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Plasmid Files

mgfp5

GFP variant with folding enhancement mutations, suitable for expression in plant cells due to removal of a cryptic intron.

 
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 600 400 200 mgfp5 End (717) PspFI (683) BpuEI (682) AlwNI - BseYI (679) MspA1I - PvuII (676) EcoP15I (649) BsaI (638) BstYI (627) BstBI (622) BtgZI (581) MfeI (560) BsaBI * (542) BclI * (537) BbsI (521) BstZ17I (450) AccI (449) PflMI (420) MmeI (401) NmeAIII (390) BspDI - ClaI (383) AflII (373) HincII (360) AcuI (350) PmlI (325) AflIII (322) BsgI (298) Bsu36I (266) BciVI (265) HaeII (260) NdeI (230) MscI (171) BtgI - NcoI - StyI (166) BaeGI - Bme1580I - Bsp1286I (74) BpmI (47) Start (0) mgfp5 717 bp
End  (717)
0 sites
PspFI  (683)
1 site
C C C A G C G G G T C G
BpuEI  (682)
1 site
C T T G A G ( N ) 14 N N G A A C T C ( N ) 14

Sticky ends from different BpuEI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
AlwNI  (679)
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.
BseYI  (679)
1 site
C C C A G C G G G T C G

After cleavage, BseYI can remain bound to DNA and alter its
electrophoretic mobility.
MspA1I  (676)
1 site
C M G C K G G K C G M C
PvuII  (676)
1 site
C A G C T G G T C G A C
EcoP15I  (649)
1 site
C A G C A G ( N ) 25 G T C G T C ( N ) 25 N N

Efficient cleavage requires two inversely oriented copies of the
EcoP15I recognition sequence.
Sticky ends from different EcoP15I sites may not be compatible.
EcoP15I requires ATP for activity.
BsaI  (638)
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.
BstYI  (627)
1 site
R G A T C Y Y C T A G R
BstBI  (622)
1 site
T T C G A A A A G C T T
BtgZI  (581)
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.
MfeI  (560)
1 site
C A A T T G G T T A A C
BsaBI  (542)
1 site
G A T N N N N A T C C T A N N N N T A G
* Blocked by Dam methylation.
BclI  (537)
1 site
T G A T C A A C T A G T
* Blocked by Dam methylation.
BclI is typically used at 50-55°C, but is 50% active at 37°C.
BbsI  (521)
1 site
G A A G A C N N C T T C T G N N ( N ) 4

Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
BstZ17I  (450)
1 site
G T A T A C C A T A T G
AccI  (449)
1 site
G T M K A C C A K M T G

Efficient cleavage with AccI requires ≥13 bp on each side of the
recognition sequence.
Sticky ends from different AccI sites may not be compatible.
PflMI  (420)
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.
MmeI  (401)
1 site
T C C R A C ( N ) 18 N N A G G Y T G ( N ) 18

Efficient cleavage requires at least two copies of the MmeI
recognition sequence.
Sticky ends from different MmeI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
NmeAIII  (390)
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).
BspDI  (383)
1 site
A T C G A T T A G C T A
ClaI  (383)
1 site
A T C G A T T A G C T A
AflII  (373)
1 site
C T T A A G G A A T T C

The sticky ends produced by AflII are hard to ligate.
HincII  (360)
1 site
G T Y R A C C A R Y T G
AcuI  (350)
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).
PmlI  (325)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
AflIII  (322)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
BsgI  (298)
1 site
G T G C A G ( N ) 14 N N C A C G T C ( N ) 14

Efficient cleavage requires at least two copies of the BsgI
recognition sequence.
Sticky ends from different BsgI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
Bsu36I  (266)
1 site
C C T N A G G G G A N T C C

Sticky ends from different Bsu36I sites may not be compatible.
BciVI  (265)
1 site
G T A T C C ( N ) 5 N C A T A G G ( N ) 5

The 1-base overhangs produced by BciVI may be hard to ligate.
Sticky ends from different BciVI sites may not be compatible.
HaeII  (260)
1 site
R G C G C Y Y C G C G R
NdeI  (230)
1 site
C A T A T G G T A T A C

Prolonged incubation with NdeI may lead to removal of additional
nucleotides.
MscI  (171)
1 site
T G G C C A A C C G G T
BtgI  (166)
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  (166)
1 site
C C A T G G G G T A C C
StyI  (166)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
BaeGI  (74)
1 site
G K G C M C C M C G K G

Sticky ends from different BaeGI sites may not be compatible.
Bme1580I  (74)
1 site
G K G C M C C M C G K G

Sticky ends from different Bme1580I sites may not be compatible.
Bsp1286I  (74)
1 site
G D G C H C C H C G D G

Sticky ends from different Bsp1286I sites may not be compatible.
BpmI  (47)
1 site
C T G G A G ( N ) 14 N N G A C C T C ( N ) 14

Efficient cleavage requires at least two copies of the BpmI
recognition sequence.
Sticky ends from different BpmI sites may not be compatible.
After cleavage, BpmI can remain bound to DNA and alter its
electrophoretic mobility.
BpmI quickly loses activity at 37°C.
Start  (0)
0 sites
mgfp5
1 .. 717  =  717 bp
238 amino acids  =  26.8 kDa
Product: GFP with folding enhancement mutations
suitable for expression in plants due to removal of a
cryptic intron
mgfp5
1 .. 717  =  717 bp
238 amino acids  =  26.8 kDa
Product: GFP with folding enhancement mutations
suitable for expression in plants due to removal of a
cryptic intron
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