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Plasmid Files

lentiGuide-Puro

Zhang lab lentiviral vector, also known as pLentiGuide-Puro, for expressing a single guide RNA (sgRNA) in Cas9-expressing cells. Use together with lentiCas9‑Blast.

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lentiGuide-Puro Sequence and MaplentiGuide-Puro.dna
Map and Sequence File   
Sequence Author:  Zhang Lab / Addgene #52963
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 BsmBI (9882) BsmBI (7997) polIII terminator lentiGuide-Puro 10,183 bp
BsmBI  (9882)
2 sites
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.
BsmBI  (7997)
2 sites
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.
filler
7998 .. 9882  =  1885 bp
digest with BsmBI to replace the filler with an
sgRNA
filler
7998 .. 9882  =  1885 bp
digest with BsmBI to replace the filler with an
sgRNA
EF-1α promoter
87 .. 1265  =  1179 bp
strong constitutive promoter for human elongation
factor EF-1α
EF-1α promoter
87 .. 1265  =  1179 bp
strong constitutive promoter for human elongation
factor EF-1α
AmpR
3953 .. 4813  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   3953 .. 4021  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3953 .. 4813  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   4022 .. 4813  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3953 .. 4813  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
PuroR
1278 .. 1877  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
1278 .. 1877  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
WPRE
1893 .. 2481  =  589 bp
woodchuck hepatitis virus posttranscriptional
regulatory element
WPRE
1893 .. 2481  =  589 bp
woodchuck hepatitis virus posttranscriptional
regulatory element
ori
4984 .. 5572  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
4984 .. 5572  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
3367 .. 3822  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
3367 .. 3822  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
U6 promoter
7753 .. 7993  =  241 bp
RNA polymerase III promoter for human U6 snRNA
U6 promoter
7753 .. 7993  =  241 bp
RNA polymerase III promoter for human U6 snRNA
3' LTR (ΔU3)
2553 .. 2786  =  234 bp
self-inactivating 3' long terminal repeat (LTR) from
HIV-1
3' LTR (ΔU3)
2553 .. 2786  =  234 bp
self-inactivating 3' long terminal repeat (LTR) from
HIV-1
RRE
7113 .. 7346  =  234 bp
The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to
the cytoplasm.
RRE
7113 .. 7346  =  234 bp
The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to
the cytoplasm.
RSV promoter
6041 .. 6267  =  227 bp
Rous sarcoma virus enhancer/promoter
RSV promoter
6041 .. 6267  =  227 bp
Rous sarcoma virus enhancer/promoter
5' LTR (truncated)
6268 .. 6448  =  181 bp
truncated 5' long terminal repeat (LTR) from HIV-1
5' LTR (truncated)
6268 .. 6448  =  181 bp
truncated 5' long terminal repeat (LTR) from HIV-1
SV40 ori
3025 .. 3160  =  136 bp
SV40 origin of replication
SV40 ori
3025 .. 3160  =  136 bp
SV40 origin of replication
HIV-1 Ψ
6495 .. 6620  =  126 bp
packaging signal of human immunodeficiency virus
type 1
HIV-1 Ψ
6495 .. 6620  =  126 bp
packaging signal of human immunodeficiency virus
type 1
SV40 poly(A) signal
2864 .. 2985  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2864 .. 2985  =  122 bp
SV40 polyadenylation signal
cPPT/CTS
10,014 .. 10,131  =  118 bp
central polypurine tract and central termination
sequence of HIV-1
cPPT/CTS
10,014 .. 10,131  =  118 bp
central polypurine tract and central termination
sequence of HIV-1
AmpR promoter
3848 .. 3952  =  105 bp
AmpR promoter
3848 .. 3952  =  105 bp
gRNA scaffold
9883 .. 9958  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system
gRNA scaffold
9883 .. 9958  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system
polIII terminator
9959 .. 9964  =  6 bp
RNA polymerase III transcription terminator
polIII terminator
9959 .. 9964  =  6 bp
RNA polymerase III transcription terminator
EF-1α intron A
318 .. 1256  =  939 bp
intron upstream of the start codon of human EF-1α
EF-1α intron A
318 .. 1256  =  939 bp
intron upstream of the start codon of human EF-1α
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