pLenti6.4 R4R2 V5-DEST

Lentiviral Gateway® destination vector for a micro-RNA (miRNA) expression cassette.

Sequence Author: Thermo Fisher (Invitrogen)

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HomePlasmidsGateway® Cloning VectorspLenti6.4 R4R2 V5-DEST
PciI (8512) AhdI (7624) DraIII (6484) AvrII (6043) SfiI (5996) PmlI (4889) PGK promoter EcoNI (942) MfeI (961) BstAPI (1225) AleI (1349) KflI (1706) HpaI (1936) lac UV5 promoter EcoRI (2396) PasI (2628) BstZ17I (2931) TspMI - XmaI (3265) SmaI - SrfI (3267) BfuAI - BspMI (3511) PstI (3522) PaeR7I - PspXI - XhoI (3679) PspOMI (3691) ApaI (3695) BstBI (3702) MluI (3746) PfoI (4045) SpeI (4624) pLenti6.4/R4R2/V5-DEST 8922 bp
PciI  (8512)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
AhdI  (7624)
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  (6484)
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.
AvrII  (6043)
1 site		 C C T A G G G G A T C C
SfiI  (5996)
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  (4889)
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.
HpaI  (1936)
1 site		 G T T A A C C A A T T G
EcoRI  (2396)
1 site		 G A A T T C C T T A A G
PasI  (2628)
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  (2931)
1 site		 G T A T A C C A T A T G
TspMI  (3265)
1 site		 C C C G G G G G G C C C
XmaI  (3265)
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  (3267)
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  (3267)
1 site		 G C C C G G G C C G G G C C C G
BfuAI  (3511)
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  (3511)
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  (3522)
1 site		 C T G C A G G A C G T C
PaeR7I  (3679)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (3679)
1 site		 V C T C G A G B B G A G C T C V
XhoI  (3679)
1 site		 C T C G A G G A G C T C
PspOMI  (3691)
1 site		 G G G C C C C C C G G G
ApaI  (3695)
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  (3702)
1 site		 T T C G A A A A G C T T
MluI  (3746)
1 site		 A C G C G T T G C G C A
PfoI  (4045)
1 site		 T C C N G G A A G G N C C T
Sticky ends from different PfoI sites may not be compatible.
SpeI  (4624)
1 site		 A C T A G T T G A T C A
AmpR
6837 .. 7697  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   6837 .. 6905  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6837 .. 7697  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   6906 .. 7697  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6837 .. 7697  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
CmR
2183 .. 2842  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
2183 .. 2842  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
WPRE
3772 .. 4360  =  589 bp
woodchuck posttranscriptional regulatory element
WPRE
3772 .. 4360  =  589 bp
woodchuck posttranscriptional regulatory element
ori
7868 .. 8456  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
7868 .. 8456  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
PGK promoter
4379 .. 4878  =  500 bp
mouse phosphoglycerate kinase 1 promoter
PGK promoter
4379 .. 4878  =  500 bp
mouse phosphoglycerate kinase 1 promoter
f1 ori
6251 .. 6706  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
6251 .. 6706  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
BSD
4958 .. 5356  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
BSD
4958 .. 5356  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
ccdB
3184 .. 3489  =  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
3184 .. 3489  =  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)
5443 .. 5676  =  234 bp
self-inactivating 3' long terminal repeat (LTR) from HIV-1
3' LTR (ΔU3)
5443 .. 5676  =  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
SV40 ori
5909 .. 6044  =  136 bp
SV40 origin of replication
SV40 ori
5909 .. 6044  =  136 bp
SV40 origin of replication
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
attR2
3530 .. 3654  =  125 bp
recombination site for the Gateway® LR reaction
attR2
3530 .. 3654  =  125 bp
recombination site for the Gateway® LR reaction
attR4
1951 .. 2074  =  124 bp
recombination site for the Gateway® LR reaction
attR4
1951 .. 2074  =  124 bp
recombination site for the Gateway® LR reaction
SV40 poly(A) signal
5748 .. 5869  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
5748 .. 5869  =  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
6732 .. 6836  =  105 bp
AmpR promoter
6732 .. 6836  =  105 bp
EM7 promoter
4892 .. 4939  =  48 bp
synthetic bacterial promoter
EM7 promoter
4892 .. 4939  =  48 bp
synthetic bacterial promoter
V5 tag
3707 .. 3748  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
V5 tag
3707 .. 3748  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
lac UV5 promoter
2099 .. 2129  =  31 bp
3 segments
   Segment 1:  -35  
   2099 .. 2104  =  6 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
2099 .. 2129  =  31 bp
3 segments
   Segment 2:  
   2105 .. 2122  =  18 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
2099 .. 2129  =  31 bp
3 segments
   Segment 3:  -10  
   2123 .. 2129  =  7 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
2099 .. 2129  =  31 bp
3 segments
E. coli lac promoter with an "up" mutation
ORF:  3184 .. 3489  =  306 bp
ORF:  101 amino acids  =  11.7 kDa
ORF:  3763 .. 4602  =  840 bp
ORF:  279 amino acids  =  30.4 kDa
ORF:  5770 .. 6006  =  237 bp
ORF:  78 amino acids  =  8.6 kDa
ORF:  8854 .. 177  =  246 bp
ORF:  81 amino acids  =  8.9 kDa
ORF:  953 .. 1963  =  1011 bp
ORF:  336 amino acids  =  38.5 kDa
ORF:  2183 .. 2842  =  660 bp
ORF:  219 amino acids  =  25.7 kDa
ORF:  3875 .. 4171  =  297 bp
ORF:  98 amino acids  =  10.2 kDa
ORF:  4958 .. 5356  =  399 bp
ORF:  132 amino acids  =  13.7 kDa
ORF:  4182 .. 4496  =  315 bp
ORF:  104 amino acids  =  11.2 kDa
ORF:  4659 .. 4928  =  270 bp
ORF:  89 amino acids  =  9.7 kDa
ORF:  5292 .. 5522  =  231 bp
ORF:  76 amino acids  =  8.5 kDa
ORF:  6837 .. 7697  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  868 .. 1248  =  381 bp
ORF:  126 amino acids  =  13.3 kDa
ORF:  8781 .. 158  =  300 bp
ORF:  99 amino acids  =  10.9 kDa
ORF:  4250 .. 4486  =  237 bp
ORF:  78 amino acids  =  8.3 kDa
ORF:  4904 .. 5260  =  357 bp
ORF:  118 amino acids  =  12.8 kDa
ORF:  7301 .. 7567  =  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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