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Plasmid Files

pL-CRISPR.SFFV.GFP

Lentiviral CRISPR/Cas9 vector with a GFP marker driven by the SFFV promoter, for genome editing in mammalian cells.

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pL-CRISPR.SFFV.GFP.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Ebert Lab / Addgene #57827
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BsmBI (1961) BsmBI (3846) pL-CRISPR.SFFV.GFP 13,735 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  (3846)
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
4636 .. 8736  =  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
4636 .. 8736  =  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
spacer
1962 .. 3846  =  1885 bp
spacer
1962 .. 3846  =  1885 bp
AmpR
11,652 .. 12,512  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   11,652 .. 11,720  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
11,652 .. 12,512  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   11,721 .. 12,512  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
11,652 .. 12,512  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
EGFP
8857 .. 9573  =  717 bp
239 amino acids  =  26.9 kDa
Product: enhanced GFP
mammalian codon-optimized
EGFP
8857 .. 9573  =  717 bp
239 amino acids  =  26.9 kDa
Product: enhanced GFP
mammalian codon-optimized
WPRE
9592 .. 10,180  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
WPRE
9592 .. 10,180  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
ori
12,683 .. 13,271  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
12,683 .. 13,271  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
11,066 .. 11,521  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
11,066 .. 11,521  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
SFFV promoter
4145 .. 4546  =  402 bp
spleen focus-forming virus long terminal repeat (LTR) promoter
SFFV promoter
4145 .. 4546  =  402 bp
spleen focus-forming virus long terminal repeat (LTR) promoter
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)
10,252 .. 10,485  =  234 bp
self-inactivating 3' long terminal repeat (LTR) from HIV-1
3' LTR (ΔU3)
10,252 .. 10,485  =  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
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
10,724 .. 10,859  =  136 bp
SV40 origin of replication
SV40 ori
10,724 .. 10,859  =  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
10,563 .. 10,684  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
10,563 .. 10,684  =  122 bp
SV40 polyadenylation signal
cPPT/CTS
3970 .. 4087  =  118 bp
central polypurine tract and central termination sequence of HIV-1
cPPT/CTS
3970 .. 4087  =  118 bp
central polypurine tract and central termination sequence of HIV-1
AmpR promoter
11,547 .. 11,651  =  105 bp
AmpR promoter
11,547 .. 11,651  =  105 bp
gRNA scaffold
3847 .. 3922  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system
gRNA scaffold
3847 .. 3922  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system
P2A
8800 .. 8856  =  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
8800 .. 8856  =  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
8737 .. 8784  =  48 bp
16 amino acids  =  1.7 kDa
Product: bipartite nuclear localization signal from nucleoplasmin
nucleoplasmin NLS
8737 .. 8784  =  48 bp
16 amino acids  =  1.7 kDa
Product: bipartite nuclear localization signal from nucleoplasmin
FLAG
4561 .. 4584  =  24 bp
8 amino acids  =  1.0 kDa
Product: FLAG® epitope tag, followed by an enterokinase cleavage site
FLAG
4561 .. 4584  =  24 bp
8 amino acids  =  1.0 kDa
Product: FLAG® epitope tag, followed by an enterokinase cleavage site
SV40 NLS
4591 .. 4611  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
4591 .. 4611  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
polIII terminator
3923 .. 3928  =  6 bp
RNA polymerase III transcription terminator
polIII terminator
3923 .. 3928  =  6 bp
RNA polymerase III transcription terminator
ATG
4558 .. 4560  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
4558 .. 4560  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ORF:  955 .. 1830  =  876 bp
ORF:  291 amino acids  =  33.6 kDa
ORF:  4165 .. 4416  =  252 bp
ORF:  83 amino acids  =  9.2 kDa
ORF:  4558 .. 9576  =  5019 bp
ORF:  1672 amino acids  =  192.2 kDa
ORF:  9682 .. 10,191  =  510 bp
ORF:  169 amino acids  =  18.5 kDa
ORF:  10,585 .. 10,821  =  237 bp
ORF:  78 amino acids  =  8.6 kDa
ORF:  13,669 .. 179  =  246 bp
ORF:  81 amino acids  =  8.9 kDa
ORF:  3950 .. 4219  =  270 bp
ORF:  89 amino acids  =  10.4 kDa
ORF:  9695 .. 10,225  =  531 bp
ORF:  176 amino acids  =  18.7 kDa
ORF:  11,652 .. 12,512  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  13,596 .. 160  =  300 bp
ORF:  99 amino acids  =  10.9 kDa
ORF:  12,116 .. 12,382  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  2140 .. 2535  =  396 bp
ORF:  131 amino acids  =  14.0 kDa
ORF:  4096 .. 4380  =  285 bp
ORF:  94 amino acids  =  10.3 kDa
ORF:  4501 .. 7152  =  2652 bp
ORF:  883 amino acids  =  92.8 kDa
ORF:  870 .. 1250  =  381 bp
ORF:  126 amino acids  =  13.3 kDa
ORF:  5982 .. 6275  =  294 bp
ORF:  97 amino acids  =  10.8 kDa
ORF:  6546 .. 6995  =  450 bp
ORF:  149 amino acids  =  17.3 kDa
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