pLX304

Lentiviral Gateway® destination vector with a blasticidin resistance marker and a V5 epitope tag, for strong expression and improved plasmid yield and viral titer.

Sequence Author: Root Lab

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cPPT/CTS XbaI * (9171) KpnI (8748) Acc65I (8744) AscI (8576) BlpI (8567) SpeI (8487) XcmI (8259) KflI (8235) AleI (7878) BstAPI (7754) MfeI (7490) EcoNI (7471) SphI (6622) AflIII - PciI (6119) PaeR7I - PspXI - XhoI (9368) NdeI (238) BstZ17I (1576) SrfI (1912) BstXI (2029) BfuAI - BspMI (2156) V5 tag NheI (2352) BmtI (2356) AfeI (2357) AgeI (2362) SacII (2887) SfiI (3603) StuI (3649) AvrII (3650) DraIII (4091) pLX304 9377 bp
XbaI  (9171)
1 site
T C T A G A A G A T C T
* Blocked by Dam methylation.
KpnI  (8748)
1 site
G G T A C C C C A T G G
Acc65I  (8744)
1 site
G G T A C C C C A T G G
AscI  (8576)
1 site
G G C G C G C C C C G C G C G G
BlpI  (8567)
1 site
G C T N A G C C G A N T C G

Sticky ends from different BlpI sites may not be compatible.
SpeI  (8487)
1 site
A C T A G T T G A T C A
XcmI  (8259)
1 site
C C A N N N N N N N N N T G G G G T N N N N N N N N N A C C

