pTurboRFP-N

Vector for fusing TurboRFP to the C-terminus of a partner protein.

Sequence Author: Evrogen

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AflIII - PciI (4651) EcoO109I (3831) PfoI (3508) RsrII (3249) BsrDI (2966) PflFI - Tth111I (2851) FspI (2835) PluTI (2736) SfoI (2734) NarI (2733) KasI (2732) BspDI * - ClaI * (2573) StuI (2554) BseRI (2551) 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) AccI (640) Acc65I (645) KpnI (649) SacII (652) PspOMI (653) TspMI - XmaI (656) ApaI (657) SmaI (658) BamHI (660) AgeI (666) BclI * (689) BsrGI (716) BbsI (974) BsgI (1171) PshAI (1306) ScaI (1339) NotI (1377) XbaI * (1387) MfeI (1483) HpaI (1496) AflII (1615) CsiI - SexAI * (2322) pTurboRFP-N 4709 bp
AflIII  (4651)
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  (4651)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
EcoO109I  (3831)
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.
PfoI  (3508)
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.
RsrII  (3249)
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  (2966)
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  (2851)
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  (2851)
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  (2835)
1 site
T G C G C A A C G C G T
PluTI  (2736)
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  (2734)
1 site
G G C G C C C C G C G G
NarI  (2733)
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  (2732)
1 site
G G C G C C C C G C G G
BspDI  (2573)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (2573)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
StuI  (2554)
1 site
A G G C C T T C C G G A
BseRI  (2551)
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.
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
AccI  (640)
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.
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
BclI  (689)
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.
BsrGI  (716)
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  (974)
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.
BsgI  (1171)
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).
PshAI  (1306)
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.
ScaI  (1339)
1 site
A G T A C T T C A T G A
NotI  (1377)
1 site
G C G G C C G C C G C C G G C G
XbaI  (1387)
1 site
T C T A G A A G A T C T
* Blocked by Dam methylation.
MfeI  (1483)
1 site
C A A T T G G T T A A C
HpaI  (1496)
1 site
G T T A A C C A A T T G
AflII  (1615)
1 site
C T T A A G G A A T T C
CsiI  (2322)
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  (2322)
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
2605 .. 3399  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin)
NeoR/KanR
2605 .. 3399  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin)
TurboRFP
679 .. 1374  =  696 bp
231 amino acids  =  26.1 kDa
Product: red fluorescent protein from Entacmaea quadricolor
mammalian codon-optimized
TurboRFP
679 .. 1374  =  696 bp
231 amino acids  =  26.1 kDa
Product: red fluorescent protein from Entacmaea quadricolor
mammalian codon-optimized
ori
4007 .. 4595  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
4007 .. 4595  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
1625 .. 2080  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
1625 .. 2080  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
SV40 promoter
2213 .. 2570  =  358 bp
SV40 enhancer and early promoter
SV40 promoter
2213 .. 2570  =  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
1497 .. 1618  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1497 .. 1618  =  122 bp
SV40 polyadenylation signal
AmpR promoter
2107 .. 2211  =  105 bp
AmpR promoter
2107 .. 2211  =  105 bp
MCS
591 .. 671  =  81 bp
multiple cloning site of fluorescent protein plasmids
MCS
591 .. 671  =  81 bp
multiple cloning site of fluorescent protein plasmids
HSV TK poly(A) signal
3631 .. 3678  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
HSV TK poly(A) signal
3631 .. 3678  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
SV40 ori
2421 .. 2556  =  136 bp
SV40 origin of replication
SV40 ori
2421 .. 2556  =  136 bp
SV40 origin of replication
ORF:  679 .. 1374  =  696 bp
ORF:  231 amino acids  =  26.1 kDa
ORF:  2605 .. 3399  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  2777 .. 3163  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  3420 .. 3869  =  450 bp
ORF:  149 amino acids  =  16.3 kDa
ORF:  447 .. 722  =  276 bp
ORF:  91 amino acids  =  9.8 kDa
ORF:  780 .. 1025  =  246 bp
ORF:  81 amino acids  =  9.1 kDa
ORF:  2914 .. 3450  =  537 bp
ORF:  178 amino acids  =  19.8 kDa
ORF:  3625 .. 3858  =  234 bp
ORF:  77 amino acids  =  8.6 kDa
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