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Plasmid Files

pLenti6/UbC/V5-DEST

Lentiviral Gateway® destination vector for C-terminal tagging of a protein with a V5 epitope tag.

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pLenti6 UbC V5-DEST Sequence and MappLenti6 UbC V5-DEST.dna
Map and Sequence File   
Sequence Author:  Invitrogen (Life Technologies)
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 PciI (8909) AhdI (8021) NaeI (6775) NgoMIV (6773) SfiI (6393) BlpI (5750) EM7 promoter PmlI (5286) BclI * (5277) SexAI * (4993) AgeI (4874) MluI (4868) BstBI (4824) ApaI (4817) PspOMI (4813) XbaI (4807) PaeR7I - PspXI - XhoI (4801) MfeI (961) AleI (1349) AfeI (1858) ZraI (2645) AatII (2647) AarI (2693) BspEI (3514) BsmBI (3742) PasI - PflMI * (3750) BstZ17I (4053) SrfI (4389) SalI (4646) pLenti6/UbC/V5-DEST 9319 bp
PciI  (8909)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AhdI  (8021)
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.
NaeI  (6775)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NaeI
recognition sequence.
NgoMIV  (6773)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NgoMIV
recognition sequence.
SfiI  (6393)
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.
BlpI  (5750)
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.
PmlI  (5286)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
BclI  (5277)
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.
SexAI  (4993)
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.
AgeI  (4874)
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.
MluI  (4868)
1 site
A C G C G T T G C G C A
BstBI  (4824)
1 site
T T C G A A A A G C T T
ApaI  (4817)
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  (4813)
1 site
G G G C C C C C C G G G
XbaI  (4807)
1 site
T C T A G A A G A T C T
PaeR7I  (4801)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (4801)
1 site
V C T C G A G B B G A G C T C V
XhoI  (4801)
1 site
C T C G A G G A G C T C
MfeI  (961)
1 site
C A A T T G G T T A A C
AleI  (1349)
1 site
C A C N N N N G T G G T G N N N N C A C
AfeI  (1858)
1 site
A G C G C T T C G C G A
ZraI  (2645)
1 site
G A C G T C C T G C A G
AatII  (2647)
1 site
G A C G T C C T G C A G
AarI  (2693)
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 AarI
recognition sequence.
Sticky ends from different AarI sites may not be compatible.
After cleavage, AarI can remain bound to DNA and alter its
electrophoretic mobility.
BspEI  (3514)
1 site
T C C G G A A G G C C T
BsmBI  (3742)
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.
PasI  (3750)
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.
PflMI  (3750)
1 site
C C A N N N N N T G G G G T N N N N N A C C
* Blocked by Dcm methylation.
Sticky ends from different PflMI sites may not be compatible.
BstZ17I  (4053)
1 site
G T A T A C C A T A T G
SrfI  (4389)
1 site
G C C C G G G C C G G G C C C G
SalI  (4646)
1 site
G T C G A C C A G C T G
UbC promoter
1803 .. 3012  =  1210 bp
human ubiquitin C promoter
UbC promoter
1803 .. 3012  =  1210 bp
human ubiquitin C promoter
AmpR
7234 .. 8094  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   7234 .. 7302  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
7234 .. 8094  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   7303 .. 8094  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
7234 .. 8094  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
CmR
3305 .. 3964  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
3305 .. 3964  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
ori
8265 .. 8853  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
8265 .. 8853  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
6648 .. 7103  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
6648 .. 7103  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
BSD
5355 .. 5753  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
BSD
5355 .. 5753  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
SV40 promoter
4912 .. 5241  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
4912 .. 5241  =  330 bp
SV40 enhancer and early promoter
ccdB
4306 .. 4611  =  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
4306 .. 4611  =  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.
RRE
1075 .. 1308  =  234 bp
The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to
the cytoplasm.
RRE
1075 .. 1308  =  234 bp
The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to
the cytoplasm.
3' LTR (ΔU3)
5840 .. 6073  =  234 bp
self-inactivating 3' long terminal repeat (LTR) from
HIV-1
3' LTR (ΔU3)
5840 .. 6073  =  234 bp
self-inactivating 3' long terminal repeat (LTR) from
HIV-1
RSV promoter
1 .. 229  =  229 bp
Rous sarcoma virus enhancer/promoter
RSV promoter
1 .. 229  =  229 bp
Rous sarcoma virus enhancer/promoter
5' LTR (truncated)
230 .. 410  =  181 bp
truncated 5' long terminal repeat (LTR) from HIV-1
5' LTR (truncated)
230 .. 410  =  181 bp
truncated 5' long terminal repeat (LTR) from HIV-1
HIV-1 Ψ
457 .. 582  =  126 bp
packaging signal of human immunodeficiency virus
type 1
HIV-1 Ψ
457 .. 582  =  126 bp
packaging signal of human immunodeficiency virus
type 1
attR1
3072 .. 3196  =  125 bp
recombination site for the Gateway® LR reaction
attR1
3072 .. 3196  =  125 bp
recombination site for the Gateway® LR reaction
attR2
4652 .. 4776  =  125 bp
recombination site for the Gateway® LR reaction
attR2
4652 .. 4776  =  125 bp
recombination site for the Gateway® LR reaction
SV40 poly(A) signal
6145 .. 6266  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
6145 .. 6266  =  122 bp
SV40 polyadenylation signal
AmpR promoter
7129 .. 7233  =  105 bp
AmpR promoter
7129 .. 7233  =  105 bp
EM7 promoter
5289 .. 5336  =  48 bp
synthetic bacterial promoter
EM7 promoter
5289 .. 5336  =  48 bp
synthetic bacterial promoter
V5 tag
4829 .. 4870  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
V5 tag
4829 .. 4870  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
lac UV5 promoter
3221 .. 3251  =  31 bp
   Segment 1:  -35  
   3221 .. 3226  =  6 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
3221 .. 3251  =  31 bp
   Segment 2:  
   3227 .. 3244  =  18 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
3221 .. 3251  =  31 bp
   Segment 3:  -10  
   3245 .. 3251  =  7 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
3221 .. 3251  =  31 bp
3 segments
E. coli lac promoter with an "up" mutation
SV40 ori
5092 .. 5227  =  136 bp
SV40 origin of replication
SV40 ori
5092 .. 5227  =  136 bp
SV40 origin of replication
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