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Plasmid Files

pGL3-U6-sgRNA-PGK-puromycin

Plasmid for mammalian cell expression of a single guide RNA (sgRNA) together with a puromycin resistance marker.

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pGL3-U6-sgRNA-PGK-puromycin Sequence and MappGL3-U6-sgRNA-PGK-puromycin.dna
Map and Sequence File   
Sequence Author:  Huang Lab / Addgene #51133
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 BsaI (318) BsaI (343) polIII terminator cPPT/CTS pGL3-U6-sgRNA-PGK-puromycin 4952 bp
BsaI  (318)
2 sites
G G T C T C N C C A G A G N ( N ) 4

Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
BsaI  (343)
2 sites
G G T C T C N C C A G A G N ( N ) 4

Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
AmpR
3214 .. 4074  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   3214 .. 4005  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3214 .. 4074  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   4006 .. 4074  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3214 .. 4074  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
PuroR
1169 .. 1768  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
1169 .. 1768  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
ori
2455 .. 3043  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
2455 .. 3043  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
hPGK promoter
637 .. 1147  =  511 bp
human phosphoglycerate kinase 1 promoter
hPGK promoter
637 .. 1147  =  511 bp
human phosphoglycerate kinase 1 promoter
f1 ori
4206 .. 4661  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
4206 .. 4661  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
U6 promoter
72 .. 312  =  241 bp
RNA polymerase III promoter for human U6 snRNA
U6 promoter
72 .. 312  =  241 bp
RNA polymerase III promoter for human U6 snRNA
SV40 poly(A) signal
1915 .. 2036  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1915 .. 2036  =  122 bp
SV40 polyadenylation signal
cPPT/CTS
471 .. 588  =  118 bp
central polypurine tract and central termination
sequence of HIV-1
cPPT/CTS
471 .. 588  =  118 bp
central polypurine tract and central termination
sequence of HIV-1
AmpR promoter
4075 .. 4179  =  105 bp
AmpR promoter
4075 .. 4179  =  105 bp
pause site
4854 .. 4945  =  92 bp
RNA polymerase II transcriptional pause signal from
the human α2 globin gene
pause site
4854 .. 4945  =  92 bp
RNA polymerase II transcriptional pause signal from
the human α2 globin gene
gRNA scaffold
344 .. 419  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system
gRNA scaffold
344 .. 419  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system
poly(A) signal
4792 .. 4840  =  49 bp
synthetic polyadenylation signal
poly(A) signal
4792 .. 4840  =  49 bp
synthetic polyadenylation signal
polIII terminator
420 .. 425  =  6 bp
RNA polymerase III transcription terminator
polIII terminator
420 .. 425  =  6 bp
RNA polymerase III transcription terminator
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