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Plasmid Files

mKate2

Monomeric far-red fluorescent protein.

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mKate2 Sequence and MapmKate2.dna
Map and Sequence File   
Sequence Author:  Evrogen
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 600 400 200 End (699) BmrI (694) AlwNI (665) MscI (658) EaeI (656) DraIII (648) PshAI (631) AccI (578) BsaHI (575) SmlI (562) BpuEI (547) BstYI (538) AcuI (508) MspA1I (451) StuI (427) PspFI (423) BseYI (419) MmeI (410) NmeAIII (396) PfoI * (332) BpmI (316) BmeT110I (257) AvaI - BsoBI (256) PasI (222) TsoI (166) BsiEI (140) BsiHKAI (82) ApaLI (78) BsrGI - TatI (41) MslI (31) NspI (30) Start (0) mKate2 699 bp
End  (699)
0 sites
BmrI  (694)
1 site
A C T G G G ( N ) 4 N T G A C C C ( N ) 4

The 1-base overhangs produced by BmrI may be hard to ligate.
Sticky ends from different BmrI sites may not be compatible.
Unlike most restriction enzymes, BmrI can cleave DNA in the
absence of magnesium.
AlwNI  (665)
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.
MscI  (658)
1 site
T G G C C A A C C G G T
EaeI  (656)
1 site
Y G G C C R R C C G G Y
DraIII  (648)
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.
PshAI  (631)
1 site
G A C N N N N G T C C T G N N N N C A G

PshAI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
AccI  (578)
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.
BsaHI  (575)
1 site
G R C G Y C C Y G C R G

BsaHI is typically used at 37°C, but is even more active at 60°C.
SmlI  (562)
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.
BpuEI  (547)
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).
BstYI  (538)
1 site
R G A T C Y Y C T A G R
AcuI  (508)
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).
MspA1I  (451)
1 site
C M G C K G G K C G M C
StuI  (427)
1 site
A G G C C T T C C G G A
PspFI  (423)
1 site
C C C A G C G G G T C G
BseYI  (419)
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.
MmeI  (410)
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).
NmeAIII  (396)
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).
PfoI  (332)
1 site
T C C N G G A A G G N C C T
* Blocked by Dcm methylation.
Sticky ends from different PfoI sites may not be compatible.
BpmI  (316)
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.
BmeT110I  (257)
1 site
C Y C G R G G R G C Y C
AvaI  (256)
1 site
C Y C G R G G R G C Y C

Sticky ends from different AvaI sites may not be compatible.
BsoBI  (256)
1 site
C Y C G R G G R G C Y C

Sticky ends from different BsoBI sites may not be compatible.
BsoBI is typically used at 37°C, but can be used at temperatures
up to 65°C.
PasI  (222)
1 site
C C C W G G G G G G W C C C

Sticky ends from different PasI sites may not be compatible.
TsoI  (166)
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).
BsiEI  (140)
1 site
C G R Y C G G C Y R G C

Sticky ends from different BsiEI sites may not be compatible.
BsiHKAI  (82)
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  (78)
1 site
G T G C A C C A C G T G
BsrGI  (41)
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  (41)
1 site
W G T A C W W C A T G W
MslI  (31)
1 site
C A Y N N N N R T G G T R N N N N Y A C
NspI  (30)
1 site
R C A T G Y Y G T A C R
Start  (0)
0 sites
mKate2
1 .. 699  =  699 bp
232 amino acids  =  26.1 kDa
Product: monomeric far-red fluorescent protein
(Shcherbo et al., 2009)
mammalian codon-optimized
mKate2
1 .. 699  =  699 bp
232 amino acids  =  26.1 kDa
Product: monomeric far-red fluorescent protein
(Shcherbo et al., 2009)
mammalian codon-optimized
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