pEF1alpha-IRES-ZsGreen1

IRES-containing bicistronic vector for expressing a gene together with the ZsGreen1 fluorescent protein.

Sequence Author: Clontech (TaKaRa)

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AseI (7) ApaLI (5668) BsaI (5053) PfoI (4839) RsrII (4580) FspI (4166) BspDI * - ClaI * (3904) SfiI (3839) CsiI - SexAI * (3653) BtsI - BtsαI (2903) AleI (141) AgeI (242) FseI (872) BbvCI (1070) NheI (1348) BmtI (1352) AfeI (1353) EcoRI (1386) TspMI - XmaI (1413) SmaI (1415) BamHI (1417) PmlI (1740) PaqCI (1763) PflMI (1877) BclI * (2093) SgrAI (2098) AhdI (2190) BsrGI (2561) NotI (2708) XbaI * (2718) HpaI (2827) pEF1 α- IRES-ZsGreen1 6040 bp
AseI  (7)
1 site
A T T A A T T A A T T A
ApaLI  (5668)
1 site
G T G C A C C A C G T G
BsaI  (5053)
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  (4839)
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  (4580)
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.
FspI  (4166)
1 site
T G C G C A A C G C G T
BspDI  (3904)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (3904)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
SfiI  (3839)
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  (3653)
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  (3653)
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.
BtsI  (2903)
1 site
G C A G T G N N C G T C A C
BtsαI  (2903)
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.
AleI  (141)
1 site
C A C N N N N G T G G T G N N N N C A C
AgeI  (242)
1 site
A C C G G T T G G C C A
FseI  (872)
1 site
G G C C G G C C C C G G C C G G

FseI gradually loses activity when stored at -20°C.
BbvCI  (1070)
1 site
C C T C A G C G G A G T C G
NheI  (1348)
1 site
G C T A G C C G A T C G
BmtI  (1352)
1 site
G C T A G C C G A T C G
AfeI  (1353)
1 site
A G C G C T T C G C G A
EcoRI  (1386)
1 site
G A A T T C C T T A A G
TspMI  (1413)
1 site
C C C G G G G G G C C C
XmaI  (1413)
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.
SmaI  (1415)
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  (1417)
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.
PmlI  (1740)
1 site
C A C G T G G T G C A C
PaqCI  (1763)
1 site
C A C C T G C ( N ) 4 G T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the PaqCI recognition sequence.
Sticky ends from different PaqCI sites may not be compatible.
Cleavage can be improved with PaqCI Activator.
PflMI  (1877)
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.
BclI  (2093)
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.
SgrAI  (2098)
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.
AhdI  (2190)
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.
BsrGI  (2561)
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  (2708)
1 site
G C G G C C G C C G C C G G C G
XbaI  (2718)
1 site
T C T A G A A G A T C T
* Blocked by Dam methylation.
HpaI  (2827)
1 site
G T T A A C C A A T T G
EF-1α promoter
165 .. 1346  =  1182 bp
strong constitutive promoter for human elongation factor EF-1α
EF-1α promoter
165 .. 1346  =  1182 bp
strong constitutive promoter for human elongation factor EF-1α
NeoR/KanR
3936 .. 4730  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin)
NeoR/KanR
3936 .. 4730  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin)
ATG
1999 .. 2001  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon for translation from IRES2
ATG
1999 .. 2001  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon for translation from IRES2
ZsGreen1
2011 .. 2706  =  696 bp
231 amino acids  =  26.1 kDa
Product: Zoanthus green fluorescent protein
mammalian codon-optimized
ZsGreen1
2011 .. 2706  =  696 bp
231 amino acids  =  26.1 kDa
Product: Zoanthus green fluorescent protein
mammalian codon-optimized
ori
5338 .. 5926  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
5338 .. 5926  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
2956 .. 3411  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
2956 .. 3411  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
SV40 promoter
3544 .. 3901  =  358 bp
SV40 enhancer and early promoter
SV40 promoter
3544 .. 3901  =  358 bp
SV40 enhancer and early promoter
SV40 poly(A) signal
2828 .. 2949  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2828 .. 2949  =  122 bp
SV40 polyadenylation signal
AmpR promoter
3438 .. 3542  =  105 bp
AmpR promoter
3438 .. 3542  =  105 bp
MCS
1366 .. 1422  =  57 bp
multiple cloning site of fluorescent protein plasmids
MCS
1366 .. 1422  =  57 bp
multiple cloning site of fluorescent protein plasmids
HSV TK poly(A) signal
4962 .. 5009  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
HSV TK poly(A) signal
4962 .. 5009  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
EF-1α intron A
395 .. 1337  =  943 bp
intron upstream of the start codon
EF-1α intron A
395 .. 1337  =  943 bp
intron upstream of the start codon
IRES2
1424 .. 2010  =  587 bp
3 segments
   Segment 1:  
   1424 .. 1998  =  575 bp
internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)
IRES2
1424 .. 2010  =  587 bp
3 segments
   Segment 2:  ATG  
   1999 .. 2001  =  3 bp
internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)
IRES2
1424 .. 2010  =  587 bp
3 segments
   Segment 3:  
   2002 .. 2010  =  9 bp
internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)
IRES2
1424 .. 2010  =  587 bp
3 segments
internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)
SV40 ori
3752 .. 3887  =  136 bp
SV40 origin of replication
SV40 ori
3752 .. 3887  =  136 bp
SV40 origin of replication
ORF:  724 .. 1161  =  438 bp
ORF:  145 amino acids  =  15.9 kDa
ORF:  1999 .. 2706  =  708 bp
ORF:  235 amino acids  =  26.5 kDa
ORF:  4108 .. 4494  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  803 .. 1036  =  234 bp
ORF:  77 amino acids  =  7.7 kDa
ORF:  4751 .. 5200  =  450 bp
ORF:  149 amino acids  =  16.3 kDa
ORF:  3936 .. 4730  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  439 .. 1086  =  648 bp
ORF:  215 amino acids  =  23.7 kDa
ORF:  2388 .. 2681  =  294 bp
ORF:  97 amino acids  =  10.9 kDa
ORF:  4245 .. 4781  =  537 bp
ORF:  178 amino acids  =  19.8 kDa
ORF:  4956 .. 5189  =  234 bp
ORF:  77 amino acids  =  8.6 kDa
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Download pEF1alpha-IRES-ZsGreen1.dna file

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