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Plasmid Files

YPet (humanized)

Yellow fluorescent protein optimized for FRET with CyPet and for expression in mammalian cells.

 
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 600 400 200 YPet End (720) BanII - SacI (710) Eco53kI (708) NaeI (684) NgoMIV (682) EcoO109I - PpuMI (630) EcoNI (625) Bpu10I (604) MspA1I - PvuII (534) BstYI (500) BsrGI (452) AflIII (449) PflMI (423) BsaWI (365) NmeAIII (356) SmaI (328) TspMI - XmaI (326) BsaAI (279) KpnI (225) Acc65I (221) Bts α I (214) PasI (192) BstEII (183) ApaLI (144) BsaHI (110) BmgBI (66) BstXI (48) SgrAI (28) BspQI - EarI - SapI (11) Start (0) YPet (humanized) 720 bp
End  (720)
0 sites
BanII  (710)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
SacI  (710)
1 site
G A G C T C C T C G A G
Eco53kI  (708)
1 site
G A G C T C C T C G A G
NaeI  (684)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NaeI
recognition sequence.
NgoMIV  (682)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NgoMIV
recognition sequence.
EcoO109I  (630)
1 site
R G G N C C Y Y C C N G G R

Sticky ends from different EcoO109I sites may not be compatible.
PpuMI  (630)
1 site
R G G W C C Y Y C C W G G R

Sticky ends from different PpuMI sites may not be compatible.
EcoNI  (625)
1 site
C C T N N N N N A G G G G A N N N N N T C C

The 1-base overhangs produced by EcoNI may be hard to ligate.
Sticky ends from different EcoNI 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.
MspA1I  (534)
1 site
C M G C K G G K C G M C
PvuII  (534)
1 site
C A G C T G G T C G A C
BstYI  (500)
1 site
R G A T C Y Y C T A G R
BsrGI  (452)
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.
AflIII  (449)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
PflMI  (423)
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.
BsaWI  (365)
1 site
W C C G G W W G G C C W

Efficient cleavage requires at least two copies of the BsaWI
recognition sequence.
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).
SmaI  (328)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
TspMI  (326)
1 site
C C C G G G G G G C C C
XmaI  (326)
1 site
C C C G G G G G G C C C

Efficient cleavage requires at least two copies of the XmaI
recognition sequence.
Full cleavage with XmaI may require a long incubation.
BsaAI  (279)
1 site
Y A C G T R R T G C A Y
KpnI  (225)
1 site
G G T A C C C C A T G G
Acc65I  (221)
1 site
G G T A C C C C A T G G
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.
PasI  (192)
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.
BstEII  (183)
1 site
G G T N A C C C C A N T G G

Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
ApaLI  (144)
1 site
G T G C A C C A C G T G
BsaHI  (110)
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.
BmgBI  (66)
1 site
C A C G T C G T G C A G

This recognition sequence is asymmetric, so ligating blunt ends
generated by BmgBI will not always regenerate a BmgBI site.
BstXI  (48)
1 site
C C A N N N N N N T G G G G T N N N N N N A C C

Sticky ends from different BstXI sites may not be compatible.
SgrAI  (28)
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.
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
YPet
1 .. 720  =  720 bp
239 amino acids  =  26.9 kDa
   Segment 1:  
   1 .. 3  =  3 bp
   1 amino acid  =  149.2 Da
Product: yellow fluorescent protein optimized for
FRET with CyPet (Nguyen and Daugherty, 2005)
mammalian codon-optimized
YPet
1 .. 720  =  720 bp
239 amino acids  =  26.9 kDa
   Segment 2:  1a  
   4 .. 6  =  3 bp
   1 amino acid  =  117.2 Da
Product: yellow fluorescent protein optimized for
FRET with CyPet (Nguyen and Daugherty, 2005)
mammalian codon-optimized
YPet
1 .. 720  =  720 bp
239 amino acids  =  26.9 kDa
   Segment 3:  
   7 .. 720  =  714 bp
   237 amino acids  =  26.7 kDa
Product: yellow fluorescent protein optimized for
FRET with CyPet (Nguyen and Daugherty, 2005)
mammalian codon-optimized
YPet
1 .. 720  =  720 bp
239 amino acids  =  26.9 kDa
3 segments
Product: yellow fluorescent protein optimized for
FRET with CyPet (Nguyen and Daugherty, 2005)
mammalian codon-optimized
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