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ratiometric pHluorin

GFP mutant that responds to a drop in pH with a reversible decrease in excitation at 395 nm and a corresponding increase in excitation at 475 nm.

 
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ratiometric pHluorin.dna
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600 400 200 ratiometric pHluorin End (717) GsaI - PspFI (683) BpuEI (682) AlwNI - BseYI - CaiI (679) MspA1I - PvuII (676) SmlI - SmoI (661) AasI - DrdI - DseDI (652) EcoP15I (649) BsaI - Bso31I - BspTNI - Eco31I (638) BstX2I - BstYI - MflI * - PsuI - XhoII (627) AsuII - Bpu14I - Bsp119I - BspT104I - BstBI - Csp45I - NspV - SfuI (622) Acc36I - BfuAI - BspMI - BveI (608) BsgI (586) BtgZI (581) MfeI - MunI (560) KpnI (482) Acc65I - AccB1I - Asp718I - BanI - BshNI - BspT107I (478) AdeI - DraIII (447) Alw21I - Bbv12I - BsiHKAI (443) DraI (390) AcuI - Eco57I (350) AcvI - BbrPI - BsaAI - BstBAI - Eco72I - PmaCI - PmlI - Ppu21I - PspCI (325) AflIII (322) FauNDI - NdeI (230) BalI - MlsI - MluNI - MscI - Msp20I (171) AcoI - CfrI - EaeI (169) Bsp19I - BssT1I - BstDSI - BtgI - Eco130I - EcoT14I - ErhI - NcoI - StyI (166) BaeGI - Bme1580I - BseSI - BstSLI (74) BpmI - GsuI (47) Start (0) ratiometric pHluorin 717 bp
End  (717)
0 sites
GsaI  (683)
1 site
C C C A G C G G G T C G
PspFI  (683)
1 site
C C C A G C G G G T C G
BpuEI  (682)
1 site
C T T G A G ( N ) 14 N N G A A C T C ( N ) 14

Sticky ends from different BpuEI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
AlwNI  (679)
1 site
C A G N N N C T G G T C N N N G A C

Sticky ends from different AlwNI sites may not be compatible.
BseYI  (679)
1 site
C C C A G C G G G T C G

After cleavage, BseYI can remain bound to DNA and alter its electrophoretic mobility.
CaiI  (679)
1 site
C A G N N N C T G G T C N N N G A C

Sticky ends from different CaiI sites may not be compatible.
MspA1I  (676)
1 site
C M G C K G G K C G M C
PvuII  (676)
1 site
C A G C T G G T C G A C
SmlI  (661)
1 site
C T Y R A G G A R Y T C

Efficient cleavage requires at least two copies of the SmlI recognition sequence.
Sticky ends from different SmlI sites may not be compatible.
SmoI  (661)
1 site
C T Y R A G G A R Y T C

Sticky ends from different SmoI sites may not be compatible.
AasI  (652)
1 site
G A C N N N N N N G T C C T G N N N N N N C A G

Sticky ends from different AasI sites may not be compatible.
DrdI  (652)
1 site
G A C N N N N N N G T C C T G N N N N N N C A G

Sticky ends from different DrdI sites may not be compatible.
DseDI  (652)
1 site
G A C N N N N N N G T C C T G N N N N N N C A G

Sticky ends from different DseDI sites may not be compatible.
EcoP15I  (649)
1 site
C A G C A G ( N ) 25 G T C G T C ( N ) 25 N N

Efficient cleavage requires two inversely oriented copies of the EcoP15I recognition sequence.
Sticky ends from different EcoP15I sites may not be compatible.
EcoP15I requires ATP for activity.
BsaI  (638)
1 site
G G T C T C N C C A G A G N ( N ) 4

Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
Bso31I  (638)
1 site
G G T C T C N C C A G A G N ( N ) 4

Sticky ends from different Bso31I sites may not be compatible.
BspTNI  (638)
1 site
G G T C T C N C C A G A G N ( N ) 4

Sticky ends from different BspTNI sites may not be compatible.
Eco31I  (638)
1 site
G G T C T C N C C A G A G N ( N ) 4

