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Plasmid Files

pL-CRISPR.EFS.GFP

Lentiviral CRISPR/Cas9 vector with a GFP marker driven by the EF-1α core promoter, for genome editing in mammalian cells.

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pL-CRISPR.EFS.GFP Sequence and MappL-CRISPR.EFS.GFP.dna
Map and Sequence File   
Sequence Author:  Ebert Lab / Addgene #57818
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 BsmBI (1961) BsmBI (1983) polIII terminator ATG FLAG SV40 NLS pL-CRISPR.EFS.GFP 11,707 bp
BsmBI  (1961)
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  (1983)
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.
Cas9
2608 .. 6708  =  4101 bp
1367 amino acids  =  158.3 kDa
Product: Cas9 (Csn1) endonuclease from the
Streptococcus pyogenes Type II CRISPR/Cas system
generates RNA-guided double strand breaks in DNA
Cas9
2608 .. 6708  =  4101 bp
1367 amino acids  =  158.3 kDa
Product: Cas9 (Csn1) endonuclease from the
Streptococcus pyogenes Type II CRISPR/Cas system
generates RNA-guided double strand breaks in DNA
AmpR
9624 .. 10,484  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   9624 .. 9692  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
9624 .. 10,484  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   9693 .. 10,484  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
9624 .. 10,484  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
EGFP
6829 .. 7545  =  717 bp
239 amino acids  =  26.9 kDa
Product: enhanced GFP
mammalian codon-optimized
EGFP
6829 .. 7545  =  717 bp
239 amino acids  =  26.9 kDa
Product: enhanced GFP
mammalian codon-optimized
WPRE
7564 .. 8152  =  589 bp
woodchuck hepatitis virus posttranscriptional
regulatory element
WPRE
7564 .. 8152  =  589 bp
woodchuck hepatitis virus posttranscriptional
regulatory element
ori
10,655 .. 11,243  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
10,655 .. 11,243  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
9038 .. 9493  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
9038 .. 9493  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
U6 promoter
1717 .. 1957  =  241 bp
RNA polymerase III promoter for human U6 snRNA
U6 promoter
1717 .. 1957  =  241 bp
RNA polymerase III promoter for human U6 snRNA
RRE
1077 .. 1310  =  234 bp
The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to
the cytoplasm.
RRE
1077 .. 1310  =  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)
8224 .. 8457  =  234 bp
self-inactivating 3' long terminal repeat (LTR) from
HIV-1
3' LTR (ΔU3)
8224 .. 8457  =  234 bp
self-inactivating 3' long terminal repeat (LTR) from
HIV-1
RSV promoter
5 .. 231  =  227 bp
Rous sarcoma virus enhancer/promoter
RSV promoter
5 .. 231  =  227 bp
Rous sarcoma virus enhancer/promoter
EF-1α core promoter
2293 .. 2504  =  212 bp
core promoter for human elongation factor EF-1α
EF-1α core promoter
2293 .. 2504  =  212 bp
core promoter for human elongation factor EF-1α
5' LTR (truncated)
232 .. 412  =  181 bp
truncated 5' long terminal repeat (LTR) from HIV-1
5' LTR (truncated)
232 .. 412  =  181 bp
truncated 5' long terminal repeat (LTR) from HIV-1
SV40 ori
8696 .. 8831  =  136 bp
SV40 origin of replication
SV40 ori
8696 .. 8831  =  136 bp
SV40 origin of replication
HIV-1 Ψ
459 .. 584  =  126 bp
packaging signal of human immunodeficiency virus
type 1
HIV-1 Ψ
459 .. 584  =  126 bp
packaging signal of human immunodeficiency virus
type 1
SV40 poly(A) signal
8535 .. 8656  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
8535 .. 8656  =  122 bp
SV40 polyadenylation signal
cPPT/CTS
2107 .. 2224  =  118 bp
central polypurine tract and central termination
sequence of HIV-1
cPPT/CTS
2107 .. 2224  =  118 bp
central polypurine tract and central termination
sequence of HIV-1
AmpR promoter
9519 .. 9623  =  105 bp
AmpR promoter
9519 .. 9623  =  105 bp
gRNA scaffold
1984 .. 2059  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system
gRNA scaffold
1984 .. 2059  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system
P2A
6772 .. 6828  =  57 bp
19 amino acids  =  2.0 kDa
Product: 2A peptide from porcine teschovirus-1
polyprotein
Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides.
P2A
6772 .. 6828  =  57 bp
19 amino acids  =  2.0 kDa
Product: 2A peptide from porcine teschovirus-1
polyprotein
Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides.
nucleoplasmin NLS
6709 .. 6756  =  48 bp
16 amino acids  =  1.7 kDa
Product: bipartite nuclear localization signal from
nucleoplasmin
nucleoplasmin NLS
6709 .. 6756  =  48 bp
16 amino acids  =  1.7 kDa
Product: bipartite nuclear localization signal from
nucleoplasmin
FLAG
2533 .. 2556  =  24 bp
8 amino acids  =  1.0 kDa
Product: FLAG® epitope tag, followed by an
enterokinase cleavage site
FLAG
2533 .. 2556  =  24 bp
8 amino acids  =  1.0 kDa
Product: FLAG® epitope tag, followed by an
enterokinase cleavage site
SV40 NLS
2563 .. 2583  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T
antigen
SV40 NLS
2563 .. 2583  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T
antigen
polIII terminator
2060 .. 2065  =  6 bp
RNA polymerase III transcription terminator
polIII terminator
2060 .. 2065  =  6 bp
RNA polymerase III transcription terminator
ATG
2530 .. 2532  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
2530 .. 2532  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
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