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Plasmid Files

pgRNA-humanized

Lentiviral vector for expressing a guide RNA (gRNA) together with a puromycin resistance marker and mCherry.

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pgRNA-humanized Sequence and MappgRNA-humanized.dna
Map and Sequence File   
Sequence Author:  Qi Lab / Addgene #44248
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 loxP BstXI (325) gRNA scaffold polIII terminator XhoI (432) pgRNA-humanized 8308 bp
BstXI  (325)
1 site
C C A N N N N N N T G G G G T N N N N N N A C C

Sticky ends from different BstXI sites may not be compatible.
XhoI  (432)
1 site
C T C G A G G A G C T C
AmpR
4699 .. 5559  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   4699 .. 5490  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
4699 .. 5559  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   5491 .. 5559  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
4699 .. 5559  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
mCherry
1762 .. 2472  =  711 bp
236 amino acids  =  26.7 kDa
Product: monomeric derivative of DsRed fluorescent
protein (Shaner et al., 2004)
mammalian codon-optimized
mCherry
1762 .. 2472  =  711 bp
236 amino acids  =  26.7 kDa
Product: monomeric derivative of DsRed fluorescent
protein (Shaner et al., 2004)
mammalian codon-optimized
PuroR
1090 .. 1686  =  597 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
1090 .. 1686  =  597 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
WPRE
2581 .. 3169  =  589 bp
woodchuck hepatitis virus posttranscriptional
regulatory element
WPRE
2581 .. 3169  =  589 bp
woodchuck hepatitis virus posttranscriptional
regulatory element
ori
3940 .. 4528  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
3940 .. 4528  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
CMV enhancer
549 .. 852  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
549 .. 852  =  304 bp
human cytomegalovirus immediate early enhancer
RRE
7387 .. 7620  =  234 bp
The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to
the cytoplasm.
RRE
7387 .. 7620  =  234 bp
The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to
the cytoplasm.
CMV promoter
853 .. 1056  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
853 .. 1056  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
LTR (truncated)
3698 .. 3878  =  181 bp
truncated long terminal repeat (LTR) from HIV-1
LTR (truncated)
3698 .. 3878  =  181 bp
truncated long terminal repeat (LTR) from HIV-1
LTR (truncated)
6536 .. 6716  =  181 bp
truncated long terminal repeat (LTR) from HIV-1
LTR (truncated)
6536 .. 6716  =  181 bp
truncated long terminal repeat (LTR) from HIV-1
HIV-1 Ψ
6763 .. 6888  =  126 bp
packaging signal of human immunodeficiency virus
type 1
HIV-1 Ψ
6763 .. 6888  =  126 bp
packaging signal of human immunodeficiency virus
type 1
cPPT/CTS
8147 .. 8264  =  118 bp
central polypurine tract and central termination
sequence of HIV-1
cPPT/CTS
8147 .. 8264  =  118 bp
central polypurine tract and central termination
sequence of HIV-1
AmpR promoter
5560 .. 5664  =  105 bp
AmpR promoter
5560 .. 5664  =  105 bp
U6 promoter
243 .. 327  =  85 bp
RNA polymerase III promoter for mouse U6 snRNA
U6 promoter
243 .. 327  =  85 bp
RNA polymerase III promoter for mouse U6 snRNA
gRNA scaffold
349 .. 424  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system
gRNA scaffold
349 .. 424  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system
T2A
1696 .. 1749  =  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
1696 .. 1749  =  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.
loxP
209 .. 242  =  34 bp
Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT).
loxP
209 .. 242  =  34 bp
Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT).
loxP
2492 .. 2525  =  34 bp
Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT).
loxP
2492 .. 2525  =  34 bp
Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT).
polIII terminator
425 .. 430  =  6 bp
RNA polymerase III transcription terminator
polIII terminator
425 .. 430  =  6 bp
RNA polymerase III transcription terminator
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