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Plasmid Files

pCLIP-gRNA-EFS-tRFP

Lentiviral CRISPR vector encoding a guide RNA (gRNA), plus the red fluorescent protein TurboRFP expressed from the EF-1α core promoter.

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pCLIP-gRNA-EFS-tRFP Sequence and MappCLIP-gRNA-EFS-tRFP.dna
Map and Sequence File   
Sequence Author:  transOMIC
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 polIII terminator BsmBI (2505) BsmBI (4390) pCLIP-gRNA-EFS-tRFP 8762 bp
BsmBI  (2505)
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  (4390)
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.
Stuffer
2506 .. 4390  =  1885 bp
Stuffer
2506 .. 4390  =  1885 bp
AmpR
7767 .. 8627  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   7767 .. 8558  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
7767 .. 8627  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   8559 .. 8627  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
7767 .. 8627  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
TurboRFP
4759 .. 5454  =  696 bp
231 amino acids  =  26.1 kDa
Product: red fluorescent protein from Entacmaea
quadricolor

mammalian codon-optimized
TurboRFP
4759 .. 5454  =  696 bp
231 amino acids  =  26.1 kDa
Product: red fluorescent protein from Entacmaea
quadricolor

mammalian codon-optimized
WPRE
5468 .. 6056  =  589 bp
woodchuck hepatitis virus posttranscriptional
regulatory element
WPRE
5468 .. 6056  =  589 bp
woodchuck hepatitis virus posttranscriptional
regulatory element
ori
7011 .. 7596  =  586 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
7011 .. 7596  =  586 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
CMV enhancer
27 .. 404  =  378 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
27 .. 404  =  378 bp
human cytomegalovirus immediate early enhancer
SV40
6593 .. 6922  =  330 bp
SV40 enhancer and early promoter
SV40
6593 .. 6922  =  330 bp
SV40 enhancer and early promoter
5' LTR (truncated)
411 .. 668  =  258 bp
truncated 5' long terminal repeat (LTR) from HIV-1
5' LTR (truncated)
411 .. 668  =  258 bp
truncated 5' long terminal repeat (LTR) from HIV-1
U6 promoter
2261 .. 2501  =  241 bp
RNA polymerase III promoter for human U6 snRNA
U6 promoter
2261 .. 2501  =  241 bp
RNA polymerase III promoter for human U6 snRNA
RRE
1333 .. 1566  =  234 bp
The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to
the cytoplasm.
RRE
1333 .. 1566  =  234 bp
The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to
the cytoplasm.
3' LTR (ΔU3)
6264 .. 6497  =  234 bp
self-inactivating 3' long terminal repeat (LTR) from
HIV-1
3' LTR (ΔU3)
6264 .. 6497  =  234 bp
self-inactivating 3' long terminal repeat (LTR) from
HIV-1
EF-1α core promoter
4528 .. 4739  =  212 bp
core promoter for human elongation factor EF-1α
EF-1α core promoter
4528 .. 4739  =  212 bp
core promoter for human elongation factor EF-1α
HIV-1 Ψ
715 .. 840  =  126 bp
packaging signal of human immunodeficiency virus
type 1
HIV-1 Ψ
715 .. 840  =  126 bp
packaging signal of human immunodeficiency virus
type 1
cPPT/CTS
2093 .. 2210  =  118 bp
central polypurine tract and central termination
sequence of HIV-1
cPPT/CTS
2093 .. 2210  =  118 bp
central polypurine tract and central termination
sequence of HIV-1
AmpR promoter
8628 .. 8732  =  105 bp
AmpR promoter
8628 .. 8732  =  105 bp
gRNA scaffold
4391 .. 4466  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system
gRNA scaffold
4391 .. 4466  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system
polIII terminator
4467 .. 4472  =  6 bp
RNA polymerase III transcription terminator
polIII terminator
4467 .. 4472  =  6 bp
RNA polymerase III transcription terminator
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