pLSC-5

Zhang lab lentiviral vector for expressing a single guide RNA (sgRNA) together with a split catalytically inactive Cas9 that is chemically inducible.

Sequence Author: Zhang Lab / Addgene #62889

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CMV enhancer cPPT/CTS BsmBI (2852) BsmBI (2872) polIII terminator EF-1α core promoter SV40 NLS pLSC-5 13,792 bp
BsmBI  (2852)
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-v2 is an improved version of BsmBI.
BsmBI  (2872)
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-v2 is an improved version of BsmBI.
ATG
3235 .. 3237  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
3235 .. 3237  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
SV40 NLS
3241 .. 3261  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
3241 .. 3261  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
FKBP
3286 .. 3606  =  321 bp
107 amino acids  =  11.8 kDa
Product: human FK506-binding protein FKBP12
FKBP
3286 .. 3606  =  321 bp
107 amino acids  =  11.8 kDa
Product: human FK506-binding protein FKBP12
dCas9(C)
3652 .. 6036  =  2385 bp
795 amino acids  =  92.0 kDa
Product: C-terminal portion of catalytically inactive Streptococcus pyogenes dCas9 (Zetsche et al., 2015)
dCas9(C)
3652 .. 6036  =  2385 bp
795 amino acids  =  92.0 kDa
Product: C-terminal portion of catalytically inactive Streptococcus pyogenes dCas9 (Zetsche et al., 2015)
nucleoplasmin NLS
6037 .. 6084  =  48 bp
16 amino acids  =  1.7 kDa
Product: bipartite nuclear localization signal from nucleoplasmin
nucleoplasmin NLS
6037 .. 6084  =  48 bp
16 amino acids  =  1.7 kDa
Product: bipartite nuclear localization signal from nucleoplasmin
P2A
6094 .. 6150  =  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
6094 .. 6150  =  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.
dCas9(N)
6175 .. 7890  =  1716 bp
572 amino acids  =  66.3 kDa
Product: N-terminal portion of catalytically inactive Streptococcus pyogenes dCas9 (Zetsche et al., 2015)
dCas9(N)
6175 .. 7890  =  1716 bp
572 amino acids  =  66.3 kDa
Product: N-terminal portion of catalytically inactive Streptococcus pyogenes dCas9 (Zetsche et al., 2015)
FRB
7936 .. 8205  =  270 bp
90 amino acids  =  10.9 kDa
Product: FKBP-rapamycin binding domain of human FRAP
FRB
7936 .. 8205  =  270 bp
90 amino acids  =  10.9 kDa
Product: FKBP-rapamycin binding domain of human FRAP
P2A
8215 .. 8271  =  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
8215 .. 8271  =  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.
PuroR
8272 .. 8868  =  597 bp
198 amino acids  =  21.4 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
8272 .. 8868  =  597 bp
198 amino acids  =  21.4 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
AmpR
12,799 .. 13,659  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   12,799 .. 13,590  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
12,799 .. 13,659  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   13,591 .. 13,659  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
12,799 .. 13,659  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
WPRE
8884 .. 9472  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
WPRE
8884 .. 9472  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
ori
12,040 .. 12,628  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
12,040 .. 12,628  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
10,079 .. 10,507  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
10,079 .. 10,507  =  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
10,964 .. 11,338  =  375 bp
124 amino acids  =  13.8 kDa
Product: antibiotic-binding protein
confers resistance to bleomycin, phleomycin, and Zeocin™
BleoR
10,964 .. 11,338  =  375 bp
124 amino acids  =  13.8 kDa
Product: antibiotic-binding protein
confers resistance to bleomycin, phleomycin, and Zeocin™
SV40 promoter
10,521 .. 10,850  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
10,521 .. 10,850  =  330 bp
SV40 enhancer and early promoter
U6 promoter
2608 .. 2848  =  241 bp
RNA polymerase III promoter for human U6 snRNA
U6 promoter
2608 .. 2848  =  241 bp
RNA polymerase III promoter for human U6 snRNA
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.
3' LTR (ΔU3)
9544 .. 9777  =  234 bp
self-inactivating 3' long terminal repeat (LTR) from HIV-1
3' LTR (ΔU3)
9544 .. 9777  =  234 bp
self-inactivating 3' long terminal repeat (LTR) from HIV-1
bGH poly(A) signal
9809 .. 10,033  =  225 bp
bovine growth hormone polyadenylation signal
bGH poly(A) signal
9809 .. 10,033  =  225 bp
bovine growth hormone polyadenylation signal
EF-1α core promoter
3010 .. 3221  =  212 bp
core promoter for human elongation factor EF-1α
EF-1α core promoter
3010 .. 3221  =  212 bp
core promoter for human elongation factor EF-1α
CMV promoter
619 .. 817  =  199 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
619 .. 817  =  199 bp
human cytomegalovirus (CMV) immediate early promoter
5' LTR (truncated)
835 .. 1015  =  181 bp
truncated 5' long terminal repeat (LTR) from HIV-1
5' LTR (truncated)
835 .. 1015  =  181 bp
truncated 5' 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
11,468 .. 11,589  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
11,468 .. 11,589  =  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
13,660 .. 13,764  =  105 bp
AmpR promoter
13,660 .. 13,764  =  105 bp
gRNA scaffold
2873 .. 2948  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system
gRNA scaffold
2873 .. 2948  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system
EM7 promoter
10,898 .. 10,945  =  48 bp
synthetic bacterial promoter
EM7 promoter
10,898 .. 10,945  =  48 bp
synthetic bacterial promoter
polIII terminator
2949 .. 2954  =  6 bp
RNA polymerase III transcription terminator
polIII terminator
2949 .. 2954  =  6 bp
RNA polymerase III transcription terminator
SV40 ori
10,701 .. 10,836  =  136 bp
SV40 origin of replication
SV40 ori
10,701 .. 10,836  =  136 bp
SV40 origin of replication
ORF:  1558 .. 2445  =  888 bp
ORF:  295 amino acids  =  33.8 kDa
ORF:  3235 .. 8868  =  5634 bp
ORF:  1877 amino acids  =  212.8 kDa
ORF:  8974 .. 9483  =  510 bp
ORF:  169 amino acids  =  18.5 kDa
ORF:  10,030 .. 10,296  =  267 bp
ORF:  88 amino acids  =  9.3 kDa
ORF:  8249 .. 8563  =  315 bp
ORF:  104 amino acids  =  10.9 kDa
ORF:  8987 .. 9517  =  531 bp
ORF:  176 amino acids  =  18.7 kDa
ORF:  10,964 .. 11,338  =  375 bp
ORF:  124 amino acids  =  13.8 kDa
ORF:  12,929 .. 13,195  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  2962 .. 4452  =  1491 bp
ORF:  496 amino acids  =  52.9 kDa
ORF:  10,957 .. 11,244  =  288 bp
ORF:  95 amino acids  =  10.1 kDa
ORF:  12,799 .. 13,659  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  1473 .. 1853  =  381 bp
ORF:  126 amino acids  =  13.3 kDa
ORF:  2907 .. 3305  =  399 bp
ORF:  132 amino acids  =  14.1 kDa
ORF:  3846 .. 4295  =  450 bp
ORF:  149 amino acids  =  17.3 kDa
ORF:  6021 .. 6299  =  279 bp
ORF:  92 amino acids  =  10.0 kDa
ORF:  7521 .. 7814  =  294 bp
ORF:  97 amino acids  =  10.8 kDa
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