pLenti7.3 V5-DEST

Lentiviral Gateway® destination vector for expression of a C-terminally V5-tagged protein together with Emerald GFP.

Sequence Author: Thermo Fisher (Invitrogen)

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HomePlasmidsGateway® Cloning VectorspLenti7.3 V5-DEST
KasI (413) PciI (9220) AhdI (8332) DraIII (7192) SfiI (6704) BclI * (5345) CsiI - SexAI * (5061) NarI (414) SfoI (415) PluTI (417) NruI * (609) EcoNI (942) MfeI (961) KflI - PpuMI (1706) HpaI (1800) cPPT/CTS BspDI - ClaI (1923) NdeI (2117) SnaBI (2223) SpeI (2534) BspEI (3010) PasI (3246) BstZ17I (3549) SrfI (3885) BmgBI (3919) BfuAI - BspMI (4129) SalI (4142) PaeR7I - PspXI - XhoI (4297) PspOMI (4309) ApaI (4313) BstBI (4320) V5 tag MluI (4364) pLenti7.3/V5-DEST 9630 bp
KasI  (413)
1 site		 G G C G C C C C G C G G
PciI  (9220)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
AhdI  (8332)
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  (7192)
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.
SfiI  (6704)
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.
BclI  (5345)
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  (5061)
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  (5061)
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.
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.
NruI  (609)
1 site		 T C G C G A A G C G C T
* Blocked by Dam methylation.
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
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.
PpuMI  (1706)
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.
HpaI  (1800)
1 site		 G T T A A C C A A T T G
BspDI  (1923)
1 site		 A T C G A T T A G C T A
ClaI  (1923)
1 site		 A T C G A T T A G C T A
NdeI  (2117)
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  (2223)
1 site		 T A C G T A A T G C A T
SpeI  (2534)
1 site		 A C T A G T T G A T C A
BspEI  (3010)
1 site		 T C C G G A A G G C C T
PasI  (3246)
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.
BstZ17I  (3549)
1 site		 G T A T A C C A T A T G
SrfI  (3885)
1 site		 G C C C G G G C C G G G C C C G
BmgBI  (3919)
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  (4129)
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  (4129)
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  (4142)
1 site		 G T C G A C C A G C T G
PaeR7I  (4297)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (4297)
1 site		 V C T C G A G B B G A G C T C V
XhoI  (4297)
1 site		 C T C G A G G A G C T C
PspOMI  (4309)
1 site		 G G G C C C C C C G G G
ApaI  (4313)
1 site		 G G G C C C C C C G G G
ApaI can be used between 25°C and 37°C.
BstBI  (4320)
1 site		 T T C G A A A A G C T T
MluI  (4364)
1 site		 A C G C G T T G C G C A
AmpR
7545 .. 8405  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   7545 .. 7613  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
7545 .. 8405  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   7614 .. 8405  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
7545 .. 8405  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
EmGFP
5360 .. 6079  =  720 bp
239 amino acids  =  26.9 kDa
3 segments
   Segment 1:  
   5360 .. 5362  =  3 bp
   1 amino acid  =  149.2 Da
Product: Emerald GFP
mammalian codon-optimized
EmGFP
5360 .. 6079  =  720 bp
239 amino acids  =  26.9 kDa
3 segments
   Segment 2:  1a  
   5363 .. 5365  =  3 bp
   1 amino acid  =  117.1 Da
Product: Emerald GFP
mammalian codon-optimized
EmGFP
5360 .. 6079  =  720 bp
239 amino acids  =  26.9 kDa
3 segments
   Segment 3:  
   5366 .. 6079  =  714 bp
   237 amino acids  =  26.7 kDa
Product: Emerald GFP
mammalian codon-optimized
EmGFP
5360 .. 6079  =  720 bp
239 amino acids  =  26.9 kDa
3 segments
Product: Emerald GFP
mammalian codon-optimized
CmR
2801 .. 3460  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
2801 .. 3460  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
WPRE
4390 .. 4978  =  589 bp
woodchuck posttranscriptional regulatory element
WPRE
4390 .. 4978  =  589 bp
woodchuck posttranscriptional regulatory element
ori
8576 .. 9164  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
8576 .. 9164  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
6959 .. 7414  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
6959 .. 7414  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
SV40 promoter
4993 .. 5309  =  317 bp
SV40 enhancer and early promoter
SV40 promoter
4993 .. 5309  =  317 bp
SV40 enhancer and early promoter
ccdB
3802 .. 4107  =  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
3802 .. 4107  =  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
1944 .. 2247  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
1944 .. 2247  =  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)
6151 .. 6384  =  234 bp
self-inactivating 3' long terminal repeat (LTR) from HIV-1
3' LTR (ΔU3)
6151 .. 6384  =  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
2248 .. 2451  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
2248 .. 2451  =  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
2568 .. 2692  =  125 bp
recombination site for the Gateway® LR reaction
attR1
2568 .. 2692  =  125 bp
recombination site for the Gateway® LR reaction
attR2
4148 .. 4272  =  125 bp
recombination site for the Gateway® LR reaction
attR2
4148 .. 4272  =  125 bp
recombination site for the Gateway® LR reaction
SV40 poly(A) signal
6456 .. 6577  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
6456 .. 6577  =  122 bp
SV40 polyadenylation signal
cPPT/CTS
1804 .. 1921  =  118 bp
central polypurine tract and central termination sequence of HIV-1
cPPT/CTS
1804 .. 1921  =  118 bp
central polypurine tract and central termination sequence of HIV-1
AmpR promoter
7440 .. 7544  =  105 bp
AmpR promoter
7440 .. 7544  =  105 bp
V5 tag
4325 .. 4366  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
V5 tag
4325 .. 4366  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
lac UV5 promoter
2717 .. 2747  =  31 bp
3 segments
   Segment 1:  -35  
   2717 .. 2722  =  6 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
2717 .. 2747  =  31 bp
3 segments
   Segment 2:  
   2723 .. 2740  =  18 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
2717 .. 2747  =  31 bp
3 segments
   Segment 3:  -10  
   2741 .. 2747  =  7 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
2717 .. 2747  =  31 bp
3 segments
E. coli lac promoter with an "up" mutation
SV40 ori
5160 .. 5295  =  136 bp
SV40 origin of replication
SV40 ori
5160 .. 5295  =  136 bp
SV40 origin of replication
ORF:  3802 .. 4107  =  306 bp
ORF:  101 amino acids  =  11.7 kDa
ORF:  4381 .. 5139  =  759 bp
ORF:  252 amino acids  =  27.2 kDa
ORF:  6478 .. 6714  =  237 bp
ORF:  78 amino acids  =  8.6 kDa
ORF:  9562 .. 177  =  246 bp
ORF:  81 amino acids  =  8.9 kDa
ORF:  953 .. 1963  =  1011 bp
ORF:  336 amino acids  =  38.5 kDa
ORF:  2801 .. 3460  =  660 bp
ORF:  219 amino acids  =  25.7 kDa
ORF:  4493 .. 4789  =  297 bp
ORF:  98 amino acids  =  10.2 kDa
ORF:  4985 .. 5257  =  273 bp
ORF:  90 amino acids  =  10.3 kDa
ORF:  5360 .. 6079  =  720 bp
ORF:  239 amino acids  =  26.9 kDa
ORF:  4800 .. 5054  =  255 bp
ORF:  84 amino acids  =  8.9 kDa
ORF:  7545 .. 8405  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  868 .. 1248  =  381 bp
ORF:  126 amino acids  =  13.3 kDa
ORF:  9489 .. 158  =  300 bp
ORF:  99 amino acids  =  10.9 kDa
ORF:  8009 .. 8275  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
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Download pLenti7.3 V5-DEST.dna file

