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Plasmid Files

mKalama1

Photostable monomeric blue variant of EGFP.

 
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 600 400 200 mKalama1 End (720) BsrGI (710) BlpI (622) Bpu10I (604) AarI - BfuAI - BspMI (603) NmeAIII (356) PfoI * (281) EciI (249) Bts α I (214) DrdI (192) BssS α I (181) TaqII (130) BtgI (117) BseRI (31) Start (0) mKalama1 720 bp
End  (720)
0 sites
BsrGI  (710)
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.
BlpI  (622)
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.
Bpu10I  (604)
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.
AarI  (603)
1 site
C A C C T G C ( N ) 4 G T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the AarI
recognition sequence.
Sticky ends from different AarI sites may not be compatible.
After cleavage, AarI can remain bound to DNA and alter its
electrophoretic mobility.
BfuAI  (603)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BfuAI
recognition sequence.
Sticky ends from different BfuAI sites may not be compatible.
BfuAI is typically used at 50°C, but is 50% active at 37°C.
BspMI  (603)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BspMI
recognition sequence.
Sticky ends from different BspMI sites may not be compatible.
NmeAIII  (356)
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  (281)
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.
EciI  (249)
1 site
G G C G G A ( N ) 9 N N C C G C C T ( N ) 9

Sticky ends from different EciI sites may not be compatible.
BtsαI  (214)
1 site
G C A G T G N N C G T C A C

Sticky ends from different BtsI sites may not be compatible.
DrdI  (192)
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.
BssSαI  (181)
1 site
C A C G A G G T G C T C
TaqII  (130)
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.
BtgI  (117)
1 site
C C R Y G G G G Y R C C

Sticky ends from different BtgI sites may not be compatible.
BseRI  (31)
1 site
G A G G A G ( N ) 8 N N C T C C T C ( N ) 8

Sticky ends from different BseRI sites may not be compatible.
BseRI quickly loses activity at 37°C.
Prolonged incubation with BseRI may lead to degradation of the
DNA.
Start  (0)
0 sites
mKalama1
1 .. 720  =  720 bp
239 amino acids  =  26.8 kDa
   Segment 1:  
   1 .. 3  =  3 bp
   1 amino acid  =  149.2 Da
Product: photostable monomeric blue variant of GFP
(Ai et al., 2007)
mammalian codon-optimized
mKalama1
1 .. 720  =  720 bp
239 amino acids  =  26.8 kDa
   Segment 2:  1a  
   4 .. 6  =  3 bp
   1 amino acid  =  117.2 Da
Product: photostable monomeric blue variant of GFP
(Ai et al., 2007)
mammalian codon-optimized
mKalama1
1 .. 720  =  720 bp
239 amino acids  =  26.8 kDa
   Segment 3:  
   7 .. 720  =  714 bp
   237 amino acids  =  26.6 kDa
Product: photostable monomeric blue variant of GFP
(Ai et al., 2007)
mammalian codon-optimized
mKalama1
1 .. 720  =  720 bp
239 amino acids  =  26.8 kDa
3 segments
Product: photostable monomeric blue variant of GFP
(Ai et al., 2007)
mammalian codon-optimized
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