pLenti6 V5-DEST

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

Sequence Author: Thermo Fisher (Invitrogen)

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PciI (8277) AhdI (7389) DraIII (6249) NaeI (6143) NgoMIV (6141) SfiI (5761) SV40 poly(A) signal KpnI (5141) Acc65I (5137) BlpI (5118) EM7 promoter PmlI (4654) BclI * (4645) CsiI - SexAI * (4361) KasI (413) NarI (414) SfoI (415) PluTI (417) EcoNI (942) MfeI (961) AleI (1349) KflI (1706) BspDI - ClaI (1795) NdeI (1989) SnaBI (2095) SpeI (2406) BspEI (2882) PasI - PflMI * (3118) BstZ17I (3421) SrfI (3757) BmgBI (3791) BfuAI - BspMI (4001) SalI (4014) PaeR7I - PspXI - XhoI (4169) PspOMI (4181) ApaI (4185) SacII (4188) BstBI (4192) MluI (4236) AgeI (4242) pLenti6/V5-DEST 8687 bp
PciI  (8277)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AhdI  (7389)
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.
DraIII  (6249)
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.
NaeI  (6143)
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  (6141)
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  (5761)
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.
KpnI  (5141)
1 site
G G T A C C C C A T G G
Acc65I  (5137)
1 site
G G T A C C C C A T G G
BlpI  (5118)
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  (4654)
1 site
C A C G T G G T G C A C
BclI  (4645)
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.
CsiI  (4361)
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  (4361)
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.
KasI  (413)
1 site
G G C G C C C C G C G G
NarI  (414)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the NarI recognition sequence.
SfoI  (415)
1 site
G G C G C C C C G C G G
PluTI  (417)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the PluTI recognition sequence.
EcoNI  (942)
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.
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
KflI  (1706)
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.
BspDI  (1795)
1 site
A T C G A T T A G C T A
ClaI  (1795)
1 site
A T C G A T T A G C T A
NdeI  (1989)
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.
SnaBI  (2095)
1 site
T A C G T A A T G C A T
SpeI  (2406)
1 site
A C T A G T T G A T C A
BspEI  (2882)
1 site
T C C G G A A G G C C T
PasI  (3118)
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  (3118)
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  (3421)
1 site
G T A T A C C A T A T G
SrfI  (3757)
1 site
G C C C G G G C C G G G C C C G
BmgBI  (3791)
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.
BfuAI  (4001)
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  (4001)
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.
SalI  (4014)
1 site
G T C G A C C A G C T G
PaeR7I  (4169)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (4169)
1 site
V C T C G A G B B G A G C T C V
XhoI  (4169)
1 site
C T C G A G G A G C T C
PspOMI  (4181)
1 site
G G G C C C C C C G G G
ApaI  (4185)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
SacII  (4188)
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.
BstBI  (4192)
1 site
T T C G A A A A G C T T
MluI  (4236)
1 site
A C G C G T T G C G C A
AgeI  (4242)
1 site
A C C G G T T G G C C A
AmpR
6602 .. 7462  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   6602 .. 6670  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6602 .. 7462  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   6671 .. 7462  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6602 .. 7462  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
CmR
2673 .. 3332  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
2673 .. 3332  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
ori
7633 .. 8221  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
7633 .. 8221  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
6016 .. 6471  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
6016 .. 6471  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
BSD
4723 .. 5121  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
BSD
4723 .. 5121  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
SV40 promoter
4280 .. 4609  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
4280 .. 4609  =  330 bp
SV40 enhancer and early promoter
ccdB
3674 .. 3979  =  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
3674 .. 3979  =  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
1816 .. 2119  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
1816 .. 2119  =  304 bp
human cytomegalovirus immediate early enhancer
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)
5208 .. 5441  =  234 bp
self-inactivating 3' long terminal repeat (LTR) from HIV-1
3' LTR (ΔU3)
5208 .. 5441  =  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
CMV promoter
2120 .. 2323  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
2120 .. 2323  =  204 bp
human cytomegalovirus (CMV) immediate early 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
2440 .. 2564  =  125 bp
recombination site for the Gateway® LR reaction
attR1
2440 .. 2564  =  125 bp
recombination site for the Gateway® LR reaction
attR2
4020 .. 4144  =  125 bp
recombination site for the Gateway® LR reaction
attR2
4020 .. 4144  =  125 bp
recombination site for the Gateway® LR reaction
SV40 poly(A) signal
5513 .. 5634  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
5513 .. 5634  =  122 bp
SV40 polyadenylation signal
AmpR promoter
6497 .. 6601  =  105 bp
AmpR promoter
6497 .. 6601  =  105 bp
EM7 promoter
4657 .. 4704  =  48 bp
synthetic bacterial promoter
EM7 promoter
4657 .. 4704  =  48 bp
synthetic bacterial promoter
V5 tag
4197 .. 4238  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
V5 tag
4197 .. 4238  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
lac UV5 promoter
2589 .. 2619  =  31 bp
3 segments
   Segment 1:  -35  
   2589 .. 2594  =  6 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
2589 .. 2619  =  31 bp
3 segments
   Segment 2:  
   2595 .. 2612  =  18 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
2589 .. 2619  =  31 bp
3 segments
   Segment 3:  -10  
   2613 .. 2619  =  7 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
2589 .. 2619  =  31 bp
3 segments
E. coli lac promoter with an "up" mutation
SV40 ori
4460 .. 4595  =  136 bp
SV40 origin of replication
SV40 ori
4460 .. 4595  =  136 bp
SV40 origin of replication
ORF:  4723 .. 5121  =  399 bp
ORF:  132 amino acids  =  13.7 kDa
ORF:  953 .. 1801  =  849 bp
ORF:  282 amino acids  =  32.5 kDa
ORF:  3674 .. 3979  =  306 bp
ORF:  101 amino acids  =  11.7 kDa
ORF:  5057 .. 5287  =  231 bp
ORF:  76 amino acids  =  8.5 kDa
ORF:  6602 .. 7462  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  2673 .. 3332  =  660 bp
ORF:  219 amino acids  =  25.7 kDa
ORF:  5535 .. 5771  =  237 bp
ORF:  78 amino acids  =  8.6 kDa
ORF:  8619 .. 177  =  246 bp
ORF:  81 amino acids  =  8.9 kDa
ORF:  8546 .. 158  =  300 bp
ORF:  99 amino acids  =  10.9 kDa
ORF:  868 .. 1248  =  381 bp
ORF:  126 amino acids  =  13.3 kDa
ORF:  4669 .. 5025  =  357 bp
ORF:  118 amino acids  =  12.8 kDa
ORF:  7066 .. 7332  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
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