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Plasmid Files

super-ecliptic pHluorin

Red-shifted ecliptic pHluorin that is excited strongly by blue light at neutral pH but minimally fluorescent at acidic pH.

 
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 600 400 200 superecliptic pHluorin End (717) PspFI (683) BpuEI (682) AlwNI - BseYI (679) MspA1I - PvuII (676) SmlI (661) DrdI (652) EcoP15I (649) BsaI (638) BstYI (627) BstBI (622) BtgZI (581) ApaI - BanII (575) PspOMI (571) MfeI (560) DraIII (447) SexAI * (443) DraI (390) AcuI (350) BsaAI - PmlI (325) AflIII (322) NdeI (230) MscI (171) EaeI (169) BtgI - NcoI - StyI (166) BpmI (47) Start (0) super-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).
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.
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.
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.
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.
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

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).
BsaAI  (325)
1 site
Y A C G T R R T G C A Y
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.
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.
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.
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
superecliptic pHluorin
1 .. 717  =  717 bp
238 amino acids  =  26.9 kDa
Product: pH-sensitive mutant of green fluorescent
protein (Ng et al., 2002)
excited strongly by blue light at neutral pH but
minimally fluorescent at acidic pH
superecliptic pHluorin
1 .. 717  =  717 bp
238 amino acids  =  26.9 kDa
Product: pH-sensitive mutant of green fluorescent
protein (Ng et al., 2002)
excited strongly by blue light at neutral pH but
minimally fluorescent at acidic pH
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