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Plasmid Files

d2EYFP

EYFP destabilized by residues 422-461 of mouse ornithine decarboxylase, giving an in vivo half-life of ~2 hours.

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d2EYFP Sequence and Mapd2EYFP.dna
Map and Sequence File   
Sequence Author:  Clontech
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 800 600 400 200 EYFP destabilizati... d2EYFP End (846) BstAPI (820) BsrBI (794) TsoI (739) BtgI - NcoI - StyI (726) BlpI (721) HindIII (718) BsrGI (710) BstYI (500) PfoI * (281) Bts α I (214) BssS α I (181) BsrFI (151) BtgZI (122) BanI (36) Start (0) d2EYFP 846 bp
End  (846)
0 sites
BstAPI  (820)
1 site
G C A N N N N N T G C C G T N N N N N A C G

Sticky ends from different BstAPI sites may not be compatible.
BsrBI  (794)
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.
TsoI  (739)
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).
BtgI  (726)
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  (726)
1 site
C C A T G G G G T A C C
StyI  (726)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
BlpI  (721)
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.
HindIII  (718)
1 site
A A G C T T T T C G A A
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.
BstYI  (500)
1 site
R G A T C Y Y C T A G R
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.
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.
BssSαI  (181)
1 site
C A C G A G G T G C T C
BsrFI  (151)
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.
BtgZI  (122)
1 site
G C G A T G ( N ) 10 C G C T A C ( N ) 10 ( N ) 4

Sticky ends from different BtgZI sites may not be compatible.
After cleavage, BtgZI can remain bound to DNA and alter its
electrophoretic mobility.
BtgZI is typically used at 60°C, but is 75% active at 37°C.
BanI  (36)
1 site
G G Y R C C C C R Y G G

Sticky ends from different BanI sites may not be compatible.
Start  (0)
0 sites
d2EYFP
1 .. 846  =  846 bp
281 amino acids  =  31.5 kDa
   Segment 1:  
   1 .. 3  =  3 bp
   1 amino acid  =  149.2 Da
Product: EYFP destabilized by residues 422-461 of
mouse ornithine decarboxylase, giving an in vivo
half-life of ~2 hours
mammalian codon-optimized
d2EYFP
1 .. 846  =  846 bp
281 amino acids  =  31.5 kDa
   Segment 2:  1a  
   4 .. 6  =  3 bp
   1 amino acid  =  117.2 Da
Product: EYFP destabilized by residues 422-461 of
mouse ornithine decarboxylase, giving an in vivo
half-life of ~2 hours
mammalian codon-optimized
d2EYFP
1 .. 846  =  846 bp
281 amino acids  =  31.5 kDa
   Segment 3:  EYFP  
   7 .. 717  =  711 bp
   237 amino acids  =  26.8 kDa
Product: EYFP destabilized by residues 422-461 of
mouse ornithine decarboxylase, giving an in vivo
half-life of ~2 hours
mammalian codon-optimized
d2EYFP
1 .. 846  =  846 bp
281 amino acids  =  31.5 kDa
   Segment 4:  
   718 .. 723  =  6 bp
   2 amino acids  =  259.3 Da
Product: EYFP destabilized by residues 422-461 of
mouse ornithine decarboxylase, giving an in vivo
half-life of ~2 hours
mammalian codon-optimized
d2EYFP
1 .. 846  =  846 bp
281 amino acids  =  31.5 kDa
   Segment 5:  destabilization domain  
   724 .. 846  =  123 bp
   40 amino acids  =  4.2 kDa
Product: EYFP destabilized by residues 422-461 of
mouse ornithine decarboxylase, giving an in vivo
half-life of ~2 hours
mammalian codon-optimized
d2EYFP
1 .. 846  =  846 bp
281 amino acids  =  31.5 kDa
5 segments
Product: EYFP destabilized by residues 422-461 of
mouse ornithine decarboxylase, giving an in vivo
half-life of ~2 hours
mammalian codon-optimized
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