ecliptic pHluorin

GFP mutant that responds to a drop in pH with a gradual loss (eclipsing) of excitation at 475 nm, accompanied by a reduction of excitation at 395 nm.
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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) BtgZI (581) ApaI - BanII (575) PspOMI (571) MfeI (560) DraIII (447) CsiI - SexAI * (443) DraI (390) AcuI (350) PmlI - BsaAI (325) AflIII (322) NdeI (230) MscI (171) EaeI (169) BtgI - StyI - NcoI (166) BpmI (47) Start (0) ecliptic pHluorin ecliptic 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
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.
ApaI  (575)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
BanII  (575)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
PspOMI  (571)
1 site
G G G C C C C C C G G G
MfeI  (560)
1 site
C A A T T G G T T A A C
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.
CsiI  (443)
1 site
A C C W G G T T G G W C C A

Sticky ends from different CsiI sites may not be compatible.
SexAI  (443)
1 site
A C C W G G T T G G W C C A
* Blocked by Dcm methylation.
Sticky ends from different SexAI 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
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
ecliptic pHluorin
1 .. 717  =  717 bp
238 amino acids  =  27.0 kDa
Product: pH-sensitive mutant of green fluorescent protein (Miesenböck et al., 1998)
responds to a drop in pH with a gradual loss (eclipsing) of excitation at 475 nm, accompanied by a reduction of excitation at 395 nm
ecliptic pHluorin
1 .. 717  =  717 bp
238 amino acids  =  27.0 kDa
Product: pH-sensitive mutant of green fluorescent protein (Miesenböck et al., 1998)
responds to a drop in pH with a gradual loss (eclipsing) of excitation at 475 nm, accompanied by a reduction of excitation at 395 nm
ORF:  1 .. 717  =  717 bp
ORF:  238 amino acids  =  27.0 kDa
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