Resources
Plasmid Files

pLenti6/BLOCK-iT™-DEST

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

To see this sequence with restriction sites, features, and translations, please download
 SnapGene or the free  SnapGene Viewer.

pLenti6 BLOCK-iT-DEST Sequence and MappLenti6 BLOCK-iT-DEST.dna
Map and Sequence File   
Sequence Author:  Invitrogen (Life Technologies)
Download Free Trial Get SnapGene Viewer

 KasI (413) PciI (8265) DraIII (6237) NaeI (6131) NgoMIV (6129) SfiI (5749) KpnI (5129) Acc65I (5125) BlpI (5106) EM7 promoter PmlI (4642) BclI * (4633) 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

PmlI gradually loses activity when stored at -20°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.
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

AgeI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
AmpR
6590 .. 7450  =  861 bp
286 amino acids  =  31.6 kDa
   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
   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
Try SnapGene and create your own beautiful maps

Individual Sequences & Maps

SnapGene offers the fastest and easiest way to plan, visualize, and document your molecular biology procedures.

Priced accessibly so that everyone in your lab can have a license.

Learn More...

SnapGene Viewer is a versatile tool for creating and sharing richly annotated sequence files. It opens many common file formats.

Free! Because there should be no barriers to seeing your data.

Learn More...

The map, notes, and annotations on this page and in the sequence/map file are copyrighted material. This material may be used without restriction by academic, nonprofit, and governmental entities, except that the source must be cited as "www.snapgene.com/resources". Commercial entities must contact GSL Biotech LLC for permission and terms of use.

Copyright © 2016 GSL Biotech LLC | Site Map | Privacy | Legal Disclaimers   Subscribe to Our Newsletter