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Plasmid Files

hrGFP

Humanized Renilla reniformis green fluorescent protein.

 
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 600 400 200 hrGFP End (720) TaqII (702) PasI (693) BsaI (649) BtgZI (648) XmnI (577) BspQI - EarI - MslI - SapI (550) FspI - FspAI (544) NspI (543) BsrGI (461) XcmI (453) BseRI (370) EcoNI (352) NaeI (279) NgoMIV (277) BanII - Bsp1286I (199) Bpu10I (193) HaeII - PluTI (167) SfoI (165) BsaHI - NarI (164) BanI - KasI (163) BtgI - NcoI (95) PmlI (87) AflIII (86) BspHI * (44) AcuI (40) SbfI (38) AlwNI (18) Start (0) hrGFP 720 bp
End  (720)
0 sites
TaqII  (702)
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.
PasI  (693)
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.
BsaI  (649)
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.
BtgZI  (648)
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.
XmnI  (577)
1 site
G A A N N N N T T C C T T N N N N A A G
BspQI  (550)
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  (550)
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.
MslI  (550)
1 site
C A Y N N N N R T G G T R N N N N Y A C
SapI  (550)
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.
FspI  (544)
1 site
T G C G C A A C G C G T
FspAI  (544)
1 site
R T G C G C A Y Y A C G C G T R
NspI  (543)
1 site
R C A T G Y Y G T A C R
BsrGI  (461)
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.
XcmI  (453)
1 site
C C A N N N N N N N N N T G G G G T N N N N N N N N N A C C

The 1-base overhangs produced by XcmI may be hard to ligate.
Sticky ends from different XcmI sites may not be compatible.
BseRI  (370)
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.
EcoNI  (352)
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.
NaeI  (279)
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  (277)
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.
BanII  (199)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
Bsp1286I  (199)
1 site
G D G C H C C H C G D G

Sticky ends from different Bsp1286I sites may not be compatible.
Bpu10I  (193)
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.
HaeII  (167)
1 site
R G C G C Y Y C G C G R
PluTI  (167)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the PluTI
recognition sequence.
SfoI  (165)
1 site
G G C G C C C C G C G G
BsaHI  (164)
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.
NarI  (164)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the NarI
recognition sequence.
BanI  (163)
1 site
G G Y R C C C C R Y G G

Sticky ends from different BanI sites may not be compatible.
KasI  (163)
1 site
G G C G C C C C G C G G
BtgI  (95)
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  (95)
1 site
C C A T G G G G T A C C
PmlI  (87)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
AflIII  (86)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
BspHI  (44)
1 site
T C A T G A A G T A C T
* Blocked by Dam methylation.
AcuI  (40)
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).
SbfI  (38)
1 site
C C T G C A G G G G A C G T C C
AlwNI  (18)
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.
Start  (0)
0 sites
hrGFP
1 .. 720  =  720 bp
239 amino acids  =  27.3 kDa
Product: humanized Renilla green fluorescent
protein
mammalian codon-optimized
hrGFP
1 .. 720  =  720 bp
239 amino acids  =  27.3 kDa
Product: humanized Renilla green fluorescent
protein
mammalian codon-optimized
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