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Plasmid Files

pLP/VSVG

Lentiviral packaging plasmid for expression of the vesicular stomatitis virus G glycoprotein.

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pLP VSVG Sequence and MappLP VSVG.dna
Map and Sequence File   
Sequence Author:  Invitrogen (Life Technologies)
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 ApaI (5757) PspOMI (5753) Bsu36I (5747) XmnI (5417) ScaI (5298) PvuI (5188) FspI (5040) AhdI (4818) PspFI (4233) BseYI (4229) HaeII (4173) EagI - NotI (3797) AccI (3680) MscI * (3642) BspQI - SapI (3289) BsrGI (37) HincII (78) BtgZI (428) SnaBI (434) BsmBI (683) HindIII (753) KflI - PpuMI (807) PmlI (1323) DraIII (1792) PflMI (1795) PasI (1984) EcoNI (1988) BstAPI (2046) XcmI (2094) PstI (2134) AgeI (2327) BclI * (2430) BmgBI (2488) Bpu10I (2598) BstBI (2607) pLP/VSVG 5821 bp
ApaI  (5757)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
PspOMI  (5753)
1 site
G G G C C C C C C G G G
Bsu36I  (5747)
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.
XmnI  (5417)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (5298)
1 site
A G T A C T T C A T G A
PvuI  (5188)
1 site
C G A T C G G C T A G C
FspI  (5040)
1 site
T G C G C A A C G C G T
AhdI  (4818)
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.
PspFI  (4233)
1 site
C C C A G C G G G T C G
BseYI  (4229)
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.
HaeII  (4173)
1 site
R G C G C Y Y C G C G R
EagI  (3797)
1 site
C G G C C G G C C G G C
NotI  (3797)
1 site
G C G G C C G C C G C C G G C G
AccI  (3680)
1 site
G T M K A C C A K M T G

Efficient cleavage with AccI requires ≥13 bp on each side of the
recognition sequence.
Sticky ends from different AccI sites may not be compatible.
MscI  (3642)
1 site
T G G C C A A C C G G T
* Blocked by Dcm methylation.
BspQI  (3289)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different BspQI sites may not be compatible.
SapI  (3289)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different SapI sites may not be compatible.
SapI gradually settles in solution, so a tube of SapI should be
mixed before removing an aliquot.
BsrGI  (37)
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.
HincII  (78)
1 site
G T Y R A C C A R Y T G
BtgZI  (428)
1 site
G C G A T G ( N ) 10 C G C T A C ( N ) 10 ( N ) 4

Sticky ends from different BtgZI sites may not be compatible.
After cleavage, BtgZI can remain bound to DNA and alter its
electrophoretic mobility.
BtgZI is typically used at 60°C, but is 75% active at 37°C.
SnaBI  (434)
1 site
T A C G T A A T G C A T
BsmBI  (683)
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.
HindIII  (753)
1 site
A A G C T T T T C G A A
KflI  (807)
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  (807)
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.
PmlI  (1323)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
DraIII  (1792)
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.
PflMI  (1795)
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.
PasI  (1984)
1 site
C C C W G G G G G G W C C C

Sticky ends from different PasI sites may not be compatible.
EcoNI  (1988)
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.
BstAPI  (2046)
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.
XcmI  (2094)
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.
PstI  (2134)
1 site
C T G C A G G A C G T C
AgeI  (2327)
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.
BclI  (2430)
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.
BmgBI  (2488)
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.
Bpu10I  (2598)
1 site
C C T N A G C G G A N T C G

Efficient cleavage requires at least two copies of the Bpu10I
recognition sequence.
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.
BstBI  (2607)
1 site
T T C G A A A A G C T T
VSV-G
1346 .. 2881  =  1536 bp
511 amino acids  =  57.5 kDa
   Segment 1:  signal peptide  
   1346 .. 1393  =  48 bp
   16 amino acids  =  1.8 kDa
Product: vesicular stomatitis virus G glycoprotein
Indiana strain
VSV-G
1346 .. 2881  =  1536 bp
511 amino acids  =  57.5 kDa
   Segment 2:  
   1394 .. 2881  =  1488 bp
   495 amino acids  =  55.7 kDa
Product: vesicular stomatitis virus G glycoprotein
Indiana strain
VSV-G
1346 .. 2881  =  1536 bp
511 amino acids  =  57.5 kDa
2 segments
Product: vesicular stomatitis virus G glycoprotein
Indiana strain
AmpR
4745 .. 5605  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   4745 .. 5536  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
4745 .. 5605  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   5537 .. 5605  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
4745 .. 5605  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
3986 .. 4574  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
3986 .. 4574  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
β-globin intron
796 .. 1271  =  476 bp
internally truncated intron from human β-globin
β-globin intron
796 .. 1271  =  476 bp
internally truncated intron from human β-globin
β-globin poly(A)
3082 .. 3476  =  395 bp
human β-globin polyadenylation signal
β-globin poly(A)
3082 .. 3476  =  395 bp
human β-globin polyadenylation signal
CMV enhancer
79 .. 458  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
79 .. 458  =  380 bp
human cytomegalovirus immediate early enhancer
CMV promoter
459 .. 662  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
459 .. 662  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
AmpR promoter
5606 .. 5710  =  105 bp
AmpR promoter
5606 .. 5710  =  105 bp
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