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Plasmid Files

Medium-FT

Blue-to-red fluorescent timer derivative of mCherry with a medium chromophore maturation rate.

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Medium-FT Sequence and MapMedium-FT.dna
Map and Sequence File   
Sequence Author:  Verkhusha Lab
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 600 400 200 End (711) BsrGI - TatI (701) NmeAIII (689) XcmI (687) BsrFI - SgrAI (682) BsaBI (613) MslI (607) HincII (600) EcoP15I (549) DrdI (541) BbvCI - Bpu10I (539) AlwNI (507) BsrBI (459) StuI (448) PspFI (444) BseYI - PflMI (440) BbsI (436) NcoI (434) MmeI (431) PstI - SbfI (356) SfcI (352) BsaAI (231) EcoO109I * (224) AhdI (195) TaqII (151) BssS α I (88) BanII (76) BsiHKAI (65) ApaLI (61) ApoI - EcoRI (43) MscI (31) Start (0) Medium-FT 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
NmeAIII  (689)
1 site
G C C G A G ( N ) 18-19 N N C G G C T C ( N ) 18-19

Efficient cleavage requires at least two copies of the NmeAIII
recognition sequence.
Sticky ends from different NmeAIII sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
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.
BsaBI  (613)
1 site
G A T N N N N A T C C T A N N N N T A G
MslI  (607)
1 site
C A Y N N N N R T G G T R N N N N Y A C
HincII  (600)
1 site
G T Y R A C C A R Y T G
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.
DrdI  (541)
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.
BbvCI  (539)
1 site
C C T C A G C G G A G T C G
Bpu10I  (539)
1 site
C C T N A G C G G A N T C G

Efficient cleavage requires at least two copies of the Bpu10I
recognition sequence.
This recognition sequence is asymmetric, so ligating sticky ends
generated by Bpu10I will not always regenerate a Bpu10I site.
Sticky ends from different Bpu10I sites may not be compatible.
AlwNI  (507)
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.
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.
StuI  (448)
1 site
A G G C C T T C C G G A
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.
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
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.
BsaAI  (231)
1 site
Y A C G T R R T G C A Y
EcoO109I  (224)
1 site
R G G N C C Y Y C C N G G R
* Blocked by Dcm methylation.
Sticky ends from different EcoO109I sites may not be compatible.
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.
TaqII  (151)
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.
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.
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
ApoI  (43)
1 site
R A A T T Y Y T T A A R

ApoI is typically used at 50°C, but is 50% active at 37°C.
EcoRI  (43)
1 site
G A A T T C C T T A A G
MscI  (31)
1 site
T G G C C A A C C G G T
Start  (0)
0 sites
Medium-FT
1 .. 711  =  711 bp
236 amino acids  =  26.7 kDa
Product: blue-to-red fluorescent timer derivative of
mCherry (Subach et al., 2009)
mammalian codon-optimized
Medium-FT
1 .. 711  =  711 bp
236 amino acids  =  26.7 kDa
Product: blue-to-red fluorescent timer derivative of
mCherry (Subach et al., 2009)
mammalian codon-optimized
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