pLVX-IRES-tdTomato

Lentiviral bicistronic expression vector for constitutively co-expressing a protein of interest together with tdTomato.

Sequence Author: Clontech (TaKaRa)

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SgrDI (8572) SspI (8455) PvuI (8021) DrdI (6866) BmtI (6375) NheI (6371) Bsu36I (5581) PfoI (5134) MluI (4848) BsrGI (4837) KasI (637) NarI (638) SfoI (639) PluTI (641) BssHII (711) NruI * (833) FseI (1150) EcoNI (1170) MfeI (1189) BbvCI - Bpu10I (1424) BstAPI (1453) KflI - PpuMI (1934) cPPT/CTS BspDI - ClaI (2180) NdeI (2374) SnaBI (2480) BsmBI - Esp3I (2729) EcoRI (2803) PaeR7I - XhoI (2809) SpeI (2815) XbaI (2821) NotI (2828) BamHI (2835) PspOMI (2956) ApaI (2960) AvrII (2994) BfuAI - BspMI - PaqCI (3182) BmgBI (3385) pLVX-IRES-tdTomato 8892 bp
SgrDI  (8572)
1 site
C G T C G A C G G C A G C T G C
SspI  (8455)
1 site
A A T A T T T T A T A A
PvuI  (8021)
1 site
C G A T C G G C T A G C
DrdI  (6866)
1 site
G A C N N N N N N G T C C T G N N N N N N C A G

Sticky ends from different DrdI sites may not be compatible.
BmtI  (6375)
1 site
G C T A G C C G A T C G
NheI  (6371)
1 site
G C T A G C C G A T C G
Bsu36I  (5581)
1 site
C C T N A G G G G A N T C C

Sticky ends from different Bsu36I sites may not be compatible.
PfoI  (5134)
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.
MluI  (4848)
1 site
A C G C G T T G C G C A
BsrGI  (4837)
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.
KasI  (637)
1 site
G G C G C C C C G C G G
NarI  (638)
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.
SfoI  (639)
1 site
G G C G C C C C G C G G
PluTI  (641)
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.
BssHII  (711)
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.
NruI  (833)
1 site
T C G C G A A G C G C T
* Blocked by Dam methylation.
FseI  (1150)
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.
EcoNI  (1170)
1 site
C C T N N N N N A G G G G A N N N N N T C C

The 1-base overhangs produced by EcoNI may be hard to ligate.
Sticky ends from different EcoNI sites may not be compatible.
MfeI  (1189)
1 site
C A A T T G G T T A A C
BbvCI  (1424)
1 site
C C T C A G C G G A G T C G
Bpu10I  (1424)
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.
BstAPI  (1453)
1 site
G C A N N N N N T G C C G T N N N N N A C G

Sticky ends from different BstAPI sites may not be compatible.
KflI  (1934)
1 site
G G G W C C C C C C W G G G

Sticky ends from different KflI sites may not be compatible.
PpuMI  (1934)
1 site
R G G W C C Y Y C C W G G R

Sticky ends from different PpuMI sites may not be compatible.
BspDI  (2180)
1 site
A T C G A T T A G C T A
ClaI  (2180)
1 site
A T C G A T T A G C T A
NdeI  (2374)
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  (2480)
1 site
T A C G T A A T G C A T
BsmBI  (2729)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different BsmBI sites may not be compatible.
BsmBI-v2 is an improved version of BsmBI.
Esp3I  (2729)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different Esp3I sites may not be compatible.
EcoRI  (2803)
1 site
G A A T T C C T T A A G
PaeR7I  (2809)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (2809)
1 site
C T C G A G G A G C T C
SpeI  (2815)
1 site
A C T A G T T G A T C A
XbaI  (2821)
1 site
T C T A G A A G A T C T
NotI  (2828)
1 site
G C G G C C G C C G C C G G C G
BamHI  (2835)
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.
PspOMI  (2956)
1 site
G G G C C C C C C G G G
ApaI  (2960)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
AvrII  (2994)
1 site
C C T A G G G G A T C C
BfuAI  (3182)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BfuAI recognition sequence.
Sticky ends from different BfuAI sites may not be compatible.
BfuAI is typically used at 50°C, but is 50% active at 37°C.
BspMI  (3182)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BspMI recognition sequence.
Sticky ends from different BspMI sites may not be compatible.
PaqCI  (3182)
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.
BmgBI  (3385)
1 site
C A C G T C G T G C A G

