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Plasmid Files

pLSC-5

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

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pLSC-5 Sequence and MappLSC-5.dna
Map and Sequence File   
Sequence Author:  Zhang Lab / Addgene #62889
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 CMV enhancer cPPT/CTS BsmBI (2852) BsmBI (2872) polIII terminator EF-1 α 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  (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.
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)
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)
AmpR
12,799 .. 13,659  =  861 bp
286 amino acids  =  31.6 kDa
   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
   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
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
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
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
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
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
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.
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.
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
EM7 promoter
10,898 .. 10,945  =  48 bp
synthetic bacterial promoter
EM7 promoter
10,898 .. 10,945  =  48 bp
synthetic bacterial promoter
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
polIII terminator
2949 .. 2954  =  6 bp
RNA polymerase III transcription terminator
polIII terminator
2949 .. 2954  =  6 bp
RNA polymerase III transcription terminator
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 ori
10,701 .. 10,836  =  136 bp
SV40 origin of replication
SV40 ori
10,701 .. 10,836  =  136 bp
SV40 origin of replication
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