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Plasmid Files

pLenti-Guide-Puro

3rd generation lentiviral vector with a puromycin resistance marker, for expressing a guide RNA (gRNA) under control of the U6 promoter.

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pLenti-Guide-Puro.dna
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Sequence Author:  OriGene
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CF3 Primer (2745 .. 2766) BamHI (2951) BsmBI (3001) pLenti-Guide-Puro 7460 bp
BamHI  (2951)
1 site
G G A T C C C C T A G G

After cleavage, BamHI-HF™ (but not the original BamHI) can remain bound to DNA and alter its electrophoretic mobility.
BsmBI  (3001)
1 site
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.
CF3 Primer
22-mer  /  45% GC
1 binding site
2745 .. 2766  =  22 annealed bases
Tm  =  56°C
primer for sequencing the inserted gRNA
CmR
6668 .. 7327  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
6668 .. 7327  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
PuroR
3814 .. 4413  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
3814 .. 4413  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
WPRE
4644 .. 5232  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
WPRE
4644 .. 5232  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
ori
6003 .. 6591  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
6003 .. 6591  =  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
CMV enhancer
3128 .. 3507  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
3128 .. 3507  =  380 bp
human cytomegalovirus immediate early enhancer
U6 promoter
2707 .. 2947  =  241 bp
RNA polymerase III promoter for human U6 snRNA
U6 promoter
2707 .. 2947  =  241 bp
RNA polymerase III promoter for human U6 snRNA
RRE
1683 .. 1916  =  234 bp
The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm.
RRE
1683 .. 1916  =  234 bp
The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm.
CMV promoter
3508 .. 3711  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
3508 .. 3711  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
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)
5761 .. 5941  =  181 bp
truncated 5' long terminal repeat (LTR) from HIV-1
5' LTR (truncated)
5761 .. 5941  =  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
2443 .. 2560  =  118 bp
central polypurine tract and central termination sequence of HIV-1
cPPT/CTS
2443 .. 2560  =  118 bp
central polypurine tract and central termination sequence of HIV-1
AmpR promoter
7328 .. 7432  =  105 bp
AmpR promoter
7328 .. 7432  =  105 bp
gRNA scaffold
3001 .. 3076  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system
gRNA scaffold
3001 .. 3076  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system
T7 promoter
3737 .. 3755  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
3737 .. 3755  =  19 bp
promoter for bacteriophage T7 RNA polymerase
ORF:  1561 .. 2448  =  888 bp
ORF:  295 amino acids  =  33.8 kDa
ORF:  3667 .. 4413  =  747 bp
ORF:  248 amino acids  =  27.2 kDa
ORF:  4747 .. 5787  =  1041 bp
ORF:  346 amino acids  =  38.3 kDa
ORF:  4734 .. 5288  =  555 bp
ORF:  184 amino acids  =  20.5 kDa
ORF:  1395 .. 1856  =  462 bp
ORF:  153 amino acids  =  16.6 kDa
ORF:  6668 .. 7327  =  660 bp
ORF:  219 amino acids  =  25.7 kDa
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