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Plasmid Files

AcGFP1

Aequorea coerulescens green fluorescent protein.

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AcGFP1 Sequence and MapAcGFP1.dna
Map and Sequence File   
Sequence Author:  Clontech
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 600 400 200 AcGFP1 End (720) PflMI (699) BtgI (693) EcoO109I - PpuMI (630) StyI (626) EcoNI (625) TsoI (610) MspA1I - PvuII (534) BstYI (500) EciI (493) BspHI (458) DraIII (450) BssHII (327) PfoI * (281) Bsu36I (269) Bts α I (214) Bpu10I (193) BstEII (183) AleI (181) AcuI - Eco57MI (154) BsrBI (92) MscI (74) PluTI (17) SfoI (15) BsaHI - NarI (14) KasI (13) Start (0) AcGFP1 720 bp
End  (720)
0 sites
PflMI  (699)
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.
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.
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.
StyI  (626)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI 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.
TsoI  (610)
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).
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
EciI  (493)
1 site
G G C G G A ( N ) 9 N N C C G C C T ( N ) 9

Sticky ends from different EciI sites may not be compatible.
BspHI  (458)
1 site
T C A T G A A G T A C T
DraIII  (450)
1 site
C A C N N N G T G G T G N N N C A C

Sticky ends from different DraIII sites may not be compatible.
BssHII  (327)
1 site
G C G C G C C G C G C G

BssHII is typically used at 50°C, but is 75% active at 37°C.
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.
Bsu36I  (269)
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.
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.
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.
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.
AleI  (181)
1 site
C A C N N N N G T G G T G N N N N C A C
AcuI  (154)
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).
Eco57MI  (154)
1 site
C T G R A G ( N ) 14 N N G A C Y T C ( N ) 14

Sticky ends from different Eco57MI sites may not be compatible.
After cleavage, Eco57MI can remain bound to DNA and alter its
electrophoretic mobility.
For full activity, add fresh S-adenosylmethionine (SAM).
BsrBI  (92)
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.
MscI  (74)
1 site
T G G C C A A C C G G T
PluTI  (17)
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  (15)
1 site
G G C G C C C C G C G G
BsaHI  (14)
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  (14)
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.
KasI  (13)
1 site
G G C G C C C C G C G G
Start  (0)
0 sites
AcGFP1
1 .. 720  =  720 bp
239 amino acids  =  26.9 kDa
Product: Aequorea coerulescens GFP
mammalian codon-optimized
AcGFP1
1 .. 720  =  720 bp
239 amino acids  =  26.9 kDa
Product: Aequorea coerulescens GFP
mammalian codon-optimized
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