
C
|
Q
|
310 |
|
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PreScission | ||||||||||
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P
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PreScission | ||||||||||
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His(8) | |||||||||
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SCHEMATIC
Perspective:
Wild-type
Construct
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DIAGRAMS
MUTATIONS
Position | Wild-type | Mutated |
---|---|---|
159 | | N | A |
DELETIONS
MODIFICATIONS
AUXILIARY PROTEINS
Protein_type: signal
Name: hemagglutinin
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: M4 muscarinic receptor (22AA)
Sequence: None
Deletions:
Positions: N-term_2 None:None
Presence in Crystal:NO
Remarks: None
Protein_type: prot_cleavage
Name: PreScission site (3C)
Sequence: None
Deletions:
Positions: N-term_3 None:None
Presence in Crystal:YES
Remarks: None
Protein_type: fusion
Name: BRIL (b562RIL)
Sequence: None
Deletions:
Positions: Within Receptor_4 212:220
Presence in Crystal:YES
Remarks: None
Protein_type: linker
Name: A2A-Receptor (8AA)
Sequence: None
Deletions:
Positions: Within Receptor_5 221:228
Presence in Crystal:YES
Remarks: None
Protein_type: prot_cleavage
Name: PreScission site (3C)
Sequence: None
Deletions:
Positions: C-term_6 311:311
Presence in Crystal:YES
Remarks: None
Protein_type: tag
Name: His(8)
Sequence: None
Deletions:
Positions: C-term_7 None:None
Presence in Crystal:NO
Remarks: None
EXPRESSION
Expression method | Host cell type | Host cell | Remarks |
---|---|---|---|
Baculovirus | Sf9 | Insect | Bac-to-Bac |
SOLUBILIZATION
Solvent type | Item | Concentration |
---|---|---|
detergent | DDM | 1.0 %w/v |
additive | CHS | 0.03 %w/v |
Remarks:
PURIFICATION
Purification type | Description |
---|---|
Alkylation by Iodoacetamide | None |
Proteolysis by HRV 3C protease | None |
detergent DDM was then gradually exchanged with 0.1%LMNG | None |
Remarks:None
CRYSTALLIZATION
METHOD: in meso/LCP
TYPE: lipidic cubic phase (LCP) (monoolein (9.9 MAG))
PROTEIN CONCENTRATION: NA mg/ml
Temperature: 20°C
ph : 7.0-8.0
Remarks:
Chemical lists
Type | Item | Concentration |
---|---|---|
Detergent | ||
LMNG | 0.01 %w/v | |
Detergent additive | ||
CHS | 0.001 %w/v | |
Additional | ||
PEG 300 | 28-38 %v/v | |
HEPES pH 7.0-8.0 | 100.0 mM | |
NH4F | 500-700 mM |