pLenti6.2 C-Lumio V5-DEST

Lentiviral Gateway® destination vector for C-terminal tagging of a protein with the Lumio™ tetracysteine tag.

Sequence Author: Thermo Fisher (Invitrogen)

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HomePlasmidsGateway® Cloning VectorspLenti6.2 C-Lumio V5-DEST
PciI (8425) AhdI (7537) DraIII (6397) NaeI (6291) NgoMIV (6289) AvrII (5956) SfiI (5909) PmlI (4802) PGK promoter EcoNI (942) MfeI (961) BstAPI (1225) AleI (1349) KflI (1706) BspDI - ClaI (1795) CMV enhancer NdeI (1989) SnaBI (2095) EcoRV (2435) lac UV5 promoter PasI - PflMI * (3118) BstZ17I (3421) TspMI - XmaI (3755) SmaI - SrfI (3757) BfuAI - BspMI (4001) SalI (4014) HpaI (4154) MluI (4197) tetracysteine tag pLenti6.2/C-Lumio™/V5-DEST 8835 bp
PciI  (8425)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
AhdI  (7537)
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  (6397)
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  (6291)
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  (6289)
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.
AvrII  (5956)
1 site		 C C T A G G G G A T C C
SfiI  (5909)
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.
PmlI  (4802)
1 site		 C A C G T G G T G C A C
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
BstAPI  (1225)
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.
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
EcoRV  (2435)
1 site		 G A T A T C C T A T A G
EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage.
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
TspMI  (3755)
1 site		 C C C G G G G G G C C C
XmaI  (3755)
1 site		 C C C G G G G G G C C C
Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
SmaI  (3757)
1 site		 C C C G G G G G G C C C
SmaI can be used at 37°C for brief incubations.
SrfI  (3757)
1 site		 G C C C G G G C C G G G C C C G
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
HpaI  (4154)
1 site		 G T T A A C C A A T T G
MluI  (4197)
1 site		 A C G C G T T G C G C A
AmpR
6750 .. 7610  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   6750 .. 6818  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6750 .. 7610  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   6819 .. 7610  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6750 .. 7610  =  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
7781 .. 8369  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
7781 .. 8369  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
PGK promoter
4292 .. 4791  =  500 bp
mouse phosphoglycerate kinase 1 promoter
PGK promoter
4292 .. 4791  =  500 bp
mouse phosphoglycerate kinase 1 promoter
f1 ori
6164 .. 6619  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
6164 .. 6619  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
BSD
4871 .. 5269  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
BSD
4871 .. 5269  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
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)
5356 .. 5589  =  234 bp
self-inactivating 3' long terminal repeat (LTR) from HIV-1
3' LTR (ΔU3)
5356 .. 5589  =  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
5661 .. 5782  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
5661 .. 5782  =  122 bp
SV40 polyadenylation signal
AmpR promoter
6645 .. 6749  =  105 bp
AmpR promoter
6645 .. 6749  =  105 bp
V5 tag
4158 .. 4199  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
V5 tag
4158 .. 4199  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
tetracysteine tag
4218 .. 4235  =  18 bp
6 amino acids  =  584.8 Da
Product: tetracysteine peptide that binds biarsenical labeling reagents
tetracysteine tag
4218 .. 4235  =  18 bp
6 amino acids  =  584.8 Da
Product: tetracysteine peptide that binds biarsenical labeling reagents
EM7 promoter
4805 .. 4852  =  48 bp
synthetic bacterial promoter
EM7 promoter
4805 .. 4852  =  48 bp
synthetic bacterial promoter
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
ORF:  4147 .. 4515  =  369 bp
ORF:  122 amino acids  =  12.9 kDa
ORF:  5683 .. 5919  =  237 bp
ORF:  78 amino acids  =  8.6 kDa
ORF:  8767 .. 177  =  246 bp
ORF:  81 amino acids  =  8.9 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:  4871 .. 5269  =  399 bp
ORF:  132 amino acids  =  13.7 kDa
ORF:  2673 .. 3332  =  660 bp
ORF:  219 amino acids  =  25.7 kDa
ORF:  4572 .. 4841  =  270 bp
ORF:  89 amino acids  =  9.7 kDa
ORF:  5205 .. 5435  =  231 bp
ORF:  76 amino acids  =  8.5 kDa
ORF:  6750 .. 7610  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  868 .. 1248  =  381 bp
ORF:  126 amino acids  =  13.3 kDa
ORF:  8694 .. 158  =  300 bp
ORF:  99 amino acids  =  10.9 kDa
ORF:  4817 .. 5173  =  357 bp
ORF:  118 amino acids  =  12.8 kDa
ORF:  7214 .. 7480  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
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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

Plasmid Sets

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I.M.A.G.E. Consortium Plasmids
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Coronavirus Resources
Insect Cell Vectors
pGEX Vectors (GE Healthcare)
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CRISPR Plasmids
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Plant Vectors
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Fluorescent Protein Genes & Plasmids
Lucigen Vectors
Qiagen Vectors
Yeast Plasmids
Gateway® Cloning Vectors
Mammalian Expression Vectors
Structural Genomics Vectors

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