The 1-base overhangs produced by XcmI may be hard to ligate.
Sticky ends from different XcmI sites may not be compatible.
KflI  (8235)
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.
AleI  (7878)
1 site
C A C N N N N G T G G T G N N N N C A C
BstAPI  (7754)
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.
MfeI  (7490)
1 site
C A A T T G G T T A A C
EcoNI  (7471)
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.
SphI  (6622)
1 site
G C A T G C C G T A C G
AflIII  (6119)
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  (6119)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
PaeR7I  (9368)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (9368)
1 site
V C T C G A G B B G A G C T C V
XhoI  (9368)
1 site
C T C G A G G A G C T C
NdeI  (238)
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.
BstZ17I  (1576)
1 site
G T A T A C C A T A T G
SrfI  (1912)
1 site
G C C C G G G C C G G G C C C G
BstXI  (2029)
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.
BfuAI  (2156)
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  (2156)
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.
NheI  (2352)
1 site
G C T A G C C G A T C G
BmtI  (2356)
1 site
G C T A G C C G A T C G
AfeI  (2357)
1 site
A G C G C T T C G C G A
AgeI  (2362)
1 site
A C C G G T T G G C C A
SacII  (2887)
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.
SfiI  (3603)
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.
StuI  (3649)
1 site
A G G C C T T C C G G A
AvrII  (3650)
1 site
C C T A G G G G A T C C
DraIII  (4091)
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.
AmpR
4444 .. 5304  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   4444 .. 4512  =  69 bp
   23 amino acids  =  2.6 kDa
AmpR
4444 .. 5304  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   4513 .. 5304  =  792 bp
   263 amino acids  =  28.9 kDa
AmpR
4444 .. 5304  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
CmR
828 .. 1487  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
828 .. 1487  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
WPRE
2384 .. 2972  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
WPRE
2384 .. 2972  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
ori
5475 .. 6063  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
5475 .. 6063  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
hPGK promoter
8241 .. 8741  =  501 bp
human phosphoglycerate kinase 1 (PGK1) promoter
hPGK promoter
8241 .. 8741  =  501 bp
human phosphoglycerate kinase 1 (PGK1) promoter
f1 ori
3858 .. 4313  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
3858 .. 4313  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
BSD
8753 .. 9148  =  396 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
BSD
8753 .. 9148  =  396 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
ccdB
1829 .. 2134  =  306 bp
101 amino acids  =  11.7 kDa
Product: CcdB, a bacterial toxin that poisons DNA gyrase
Plasmids containing the ccdB gene cannot be propagated in standard E. coli strains.
ccdB
1829 .. 2134  =  306 bp
101 amino acids  =  11.7 kDa
Product: CcdB, a bacterial toxin that poisons DNA gyrase
Plasmids containing the ccdB gene cannot be propagated in standard E. coli strains.
CMV enhancer
65 .. 368  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
65 .. 368  =  304 bp
human cytomegalovirus immediate early enhancer
3' LTR (ΔU3)
3044 .. 3277  =  234 bp
self-inactivating 3' long terminal repeat (LTR) from HIV-1
3' LTR (ΔU3)
3044 .. 3277  =  234 bp
self-inactivating 3' long terminal repeat (LTR) from HIV-1
RRE
7604 .. 7837  =  234 bp
The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm.
RRE
7604 .. 7837  =  234 bp
The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm.
RSV promoter
6530 .. 6758  =  229 bp
Rous sarcoma virus enhancer/promoter
RSV promoter
6530 .. 6758  =  229 bp
Rous sarcoma virus enhancer/promoter
CMV promoter
369 .. 572  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
369 .. 572  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
5' LTR (truncated)
6759 .. 6939  =  181 bp
truncated 5' long terminal repeat (LTR) from HIV-1
5' LTR (truncated)
6759 .. 6939  =  181 bp
truncated 5' long terminal repeat (LTR) from HIV-1
SV40 ori
3516 .. 3651  =  136 bp
SV40 origin of replication
SV40 ori
3516 .. 3651  =  136 bp
SV40 origin of replication
HIV-1 Ψ
6986 .. 7111  =  126 bp
packaging signal of human immunodeficiency virus type 1
HIV-1 Ψ
6986 .. 7111  =  126 bp
packaging signal of human immunodeficiency virus type 1
attR1
595 .. 719  =  125 bp
recombination site for the Gateway® LR reaction
attR1
595 .. 719  =  125 bp
recombination site for the Gateway® LR reaction
attR2
2175 .. 2299  =  125 bp
recombination site for the Gateway® LR reaction
attR2
2175 .. 2299  =  125 bp
recombination site for the Gateway® LR reaction
SV40 poly(A) signal
3355 .. 3476  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
3355 .. 3476  =  122 bp
SV40 polyadenylation signal
cPPT/CTS
9208 .. 9325  =  118 bp
central polypurine tract and central termination sequence of HIV-1
cPPT/CTS
9208 .. 9325  =  118 bp
central polypurine tract and central termination sequence of HIV-1
AmpR promoter
4339 .. 4443  =  105 bp
AmpR promoter
4339 .. 4443  =  105 bp
V5 tag
2301 .. 2342  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
V5 tag
2301 .. 2342  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
lac UV5 promoter
744 .. 774  =  31 bp
3 segments
   Segment 1:  -35  
   744 .. 749  =  6 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
744 .. 774  =  31 bp
3 segments
   Segment 2:  
   750 .. 767  =  18 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
744 .. 774  =  31 bp
3 segments
   Segment 3:  -10  
   768 .. 774  =  7 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
744 .. 774  =  31 bp
3 segments
E. coli lac promoter with an "up" mutation
stop codons
2343 .. 2351  =  9 bp
stop codons
2343 .. 2351  =  9 bp
ORF:  4444 .. 5304  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  1829 .. 2134  =  306 bp
ORF:  101 amino acids  =  11.7 kDa
ORF:  2474 .. 2983  =  510 bp
ORF:  169 amino acids  =  18.5 kDa
ORF:  3377 .. 3613  =  237 bp
ORF:  78 amino acids  =  8.6 kDa
ORF:  6461 .. 6706  =  246 bp
ORF:  81 amino acids  =  8.9 kDa
ORF:  8753 .. 9163  =  411 bp
ORF:  136 amino acids  =  14.2 kDa
ORF:  828 .. 1487  =  660 bp
ORF:  219 amino acids  =  25.7 kDa
ORF:  2487 .. 3017  =  531 bp
ORF:  176 amino acids  =  18.7 kDa
ORF:  7482 .. 8465  =  984 bp
ORF:  327 amino acids  =  36.7 kDa
ORF:  8523 .. 8972  =  450 bp
ORF:  149 amino acids  =  15.8 kDa
ORF:  4908 .. 5174  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  7397 .. 7777  =  381 bp
ORF:  126 amino acids  =  13.3 kDa
ORF:  8210 .. 9055  =  846 bp
ORF:  281 amino acids  =  30.3 kDa
ORF:  6388 .. 6687  =  300 bp
ORF:  99 amino acids  =  10.9 kDa
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Download pLX304.dna file

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Individual Sequences & Maps

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