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pLX304-V5-Blast-Empty

Empty control vector for the ORF collection in the pLX304 lentiviral vector.

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pLX304-V5-Blast-Empty Sequence and MappLX304-V5-Blast-Empty.dna
Map and Sequence File   
Sequence Author:  transOMIC
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 PaeR7I - PspXI - XhoI (7662) XbaI * (7465) EcoRI (7461) BsaBI * (7345) KpnI (7042) Acc65I (7038) BspEI (6975) AscI (6870) BlpI (6861) BsmBI - SpeI (6781) XcmI (6553) KflI (6529) AleI (6172) BstAPI (6048) BbvCI - Bpu10I (6019) MfeI (5784) EcoNI (5765) NotI (5743) SphI (4916) AflIII - PciI (4413) TspMI - XmaI (1) SmaI (3) CMV enhancer NdeI (238) NheI (646) BmtI (650) AfeI (651) AgeI (656) PflMI (919) SacII (1181) SfiI (1897) StuI (1943) AvrII (1944) DraIII (2385) ScaI (3044) pLX304-V5-Blast-Empty 7671 bp
PaeR7I  (7662)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (7662)
1 site
V C T C G A G B B G A G C T C V
XhoI  (7662)
1 site
C T C G A G G A G C T C
XbaI  (7465)
1 site
T C T A G A A G A T C T
* Blocked by Dam methylation.
EcoRI  (7461)
1 site
G A A T T C C T T A A G
BsaBI  (7345)
1 site
G A T N N N N A T C C T A N N N N T A G
* Blocked by Dam methylation.
KpnI  (7042)
1 site
G G T A C C C C A T G G
Acc65I  (7038)
1 site
G G T A C C C C A T G G
BspEI  (6975)
1 site
T C C G G A A G G C C T
AscI  (6870)
1 site
G G C G C G C C C C G C G C G G
BlpI  (6861)
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.
BsmBI  (6781)
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.
SpeI  (6781)
1 site
A C T A G T T G A T C A
XcmI  (6553)
1 site
C C A N N N N N N N N N T G G G G T N N N N N N N N N A C C

The 1-base overhangs produced by XcmI may be hard to ligate.
Sticky ends from different XcmI sites may not be compatible.
KflI  (6529)
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.
AleI  (6172)
1 site
C A C N N N N G T G G T G N N N N C A C
BstAPI  (6048)
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.
BbvCI  (6019)
1 site
C C T C A G C G G A G T C G
Bpu10I  (6019)
1 site
C C T N A G C G G A N T C G

Efficient cleavage requires at least two copies of the Bpu10I
recognition sequence.
This recognition sequence is asymmetric, so ligating sticky ends
generated by Bpu10I will not always regenerate a Bpu10I site.
Sticky ends from different Bpu10I sites may not be compatible.
MfeI  (5784)
1 site
C A A T T G G T T A A C
EcoNI  (5765)
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.
NotI  (5743)
1 site
G C G G C C G C C G C C G G C G
SphI  (4916)
1 site
G C A T G C C G T A C G
AflIII  (4413)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
PciI  (4413)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
TspMI  (1)
1 site
C C C G G G G G G C C C
XmaI  (1)
1 site
C C C G G G G G G C C C

Efficient cleavage requires at least two copies of the XmaI
recognition sequence.
Full cleavage with XmaI may require a long incubation.
SmaI  (3)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
NdeI  (238)
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.
NheI  (646)
1 site
G C T A G C C G A T C G
BmtI  (650)
1 site
G C T A G C C G A T C G
AfeI  (651)
1 site
A G C G C T T C G C G A
AgeI  (656)
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.
PflMI  (919)
1 site
C C A N N N N N T G G G G T N N N N N A C C

Sticky ends from different PflMI sites may not be compatible.
SacII  (1181)
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.
SfiI  (1897)
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.
StuI  (1943)
1 site
A G G C C T T C C G G A
AvrII  (1944)
1 site
C C T A G G G G A T C C
DraIII  (2385)
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.
ScaI  (3044)
1 site
A G T A C T T C A T G A
AmpR
2738 .. 3598  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   2738 .. 2806  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2738 .. 3598  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   2807 .. 3598  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2738 .. 3598  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
WPRE
678 .. 1266  =  589 bp
woodchuck hepatitis virus posttranscriptional
regulatory element
WPRE
678 .. 1266  =  589 bp
woodchuck hepatitis virus posttranscriptional
regulatory element
ori
3769 .. 4357  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
3769 .. 4357  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
hPGK promoter
6535 .. 7035  =  501 bp
human phosphoglycerate kinase 1 promoter
hPGK promoter
6535 .. 7035  =  501 bp
human phosphoglycerate kinase 1 promoter
f1 ori
2152 .. 2607  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
2152 .. 2607  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
BSD
7047 .. 7442  =  396 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
BSD
7047 .. 7442  =  396 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
CMV enhancer
65 .. 368  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
65 .. 368  =  304 bp
human cytomegalovirus immediate early enhancer
3' LTR (ΔU3)
1338 .. 1571  =  234 bp
self-inactivating 3' long terminal repeat (LTR) from
HIV-1
3' LTR (ΔU3)
1338 .. 1571  =  234 bp
self-inactivating 3' long terminal repeat (LTR) from
HIV-1
RRE
5898 .. 6131  =  234 bp
The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to
the cytoplasm.
RRE
5898 .. 6131  =  234 bp
The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to
the cytoplasm.
RSV promoter
4826 .. 5052  =  227 bp
Rous sarcoma virus enhancer/promoter
RSV promoter
4826 .. 5052  =  227 bp
Rous sarcoma virus enhancer/promoter
CMV promoter
369 .. 572  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
369 .. 572  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
5' LTR (truncated)
5053 .. 5233  =  181 bp
truncated 5' long terminal repeat (LTR) from HIV-1
5' LTR (truncated)
5053 .. 5233  =  181 bp
truncated 5' long terminal repeat (LTR) from HIV-1
SV40 ori
1810 .. 1945  =  136 bp
SV40 origin of replication
SV40 ori
1810 .. 1945  =  136 bp
SV40 origin of replication
HIV-1 Ψ
5280 .. 5405  =  126 bp
packaging signal of human immunodeficiency virus
type 1
HIV-1 Ψ
5280 .. 5405  =  126 bp
packaging signal of human immunodeficiency virus
type 1
SV40 poly(A) signal
1649 .. 1770  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1649 .. 1770  =  122 bp
SV40 polyadenylation signal
cPPT/CTS
7502 .. 7619  =  118 bp
central polypurine tract and central termination
sequence of HIV-1
cPPT/CTS
7502 .. 7619  =  118 bp
central polypurine tract and central termination
sequence of HIV-1
AmpR promoter
2633 .. 2737  =  105 bp
AmpR promoter
2633 .. 2737  =  105 bp
V5 tag
595 .. 636  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
V5 tag
595 .. 636  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
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