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Plasmid Files

CyPet (humanized)

Cyan fluorescent protein optimized for FRET with YPet and for expression in mammalian cells.

 
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 600 400 200 CyPet End (720) BtgI (693) NaeI (684) NgoMIV (682) EcoO109I - PpuMI (630) EcoNI (625) HaeII (620) BsiHKAI (610) MspA1I - PvuII (534) AflIII (449) BsaWI (365) NmeAIII (356) SmaI (328) TspMI - XmaI (326) BsaAI (279) KpnI (225) Acc65I - BanI (221) Bts α I (214) BstEII (183) BsaHI (110) BmgBI (66) BstXI (48) Start (0) CyPet (humanized) 720 bp
End  (720)
0 sites
BtgI  (693)
1 site
C C R Y G G G G Y R C C

Sticky ends from different BtgI sites may not be compatible.
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.
HaeII  (620)
1 site
R G C G C Y Y C G C G R
BsiHKAI  (610)
1 site
G W G C W C C W C G W G

Sticky ends from different BsiHKAI 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
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.
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
BanI  (221)
1 site
G G Y R C C C C R Y G G

Sticky ends from different BanI 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.
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.
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.
Start  (0)
0 sites
CyPet
1 .. 720  =  720 bp
239 amino acids  =  26.9 kDa
   Segment 1:  
   1 .. 3  =  3 bp
   1 amino acid  =  149.2 Da
Product: cyan fluorescent protein optimized for FRET
with YPet (Nguyen and Daugherty, 2005)
mammalian codon-optimized
CyPet
1 .. 720  =  720 bp
239 amino acids  =  26.9 kDa
   Segment 2:  1a  
   4 .. 6  =  3 bp
   1 amino acid  =  117.2 Da
Product: cyan fluorescent protein optimized for FRET
with YPet (Nguyen and Daugherty, 2005)
mammalian codon-optimized
CyPet
1 .. 720  =  720 bp
239 amino acids  =  26.9 kDa
   Segment 3:  
   7 .. 720  =  714 bp
   237 amino acids  =  26.6 kDa
Product: cyan fluorescent protein optimized for FRET
with YPet (Nguyen and Daugherty, 2005)
mammalian codon-optimized
CyPet
1 .. 720  =  720 bp
239 amino acids  =  26.9 kDa
3 segments
Product: cyan fluorescent protein optimized for FRET
with YPet (Nguyen and Daugherty, 2005)
mammalian codon-optimized
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