pLV hUbC-Cas9-T2A-GFP

Lentiviral vector for co-expression of human codon-optimized Cas9 and enhanced GFP (EGFP).

Sequence Author: Gersbach Lab / Addgene #53190

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AmpR promoter SV40 NLS HIV-1 Ψ ATG SV40 NLS pLV hUbC-Cas9-T2A-GFP 14,772 bp
ATG
4369 .. 4371  =  3 bp
1 amino acid  =  149.2 Da
ATG
4369 .. 4371  =  3 bp
1 amino acid  =  149.2 Da
3xFLAG
4372 .. 4437  =  66 bp
22 amino acids  =  2.7 kDa
Product: three tandem FLAG® epitope tags, followed by an enterokinase cleavage site
3xFLAG
4372 .. 4437  =  66 bp
22 amino acids  =  2.7 kDa
Product: three tandem FLAG® epitope tags, followed by an enterokinase cleavage site
SV40 NLS
4444 .. 4464  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
4444 .. 4464  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
Cas9
4474 .. 8577  =  4104 bp
1368 amino acids  =  158.4 kDa
Product: Cas9 (Csn1) endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system
generates RNA-guided double strand breaks in DNA
Cas9
4474 .. 8577  =  4104 bp
1368 amino acids  =  158.4 kDa
Product: Cas9 (Csn1) endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system
generates RNA-guided double strand breaks in DNA
SV40 NLS
8590 .. 8610  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
8590 .. 8610  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
T2A
8617 .. 8670  =  54 bp
18 amino acids  =  1.8 kDa
Product: 2A peptide from Thosea asigna virus capsid protein
Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides.
T2A
8617 .. 8670  =  54 bp
18 amino acids  =  1.8 kDa
Product: 2A peptide from Thosea asigna virus capsid protein
Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides.
EGFP
8677 .. 9390  =  714 bp
238 amino acids  =  26.8 kDa
2 segments
   Segment 1:  1a  
   8677 .. 8679  =  3 bp
   1 amino acid  =  117.1 Da
Product: enhanced GFP
mammalian codon-optimized
EGFP
8677 .. 9390  =  714 bp
238 amino acids  =  26.8 kDa
2 segments
   Segment 2:  
   8680 .. 9390  =  711 bp
   237 amino acids  =  26.7 kDa
Product: enhanced GFP
mammalian codon-optimized
EGFP
8677 .. 9390  =  714 bp
238 amino acids  =  26.8 kDa
2 segments
Product: enhanced GFP
mammalian codon-optimized
UbC promoter
3124 .. 4334  =  1211 bp
human ubiquitin C promoter
UbC promoter
3124 .. 4334  =  1211 bp
human ubiquitin C promoter
AmpR
13,779 .. 14,639  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   13,779 .. 14,570  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
13,779 .. 14,639  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   14,571 .. 14,639  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
13,779 .. 14,639  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
13,020 .. 13,608  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
13,020 .. 13,608  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
WPRE
9464 .. 10,052  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
WPRE
9464 .. 10,052  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
f1 ori
11,059 .. 11,487  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
11,059 .. 11,487  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
CMV enhancer
238 .. 617  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
238 .. 617  =  380 bp
human cytomegalovirus immediate early enhancer
BleoR
11,944 .. 12,318  =  375 bp
124 amino acids  =  13.8 kDa
Product: antibiotic-binding protein
confers resistance to bleomycin, phleomycin, and Zeocin™
BleoR
11,944 .. 12,318  =  375 bp
124 amino acids  =  13.8 kDa
