pTurboYFP-N

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

Sequence Author: Evrogen

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AflIII - PciI (4687) EcoO109I (3867) PfoI (3544) RsrII (3285) PflFI - Tth111I (2887) FspI (2871) BspDI * - ClaI * (2609) StuI (2590) BseRI (2587) SfiI (2544) 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) PspOMI (653) TspMI - XmaI (656) ApaI (657) SmaI (658) BamHI (660) AgeI (666) SgrAI (826) AleI (856) XmnI (916) PflMI (1080) BlpI (1297) BssHII (1350) BspEI (1393) NotI (1413) XbaI * (1423) MfeI (1519) HpaI (1532) AflII (1651) DraIII (1885) CsiI - SexAI * (2358) pTurboYFP-N 4745 bp
AflIII  (4687)
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  (4687)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
EcoO109I  (3867)
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  (3544)
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  (3285)
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.
PflFI  (2887)
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  (2887)
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  (2871)
1 site
T G C G C A A C G C G T
BspDI  (2609)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (2609)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
StuI  (2590)
1 site
A G G C C T T C C G G A
BseRI  (2587)
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  (2544)
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
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
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
SgrAI  (826)
1 site
C R C C G G Y G G Y G G C C R C

Efficient cleavage requires at least two copies of the SgrAI recognition sequence.
AleI  (856)
1 site
C A C N N N N G T G G T G N N N N C A C
XmnI  (916)
1 site
G A A N N N N T T C C T T N N N N A A G
PflMI  (1080)
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.
BlpI  (1297)
1 site
G C T N A G C C G A N T C G

Sticky ends from different BlpI sites may not be compatible.
BssHII  (1350)
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.
BspEI  (1393)
1 site
T C C G G A A G G C C T
NotI  (1413)
1 site
G C G G C C G C C G C C G G C G
XbaI  (1423)
1 site
T C T A G A A G A T C T
* Blocked by Dam methylation.
MfeI  (1519)
1 site
C A A T T G G T T A A C
HpaI  (1532)
1 site
G T T A A C C A A T T G
AflII  (1651)
1 site
C T T A A G G A A T T C
DraIII  (1885)
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.
CsiI  (2358)
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  (2358)
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
2641 .. 3435  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin)
NeoR/KanR
2641 .. 3435  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin)
TurboYFP
679 .. 1410  =  732 bp
243 amino acids  =  26.9 kDa
Product: enhanced derivative of PhiYFP
mammalian codon-optimized
TurboYFP
679 .. 1410  =  732 bp
243 amino acids  =  26.9 kDa
Product: enhanced derivative of PhiYFP
mammalian codon-optimized
ori
4043 .. 4631  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
4043 .. 4631  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
1661 .. 2116  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
1661 .. 2116  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
SV40 promoter
2249 .. 2606  =  358 bp
SV40 enhancer and early promoter
SV40 promoter
2249 .. 2606  =  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
1533 .. 1654  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1533 .. 1654  =  122 bp
SV40 polyadenylation signal
AmpR promoter
2143 .. 2247  =  105 bp
AmpR promoter
2143 .. 2247  =  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
3667 .. 3714  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
HSV TK poly(A) signal
3667 .. 3714  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
SV40 ori
2457 .. 2592  =  136 bp
SV40 origin of replication
SV40 ori
2457 .. 2592  =  136 bp
SV40 origin of replication
ORF:  679 .. 1410  =  732 bp
ORF:  243 amino acids  =  26.9 kDa
ORF:  2641 .. 3435  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  2813 .. 3199  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  3456 .. 3905  =  450 bp
ORF:  149 amino acids  =  16.3 kDa
ORF:  447 .. 680  =  234 bp
ORF:  77 amino acids  =  8.2 kDa
ORF:  2950 .. 3486  =  537 bp
ORF:  178 amino acids  =  19.8 kDa
ORF:  3661 .. 3894  =  234 bp
ORF:  77 amino acids  =  8.6 kDa
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