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Plasmid Files

PSmOrange2

Enhanced orange to far-red photoswitchable monomeric fluorescent protein.

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PSmOrange2 Sequence and MapPSmOrange2.dna
Map and Sequence File   
Sequence Author:  Verkhusha Lab
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 600 400 200 End (708) BsrGI - TatI (701) XcmI (687) BsrFI - SgrAI (682) AleI - MslI (643) EcoP15I (549) BsrBI (459) PspFI (444) BseYI - PflMI (440) BbsI (436) BtgI - NcoI (434) MmeI (431) PstI (356) SfcI (352) BsaAI (231) AhdI (195) BlpI (146) BssS α I (88) BsiHKAI (65) ApaLI (61) MscI (31) Start (0) PSmOrange2 708 bp
End  (708)
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
EcoP15I  (549)
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.
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.
PspFI  (444)
1 site
C C C A G C G G G T C G
BseYI  (440)
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.
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
MmeI  (431)
1 site
T C C R A C ( N ) 18 N N A G G Y T G ( N ) 18

Efficient cleavage requires at least two copies of the MmeI
recognition sequence.
Sticky ends from different MmeI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
PstI  (356)
1 site
C T G C A G G A C G T 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.
BsaAI  (231)
1 site
Y A C G T R R T G C A Y
AhdI  (195)
1 site
G A C N N N N N G T C C T G N N N N N C A G

The 1-base overhangs produced by AhdI may be hard to ligate.
Sticky ends from different AhdI sites may not be compatible.
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
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
PSmOrange2
1 .. 708  =  708 bp
236 amino acids  =  26.7 kDa
Product: enhanced orange to far-red
photoswitchable monomeric fluorescent protein
(Subach et al., 2012)
mammalian codon-optimized
PSmOrange2
1 .. 708  =  708 bp
236 amino acids  =  26.7 kDa
Product: enhanced orange to far-red
photoswitchable monomeric fluorescent protein
(Subach et al., 2012)
mammalian codon-optimized
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