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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No matches
600 400 200 End (717) PspFI (683) BpuEI (682) BseYI - AlwNI (679) PvuII - MspA1I (676) SmlI (661) DrdI (652) EcoP15I (649) BsaI (638) MflI * - BstYI (627) BstBI (622) BfuAI - BspMI (608) BsgI (586) BtgZI (581) MfeI (560) KpnI (482) BanI - Acc65I (478) DraIII (447) BsiHKAI (443) DraI (390) AcuI (350) PmlI - BsaAI (325) AflIII (322) NdeI (230) MscI (171) EaeI (169) BtgI - StyI - NcoI (166) BaeGI - Bme1580I (74) BpmI (47) Start (0) ratiometric pHluorin ratiometric pHluorin 717 bp
End  (717)
0 sites
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).
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.
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.
PvuII  (676)
1 site
C A G C T G G T C G A C
MspA1I  (676)
1 site
C M G C K G G K C G M C
SmlI  (661)
1 site
C T Y R A G G A R Y T C

Cleavage may be enhanced when more than one copy of the SmlI recognition sequence is present.
Sticky ends from different SmlI 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.
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.
MflI  (627)
1 site
R G A T C Y Y C T A G R
* Blocked by Dam methylation.
BstYI  (627)
1 site
R G A T C Y Y C T A G R
BstBI  (622)
1 site
T T C G A A A A G C T T
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.
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
KpnI  (482)
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.
Acc65I  (478)
1 site
G G T A C C C C A T G G
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.
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

Cleavage may be enhanced when more than one copy of the AcuI recognition sequence is present.
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).
PmlI  (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
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.
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.
MscI  (171)
1 site
T G G C C A A C C G G T
EaeI  (169)
1 site
Y G G C C R R C C G G Y
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.
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.
NcoI  (166)
1 site
C C A T G G G G T A C C
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.
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.
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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