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Plasmid Files

PAmKate

Monomeric photoactivatable red fluorescent protein derived from mKate.

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PAmKate Sequence and MapPAmKate.dna
Map and Sequence File   
Sequence Author:  Verkhusha Lab
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 600 400 200 End (714) PacI (711) BmrI - HindIII (703) AlwNI (674) MscI (667) EaeI (665) DraIII (657) PshAI (640) BsaI (628) AccI (587) BsaHI (584) SmlI (571) BpuEI (556) BstYI (547) AcuI (517) BglI (476) MspA1I (460) PspFI (432) BseYI (428) MmeI (419) NmeAIII (405) PfoI * (341) BpmI (325) Bsu36I (266) PasI (231) TsoI (175) BsiHKAI (91) ApaLI (87) BsrGI - TatI (50) NspI (39) BspQI - EarI - SapI (11) Start (0) PAmKate 714 bp
End  (714)
0 sites
PacI  (711)
1 site
T T A A T T A A A A T T A A T T
BmrI  (703)
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.
HindIII  (703)
1 site
A A G C T T T T C G A A
AlwNI  (674)
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  (667)
1 site
T G G C C A A C C G G T
EaeI  (665)
1 site
Y G G C C R R C C G G Y
DraIII  (657)
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  (640)
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.
BsaI  (628)
1 site
G G T C T C N C C A G A G N ( N ) 4

Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
AccI  (587)
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  (584)
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  (571)
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  (556)
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  (547)
1 site
R G A T C Y Y C T A G R
AcuI  (517)
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).
BglI  (476)
1 site
G C C N N N N N G G C C G G N N N N N C C G

Sticky ends from different BglI sites may not be compatible.
MspA1I  (460)
1 site
C M G C K G G K C G M C
PspFI  (432)
1 site
C C C A G C G G G T C G
BseYI  (428)
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  (419)
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  (405)
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  (341)
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  (325)
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.
Bsu36I  (266)
1 site
C C T N A G G G G A N T C C

Sticky ends from different Bsu36I sites may not be compatible.
PasI  (231)
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  (175)
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).
BsiHKAI  (91)
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  (87)
1 site
G T G C A C C A C G T G
BsrGI  (50)
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  (50)
1 site
W G T A C W W C A T G W
NspI  (39)
1 site
R C A T G Y Y G T A C R
BspQI  (11)
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  (11)
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  (11)
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.
Start  (0)
0 sites
PAmKate
1 .. 714  =  714 bp
237 amino acids  =  26.7 kDa
Product: monomeric photoactivatable red
fluorescent protein (Gunewardene et al., 2011)
mammalian codon-optimized
PAmKate
1 .. 714  =  714 bp
237 amino acids  =  26.7 kDa
Product: monomeric photoactivatable red
fluorescent protein (Gunewardene et al., 2011)
mammalian codon-optimized
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