Resources
Plasmid Files

pSECC

Lentiviral vector for expressing a single guide RNA (sgRNA) together with S. pyogenes Cas9 and Cre recombinase.

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pSECC Sequence and MappSECC.dna
Map and Sequence File   
Sequence Author:  Jacks Lab / Addgene #60820
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 AmpR promoter SV40 NLS BsmBI (2899) BsmBI (4784) gRNA scaffold polIII terminator ATG FLAG SV40 NLS pSECC 13,901 bp
BsmBI  (2899)
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  (4784)
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
5464 .. 9564  =  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
5464 .. 9564  =  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
filler
2900 .. 4784  =  1885 bp
digest with BsmBI to replace the filler with an
sgRNA
filler
2900 .. 4784  =  1885 bp
digest with BsmBI to replace the filler with an
sgRNA
Cre
9709 .. 10,737  =  1029 bp
342 amino acids  =  38.3 kDa
Product: site-specific recombinase
Cre is a site-specific recombinase from
bacteriophage P1. Recombination occurs at loxP
sequences.
Cre
9709 .. 10,737  =  1029 bp
342 amino acids  =  38.3 kDa
Product: site-specific recombinase
Cre is a site-specific recombinase from
bacteriophage P1. Recombination occurs at loxP
sequences.
AmpR
12,908 .. 13,768  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   12,908 .. 13,699  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
12,908 .. 13,768  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   13,700 .. 13,768  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
12,908 .. 13,768  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
WPRE
10,790 .. 11,378  =  589 bp
woodchuck hepatitis virus posttranscriptional
regulatory element
WPRE
10,790 .. 11,378  =  589 bp
woodchuck hepatitis virus posttranscriptional
regulatory element
ori
12,149 .. 12,737  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
12,149 .. 12,737  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
CMV enhancer
237 .. 616  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
237 .. 616  =  380 bp
human cytomegalovirus immediate early enhancer
U6 promoter
2655 .. 2895  =  241 bp
RNA polymerase III promoter for human U6 snRNA
U6 promoter
2655 .. 2895  =  241 bp
RNA polymerase III promoter for human U6 snRNA
RRE
1685 .. 1918  =  234 bp
The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to
the cytoplasm.
RRE
1685 .. 1918  =  234 bp
The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to
the cytoplasm.
EF-1α core promoter
5124 .. 5335  =  212 bp
core promoter for human elongation factor EF-1α
EF-1α core promoter
5124 .. 5335  =  212 bp
core promoter for human elongation factor EF-1α
CMV promoter
618 .. 816  =  199 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
618 .. 816  =  199 bp
human cytomegalovirus (CMV) immediate early
promoter
5' LTR (truncated)
834 .. 1014  =  181 bp
truncated 5' long terminal repeat (LTR) from HIV-1
5' LTR (truncated)
834 .. 1014  =  181 bp
truncated 5' long terminal repeat (LTR) from HIV-1
5' LTR (truncated)
11,907 .. 12,087  =  181 bp
truncated 5' long terminal repeat (LTR) from HIV-1
5' LTR (truncated)
11,907 .. 12,087  =  181 bp
truncated 5' long terminal repeat (LTR) from HIV-1
HIV-1 Ψ
1061 .. 1186  =  126 bp
packaging signal of human immunodeficiency virus
type 1
HIV-1 Ψ
1061 .. 1186  =  126 bp
packaging signal of human immunodeficiency virus
type 1
cPPT/CTS
2445 .. 2562  =  118 bp
central polypurine tract and central termination
sequence of HIV-1
cPPT/CTS
2445 .. 2562  =  118 bp
central polypurine tract and central termination
sequence of HIV-1
cPPT/CTS
4908 .. 5025  =  118 bp
central polypurine tract and central termination
sequence of HIV-1
cPPT/CTS
4908 .. 5025  =  118 bp
central polypurine tract and central termination
sequence of HIV-1
AmpR promoter
13,769 .. 13,873  =  105 bp
AmpR promoter
13,769 .. 13,873  =  105 bp
gRNA scaffold
4785 .. 4860  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system
gRNA scaffold
4785 .. 4860  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system
P2A
9628 .. 9684  =  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
9628 .. 9684  =  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
9565 .. 9612  =  48 bp
16 amino acids  =  1.7 kDa
Product: bipartite nuclear localization signal from
nucleoplasmin
nucleoplasmin NLS
9565 .. 9612  =  48 bp
16 amino acids  =  1.7 kDa
Product: bipartite nuclear localization signal from
nucleoplasmin
FLAG
5389 .. 5412  =  24 bp
8 amino acids  =  1.0 kDa
Product: FLAG® epitope tag, followed by an
enterokinase cleavage site
FLAG
5389 .. 5412  =  24 bp
8 amino acids  =  1.0 kDa
Product: FLAG® epitope tag, followed by an
enterokinase cleavage site
SV40 NLS
5419 .. 5439  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T
antigen
SV40 NLS
5419 .. 5439  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T
antigen
SV40 NLS
9688 .. 9708  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T
antigen
SV40 NLS
9688 .. 9708  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T
antigen
polIII terminator
4861 .. 4866  =  6 bp
RNA polymerase III transcription terminator
polIII terminator
4861 .. 4866  =  6 bp
RNA polymerase III transcription terminator
ATG
5386 .. 5388  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
5386 .. 5388  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
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