Sticky ends from different Eco31I sites may not be compatible.
BstX2I  (627)
1 site
R G A T C Y Y C T A G R
BstYI  (627)
1 site
R G A T C Y Y C T A G R
MflI  (627)
1 site
R G A T C Y Y C T A G R
* Blocked by Dam methylation.
PsuI  (627)
1 site
R G A T C Y Y C T A G R
XhoII  (627)
1 site
R G A T C Y Y C T A G R
AsuII  (622)
1 site
T T C G A A A A G C T T
Bpu14I  (622)
1 site
T T C G A A A A G C T T
Bsp119I  (622)
1 site
T T C G A A A A G C T T
BspT104I  (622)
1 site
T T C G A A A A G C T T
BstBI  (622)
1 site
T T C G A A A A G C T T
Csp45I  (622)
1 site
T T C G A A A A G C T T
NspV  (622)
1 site
T T C G A A A A G C T T
SfuI  (622)
1 site
T T C G A A A A G C T T
Acc36I  (608)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Sticky ends from different Acc36I sites may not be compatible.
BfuAI  (608)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BfuAI recognition sequence.
Sticky ends from different BfuAI sites may not be compatible.
BfuAI is typically used at 50°C, but is 50% active at 37°C.
BspMI  (608)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BspMI recognition sequence.
Sticky ends from different BspMI sites may not be compatible.
BveI  (608)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BveI recognition sequence.
Sticky ends from different BveI sites may not be compatible.
After cleavage, BveI can remain bound to DNA and alter its electrophoretic mobility.
BsgI  (586)
1 site
G T G C A G ( N ) 14 N N C A C G T C ( N ) 14

Efficient cleavage requires at least two copies of the BsgI recognition sequence.
Sticky ends from different BsgI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
BtgZI  (581)
1 site
G C G A T G ( N ) 10 C G C T A C ( N ) 10 ( N ) 4

Sticky ends from different BtgZI sites may not be compatible.
After cleavage, BtgZI can remain bound to DNA and alter its electrophoretic mobility.
BtgZI is typically used at 60°C, but is 75% active at 37°C.
MfeI  (560)
1 site
C A A T T G G T T A A C
MunI  (560)
1 site
C A A T T G G T T A A C
KpnI  (482)
1 site
G G T A C C C C A T G G
Acc65I  (478)
1 site
G G T A C C C C A T G G
AccB1I  (478)
1 site
G G Y R C C C C R Y G G

Sticky ends from different AccB1I sites may not be compatible.
Asp718I  (478)
1 site
G G T A C C C C A T G G
BanI  (478)
1 site
G G Y R C C C C R Y G G

Sticky ends from different BanI sites may not be compatible.
BshNI  (478)
1 site
G G Y R C C C C R Y G G

Sticky ends from different BshNI sites may not be compatible.
BspT107I  (478)
1 site
G G Y R C C C C R Y G G

Sticky ends from different BspT107I sites may not be compatible.
AdeI  (447)
1 site
C A C N N N G T G G T G N N N C A C

Sticky ends from different AdeI sites may not be compatible.
DraIII  (447)
1 site
C A C N N N G T G G T G N N N C A C

Sticky ends from different DraIII sites may not be compatible.
Alw21I  (443)
1 site
G W G C W C C W C G W G

Sticky ends from different Alw21I sites may not be compatible.
Bbv12I  (443)
1 site
G W G C W C C W C G W G

Sticky ends from different Bbv12I sites may not be compatible.
BsiHKAI  (443)
1 site
G W G C W C C W C G W G

Sticky ends from different BsiHKAI sites may not be compatible.
DraI  (390)
1 site
T T T A A A A A A T T T
AcuI  (350)
1 site
C T G A A G ( N ) 14 N N G A C T T C ( N ) 14

Efficient cleavage requires at least two copies of the AcuI recognition sequence.
Sticky ends from different AcuI sites may not be compatible.
After cleavage, AcuI can remain bound to DNA and alter its electrophoretic mobility.
For full activity, add fresh S-adenosylmethionine (SAM).
Eco57I  (350)
1 site
C T G A A G ( N ) 14 N N G A C T T C ( N ) 14

