pECFP-N1

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

Sequence Author: Clontech (TaKaRa)

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AflIII - PciI (4675) ApaLI (4361) EcoO109I (3855) BsaI (3746) RsrII (3273) BsrDI (2990) PflFI - Tth111I (2875) FspI (2859) MscI (2839) PluTI (2760) SfoI (2758) NarI (2757) KasI (2756) BspDI * - ClaI * (2597) StuI (2578) SfiI (2532) 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) BsrGI (1388) NotI (1401) XbaI * (1411) MfeI (1507) HpaI (1520) AflII (1639) DraIII (1873) CsiI - SexAI * (2346) pECFP-N1 4733 bp
AflIII  (4675)
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  (4675)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
ApaLI  (4361)
1 site
G T G C A C C A C G T G
EcoO109I  (3855)
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  (3746)
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.
RsrII  (3273)
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  (2990)
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  (2875)
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  (2875)
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  (2859)
1 site
T G C G C A A C G C G T
MscI  (2839)
1 site
T G G C C A A C C G G T
PluTI  (2760)
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  (2758)
1 site
G G C G C C C C G C G G
NarI  (2757)
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  (2756)
1 site
G G C G C C C C G C G G
BspDI  (2597)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (2597)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
StuI  (2578)
1 site
A G G C C T T C C G G A
SfiI  (2532)
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
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
BsrGI  (1388)
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.
NotI  (1401)
1 site
G C G G C C G C C G C C G G C G
XbaI  (1411)
1 site
T C T A G A A G A T C T
* Blocked by Dam methylation.
MfeI  (1507)
1 site
C A A T T G G T T A A C
HpaI  (1520)
1 site
G T T A A C C A A T T G
AflII  (1639)
1 site
C T T A A G G A A T T C
DraIII  (1873)
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  (2346)
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  (2346)
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
2629 .. 3423  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin)
NeoR/KanR
2629 .. 3423  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin)
ECFP
679 .. 1398  =  720 bp
239 amino acids  =  26.9 kDa
3 segments
   Segment 1:  
   679 .. 681  =  3 bp
   1 amino acid  =  149.2 Da
Product: enhanced CFP
mammalian codon-optimized
ECFP
679 .. 1398  =  720 bp
239 amino acids  =  26.9 kDa
3 segments
   Segment 2:  1a  
   682 .. 684  =  3 bp
   1 amino acid  =  117.1 Da
Product: enhanced CFP
mammalian codon-optimized
ECFP
679 .. 1398  =  720 bp
239 amino acids  =  26.9 kDa
3 segments
   Segment 3:  
   685 .. 1398  =  714 bp
   237 amino acids  =  26.7 kDa
Product: enhanced CFP
mammalian codon-optimized
ECFP
679 .. 1398  =  720 bp
239 amino acids  =  26.9 kDa
3 segments
Product: enhanced CFP
mammalian codon-optimized
ori
4031 .. 4619  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
4031 .. 4619  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
1649 .. 2104  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
1649 .. 2104  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
SV40 promoter
2237 .. 2594  =  358 bp
SV40 enhancer and early promoter
SV40 promoter
2237 .. 2594  =  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
1521 .. 1642  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1521 .. 1642  =  122 bp
SV40 polyadenylation signal
AmpR promoter
2131 .. 2235  =  105 bp
AmpR promoter
2131 .. 2235  =  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
3655 .. 3702  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
HSV TK poly(A) signal
3655 .. 3702  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
SV40 ori
2445 .. 2580  =  136 bp
SV40 origin of replication
SV40 ori
2445 .. 2580  =  136 bp
SV40 origin of replication
ORF:  679 .. 1398  =  720 bp
ORF:  239 amino acids  =  26.9 kDa
ORF:  2629 .. 3423  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  2801 .. 3187  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  3444 .. 3893  =  450 bp
ORF:  149 amino acids  =  16.3 kDa
ORF:  447 .. 722  =  276 bp
ORF:  91 amino acids  =  9.6 kDa
ORF:  2938 .. 3474  =  537 bp
ORF:  178 amino acids  =  19.8 kDa
ORF:  3649 .. 3882  =  234 bp
ORF:  77 amino acids  =  8.6 kDa
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