pLX301

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

Sequence Author: Root Lab

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cPPT/CTS XbaI * (9325) CsiI - SexAI * (9288) BstEII (8855) RsrII (8837) BsiWI (8777) PflFI - Tth111I (8763) KpnI (8706) Acc65I (8702) AscI (8534) BlpI (8525) SpeI (8445) XcmI (8217) KflI - PpuMI (8193) AleI (7836) MfeI (7448) EcoNI (7429) NruI * (7096) SphI (6580) AflIII - PciI (6077) PspFI (5777) BseYI (5773) PaeR7I - PspXI - XhoI (9522) NdeI (238) lac UV5 promoter BstZ17I (1576) SrfI (1912) BstXI (2029) BfuAI - BspMI (2156) PstI (2167) stop codons NheI (2310) BmtI (2314) AfeI (2315) AgeI (2320) SacII (2845) SfiI (3561) AvrII (3608) pLX301 9531 bp
XbaI  (9325)
1 site
T C T A G A A G A T C T
* Blocked by Dam methylation.
CsiI  (9288)
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  (9288)
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.
BstEII  (8855)
1 site
G G T N A C C C C A N T G G

Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
RsrII  (8837)
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.
BsiWI  (8777)
1 site
C G T A C G G C A T G C

BsiWI is typically used at 55°C, but is 50% active at 37°C.
PflFI  (8763)
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  (8763)
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.
KpnI  (8706)
1 site
G G T A C C C C A T G G
Acc65I  (8702)
1 site
G G T A C C C C A T G G
AscI  (8534)
1 site
G G C G C G C C C C G C G C G G
BlpI  (8525)
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  (8445)
1 site
A C T A G T T G A T C A
XcmI  (8217)
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  (8193)
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  (8193)
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.
AleI  (7836)
1 site
C A C N N N N G T G G T G N N N N C A C
MfeI  (7448)
1 site
C A A T T G G T T A A C
EcoNI  (7429)
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.
NruI  (7096)
1 site
T C G C G A A G C G C T
* Blocked by Dam methylation.
SphI  (6580)
1 site
G C A T G C C G T A C G
AflIII  (6077)
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  (6077)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
PspFI  (5777)
1 site
C C C A G C G G G T C G
BseYI  (5773)
1 site
C C C A G C G G G T C G

After cleavage, BseYI can remain bound to DNA and alter its electrophoretic mobility.
PaeR7I  (9522)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (9522)
1 site
V C T C G A G B B G A G C T C V
XhoI  (9522)
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.
PstI  (2167)
1 site
C T G C A G G A C G T C
NheI  (2310)
1 site
G C T A G C C G A T C G
BmtI  (2314)
1 site
G C T A G C C G A T C G
AfeI  (2315)
1 site
A G C G C T T C G C G A
AgeI  (2320)
1 site
A C C G G T T G G C C A
SacII  (2845)
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  (3561)
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.
AvrII  (3608)
1 site
C C T A G G G G A T C C
AmpR
4402 .. 5262  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   4402 .. 4470  =  69 bp
   23 amino acids  =  2.6 kDa
AmpR
4402 .. 5262  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   4471 .. 5262  =  792 bp
   263 amino acids  =  28.9 kDa
AmpR
4402 .. 5262  =  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
PuroR
8721 .. 9320  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
8721 .. 9320  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
WPRE
2342 .. 2930  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
WPRE
2342 .. 2930  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
ori
5433 .. 6021  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
5433 .. 6021  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
hPGK promoter
8199 .. 8699  =  501 bp
human phosphoglycerate kinase 1 (PGK1) promoter
hPGK promoter
8199 .. 8699  =  501 bp
human phosphoglycerate kinase 1 (PGK1) promoter
f1 ori
3816 .. 4271  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
3816 .. 4271  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
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)
3002 .. 3235  =  234 bp
self-inactivating 3' long terminal repeat (LTR) from HIV-1
3' LTR (ΔU3)
3002 .. 3235  =  234 bp
self-inactivating 3' long terminal repeat (LTR) from HIV-1
RRE
7562 .. 7795  =  234 bp
The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm.
RRE
7562 .. 7795  =  234 bp
The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm.
RSV promoter
6488 .. 6716  =  229 bp
Rous sarcoma virus enhancer/promoter
RSV promoter
6488 .. 6716  =  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)
6717 .. 6897  =  181 bp
truncated 5' long terminal repeat (LTR) from HIV-1
5' LTR (truncated)
6717 .. 6897  =  181 bp
truncated 5' long terminal repeat (LTR) from HIV-1
SV40 ori
3474 .. 3609  =  136 bp
SV40 origin of replication
SV40 ori
3474 .. 3609  =  136 bp
SV40 origin of replication
HIV-1 Ψ
6944 .. 7069  =  126 bp
packaging signal of human immunodeficiency virus type 1
HIV-1 Ψ
6944 .. 7069  =  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
3313 .. 3434  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
3313 .. 3434  =  122 bp
SV40 polyadenylation signal
cPPT/CTS
9362 .. 9479  =  118 bp
central polypurine tract and central termination sequence of HIV-1
cPPT/CTS
9362 .. 9479  =  118 bp
central polypurine tract and central termination sequence of HIV-1
AmpR promoter
4297 .. 4401  =  105 bp
AmpR promoter
4297 .. 4401  =  105 bp
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
2301 .. 2309  =  9 bp
stop codons
2301 .. 2309  =  9 bp
ORF:  4402 .. 5262  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  1829 .. 2134  =  306 bp
ORF:  101 amino acids  =  11.7 kDa
ORF:  2432 .. 2941  =  510 bp
ORF:  169 amino acids  =  18.5 kDa
ORF:  3335 .. 3571  =  237 bp
ORF:  78 amino acids  =  8.6 kDa
ORF:  6419 .. 6664  =  246 bp
ORF:  81 amino acids  =  8.9 kDa
ORF:  828 .. 1487  =  660 bp
ORF:  219 amino acids  =  25.7 kDa
ORF:  2445 .. 2975  =  531 bp
ORF:  176 amino acids  =  18.7 kDa
ORF:  7440 .. 8423  =  984 bp
ORF:  327 amino acids  =  36.7 kDa
ORF:  8481 .. 9320  =  840 bp
ORF:  279 amino acids  =  29.8 kDa
ORF:  6346 .. 6645  =  300 bp
ORF:  99 amino acids  =  10.9 kDa
ORF:  4866 .. 5132  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  7355 .. 7735  =  381 bp
ORF:  126 amino acids  =  13.3 kDa
ORF:  8168 .. 8722  =  555 bp
ORF:  184 amino acids  =  19.9 kDa
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Individual Sequences & Maps

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