pFusionRed-N

Vector for fusing the monomeric red fluorescent protein FusionRed to the C-terminus of a partner protein.

Sequence Author: Evrogen

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AflIII - PciI (4653) EcoO109I (3833) RsrII (3251) BsrDI (2968) PflFI - Tth111I (2853) FspI (2837) PluTI (2738) SfoI (2736) NarI (2735) KasI (2734) BspDI * - ClaI * (2575) BseRI (2553) SfiI (2510) AseI (7) NdeI (234) SnaBI (340) NheI (591) BmtI (595) AfeI (596) BglII (609) PaeR7I - XhoI (613) Eco53kI (618) SacI (620) HindIII (622) EcoRI (629) PstI (638) SalI (639) Acc65I (645) KpnI (649) SacII (652) PspOMI (653) TspMI - XmaI (656) ApaI (657) SmaI (658) BamHI (660) AgeI (666) AleI (679) BsrGI (719) BbsI (977) BsmBI - Esp3I (1108) PflMI (1275) PshAI (1309) NotI (1379) XbaI * (1389) MfeI (1485) HpaI (1498) BtsI - BtsαI (1574) AflII (1617) CsiI - SexAI * (2324) pFusionRed-N 4711 bp
AflIII  (4653)
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  (4653)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
EcoO109I  (3833)
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.
RsrII  (3251)
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  (2968)
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  (2853)
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  (2853)
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  (2837)
1 site
T G C G C A A C G C G T
PluTI  (2738)
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  (2736)
1 site
G G C G C C C C G C G G
NarI  (2735)
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  (2734)
1 site
G G C G C C C C G C G G
BspDI  (2575)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (2575)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
BseRI  (2553)
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  (2510)
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  (7)
1 site
A T T A A T T A A T T A
NdeI  (234)
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  (340)
1 site
T A C G T A A T G C A T
NheI  (591)
1 site
G C T A G C C G A T C G
BmtI  (595)
1 site
G C T A G C C G A T C G
AfeI  (596)
1 site
A G C G C T T C G C G A
BglII  (609)
1 site
A G A T C T T C T A G A
PaeR7I  (613)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (613)
1 site
C T C G A G G A G C T C
Eco53kI  (618)
1 site
G A G C T C C T C G A G
SacI  (620)
1 site
G A G C T C C T C G A G
HindIII  (622)
1 site
A A G C T T T T C G A A
EcoRI  (629)
1 site
G A A T T C C T T A A G
PstI  (638)
1 site
C T G C A G G A C G T C
SalI  (639)
1 site
G T C G A C C A G C T G
Acc65I  (645)
1 site
G G T A C C C C A T G G
KpnI  (649)
1 site
G G T A C C C C A T G G
SacII  (652)
1 site
C C G C G G G G C G C C

Efficient cleavage requires at least two copies of the SacII recognition sequence.
PspOMI  (653)
1 site
G G G C C C C C C G G G
TspMI  (656)
1 site
C C C G G G G G G C C C
XmaI  (656)
1 site
C C C G G G G G G C C C

Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
ApaI  (657)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
SmaI  (658)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
BamHI  (660)
1 site
G G A T C C C C T A G G

After cleavage, BamHI-HF® (but not the original BamHI) can remain bound to DNA and alter its electrophoretic mobility.
AgeI  (666)
1 site
A C C G G T T G G C C A
AleI  (679)
1 site
C A C N N N N G T G G T G N N N N C A C
BsrGI  (719)
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.
BbsI  (977)
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.
BsmBI  (1108)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different BsmBI sites may not be compatible.
BsmBI-v2 is an improved version of BsmBI.
Esp3I  (1108)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different Esp3I sites may not be compatible.
PflMI  (1275)
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.
PshAI  (1309)
1 site
G A C N N N N G T C C T G N N N N C A G

PshAI quickly loses activity at 37°C, but can be used at 25°C for long incubations.
NotI  (1379)
1 site
G C G G C C G C C G C C G G C G
XbaI  (1389)
1 site
T C T A G A A G A T C T
* Blocked by Dam methylation.
MfeI  (1485)
1 site
C A A T T G G T T A A C
HpaI  (1498)
1 site
G T T A A C C A A T T G
BtsI  (1574)
1 site
G C A G T G N N C G T C A C
BtsαI  (1574)
1 site
G C A G T G N N C G T C A C

Sticky ends from different BtsαI sites may not be compatible.
AflII  (1617)
1 site
C T T A A G G A A T T C
CsiI  (2324)
1 site
A C C W G G T T G G W C C A

Sticky ends from different CsiI sites may not be compatible.
SexAI  (2324)
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
2607 .. 3401  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR/KanR
2607 .. 3401  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
FusionRed
679 .. 1377  =  699 bp
232 amino acids  =  25.6 kDa
Product: monomeric red fluorescent protein optimized for use as a fusion tag (Shemiakina et al., 2012)
mammalian codon-optimized
FusionRed
679 .. 1377  =  699 bp
232 amino acids  =  25.6 kDa
Product: monomeric red fluorescent protein optimized for use as a fusion tag (Shemiakina et al., 2012)
mammalian codon-optimized
ori
4009 .. 4597  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
4009 .. 4597  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
1627 .. 2082  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
1627 .. 2082  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
SV40 promoter
2215 .. 2572  =  358 bp
SV40 enhancer and early promoter
SV40 promoter
2215 .. 2572  =  358 bp
SV40 enhancer and early promoter
CMV enhancer
61 .. 364  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
61 .. 364  =  304 bp
human cytomegalovirus immediate early enhancer
CMV promoter
365 .. 568  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
365 .. 568  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
SV40 poly(A) signal
1499 .. 1620  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1499 .. 1620  =  122 bp
SV40 polyadenylation signal
AmpR promoter
2109 .. 2213  =  105 bp
AmpR promoter
2109 .. 2213  =  105 bp
MCS
591 .. 671  =  81 bp
multiple cloning site
MCS
591 .. 671  =  81 bp
multiple cloning site
HSV TK poly(A) signal
3633 .. 3680  =  48 bp
herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)
HSV TK poly(A) signal
3633 .. 3680  =  48 bp
herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)
SV40 ori
2423 .. 2558  =  136 bp
SV40 origin of replication
SV40 ori
2423 .. 2558  =  136 bp
SV40 origin of replication
ORF:  679 .. 1377  =  699 bp
ORF:  232 amino acids  =  25.6 kDa
ORF:  2779 .. 3165  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  3422 .. 3871  =  450 bp
ORF:  149 amino acids  =  16.3 kDa
ORF:  2607 .. 3401  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  447 .. 725  =  279 bp
ORF:  92 amino acids  =  9.9 kDa
ORF:  783 .. 1028  =  246 bp
ORF:  81 amino acids  =  8.8 kDa
ORF:  2916 .. 3452  =  537 bp
ORF:  178 amino acids  =  19.8 kDa
ORF:  3627 .. 3860  =  234 bp
ORF:  77 amino acids  =  8.6 kDa
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