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lenti sgRNA(MS2)_zeo backbone

Zhang lab lentiviral vector for expressing a Zeocin™-resistance marker plus a single guide RNA (sgRNA) containing two MS2 stem loops.

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lenti sgRNA(MS2)_zeo backbone Sequence and Maplenti sgRNA(MS2)_zeo backbone.dna
Map and Sequence File   
Sequence Author:  Zhang Lab / Addgene #61427
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 CMV enhancer cPPT/CTS BsmBI (2851) BsmBI (2875) lenti sgRNA(MS2)_zeo backbone 9996 bp
BsmBI  (2851)
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  (2875)
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.
EF-1α promoter
3105 .. 4282  =  1178 bp
strong constitutive promoter for human elongation
factor EF-1α
EF-1α promoter
3105 .. 4282  =  1178 bp
strong constitutive promoter for human elongation
factor EF-1α
AmpR
9003 .. 9863  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   9003 .. 9794  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
9003 .. 9863  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   9795 .. 9863  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
9003 .. 9863  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
WPRE
4688 .. 5276  =  589 bp
woodchuck hepatitis virus posttranscriptional
regulatory element
WPRE
4688 .. 5276  =  589 bp
woodchuck hepatitis virus posttranscriptional
regulatory element
ori
8244 .. 8832  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
8244 .. 8832  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
6283 .. 6711  =  429 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
6283 .. 6711  =  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
4289 .. 4663  =  375 bp
124 amino acids  =  13.8 kDa
Product: antibiotic-binding protein
confers resistance to bleomycin, phleomycin, and
Zeocin™
BleoR
4289 .. 4663  =  375 bp
124 amino acids  =  13.8 kDa
Product: antibiotic-binding protein
confers resistance to bleomycin, phleomycin, and
Zeocin™
BleoR
7168 .. 7542  =  375 bp
124 amino acids  =  13.8 kDa
Product: antibiotic-binding protein
confers resistance to bleomycin, phleomycin, and
Zeocin™
BleoR
7168 .. 7542  =  375 bp
124 amino acids  =  13.8 kDa
Product: antibiotic-binding protein
confers resistance to bleomycin, phleomycin, and
Zeocin™
SV40 promoter
6725 .. 7054  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
6725 .. 7054  =  330 bp
SV40 enhancer and early promoter
U6 promoter
2607 .. 2847  =  241 bp
RNA polymerase III promoter for human U6 snRNA
U6 promoter
2607 .. 2847  =  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.
bGH poly(A) signal
6013 .. 6237  =  225 bp
bovine growth hormone polyadenylation signal
bGH poly(A) signal
6013 .. 6237  =  225 bp
bovine growth hormone polyadenylation signal
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
5' LTR (truncated)
5801 .. 5981  =  181 bp
truncated 5' long terminal repeat (LTR) from HIV-1
5' LTR (truncated)
5801 .. 5981  =  181 bp
truncated 5' long terminal repeat (LTR) from HIV-1
gRNA scaffold
2876 .. 3011  =  136 bp
guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system, modified by the insertion of
two MS2 stem loops
gRNA scaffold
2876 .. 3011  =  136 bp
guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system, modified by the insertion of
two MS2 stem loops
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
7672 .. 7793  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
7672 .. 7793  =  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
9864 .. 9968  =  105 bp
AmpR promoter
9864 .. 9968  =  105 bp
EM7 promoter
7102 .. 7149  =  48 bp
synthetic bacterial promoter
EM7 promoter
7102 .. 7149  =  48 bp
synthetic bacterial promoter
polIII terminator
3012 .. 3017  =  6 bp
RNA polymerase III transcription terminator
polIII terminator
3012 .. 3017  =  6 bp
RNA polymerase III transcription terminator
EF-1α intron A
3336 .. 4273  =  938 bp
intron upstream of the start codon of human EF-1α
EF-1α intron A
3336 .. 4273  =  938 bp
intron upstream of the start codon of human EF-1α
SV40 ori
6905 .. 7040  =  136 bp
SV40 origin of replication
SV40 ori
6905 .. 7040  =  136 bp
SV40 origin of replication
MS2 stem loop
2893 .. 2911  =  19 bp
stem loop that binds the bacteriophage MS2 coat
protein
MS2 stem loop
2893 .. 2911  =  19 bp
stem loop that binds the bacteriophage MS2 coat
protein
MS2 stem loop
2963 .. 2981  =  19 bp
stem loop that binds the bacteriophage MS2 coat
protein
MS2 stem loop
2963 .. 2981  =  19 bp
stem loop that binds the bacteriophage MS2 coat
protein
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