This recognition sequence is asymmetric, so ligating blunt ends generated by BmgBI will not always regenerate a BmgBI site.
tdTomato
3417 .. 4847  =  1431 bp
476 amino acids  =  54.2 kDa
Product: tandem dimeric (pseudo-monomeric) derivative of DsRed (Shaner et al., 2004)
mammalian codon-optimized
tdTomato
3417 .. 4847  =  1431 bp
476 amino acids  =  54.2 kDa
Product: tandem dimeric (pseudo-monomeric) derivative of DsRed (Shaner et al., 2004)
mammalian codon-optimized
AmpR
7578 .. 8438  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   7578 .. 8369  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
7578 .. 8438  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   8370 .. 8438  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
7578 .. 8438  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
3' LTR
1 .. 634  =  634 bp
3' long terminal repeat (LTR) from HIV-1
3' LTR
1 .. 634  =  634 bp
3' long terminal repeat (LTR) from HIV-1
3' LTR
5656 .. 6289  =  634 bp
3' long terminal repeat (LTR) from HIV-1
3' LTR
5656 .. 6289  =  634 bp
3' long terminal repeat (LTR) from HIV-1
WPRE
4861 .. 5449  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
WPRE
4861 .. 5449  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
ori
6819 .. 7407  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
6819 .. 7407  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
IRES
2843 .. 3415  =  573 bp
internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)
IRES
2843 .. 3415  =  573 bp
internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)
CMV enhancer
2201 .. 2504  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
2201 .. 2504  =  304 bp
human cytomegalovirus immediate early enhancer
RRE
1303 .. 1536  =  234 bp
The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm.
RRE
1303 .. 1536  =  234 bp
The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm.
CMV promoter
2505 .. 2708  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
2505 .. 2708  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
SV40 poly(A) signal
8591 .. 8725  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
8591 .. 8725  =  135 bp
SV40 polyadenylation signal
HIV-1 Ψ
681 .. 806  =  126 bp
packaging signal of human immunodeficiency virus type 1
HIV-1 Ψ
681 .. 806  =  126 bp
packaging signal of human immunodeficiency virus type 1
cPPT/CTS
2028 .. 2143  =  116 bp
central polypurine tract and central termination sequence of HIV-1 (lacking the first T)
cPPT/CTS
2028 .. 2143  =  116 bp
central polypurine tract and central termination sequence of HIV-1 (lacking the first T)
AmpR promoter
8439 .. 8543  =  105 bp
AmpR promoter
8439 .. 8543  =  105 bp
MCS
2803 .. 2840  =  38 bp
multiple cloning site
MCS
2803 .. 2840  =  38 bp
multiple cloning site
ORF:  4951 .. 5478  =  528 bp
ORF:  175 amino acids  =  19.0 kDa
ORF:  7708 .. 7974  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  1181 .. 2032  =  852 bp
ORF:  283 amino acids  =  32.7 kDa
ORF:  4964 .. 5260  =  297 bp
ORF:  98 amino acids  =  10.2 kDa
ORF:  3417 .. 4847  =  1431 bp
ORF:  476 amino acids  =  54.2 kDa
ORF:  1015 .. 1476  =  462 bp
ORF:  153 amino acids  =  16.6 kDa
ORF:  3351 .. 4100  =  750 bp
ORF:  249 amino acids  =  25.2 kDa
ORF:  7578 .. 8438  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
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Download pLVX-IRES-tdTomato.dna file

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