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Plasmid Files

pLenti6/capTEV™-NT-DEST1

Lentiviral Gateway® destination vector for expression of N-terminally capTEV™-tagged proteins.

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pLenti6 capTEV-NT-DEST1 Sequence and MappLenti6 capTEV-NT-DEST1.dna
Map and Sequence File   
Sequence Author:  Invitrogen (Life Technologies)
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 PciI (8677) AhdI (7789) DraIII (6649) SfiI (6161) KpnI (5541) Acc65I (5537) BlpI (5518) EM7 promoter PmlI (5054) BclI * (5045) SexAI * (4761) AgeI (4642) BstBI (4592) SacII (4588) ApaI (4585) PspOMI (4581) XbaI (4575) EcoNI (942) KflI (1706) BspDI - ClaI (1795) HpaI (1808) SpeI (1886) NdeI (2121) SnaBI (2227) BstEII (2527) BspEI (3282) BsmBI (3510) PasI (3518) BstZ17I (3821) SrfI (4157) BmgBI (4191) BfuAI - BspMI (4401) PaeR7I - PspXI - XhoI (4569) pLenti6/capTEV™-NT-DEST1 9087 bp
PciI  (8677)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AhdI  (7789)
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  (6649)
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.
SfiI  (6161)
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  (5541)
1 site
G G T A C C C C A T G G
Acc65I  (5537)
1 site
G G T A C C C C A T G G
BlpI  (5518)
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  (5054)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
BclI  (5045)
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  (4761)
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.
AgeI  (4642)
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.
BstBI  (4592)
1 site
T T C G A A A A G C T T
SacII  (4588)
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.
ApaI  (4585)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
PspOMI  (4581)
1 site
G G G C C C C C C G G G
XbaI  (4575)
1 site
T C T A G A A G A T C T
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.
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
HpaI  (1808)
1 site
G T T A A C C A A T T G
SpeI  (1886)
1 site
A C T A G T T G A T C A
NdeI  (2121)
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  (2227)
1 site
T A C G T A A T G C A T
BstEII  (2527)
1 site
G G T N A C C C C A N T G G

Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
BspEI  (3282)
1 site
T C C G G A A G G C C T
BsmBI  (3510)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different BsmBI sites may not be compatible.
PasI  (3518)
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  (3821)
1 site
G T A T A C C A T A T G
SrfI  (4157)
1 site
G C C C G G G C C G G G C C C G
BmgBI  (4191)
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.
BfuAI  (4401)
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  (4401)
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.
PaeR7I  (4569)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (4569)
1 site
V C T C G A G B B G A G C T C V
XhoI  (4569)
1 site
C T C G A G G A G C T C
AmpR
7002 .. 7862  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   7002 .. 7070  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
7002 .. 7862  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   7071 .. 7862  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
7002 .. 7862  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
CmR
3073 .. 3732  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
3073 .. 3732  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
ori
8033 .. 8621  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
8033 .. 8621  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
6416 .. 6871  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
6416 .. 6871  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
BSD
5123 .. 5521  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
BSD
5123 .. 5521  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
CMV enhancer
1872 .. 2251  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
1872 .. 2251  =  380 bp
human cytomegalovirus immediate early enhancer
SV40 promoter
4680 .. 5009  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
4680 .. 5009  =  330 bp
SV40 enhancer and early promoter
ccdB
4074 .. 4379  =  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
4074 .. 4379  =  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.
capTEV™
2510 .. 2812  =  303 bp
101 amino acids  =  10.6 kDa
   Segment 1:  BioEase™ tag  
   2510 .. 2728  =  219 bp
   73 amino acids  =  7.3 kDa
Product: composite tag for in vivo biotinylation, TEV
protease cleavage, and 6xHis tagging
capTEV™
2510 .. 2812  =  303 bp
101 amino acids  =  10.6 kDa
   Segment 2:  
   2729 .. 2740  =  12 bp
   4 amino acids  =  306.3 Da
Product: composite tag for in vivo biotinylation, TEV
protease cleavage, and 6xHis tagging
capTEV™
2510 .. 2812  =  303 bp
101 amino acids  =  10.6 kDa
   Segment 3:  TEV site  
   2741 .. 2761  =  21 bp
   7 amino acids  =  870.0 Da
Product: composite tag for in vivo biotinylation, TEV
protease cleavage, and 6xHis tagging
capTEV™
2510 .. 2812  =  303 bp
101 amino acids  =  10.6 kDa
   Segment 4:  
   2762 .. 2767  =  6 bp
   2 amino acids  =  259.3 Da
Product: composite tag for in vivo biotinylation, TEV
protease cleavage, and 6xHis tagging
capTEV™
2510 .. 2812  =  303 bp
101 amino acids  =  10.6 kDa
   Segment 5:  TEV site  
   2768 .. 2788  =  21 bp
   7 amino acids  =  870.0 Da
Product: composite tag for in vivo biotinylation, TEV
protease cleavage, and 6xHis tagging
capTEV™
2510 .. 2812  =  303 bp
101 amino acids  =  10.6 kDa
   Segment 6:  
   2789 .. 2794  =  6 bp
   2 amino acids  =  259.3 Da
Product: composite tag for in vivo biotinylation, TEV
protease cleavage, and 6xHis tagging
capTEV™
2510 .. 2812  =  303 bp
101 amino acids  =  10.6 kDa
   Segment 7:  6xHis  
   2795 .. 2812  =  18 bp
   6 amino acids  =  840.9 Da
Product: composite tag for in vivo biotinylation, TEV
protease cleavage, and 6xHis tagging
capTEV™
2510 .. 2812  =  303 bp
101 amino acids  =  10.6 kDa
7 segments
Product: composite tag for in vivo biotinylation, TEV
protease cleavage, and 6xHis tagging
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)
5608 .. 5841  =  234 bp
self-inactivating 3' long terminal repeat (LTR) from
HIV-1
3' LTR (ΔU3)
5608 .. 5841  =  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
2252 .. 2455  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
2252 .. 2455  =  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
2840 .. 2964  =  125 bp
recombination site for the Gateway® LR reaction
attR1
2840 .. 2964  =  125 bp
recombination site for the Gateway® LR reaction
attR2
4420 .. 4544  =  125 bp
recombination site for the Gateway® LR reaction
attR2
4420 .. 4544  =  125 bp
recombination site for the Gateway® LR reaction
SV40 poly(A) signal
5913 .. 6034  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
5913 .. 6034  =  122 bp
SV40 polyadenylation signal
AmpR promoter
6897 .. 7001  =  105 bp
AmpR promoter
6897 .. 7001  =  105 bp
EM7 promoter
5057 .. 5104  =  48 bp
synthetic bacterial promoter
EM7 promoter
5057 .. 5104  =  48 bp
synthetic bacterial promoter
V5 tag
4597 .. 4638  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
V5 tag
4597 .. 4638  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
lac UV5 promoter
2989 .. 3019  =  31 bp
   Segment 1:  -35  
   2989 .. 2994  =  6 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
2989 .. 3019  =  31 bp
   Segment 2:  
   2995 .. 3012  =  18 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
2989 .. 3019  =  31 bp
   Segment 3:  -10  
   3013 .. 3019  =  7 bp
E. coli lac promoter with an "up" mutation
lac UV5 promoter
2989 .. 3019  =  31 bp
3 segments
E. coli lac promoter with an "up" mutation
SV40 ori
4860 .. 4995  =  136 bp
SV40 origin of replication
SV40 ori
4860 .. 4995  =  136 bp
SV40 origin of replication
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