Product: antibiotic-binding protein
confers resistance to bleomycin, phleomycin, and Zeocin™
SV40 promoter
11,501 .. 11,830  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
11,501 .. 11,830  =  330 bp
SV40 enhancer and early promoter
RRE
1680 .. 1913  =  234 bp
The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm.
RRE
1680 .. 1913  =  234 bp
The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm.
bGH poly(A) signal
10,789 .. 11,013  =  225 bp
bovine growth hormone polyadenylation signal
bGH poly(A) signal
10,789 .. 11,013  =  225 bp
bovine growth hormone polyadenylation signal
CMV promoter
619 .. 817  =  199 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
619 .. 817  =  199 bp
human cytomegalovirus (CMV) immediate early promoter
LTR (truncated)
10,577 .. 10,757  =  181 bp
truncated long terminal repeat (LTR) from HIV-1
LTR (truncated)
10,577 .. 10,757  =  181 bp
truncated long terminal repeat (LTR) from HIV-1
LTR (truncated)
835 .. 1015  =  181 bp
truncated long terminal repeat (LTR) from HIV-1
LTR (truncated)
835 .. 1015  =  181 bp
truncated long terminal repeat (LTR) from HIV-1
HIV-1 Ψ
1062 .. 1187  =  126 bp
packaging signal of human immunodeficiency virus type 1
HIV-1 Ψ
1062 .. 1187  =  126 bp
packaging signal of human immunodeficiency virus type 1
SV40 poly(A) signal
12,448 .. 12,569  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
12,448 .. 12,569  =  122 bp
SV40 polyadenylation signal
cPPT/CTS
2440 .. 2557  =  118 bp
central polypurine tract and central termination sequence of HIV-1
cPPT/CTS
2440 .. 2557  =  118 bp
central polypurine tract and central termination sequence of HIV-1
AmpR promoter
14,640 .. 14,744  =  105 bp
AmpR promoter
14,640 .. 14,744  =  105 bp
EM7 promoter
11,878 .. 11,925  =  48 bp
synthetic bacterial promoter
EM7 promoter
11,878 .. 11,925  =  48 bp
synthetic bacterial promoter
loxP
2602 .. 2635  =  34 bp
Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT).
loxP
2602 .. 2635  =  34 bp
Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT).
loxP
9406 .. 9439  =  34 bp
Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT).
loxP
9406 .. 9439  =  34 bp
Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT).
SV40 ori
11,681 .. 11,816  =  136 bp
SV40 origin of replication
SV40 ori
11,681 .. 11,816  =  136 bp
SV40 origin of replication
ORF:  1558 .. 2445  =  888 bp
ORF:  295 amino acids  =  33.8 kDa
ORF:  4369 .. 9399  =  5031 bp
ORF:  1676 amino acids  =  193.0 kDa
ORF:  11,944 .. 12,318  =  375 bp
ORF:  124 amino acids  =  13.8 kDa
ORF:  13,909 .. 14,175  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  9554 .. 10,603  =  1050 bp
ORF:  349 amino acids  =  39.4 kDa
ORF:  9567 .. 10,079  =  513 bp
ORF:  170 amino acids  =  18.0 kDa
ORF:  11,010 .. 11,276  =  267 bp
ORF:  88 amino acids  =  9.3 kDa
ORF:  4300 .. 4953  =  654 bp
ORF:  217 amino acids  =  24.1 kDa
ORF:  5878 .. 6396  =  519 bp
ORF:  172 amino acids  =  20.1 kDa
ORF:  6460 .. 6993  =  534 bp
ORF:  177 amino acids  =  19.9 kDa
ORF:  7987 .. 8520  =  534 bp
ORF:  177 amino acids  =  20.0 kDa
ORF:  8635 .. 9426  =  792 bp
ORF:  263 amino acids  =  26.5 kDa
ORF:  1473 .. 1853  =  381 bp
ORF:  126 amino acids  =  13.3 kDa
ORF:  2622 .. 2948  =  327 bp
ORF:  108 amino acids  =  12.1 kDa
ORF:  11,937 .. 12,224  =  288 bp
ORF:  95 amino acids  =  10.1 kDa
ORF:  13,779 .. 14,639  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  3515 .. 3748  =  234 bp
ORF:  77 amino acids  =  8.4 kDa
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