pLenti6 BLOCK-iT-DEST

Lentiviral Gateway® destination vector for an shRNA cassette from a U6 entry vector.

Sequence Author: Thermo Fisher (Invitrogen)

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HomePlasmidsGateway® Cloning VectorspLenti6 BLOCK-iT-DEST
KasI (413) PciI (8265) DraIII (6237) NaeI (6131) NgoMIV (6129) SfiI (5749) KpnI (5129) Acc65I (5125) BlpI (5106) EM7 promoter PmlI (4642) BclI * (4633) CsiI - SexAI * (4349) NarI (414) SfoI (415) PluTI (417) EcoNI (942) MfeI (961) AleI (1349) KflI (1706) NheI (1797) BmtI (1801) AfeI (1802) SpeI (1827) BspDI - ClaI (1834) HpaI (1843) BfuAI - BspMI (2394) BmgBI (2608) SrfI (2642) BstZ17I (2978) BamHI (3056) BspEI (3513) PaeR7I - PspXI - XhoI (4157) XbaI (4163) PspOMI (4169) ApaI (4173) SacII (4176) MluI (4224) AgeI (4230) pLenti6/BLOCK-iT™-DEST 8675 bp
KasI  (413)
1 site		 G G C G C C C C G C G G
PciI  (8265)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
DraIII  (6237)
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  (6131)
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  (6129)
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.
SfiI  (5749)
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.
KpnI  (5129)
1 site		 G G T A C C C C A T G G
Acc65I  (5125)
1 site		 G G T A C C C C A T G G
BlpI  (5106)
1 site		 G C T N A G C C G A N T C G
Sticky ends from different BlpI sites may not be compatible.
PmlI  (4642)
1 site		 C A C G T G G T G C A C
BclI  (4633)
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  (4349)
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  (4349)
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.
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
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.
NheI  (1797)
1 site		 G C T A G C C G A T C G
BmtI  (1801)
1 site		 G C T A G C C G A T C G
AfeI  (1802)
1 site		 A G C G C T T C G C G A
SpeI  (1827)
1 site		 A C T A G T T G A T C A
BspDI  (1834)
1 site		 A T C G A T T A G C T A
ClaI  (1834)
1 site		 A T C G A T T A G C T A
HpaI  (1843)
1 site		 G T T A A C C A A T T G
BfuAI  (2394)
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  (2394)
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.
BmgBI  (2608)
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.
SrfI  (2642)
1 site		 G C C C G G G C C G G G C C C G
BstZ17I  (2978)
1 site		 G T A T A C C A T A T G
BamHI  (3056)
1 site		 G G A T C C C C T A G G
After cleavage, BamHI-HF® (but not the original BamHI) can remain bound to DNA and alter its electrophoretic mobility.
BspEI  (3513)
1 site		 T C C G G A A G G C C T
PaeR7I  (4157)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (4157)
1 site		 V C T C G A G B B G A G C T C V
XhoI  (4157)
1 site		 C T C G A G G A G C T C
XbaI  (4163)
1 site		 T C T A G A A G A T C T
PspOMI  (4169)
1 site		 G G G C C C C C C G G G
ApaI  (4173)
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  (4176)
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.
MluI  (4224)
1 site		 A C G C G T T G C G C A
AgeI  (4230)
1 site		 A C C G G T T G G C C A
AmpR
6590 .. 7450  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   6590 .. 6658  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6590 .. 7450  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   6659 .. 7450  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6590 .. 7450  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
CmR
3068 .. 3727  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
3068 .. 3727  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
ori
7621 .. 8209  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
7621 .. 8209  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
6004 .. 6459  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
6004 .. 6459  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
BSD
4711 .. 5109  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
BSD
4711 .. 5109  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
SV40 promoter
4268 .. 4597  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
4268 .. 4597  =  330 bp
SV40 enhancer and early promoter
ccdB
2421 .. 2726  =  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
2421 .. 2726  =  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)
5196 .. 5429  =  234 bp
self-inactivating 3' long terminal repeat (LTR) from HIV-1
3' LTR (ΔU3)
5196 .. 5429  =  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
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
1868 .. 1992  =  125 bp
recombination site for the Gateway® LR reaction
attR1
1868 .. 1992  =  125 bp
recombination site for the Gateway® LR reaction
attR2
4008 .. 4132  =  125 bp
recombination site for the Gateway® LR reaction
attR2
4008 .. 4132  =  125 bp
recombination site for the Gateway® LR reaction
SV40 poly(A) signal
5501 .. 5622  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
5501 .. 5622  =  122 bp
SV40 polyadenylation signal
AmpR promoter
6485 .. 6589  =  105 bp
AmpR promoter
6485 .. 6589  =  105 bp
cat promoter
3728 .. 3830  =  103 bp
promoter of the E. coli cat gene
cat promoter
3728 .. 3830  =  103 bp
promoter of the E. coli cat gene
EM7 promoter
4645 .. 4692  =  48 bp
synthetic bacterial promoter
EM7 promoter
4645 .. 4692  =  48 bp
synthetic bacterial promoter
V5 tag
4185 .. 4226  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
V5 tag
4185 .. 4226  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
SV40 ori
4448 .. 4583  =  136 bp
SV40 origin of replication
SV40 ori
4448 .. 4583  =  136 bp
SV40 origin of replication
ORF:  4711 .. 5109  =  399 bp
ORF:  132 amino acids  =  13.7 kDa
ORF:  953 .. 1801  =  849 bp
ORF:  282 amino acids  =  32.5 kDa
ORF:  5045 .. 5275  =  231 bp
ORF:  76 amino acids  =  8.5 kDa
ORF:  6590 .. 7450  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  5523 .. 5759  =  237 bp
ORF:  78 amino acids  =  8.6 kDa
ORF:  8607 .. 177  =  246 bp
ORF:  81 amino acids  =  8.9 kDa
ORF:  8534 .. 158  =  300 bp
ORF:  99 amino acids  =  10.9 kDa
ORF:  2421 .. 2726  =  306 bp
ORF:  101 amino acids  =  11.7 kDa
ORF:  3068 .. 3727  =  660 bp
ORF:  219 amino acids  =  25.7 kDa
ORF:  868 .. 1248  =  381 bp
ORF:  126 amino acids  =  13.3 kDa
ORF:  4657 .. 5013  =  357 bp
ORF:  118 amino acids  =  12.8 kDa
ORF:  7054 .. 7320  =  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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