pIRES2-ZsGreen1

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

Sequence Author: Clontech (TaKaRa)

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ApaLI (4911) BsaI (4296) PfoI (4082) RsrII (3823) FspI (3409) BspDI * - ClaI * (3147) StuI (3128) SfiI (3082) CsiI - SexAI * (2896) AseI (7) NdeI (234) SnaBI (340) NheI (591) BmtI (595) AfeI (596) BglII (609) PaeR7I - XhoI (613) Eco53kI (618) SacI (620) EcoRI (629) PstI (638) SacII (652) TspMI - XmaI (656) SmaI (658) BamHI (660) AclI (690) XmnI (879) PmlI (983) PaqCI (1006) PflMI (1120) BstXI (1253) BclI * (1336) SgrAI (1341) AhdI (1433) BsrGI (1804) Bpu10I (1946) NotI (1951) XbaI * (1961) MfeI (2057) HpaI (2070) BtsI - BtsαI (2146) AflII (2189) pIRES2-ZsGreen1 5283 bp
ApaLI  (4911)
1 site
G T G C A C C A C G T G
BsaI  (4296)
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  (4082)
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  (3823)
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  (3409)
1 site
T G C G C A A C G C G T
BspDI  (3147)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (3147)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
StuI  (3128)
1 site
A G G C C T T C C G G A
SfiI  (3082)
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  (2896)
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  (2896)
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.
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
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
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.
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.
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.
AclI  (690)
1 site
A A C G T T T T G C A A
XmnI  (879)
1 site
G A A N N N N T T C C T T N N N N A A G
PmlI  (983)
1 site
C A C G T G G T G C A C
PaqCI  (1006)
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  (1120)
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.
BstXI  (1253)
1 site
C C A N N N N N N T G G G G T N N N N N N A C C

Sticky ends from different BstXI sites may not be compatible.
BclI  (1336)
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  (1341)
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  (1433)
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  (1804)
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.
Bpu10I  (1946)
1 site
C C T N A G C G G A N T C G

Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present.
This recognition sequence is asymmetric, so ligating sticky ends generated by Bpu10I will not always regenerate a Bpu10I site.
Sticky ends from different Bpu10I sites may not be compatible.
NotI  (1951)
1 site
G C G G C C G C C G C C G G C G
XbaI  (1961)
1 site
T C T A G A A G A T C T
* Blocked by Dam methylation.
MfeI  (2057)
1 site
C A A T T G G T T A A C
HpaI  (2070)
1 site
G T T A A C C A A T T G
BtsI  (2146)
1 site
G C A G T G N N C G T C A C
BtsαI  (2146)
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.
AflII  (2189)
1 site
C T T A A G G A A T T C
NeoR/KanR
3179 .. 3973  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin)
NeoR/KanR
3179 .. 3973  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin)
ATG
1242 .. 1244  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon for translation from IRES2
ATG
1242 .. 1244  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon for translation from IRES2
ZsGreen1
1254 .. 1949  =  696 bp
231 amino acids  =  26.1 kDa
Product: Zoanthus green fluorescent protein
mammalian codon-optimized
ZsGreen1
1254 .. 1949  =  696 bp
231 amino acids  =  26.1 kDa
Product: Zoanthus green fluorescent protein
mammalian codon-optimized
ori
4581 .. 5169  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
4581 .. 5169  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
2199 .. 2654  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
2199 .. 2654  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
SV40 promoter
2787 .. 3144  =  358 bp
SV40 enhancer and early promoter
SV40 promoter
2787 .. 3144  =  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
2071 .. 2192  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2071 .. 2192  =  122 bp
SV40 polyadenylation signal
AmpR promoter
2681 .. 2785  =  105 bp
AmpR promoter
2681 .. 2785  =  105 bp
MCS
591 .. 665  =  75 bp
multiple cloning site
MCS
591 .. 665  =  75 bp
multiple cloning site
HSV TK poly(A) signal
4205 .. 4252  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
HSV TK poly(A) signal
4205 .. 4252  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
IRES2
667 .. 1253  =  587 bp
3 segments
   Segment 1:  
   667 .. 1241  =  575 bp
internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)
IRES2
667 .. 1253  =  587 bp
3 segments
   Segment 2:  ATG  
   1242 .. 1244  =  3 bp
internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)
IRES2
667 .. 1253  =  587 bp
3 segments
   Segment 3:  
   1245 .. 1253  =  9 bp
internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)
IRES2
667 .. 1253  =  587 bp
3 segments
internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)
SV40 ori
2995 .. 3130  =  136 bp
SV40 origin of replication
SV40 ori
2995 .. 3130  =  136 bp
SV40 origin of replication
ORF:  3994 .. 4443  =  450 bp
ORF:  149 amino acids  =  16.3 kDa
ORF:  3179 .. 3973  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  1242 .. 1949  =  708 bp
ORF:  235 amino acids  =  26.5 kDa
ORF:  3351 .. 3737  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  1631 .. 1924  =  294 bp
ORF:  97 amino acids  =  10.9 kDa
ORF:  3488 .. 4024  =  537 bp
ORF:  178 amino acids  =  19.8 kDa
ORF:  4199 .. 4432  =  234 bp
ORF:  77 amino acids  =  8.6 kDa
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Download pIRES2-ZsGreen1.dna file

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