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Individual Sequences & Maps

pAd BLOCK-iT-DEST
pDEST R4-R3 Vector II
pENTR11
pHELLSGATE 4
pAd CMV V5-DEST
pDEST10
pENTR1A
pHELLSGATE 8
pAd PL-DEST
pDEST14
pENTR2B
pIB V5-His-DEST
pBAD-DEST49
pDEST15
pENTR3C
pLenti4 BLOCK-iT-DEST
pcDNA-DEST40
pDEST17
pENTR4
pLenti4 V5-DEST
pcDNA-DEST47
pDEST20
pET-53-DEST
pLenti6.2 C-Lumio V5-DEST
pcDNA-DEST53
pDEST24
pET-54-DEST
pLenti6.2 N-Lumio V5-DEST
pcDNA3.1 nV5-DEST
pDEST26
pET-55-DEST
pLenti6.2 V5-DEST
pcDNA3.2 capTEV-CT V5-DEST
pDEST27
pET-56-DEST
pLenti6.3 TO V5-DEST
pcDNA3.2 capTEV-NT V5-DEST
pDEST8
pET-57-DEST
pLenti6.3 V5-DEST
pcDNA3.2 V5-DEST
pDONR221 P5-P2
pET-58-DEST
pLenti6.4 R4R2 V5-DEST
pcDNA6.2 C-EmGFP-DEST
pDONR P2r-P3
pET-59-DEST
pLenti6 BLOCK-iT-DEST
pcDNA6.2 C-YFP-DEST
pDONR P4-P1r
pET-60-DEST
pLenti6 capTEV-CT-DEST
pcDNA6.2 cGeneBLAzer-DEST
pDONR Zeo
pET-61-DEST
pLenti6 capTEV-NT-DEST1
pcDNA6.2 cLumio-DEST
pDONR201
pET-62-DEST
pLenti6 UbC V5-DEST
pcDNA6.2 EmGFP-Bsd V5-DEST
pDONR207
pET-DEST42
pLenti6 V5-DEST
pcDNA6.2 N-EmGFP-DEST
pDONR221
pET104-DEST
pLenti7.3 V5-DEST
pcDNA6.2 N-YFP-DEST
pDONR221 P1-P4
pET104.1-DEST
pMT-DEST48
pcDNA6.2 nGeneBLAzer-DEST
pDONR221 P1-P5r
pET160-DEST
pMT BioEase-DEST
pcDNA6.2 nLumio-DEST
pDONR221 P3-P2
pET161-DEST
pSCREEN-iT lacZ-DEST
pcDNA6.2 V5-DEST
pDONR221 P4r-P3r
pEXP1-DEST
pT-REx-DEST30
pcDNA6.2 V5-pL-DEST
pDONR221 P5-P4
pEXP2-DEST
pT-REx-DEST31
pCMV SPORT6
pDONR222
pEXP3-DEST
pYES-DEST52
pCMV SPORT6.1
pDONR223
pEXP4-DEST
Reading Frame Cassette A
pCOLA-2-DEST
pEF-DEST51
pHELLSGATE
Reading Frame Cassette B
pCOLA-3-DEST
pEF5 FRT V5-DEST
pHELLSGATE 12
Reading Frame Cassette C.1
pDEST R4-R3
pENTR GeneBLAzer

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