pLenti6.2 N-Lumio V5-DEST

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

Sequence Author: Thermo Fisher (Invitrogen)

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HomePlasmidsGateway® Cloning VectorspLenti6.2 N-Lumio V5-DEST
PciI (8546) AhdI (7658) DraIII (6518) NaeI (6412) NgoMIV (6410) AvrII (6077) SfiI (6030) SV40 poly(A) signal PmlI (4923) PGK promoter SpeI (4658) EcoNI (942) MfeI (961) BstAPI (1225) AleI (1349) KflI (1706) BspDI - ClaI (1795) NdeI (2055) SnaBI (2161) CMV promoter tetracysteine tag V5 tag lac UV5 promoter EcoRI (3028) PasI - PflMI * (3260) BstZ17I (3563) TspMI - XmaI (3897) SmaI - SrfI (3899) BfuAI - BspMI (4143) PstI (4154) EcoRV (4290) PaeR7I - PspXI - XhoI (4311) PspOMI (4323) ApaI (4327) SacII (4330) BstBI (4334) MluI (4378) pLenti6.2/N-Lumio™/V5-DEST 8956 bp
PciI  (8546)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
AhdI  (7658)
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  (6518)
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  (6412)
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  (6410)
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  (6077)
1 site		 C C T A G G G G A T C C
SfiI  (6030)
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  (4923)
1 site		 C A C G T G G T G C A C
SpeI  (4658)
1 site		 A C T A G T T G A T C A
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  (2055)
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  (2161)
1 site		 T A C G T A A T G C A T
EcoRI  (3028)
1 site		 G A A T T C C T T A A G
PasI  (3260)
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  (3260)
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  (3563)
1 site		 G T A T A C C A T A T G
TspMI  (3897)
1 site		 C C C G G G G G G C C C
XmaI  (3897)
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  (3899)
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  (3899)
1 site		 G C C C G G G C C G G G C C C G
BfuAI  (4143)
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  (4143)
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.
PstI  (4154)
1 site		 C T G C A G G A C G T C
EcoRV  (4290)
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.
PaeR7I  (4311)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (4311)
1 site		 V C T C G A G B B G A G C T C V
XhoI  (4311)
1 site		 C T C G A G G A G C T C
PspOMI  (4323)
1 site		 G G G C C C C C C G G G
ApaI  (4327)
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  (4330)
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  (4334)
1 site		 T T C G A A A A G C T T
MluI  (4378)
1 site		 A C G C G T T G C G C A
AmpR
6871 .. 7731  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   6871 .. 6939  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6871 .. 7731  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   6940 .. 7731  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6871 .. 7731  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
CmR
2815 .. 3474  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
2815 .. 3474  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
ori
7902 .. 8490  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
7902 .. 8490  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
PGK promoter
4413 .. 4912  =  500 bp
mouse phosphoglycerate kinase 1 promoter
PGK promoter
4413 .. 4912  =  500 bp
mouse phosphoglycerate kinase 1 promoter
f1 ori
6285 .. 6740  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
6285 .. 6740  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
BSD
4992 .. 5390  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
BSD
4992 .. 5390  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
ccdB
3816 .. 4121  =  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
3816 .. 4121  =  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
1882 .. 2185  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
1882 .. 2185  =  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)
5477 .. 5710  =  234 bp
self-inactivating 3' long terminal repeat (LTR) from HIV-1
3' LTR (ΔU3)
5477 .. 5710  =  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
2186 .. 2389  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
2186 .. 2389  =  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
2582 .. 2706  =  125 bp
recombination site for the Gateway® LR reaction
attR1
2582 .. 2706  =  125 bp
recombination site for the Gateway® LR reaction
attR2
4162 .. 4286  =  125 bp
recombination site for the Gateway® LR reaction
attR2
4162 .. 4286  =  125 bp
recombination site for the Gateway® LR reaction
SV40 poly(A) signal
5782 .. 5903  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
5782 .. 5903  =  122 bp
SV40 polyadenylation signal
AmpR promoter
6766 .. 6870  =  105 bp
AmpR promoter
6766 .. 6870  =  105 bp
ATG
2483 .. 2485  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
2483 .. 2485  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
tetracysteine tag
2495 .. 2512  =  18 bp
6 amino acids  =  584.8 Da
Product: tetracysteine peptide that binds biarsenical labeling reagents
tetracysteine tag
2495 .. 2512  =  18 bp
6 amino acids  =  584.8 Da
Product: tetracysteine peptide that binds biarsenical labeling reagents
V5 tag
2528 .. 2569  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
V5 tag
2528 .. 2569  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
EM7 promoter
4926 .. 4973  =  48 bp
synthetic bacterial promoter
EM7 promoter
4926 .. 4973  =  48 bp
synthetic bacterial promoter
lac UV5 promoter
2731 .. 2761  =  31 bp
3 segments
   Segment 1:  -35  
   2731 .. 2736  =  6 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
2731 .. 2761  =  31 bp
3 segments
   Segment 2:  
   2737 .. 2754  =  18 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
2731 .. 2761  =  31 bp
3 segments
   Segment 3:  -10  
   2755 .. 2761  =  7 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
2731 .. 2761  =  31 bp
3 segments
E. coli lac promoter with an "up" mutation
ORF:  1798 .. 2079  =  282 bp
ORF:  93 amino acids  =  9.8 kDa
ORF:  2815 .. 3474  =  660 bp
ORF:  219 amino acids  =  25.7 kDa
ORF:  4693 .. 4962  =  270 bp
ORF:  89 amino acids  =  9.7 kDa
ORF:  5326 .. 5556  =  231 bp
ORF:  76 amino acids  =  8.5 kDa
ORF:  6871 .. 7731  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  953 .. 1810  =  858 bp
ORF:  285 amino acids  =  33.0 kDa
ORF:  5804 .. 6040  =  237 bp
ORF:  78 amino acids  =  8.6 kDa
ORF:  8888 .. 177  =  246 bp
ORF:  81 amino acids  =  8.9 kDa
ORF:  3816 .. 4121  =  306 bp
ORF:  101 amino acids  =  11.7 kDa
ORF:  4992 .. 5390  =  399 bp
ORF:  132 amino acids  =  13.7 kDa
ORF:  868 .. 1248  =  381 bp
ORF:  126 amino acids  =  13.3 kDa
ORF:  8815 .. 158  =  300 bp
ORF:  99 amino acids  =  10.9 kDa
ORF:  4938 .. 5294  =  357 bp
ORF:  118 amino acids  =  12.8 kDa
ORF:  7335 .. 7601  =  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

Basic Cloning Vectors
I.M.A.G.E. Consortium Plasmids
pET & Duet Vectors (Novagen)
TA and GC Cloning Vectors
Coronavirus Resources
Insect Cell Vectors
pGEX Vectors (GE Healthcare)
TOPO Cloning Vectors
CRISPR Plasmids
Luciferase Vectors
Plant Vectors
Viral Expression & Packaging Vectors
Fluorescent Protein Genes & Plasmids
Lucigen Vectors
Qiagen Vectors
Yeast Plasmids
Gateway® Cloning Vectors
Mammalian Expression Vectors
Structural Genomics Vectors

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