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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HomePlasmidsViral Expression & Packaging 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 hepatitis virus posttranscriptional regulatory element
WPRE
3772 .. 4360  =  589 bp
woodchuck hepatitis virus 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

pAAV-AC-GFP
pLenti6.2 N-Lumio V5-DEST
pLVX-rHom-1
pRetroX-Tet3G
pAAV-AC-Myc-DDK
pLenti6.2 V5-DEST
pLVX-rHom-Nuc1
pRetroX-TetOne
pAAV-AC-RFP
pLenti6.3 TO V5-DEST
pLVX-rHom-Sec1
pRetroX-TetOne-Puro
pAAV2-EF1a-tGFP-WPRE
pLenti6.3 V5-DEST
pLVX-Tet-Off Advanced
pRetroX-Tight-Hyg
pAd BLOCK-iT-DEST
pLenti6.4 R4R2 V5-DEST
pLVX-Tet-On Advanced
pRetroX-Tight-Pur
pAd CMV V5-DEST
pLenti6 BLOCK-iT-DEST
pLVX-Tet3G
pRetroX-TRE3G
pAd PL-DEST
pLenti6 capTEV-CT-DEST
pLVX-TetOne
pRFP-C-RS
pAdenoX-CMV Linear
pLenti6 capTEV-NT-DEST1
pLVX-TetOne-Puro
pRFP-CB-shLenti
pAdenoX-DsRed-Express Linear
pLenti6 TR
pLVX-Tight-Puro
pRS
pAdenoX-PRLS Linear
pLenti6 UbC V5-DEST
pLVX-TRE3G
pRSGC1-U6-sg-HTS6-CMV-Cas9-2A-Puro
pAdenoX-PRLS-DsRed-Express Linear
pLenti6 V5-DEST
pLVX-TRE3G-Hom1
pRSGC2-U6-sg-HTS6-UbiC-Cas9-2A-Puro
pAdenoX-PRLS-ZsGreen1 Linear
pLenti7.3 V5-DEST
pLVX-TRE3G-IRES
pRSI12-U6-sh-HTS4-UbiC-TagRFP-2A-Puro
pAdenoX-Tet3G Linear
pLEX-MCS
pLVX-TRE3G-mCherry
pRSI16cb-U6-sh-13kCB18-HTS6-UbiC-TagRFP-2A-Puro
pAdenoX-ZsGreen1 Linear
pLHCX
pLVX-TRE3G-ZsGreen1
pRSI16-U6-sh-HTS6-UbiC-TagRFP-2A-Puro
pB-RS
pLKO.1
pLX301
pRSI17cb-U6-sh-13kCB18-HTS6-UbiC-TagGFP2-2A-Puro
pBABE-Hygro
pLKO.1 puro
pLX302
pRSI17-U6-sh-HTS6-UbiC-TagGFP2-2A-Puro
pBABE-Neo
pLMN-ZsGreen-Neomycin
pLX303
pRSIT12-U6Tet-sh-HTS4-CMV-TetRep-2A-TagRFP-2A-Puro
pBABE-Puro
pLMPd-Ametrine
pLX304
pRSIT16-U6Tet-sh-HTS6-CMV-TetRep-2A-TagRFP-2A-Puro
pBABE-Zeo
pLNCX2
pLX304-V5-Blast-Empty
pRSIT17-U6Tet-sh-HTS6-CMV-TetRep-2A-TagGFP2-2A-Puro
pCMV-VSV-G
pLP VSVG
pLXIN
pRSITEP-U6Tet-sh-noHTS-EF1-TetRep-2A-Puro
pGFP-B-RS
pLP1
pLXSN
pRSITUR-U6Tet-sh-HTS3-UbiC-TetRep-2A-TagRFP
pGFP-C-shLenti
pLP2
pMD2.G
pShuttle2
pGFP-V-RS
pLPCX
pMSCV16URP-U6-sh-HTS6-UbiC-TagRFP-2A-Puro
pSIREN-RetroQ
pGIPZ
pLVX-EF1alpha-IRES-mCherry
pMSCVhyg
pSIREN-RetroQ-DsRed-Express
pLenti-C-GFP
pLVX-EF1alpha-IRES-Puro
pMSCVneo
pSIREN-RetroQ-TetH
pLenti-C-mGFP
pLVX-EF1alpha-IRES-ZsGreen1
pMSCVpuro
pSIREN-RetroQ-TetP
pLenti-C-Myc-DDK
pLVX-EF1alpha-Tet3G
pQC-tTS-IN
pSIREN-RetroQ-ZsGreen1
pLenti-C-Myc-DDK-IRES-BSD
pLVX-Het-1
pQCXIH
psPAX2
pLenti-C-Myc-DDK-IRES-GFP
pLVX-Het-2
pQCXIN
pTRE3G-Hom1
pLenti-C-Myc-DDK-IRES-Neo
pLVX-Het-Mem1
pQCXIP
pTRIPZ
pLenti-C-Myc-DDK-IRES-Puro
pLVX-Het-Nuc1
pQCXIX
ptTS-Neo
pLenti-C-Myc-DDK-IRES-tRFP
pLVX-Hom-1
pRetro-Lib
pZIP-hCMV-RFP-Puro
pLenti-C-tRFP
pLVX-Hom-Mem1
pRetroQ-AcGFP1-C1
pZIP-hCMV-ZSGreen-Puro
pLenti-C-tYFP
pLVX-Hom-Nuc1
pRetroQ-AcGFP1-N1
pZIP-hEF1-alpha-RFP-Puro
pLenti-EF1a-C-mGFP
pLVX-IRES-Hyg
pRetroQ-DsRed-Monomer-C1
pZIP-hEF1-alpha-ZsGreen-Puro
pLenti-EF1a-C-Myc-DDK-IRES-Puro
pLVX-IRES-mCherry
pRetroQ-DsRed-Monomer-N1
pZIP-mCMV-RFP-Puro
pLenti-EF1a-C-tGFP
pLVX-IRES-Neo
pRetroQ-mCherry-C1
pZIP-mCMV-ZsGreen-Puro
pLenti-N-tGFP
pLVX-IRES-Puro
pRetroQ-mCherry-N1
pZIP-mEF1-alpha-RFP-Puro
pLenti-N-tRFP
pLVX-IRES-tdTomato
pRetroX-IRES-DsRedExpress
pZIP-mEF1-alpha-ZsGreen-Puro
pLenti-N-tYFP
pLVX-IRES-ZsGreen1
pRetroX-IRES-ZsGreen1
pZIP-SFFV-RFP-Puro
pLenti3.3 TR
pLVX-PTuner
pRetroX-PTuner
pZIP-SFFV-ZsGreen-Puro
pLenti4 BLOCK-iT-DEST
pLVX-PTuner Green
pRetroX-PTuner IRES
pZIP-TRE3G-ZsGreen-Puro
pLenti4 V5-DEST
pLVX-PTunerC
pRetroX-Tet-Off Advanced
pSIREN-Shuttle (linearized)
pLenti6.2 C-Lumio V5-DEST
pLVX-Puro
pRetroX-Tet-On Advanced
scramble shRNA

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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