pEGFP-C2

Vector for fusing EGFP to the N-terminus of a partner protein. For other reading frames, use pEGFP-C1 or pEGFP-C3.

Sequence Author: Clontech (TaKaRa)

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PciI (4677) ApaLI (4363) EcoO109I (3857) BsaI (3748) RsrII (3275) BsrDI (2992) PflFI - Tth111I (2877) FspI (2861) MscI (2841) PluTI (2762) SfoI (2760) NarI (2759) KasI (2758) BspDI * - ClaI * (2599) StuI (2580) SfiI (2534) AseI (7) NdeI (234) SnaBI (340) NheI (591) BmtI (595) AfeI (596) AgeI (600) BsrGI (1322) BglII (1343) PaeR7I - XhoI (1347) Eco53kI (1352) SacI (1354) HindIII (1356) EcoRI (1363) PstI (1372) SalI (1373) AccI (1374) Acc65I (1379) KpnI (1383) SacII (1386) PspOMI (1387) TspMI - XmaI (1390) ApaI (1391) SmaI (1392) BamHI (1394) XbaI * (1406) stop codons BclI * (1416) MfeI (1509) HpaI (1522) MluI (1645) DraIII (1875) CsiI - SexAI * (2348) pEGFP-C2 4735 bp
PciI  (4677)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
ApaLI  (4363)
1 site
G T G C A C C A C G T G
EcoO109I  (3857)
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  (3748)
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  (3275)
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  (2992)
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  (2877)
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  (2877)
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  (2861)
1 site
T G C G C A A C G C G T
MscI  (2841)
1 site
T G G C C A A C C G G T
PluTI  (2762)
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  (2760)
1 site
G G C G C C C C G C G G
NarI  (2759)
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  (2758)
1 site
G G C G C C C C G C G G
BspDI  (2599)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (2599)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
StuI  (2580)
1 site
A G G C C T T C C G G A
SfiI  (2534)
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
AgeI  (600)
1 site
A C C G G T T G G C C A
BsrGI  (1322)
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.
BglII  (1343)
1 site
A G A T C T T C T A G A
PaeR7I  (1347)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (1347)
1 site
C T C G A G G A G C T C
Eco53kI  (1352)
1 site
G A G C T C C T C G A G
SacI  (1354)
1 site
G A G C T C C T C G A G
HindIII  (1356)
1 site
A A G C T T T T C G A A
EcoRI  (1363)
1 site
G A A T T C C T T A A G
PstI  (1372)
1 site
C T G C A G G A C G T C
SalI  (1373)
1 site
G T C G A C C A G C T G
AccI  (1374)
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  (1379)
1 site
G G T A C C C C A T G G
KpnI  (1383)
1 site
G G T A C C C C A T G G
SacII  (1386)
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  (1387)
1 site
G G G C C C C C C G G G
TspMI  (1390)
1 site
C C C G G G G G G C C C
XmaI  (1390)
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  (1391)
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  (1392)
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  (1394)
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.
XbaI  (1406)
1 site
T C T A G A A G A T C T
* Blocked by Dam methylation.
BclI  (1416)
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.
MfeI  (1509)
1 site
C A A T T G G T T A A C
HpaI  (1522)
1 site
G T T A A C C A A T T G
MluI  (1645)
1 site
A C G C G T T G C G C A
DraIII  (1875)
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  (2348)
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  (2348)
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
2631 .. 3425  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin)
NeoR/KanR
2631 .. 3425  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin)
EGFP
613 .. 1329  =  717 bp
239 amino acids  =  26.9 kDa
3 segments
   Segment 1:  
   613 .. 615  =  3 bp
   1 amino acid  =  149.2 Da
Product: enhanced GFP
mammalian codon-optimized
EGFP
613 .. 1329  =  717 bp
239 amino acids  =  26.9 kDa
3 segments
   Segment 2:  1a  
   616 .. 618  =  3 bp
   1 amino acid  =  117.1 Da
Product: enhanced GFP
mammalian codon-optimized
EGFP
613 .. 1329  =  717 bp
239 amino acids  =  26.9 kDa
3 segments
   Segment 3:  
   619 .. 1329  =  711 bp
   237 amino acids  =  26.7 kDa
Product: enhanced GFP
mammalian codon-optimized
EGFP
613 .. 1329  =  717 bp
239 amino acids  =  26.9 kDa
3 segments
Product: enhanced GFP
mammalian codon-optimized
ori
4033 .. 4621  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
4033 .. 4621  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
1651 .. 2106  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
1651 .. 2106  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
SV40 promoter
2239 .. 2596  =  358 bp
SV40 enhancer and early promoter
SV40 promoter
2239 .. 2596  =  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
1523 .. 1644  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1523 .. 1644  =  122 bp
SV40 polyadenylation signal
AmpR promoter
2133 .. 2237  =  105 bp
AmpR promoter
2133 .. 2237  =  105 bp
MCS
1343 .. 1399  =  57 bp
multiple cloning site of fluorescent protein plasmids
MCS
1343 .. 1399  =  57 bp
multiple cloning site of fluorescent protein plasmids
HSV TK poly(A) signal
3657 .. 3704  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
HSV TK poly(A) signal
3657 .. 3704  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
stop codons
1408 .. 1418  =  11 bp
stop codons in all three reading frames
stop codons
1408 .. 1418  =  11 bp
stop codons in all three reading frames
SV40 ori
2447 .. 2582  =  136 bp
SV40 origin of replication
SV40 ori
2447 .. 2582  =  136 bp
SV40 origin of replication
ORF:  613 .. 1410  =  798 bp
ORF:  265 amino acids  =  29.9 kDa
ORF:  2803 .. 3189  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  3446 .. 3895  =  450 bp
ORF:  149 amino acids  =  16.3 kDa
ORF:  2631 .. 3425  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  2940 .. 3476  =  537 bp
ORF:  178 amino acids  =  19.8 kDa
ORF:  3651 .. 3884  =  234 bp
ORF:  77 amino acids  =  8.6 kDa
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Download pEGFP-C2.dna file

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