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Plasmid Files

pAcGFP1-N In-Fusion® Ready

Linearized vector for fusing AcGFP1 to the C-terminus of a partner protein.

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pAcGFP1-N In-Fusion Ready Sequence and MappAcGFP1-N In-Fusion Ready.dna
Map and Sequence File   
Sequence Author:  Clontech
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 AflIII - PciI (4019) ApaLI (3705) BsaI (3090) BstBI (2783) RsrII (2617) BsrDI (2334) PflFI - Tth111I (2219) FspI (2203) BspDI * - ClaI * (1941) BseRI (1919) SfiI (1876) AseI (4084) NdeI (4311) SnaBI (4417) NheI (4668) BmtI (4672) End (4688) Start (0) EcoRI (8) AgeI (20) NruI (27) AleI (204) BstEII (206) BssHII (350) EcoNI (648) PpuMI (653) PflMI (722) NotI (745) XbaI * (755) MfeI (851) HincII - HpaI (864) AflII (983) SexAI * (1690) pAcGFP1-N In-Fusion® Ready 4688 bp
AflIII  (4019)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
PciI  (4019)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
ApaLI  (3705)
1 site
G T G C A C C A C G T G
BsaI  (3090)
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.
BstBI  (2783)
1 site
T T C G A A A A G C T T
RsrII  (2617)
1 site
C G G W C C G G C C W G G C

Efficient cleavage requires at least two copies of the RsrII
recognition sequence.
Sticky ends from different RsrII sites may not be compatible.
For full activity, add fresh DTT.
BsrDI  (2334)
1 site
G C A A T G N N C G T T A C

Sticky ends from different BsrDI sites may not be compatible.
PflFI  (2219)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (2219)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
FspI  (2203)
1 site
T G C G C A A C G C G T
BspDI  (1941)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (1941)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
BseRI  (1919)
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.
SfiI  (1876)
1 site
G G C C N N N N N G G C C C C G G N N N N N C C G G

Efficient cleavage requires at least two copies of the SfiI
recognition sequence.
Sticky ends from different SfiI sites may not be compatible.
AseI  (4084)
1 site
A T T A A T T A A T T A
NdeI  (4311)
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.
SnaBI  (4417)
1 site
T A C G T A A T G C A T
NheI  (4668)
1 site
G C T A G C C G A T C G
BmtI  (4672)
1 site
G C T A G C C G A T C G
End  (4688)
0 sites
Start  (0)
0 sites
EcoRI  (8)
1 site
G A A T T C C T T A A G
AgeI  (20)
1 site
A C C G G T T G G C C A

AgeI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
NruI  (27)
1 site
T C G C G A A G C G C T
AleI  (204)
1 site
C A C N N N N G T G G T G N N N N C A C
BstEII  (206)
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.
BssHII  (350)
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.
EcoNI  (648)
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.
PpuMI  (653)
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.
PflMI  (722)
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.
NotI  (745)
1 site
G C G G C C G C C G C C G G C G
XbaI  (755)
1 site
T C T A G A A G A T C T
* Blocked by Dam methylation.
MfeI  (851)
1 site
C A A T T G G T T A A C
HincII  (864)
1 site
G T Y R A C C A R Y T G
HpaI  (864)
1 site
G T T A A C C A A T T G
AflII  (983)
1 site
C T T A A G G A A T T C

The sticky ends produced by AflII are hard to ligate.
SexAI  (1690)
1 site
A C C W G G T T G G W C C A
* Blocked by Dcm methylation.
Sticky ends from different SexAI sites may not be compatible.
NeoR/KanR
1973 .. 2767  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin)
NeoR/KanR
1973 .. 2767  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin)
AcGFP1
30 .. 743  =  714 bp
237 amino acids  =  26.6 kDa
Product: Aequorea coerulescens GFP lacking the
first two codons
mammalian codon-optimized
AcGFP1
30 .. 743  =  714 bp
237 amino acids  =  26.6 kDa
Product: Aequorea coerulescens GFP lacking the
first two codons
mammalian codon-optimized
ori
3375 .. 3963  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
3375 .. 3963  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
993 .. 1448  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
993 .. 1448  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
SV40 promoter
1581 .. 1938  =  358 bp
SV40 enhancer and early promoter
SV40 promoter
1581 .. 1938  =  358 bp
SV40 enhancer and early promoter
CMV enhancer
4138 .. 4441  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
4138 .. 4441  =  304 bp
human cytomegalovirus immediate early enhancer
CMV promoter
4442 .. 4645  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
4442 .. 4645  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
SV40 poly(A) signal
865 .. 986  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
865 .. 986  =  122 bp
SV40 polyadenylation signal
AmpR promoter
1475 .. 1579  =  105 bp
AmpR promoter
1475 .. 1579  =  105 bp
HSV TK poly(A) signal
2999 .. 3046  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
HSV TK poly(A) signal
2999 .. 3046  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
SV40 ori
1789 .. 1924  =  136 bp
SV40 origin of replication
SV40 ori
1789 .. 1924  =  136 bp
SV40 origin of replication
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