pDsRed-Monomer-N In-Fusion Ready

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

Sequence Author: Clontech (TaKaRa)

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AflIII - PciI (3986) ApaLI (3672) EcoO109I (3166) BsaI (3057) PfoI (2843) BstBI (2750) RsrII (2584) BsrDI (2301) PflFI - Tth111I (2186) MscI (2150) PluTI (2071) SfoI (2069) NarI (2068) KasI (2067) BspDI * - ClaI * (1908) SfiI (1843) CsiI - SexAI * (1657) AseI (4051) NdeI (4278) SnaBI (4384) NheI (4635) BmtI (4639) Bpu10I (4640) End (4655) Start (0) EcoRI (8) AgeI (20) FspAI (75) BstEII (167) AhdI (206) AvaI - BsoBI (288) AfeI (309) SbfI (367) BbsI (447) BglII (505) BsrGI (565) PflMI (608) AleI (654) NotI (706) XbaI * (722) MfeI (818) HincII - HpaI (831) BtsI - BtsαI (907) AflII (950) pDsRed-Monomer-N In-Fusion® Ready 4655 bp
AflIII  (3986)
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  (3986)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
ApaLI  (3672)
1 site
G T G C A C C A C G T G
EcoO109I  (3166)
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.
BsaI  (3057)
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.
PfoI  (2843)
1 site
T C C N G G A A G G N C C T

Sticky ends from different PfoI sites may not be compatible.
BstBI  (2750)
1 site
T T C G A A A A G C T T
RsrII  (2584)
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  (2301)
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  (2186)
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  (2186)
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.
MscI  (2150)
1 site
T G G C C A A C C G G T
PluTI  (2071)
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  (2069)
1 site
G G C G C C C C G C G G
NarI  (2068)
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  (2067)
1 site
G G C G C C C C G C G G
BspDI  (1908)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (1908)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
SfiI  (1843)
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.
CsiI  (1657)
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  (1657)
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.
AseI  (4051)
1 site
A T T A A T T A A T T A
NdeI  (4278)
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  (4384)
1 site
T A C G T A A T G C A T
NheI  (4635)
1 site
G C T A G C C G A T C G
BmtI  (4639)
1 site
G C T A G C C G A T C G
Bpu10I  (4640)
1 site
C C T N A G C G G A N T C G

Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present.
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.
End  (4655)
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
FspAI  (75)
1 site
R T G C G C A Y Y A C G C G T R
BstEII  (167)
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.
AhdI  (206)
1 site
G A C N N N N N G T C C T G N N N N N C A G

The 1-base overhangs produced by AhdI may be hard to ligate.
Sticky ends from different AhdI sites may not be compatible.
AvaI  (288)
1 site
C Y C G R G G R G C Y C

Sticky ends from different AvaI sites may not be compatible.
BsoBI  (288)
1 site
C Y C G R G G R G C Y C

Sticky ends from different BsoBI sites may not be compatible.
BsoBI is typically used at 37°C, but can be used at temperatures up to 65°C.
AfeI  (309)
1 site
A G C G C T T C G C G A
SbfI  (367)
1 site
C C T G C A G G G G A C G T C C
BbsI  (447)
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.
BglII  (505)
1 site
A G A T C T T C T A G A
BsrGI  (565)
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.
PflMI  (608)
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.
AleI  (654)
1 site
C A C N N N N G T G G T G N N N N C A C
NotI  (706)
1 site
G C G G C C G C C G C C G G C G
XbaI  (722)
1 site
T C T A G A A G A T C T
* Blocked by Dam methylation.
MfeI  (818)
1 site
C A A T T G G T T A A C
HincII  (831)
1 site
G T Y R A C C A R Y T G
HpaI  (831)
1 site
G T T A A C C A A T T G
BtsI  (907)
1 site
G C A G T G N N C G T C A C
BtsαI  (907)
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  (950)
1 site
C T T A A G G A A T T C
NeoR/KanR
1940 .. 2734  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin)
NeoR/KanR
1940 .. 2734  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin)
DsRed-Monomer
30 .. 704  =  675 bp
224 amino acids  =  25.3 kDa
Product: monomeric derivative of DsRed fluorescent protein (Strongin et al., 2007)
mammalian codon-optimized
DsRed-Monomer
30 .. 704  =  675 bp
224 amino acids  =  25.3 kDa
Product: monomeric derivative of DsRed fluorescent protein (Strongin et al., 2007)
mammalian codon-optimized
ori
3342 .. 3930  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3342 .. 3930  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
960 .. 1415  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
960 .. 1415  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
SV40 promoter
1548 .. 1905  =  358 bp
SV40 enhancer and early promoter
SV40 promoter
1548 .. 1905  =  358 bp
SV40 enhancer and early promoter
CMV enhancer
4105 .. 4408  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
4105 .. 4408  =  304 bp
human cytomegalovirus immediate early enhancer
CMV promoter
4409 .. 4612  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
4409 .. 4612  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
SV40 poly(A) signal
832 .. 953  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
832 .. 953  =  122 bp
SV40 polyadenylation signal
AmpR promoter
1442 .. 1546  =  105 bp
AmpR promoter
1442 .. 1546  =  105 bp
HSV TK poly(A) signal
2966 .. 3013  =  48 bp
herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)
HSV TK poly(A) signal
2966 .. 3013  =  48 bp
herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)
SV40 ori
1756 .. 1891  =  136 bp
SV40 origin of replication
SV40 ori
1756 .. 1891  =  136 bp
SV40 origin of replication
ORF:  2755 .. 3204  =  450 bp
ORF:  149 amino acids  =  16.3 kDa
ORF:  2 .. 733  =  732 bp
ORF:  243 amino acids  =  26.8 kDa  (no start codon)
ORF:  1940 .. 2734  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  3 .. 704  =  702 bp
ORF:  233 amino acids  =  26.1 kDa  (no start codon)
ORF:  2112 .. 2498  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  2249 .. 2785  =  537 bp
ORF:  178 amino acids  =  19.8 kDa
ORF:  2960 .. 3193  =  234 bp
ORF:  77 amino acids  =  8.6 kDa
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