Efficient cleavage requires at least two copies of the Eco57I recognition sequence.
Sticky ends from different Eco57I sites may not be compatible.
After cleavage, Eco57I can remain bound to DNA and alter its electrophoretic mobility.
For full activity, add fresh S-adenosylmethionine (SAM).
AcvI  (325)
1 site
C A C G T G G T G C A C
BbrPI  (325)
1 site
C A C G T G G T G C A C
BsaAI  (325)
1 site
Y A C G T R R T G C A Y
BstBAI  (325)
1 site
Y A C G T R R T G C A Y
Eco72I  (325)
1 site
C A C G T G G T G C A C
PmaCI  (325)
1 site
C A C G T G G T G C A C
PmlI  (325)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
Ppu21I  (325)
1 site
Y A C G T R R T G C A Y
PspCI  (325)
1 site
C A C G T G G T G C A C
AflIII  (322)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
FauNDI  (230)
1 site
C A T A T G G T A T A C
NdeI  (230)
1 site
C A T A T G G T A T A C

Prolonged incubation with NdeI may lead to removal of additional nucleotides.
BalI  (171)
1 site
T G G C C A A C C G G T
MlsI  (171)
1 site
T G G C C A A C C G G T
MluNI  (171)
1 site
T G G C C A A C C G G T
MscI  (171)
1 site
T G G C C A A C C G G T
Msp20I  (171)
1 site
T G G C C A A C C G G T
AcoI  (169)
1 site
Y G G C C R R C C G G Y
CfrI  (169)
1 site
Y G G C C R R C C G G Y
EaeI  (169)
1 site
Y G G C C R R C C G G Y
Bsp19I  (166)
1 site
C C A T G G G G T A C C
BssT1I  (166)
1 site
C C W W G G G G W W C C

Sticky ends from different BssT1I sites may not be compatible.
BstDSI  (166)
1 site
C C R Y G G G G Y R C C

Sticky ends from different BstDSI sites may not be compatible.
BtgI  (166)
1 site
C C R Y G G G G Y R C C

Sticky ends from different BtgI sites may not be compatible.
Eco130I  (166)
1 site
C C W W G G G G W W C C

Sticky ends from different Eco130I sites may not be compatible.
EcoT14I  (166)
1 site
C C W W G G G G W W C C

Sticky ends from different EcoT14I sites may not be compatible.
ErhI  (166)
1 site
C C W W G G G G W W C C

Sticky ends from different ErhI sites may not be compatible.
NcoI  (166)
1 site
C C A T G G G G T A C C
StyI  (166)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
BaeGI  (74)
1 site
G K G C M C C M C G K G

Sticky ends from different BaeGI sites may not be compatible.
Bme1580I  (74)
1 site
G K G C M C C M C G K G

Sticky ends from different Bme1580I sites may not be compatible.
BseSI  (74)
1 site
G K G C M C C M C G K G

Sticky ends from different BseSI sites may not be compatible.
BstSLI  (74)
1 site
G K G C M C C M C G K G

Sticky ends from different BstSLI sites may not be compatible.
BpmI  (47)
1 site
C T G G A G ( N ) 14 N N G A C C T C ( N ) 14

Efficient cleavage requires at least two copies of the BpmI recognition sequence.
Sticky ends from different BpmI sites may not be compatible.
After cleavage, BpmI can remain bound to DNA and alter its electrophoretic mobility.
BpmI quickly loses activity at 37°C.
GsuI  (47)
1 site
C T G G A G ( N ) 14 N N G A C C T C ( N ) 14

Sticky ends from different GsuI sites may not be compatible.
After cleavage, GsuI can remain bound to DNA and alter its electrophoretic mobility.
GsuI is typically used at 30°C, but is 70% active at 37°C.
For full activity, add fresh S-adenosylmethionine (SAM).
Start  (0)
0 sites
ratiometric pHluorin
1 .. 717  =  717 bp
238 amino acids  =  26.9 kDa
Product: pH-sensitive mutant of green fluorescent protein (Miesenböck et al., 1998)
responds to a drop in pH with a reversible decrease in excitation at 395 nm and a corresponding increase in excitation at 475 nm
ratiometric pHluorin
1 .. 717  =  717 bp
238 amino acids  =  26.9 kDa
Product: pH-sensitive mutant of green fluorescent protein (Miesenböck et al., 1998)
responds to a drop in pH with a reversible decrease in excitation at 395 nm and a corresponding increase in excitation at 475 nm
ORF:  1 .. 717  =  717 bp
ORF:  238 amino acids  =  26.9 kDa
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