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Plasmid Files

pLX303

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

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pLX303 Sequence and MappLX303.dna
Map and Sequence File   
Sequence Author:  Root Lab
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 cPPT/CTS XbaI * (9129) KpnI (8706) Acc65I (8702) AscI (8534) BlpI (8525) SpeI (8445) XcmI (8217) KflI (8193) AleI (7836) BstAPI (7712) MfeI (7448) EcoNI (7429) SphI (6580) AflIII - PciI (6077) PaeR7I - PspXI - XhoI (9326) NdeI (238) BstZ17I (1576) SrfI (1912) BstXI (2029) BfuAI - BspMI (2156) stop codons NheI (2310) BmtI (2314) AfeI (2315) AgeI (2320) SacII (2845) SfiI (3561) StuI (3607) AvrII (3608) DraIII (4049) pLX303 9335 bp
XbaI  (9129)
1 site
T C T A G A A G A T C T
* Blocked by Dam methylation.
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.
AleI  (7836)
1 site
C A C N N N N G T G G T G N N N N C A C
BstAPI  (7712)
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  (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.
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.
PaeR7I  (9326)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (9326)
1 site
V C T C G A G B B G A G C T C V
XhoI  (9326)
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  (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

AgeI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
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.
StuI  (3607)
1 site
A G G C C T T C C G G A
AvrII  (3608)
1 site
C C T A G G G G A T C C
DraIII  (4049)
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
4402 .. 5262  =  861 bp
286 amino acids  =  31.6 kDa
   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
   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
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
BSD
8711 .. 9106  =  396 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
BSD
8711 .. 9106  =  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)
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
9166 .. 9283  =  118 bp
central polypurine tract and central termination
sequence of HIV-1
cPPT/CTS
9166 .. 9283  =  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
   Segment 1:  -35  
   744 .. 749  =  6 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
744 .. 774  =  31 bp
   Segment 2:  
   750 .. 767  =  18 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
744 .. 774  =  31 bp
   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
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