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Plasmid Files

Fast-FT

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

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Fast-FT Sequence and MapFast-FT.dna
Map and Sequence File   
Sequence Author:  Verkhusha Lab
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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) DrdI (541) BbvCI - Bpu10I (539) AlwNI (507) BsrBI (459) StuI (448) PspFI (444) BseYI - PflMI (440) BbsI (436) BtgI - NcoI (434) PstI - SbfI (356) SfcI (352) EcoO109I * (224) BpmI (204) AhdI (195) TaqII (151) BssS α I (88) PmlI (66) BsiHKAI (65) ApaLI (61) MscI (31) Start (0) Fast-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
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
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.
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.
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.
BpmI  (204)
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.
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
PmlI  (66)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°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
Fast-FT
1 .. 711  =  711 bp
236 amino acids  =  26.8 kDa
Product: blue-to-red fluorescent timer derivative of
mCherry (Subach et al., 2009)
mammalian codon-optimized
Fast-FT
1 .. 711  =  711 bp
236 amino acids  =  26.8 kDa
Product: blue-to-red fluorescent timer derivative of
mCherry (Subach et al., 2009)
mammalian codon-optimized
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