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Plasmid Files

pHuji

pH-sensitive red fluorescent protein derived from mApple, for imaging of exocytosis and endocytosis.

 
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 600 400 200 End (711) BsrGI - TatI (701) XcmI (687) BsrFI - SgrAI (682) AleI - MslI (643) BsgI (595) MspA1I - PvuII (579) BspQI - EarI - SapI (481) BsrBI (459) PflMI (440) BbsI (436) BtgI - NcoI (434) PstI - SbfI (356) SfcI - TsoI (352) HincII - HpaI (336) TaqII (254) AccI (227) BstEII (156) BlpI (146) BssS α I (88) BanII (76) BaeGI - Bme1580I - BsiHKAI (65) ApaLI (61) MscI (31) Start (0) pHuji 711 bp
End  (711)
0 sites
BsrGI  (701)
1 site
T G T A C A A C A T G T

BsrGI is typically used at 37°C, but is even more active at 60°C.
TatI  (701)
1 site
W G T A C W W C A T G W
XcmI  (687)
1 site
C C A N N N N N N N N N T G G G G T N N N N N N N N N A C C

The 1-base overhangs produced by XcmI may be hard to ligate.
Sticky ends from different XcmI sites may not be compatible.
BsrFI  (682)
1 site
R C C G G Y Y G G C C R

Efficient cleavage requires at least two copies of the BsrFI
recognition sequence.
After cleavage, BsrFI can remain bound to DNA and alter its
electrophoretic mobility.
SgrAI  (682)
1 site
C R C C G G Y G G Y G G C C R C

Efficient cleavage requires at least two copies of the SgrAI
recognition sequence.
AleI  (643)
1 site
C A C N N N N G T G G T G N N N N C A C
MslI  (643)
1 site
C A Y N N N N R T G G T R N N N N Y A C
BsgI  (595)
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).
MspA1I  (579)
1 site
C M G C K G G K C G M C
PvuII  (579)
1 site
C A G C T G G T C G A C
BspQI  (481)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different BspQI sites may not be compatible.
EarI  (481)
1 site
C T C T T C N G A G A A G N N N N

Efficient cleavage requires at least two copies of the EarI
recognition sequence.
Sticky ends from different EarI sites may not be compatible.
SapI  (481)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different SapI sites may not be compatible.
SapI gradually settles in solution, so a tube of SapI should be
mixed before removing an aliquot.
BsrBI  (459)
1 site
C C G C T C G G C G A G

This recognition sequence is asymmetric, so ligating blunt ends
generated by BsrBI will not always regenerate a BsrBI site.
BsrBI is typically used at 37°C, but can be used at temperatures
up to 50°C.
PflMI  (440)
1 site
C C A N N N N N T G G G G T N N N N N A C C

Sticky ends from different PflMI sites may not be compatible.
BbsI  (436)
1 site
G A A G A C N N C T T C T G N N ( N ) 4

Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
BtgI  (434)
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  (434)
1 site
C C A T G G G G T A C C
PstI  (356)
1 site
C T G C A G G A C G T C
SbfI  (356)
1 site
C C T G C A G G G G A C G T C C
SfcI  (352)
1 site
C T R Y A G G A Y R T C

Sticky ends from different SfcI sites may not be compatible.
SfcI quickly loses activity at 37°C, but can be used at 25°C for long
incubations.
TsoI  (352)
1 site
T A R C C A ( N ) 9 N N A T Y G G T ( N ) 9

Sticky ends from different TsoI sites may not be compatible.
After cleavage, TsoI can remain bound to DNA and alter its
electrophoretic mobility.
For full activity, add fresh S-adenosylmethionine (SAM).
HincII  (336)
1 site
G T Y R A C C A R Y T G
HpaI  (336)
1 site
G T T A A C C A A T T G
TaqII  (254)
1 site
G A C C G A ( N ) 9 N N C T G G C T ( N ) 9

Sticky ends from different TaqII sites may not be compatible.
AccI  (227)
1 site
G T M K A C C A K M T G

Efficient cleavage with AccI requires ≥13 bp on each side of the
recognition sequence.
Sticky ends from different AccI sites may not be compatible.
BstEII  (156)
1 site
G G T N A C C C C A N T G G

Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
BlpI  (146)
1 site
G C T N A G C C G A N T C G

Sticky ends from different BlpI sites may not be compatible.
BssSαI  (88)
1 site
C A C G A G G T G C T C
BanII  (76)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
BaeGI  (65)
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  (65)
1 site
G K G C M C C M C G K G

Sticky ends from different Bme1580I sites may not be compatible.
BsiHKAI  (65)
1 site
G W G C W C C W C G W G

Sticky ends from different BsiHKAI sites may not be compatible.
ApaLI  (61)
1 site
G T G C A C C A C G T G
MscI  (31)
1 site
T G G C C A A C C G G T
Start  (0)
0 sites
pHuji
1 .. 711  =  711 bp
236 amino acids  =  27.0 kDa
Product: pH-sensitive red fluorescent protein
derived from mApple, for imaging of exocytosis and
endocytosis (Shen et al., 2014)
mammalian codon-optimized
pHuji
1 .. 711  =  711 bp
236 amino acids  =  27.0 kDa
Product: pH-sensitive red fluorescent protein
derived from mApple, for imaging of exocytosis and
endocytosis (Shen et al., 2014)
mammalian codon-optimized
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