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Plasmid Files

CRISPRi library backbone

Backbone vector for generating a CRISPR-mediated transcriptional repression (CRISPRi) library.

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CRISPRi library backbone Sequence and MapCRISPRi library backbone.dna
Map and Sequence File   
Sequence Author:  Weissman Lab / Addgene #62217
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 CMV enhancer U6 promoter BstXI (2941) BlpI (2974) polIII terminator XhoI (3058) CRISPRi library backbone 8888 bp
BstXI  (2941)
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.
BlpI  (2974)
1 site
G C T N A G C C G A N T C G

Sticky ends from different BlpI sites may not be compatible.
XhoI  (3058)
1 site
C T C G A G G A G C T C
EF-1α intron A
3339 .. 4277  =  939 bp
intron upstream of the start codon of human EF-1α
EF-1α intron A
3339 .. 4277  =  939 bp
intron upstream of the start codon of human EF-1α
AmpR
7895 .. 8755  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   7895 .. 8686  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
7895 .. 8755  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   8687 .. 8755  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
7895 .. 8755  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
TagBFP
4970 .. 5668  =  699 bp
232 amino acids  =  26.2 kDa
Product: monomeric blue fluorescent protein
mammalian codon-optimized
TagBFP
4970 .. 5668  =  699 bp
232 amino acids  =  26.2 kDa
Product: monomeric blue fluorescent protein
mammalian codon-optimized
PuroR
4298 .. 4894  =  597 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
4298 .. 4894  =  597 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
WPRE
5777 .. 6365  =  589 bp
woodchuck hepatitis virus posttranscriptional
regulatory element
WPRE
5777 .. 6365  =  589 bp
woodchuck hepatitis virus posttranscriptional
regulatory element
ori
7136 .. 7724  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
7136 .. 7724  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
CMV enhancer
247 .. 626  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
247 .. 626  =  380 bp
human cytomegalovirus immediate early enhancer
U6 promoter
2631 .. 2943  =  313 bp
modified RNA polymerase III promoter for mouse
U6 snRNA
U6 promoter
2631 .. 2943  =  313 bp
modified RNA polymerase III promoter for mouse
U6 snRNA
RRE
1695 .. 1928  =  234 bp
The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to
the cytoplasm.
RRE
1695 .. 1928  =  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
3127 .. 3338  =  212 bp
core promoter for human elongation factor EF-1α
EF-1α core promoter
3127 .. 3338  =  212 bp
core promoter for human elongation factor EF-1α
CMV promoter
628 .. 826  =  199 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
628 .. 826  =  199 bp
human cytomegalovirus (CMV) immediate early
promoter
LTR (truncated)
844 .. 1024  =  181 bp
truncated long terminal repeat (LTR) from HIV-1
LTR (truncated)
844 .. 1024  =  181 bp
truncated long terminal repeat (LTR) from HIV-1
LTR (truncated)
6894 .. 7074  =  181 bp
truncated long terminal repeat (LTR) from HIV-1
LTR (truncated)
6894 .. 7074  =  181 bp
truncated long terminal repeat (LTR) from HIV-1
HIV-1 Ψ
1071 .. 1196  =  126 bp
packaging signal of human immunodeficiency virus
type 1
HIV-1 Ψ
1071 .. 1196  =  126 bp
packaging signal of human immunodeficiency virus
type 1
cPPT/CTS
2455 .. 2572  =  118 bp
central polypurine tract and central termination
sequence of HIV-1
cPPT/CTS
2455 .. 2572  =  118 bp
central polypurine tract and central termination
sequence of HIV-1
AmpR promoter
8756 .. 8860  =  105 bp
AmpR promoter
8756 .. 8860  =  105 bp
gRNA scaffold
2965 .. 3050  =  86 bp
modified guide RNA scaffold for the Streptococcus
pyogenes
CRISPR/Cas9 system
gRNA scaffold
2965 .. 3050  =  86 bp
modified guide RNA scaffold for the Streptococcus
pyogenes
CRISPR/Cas9 system
T2A
4904 .. 4957  =  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
4904 .. 4957  =  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
5688 .. 5721  =  34 bp
Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT).
loxP
5688 .. 5721  =  34 bp
Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT).
polIII terminator
3051 .. 3056  =  6 bp
RNA polymerase III transcription terminator
polIII terminator
3051 .. 3056  =  6 bp
RNA polymerase III transcription terminator
loxP
2825 .. 2858  =  34 bp
Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT).
loxP
2825 .. 2858  =  34 bp
Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT).
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