pCW-Cas9

Lentiviral vector for doxycycline-inducible expression of FLAG®-tagged Cas9.

Sequence Author: Sabatini

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ATG AmpR promoter 3xFLAG SV40 NLS pCW-Cas9 11,885 bp
ATG
549 .. 551  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
549 .. 551  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
3xFLAG
552 .. 617  =  66 bp
22 amino acids  =  2.7 kDa
Product: three tandem FLAG® epitope tags, followed by an enterokinase cleavage site
3xFLAG
552 .. 617  =  66 bp
22 amino acids  =  2.7 kDa
Product: three tandem FLAG® epitope tags, followed by an enterokinase cleavage site
SV40 NLS
624 .. 644  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
624 .. 644  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
Cas9
669 .. 4769  =  4101 bp
1367 amino acids  =  158.3 kDa
Product: Cas9 (Csn1) endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system
generates RNA-guided double strand breaks in DNA
Cas9
669 .. 4769  =  4101 bp
1367 amino acids  =  158.3 kDa
Product: Cas9 (Csn1) endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system
generates RNA-guided double strand breaks in DNA
nucleoplasmin NLS
4770 .. 4817  =  48 bp
16 amino acids  =  1.7 kDa
Product: bipartite nuclear localization signal from nucleoplasmin
nucleoplasmin NLS
4770 .. 4817  =  48 bp
16 amino acids  =  1.7 kDa
Product: bipartite nuclear localization signal from nucleoplasmin
PuroR
5347 .. 5943  =  597 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
5347 .. 5943  =  597 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
T2A
5944 .. 5997  =  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
5944 .. 5997  =  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.
rtTA-Advanced
5998 .. 6744  =  747 bp
248 amino acids  =  27.8 kDa
8 segments
   Segment 1:  rTetR  
   5998 .. 6615  =  618 bp
   206 amino acids  =  23.2 kDa
Product: improved tetracycline-controlled transactivator
In the Tet-On® system, rtTA-Advanced binds to the Tet-responsive element and stimulates transcription only in the presence of tetracycline or doxycycline.
rtTA-Advanced
5998 .. 6744  =  747 bp
248 amino acids  =  27.8 kDa
8 segments
   Segment 2:  
   6616 .. 6621  =  6 bp
   2 amino acids  =  172.2 Da
Product: improved tetracycline-controlled transactivator
In the Tet-On® system, rtTA-Advanced binds to the Tet-responsive element and stimulates transcription only in the presence of tetracycline or doxycycline.
rtTA-Advanced
5998 .. 6744  =  747 bp
248 amino acids  =  27.8 kDa
8 segments
   Segment 3:  VP16 minimal transcriptional activation domain  
   6622 .. 6657  =  36 bp
   12 amino acids  =  1.4 kDa
Product: improved tetracycline-controlled transactivator
In the Tet-On® system, rtTA-Advanced binds to the Tet-responsive element and stimulates transcription only in the presence of tetracycline or doxycycline.
rtTA-Advanced
5998 .. 6744  =  747 bp
248 amino acids  =  27.8 kDa
8 segments
   Segment 4:  
   6658 .. 6660  =  3 bp
   1 amino acid  =  115.1 Da
Product: improved tetracycline-controlled transactivator
In the Tet-On® system, rtTA-Advanced binds to the Tet-responsive element and stimulates transcription only in the presence of tetracycline or doxycycline.
rtTA-Advanced
5998 .. 6744  =  747 bp
248 amino acids  =  27.8 kDa
8 segments
   Segment 5:  VP16 minimal transcriptional activation domain  
   6661 .. 6696  =  36 bp
   12 amino acids  =  1.4 kDa
Product: improved tetracycline-controlled transactivator
In the Tet-On® system, rtTA-Advanced binds to the Tet-responsive element and stimulates transcription only in the presence of tetracycline or doxycycline.
rtTA-Advanced
5998 .. 6744  =  747 bp
248 amino acids  =  27.8 kDa
8 segments
   Segment 6:  
   6697 .. 6699  =  3 bp
   1 amino acid  =  115.1 Da
Product: improved tetracycline-controlled transactivator
In the Tet-On® system, rtTA-Advanced binds to the Tet-responsive element and stimulates transcription only in the presence of tetracycline or doxycycline.
rtTA-Advanced
5998 .. 6744  =  747 bp
248 amino acids  =  27.8 kDa
8 segments
   Segment 7:  VP16 minimal transcriptional activation domain  
   6700 .. 6735  =  36 bp
   12 amino acids  =  1.4 kDa
Product: improved tetracycline-controlled transactivator
In the Tet-On® system, rtTA-Advanced binds to the Tet-responsive element and stimulates transcription only in the presence of tetracycline or doxycycline.
rtTA-Advanced
5998 .. 6744  =  747 bp
248 amino acids  =  27.8 kDa
8 segments
   Segment 8:  
   6736 .. 6744  =  9 bp
   2 amino acids  =  172.2 Da
Product: improved tetracycline-controlled transactivator
In the Tet-On® system, rtTA-Advanced binds to the Tet-responsive element and stimulates transcription only in the presence of tetracycline or doxycycline.
rtTA-Advanced
5998 .. 6744  =  747 bp
248 amino acids  =  27.8 kDa
8 segments
Product: improved tetracycline-controlled transactivator
In the Tet-On® system, rtTA-Advanced binds to the Tet-responsive element and stimulates transcription only in the presence of tetracycline or doxycycline.
AmpR
8091 .. 8951  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   8091 .. 8159  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
8091 .. 8951  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   8160 .. 8951  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
8091 .. 8951  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
9122 .. 9710  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
9122 .. 9710  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
hPGK promoter
4827 .. 5337  =  511 bp
human phosphoglycerate kinase 1 promoter
hPGK promoter
4827 .. 5337  =  511 bp
human phosphoglycerate kinase 1 promoter
tight TRE promoter
212 .. 526  =  315 bp
Tet-responsive promoter PTight, consisting of seven tet operator sequences followed by the minimal CMV promoter
tight TRE promoter
212 .. 526  =  315 bp
Tet-responsive promoter PTight, consisting of seven tet operator sequences followed by the minimal CMV promoter
RRE
11,251 .. 11,484  =  234 bp
The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm.
RRE
11,251 .. 11,484  =  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)
7418 .. 7651  =  234 bp
self-inactivating 3' long terminal repeat (LTR) from HIV-1
3' LTR (ΔU3)
7418 .. 7651  =  234 bp
self-inactivating 3' long terminal repeat (LTR) from HIV-1
RSV promoter
10,179 .. 10,405  =  227 bp
Rous sarcoma virus enhancer/promoter
RSV promoter
10,179 .. 10,405  =  227 bp
Rous sarcoma virus enhancer/promoter
5' LTR (truncated)
10,406 .. 10,586  =  181 bp
truncated 5' long terminal repeat (LTR) from HIV-1
5' LTR (truncated)
10,406 .. 10,586  =  181 bp
truncated 5' long terminal repeat (LTR) from HIV-1
SV40 ori
7884 .. 8019  =  136 bp
SV40 origin of replication
SV40 ori
7884 .. 8019  =  136 bp
SV40 origin of replication
HIV-1 Ψ
10,633 .. 10,758  =  126 bp
packaging signal of human immunodeficiency virus type 1
HIV-1 Ψ
10,633 .. 10,758  =  126 bp
packaging signal of human immunodeficiency virus type 1
SV40 poly(A) signal
7723 .. 7844  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
7723 .. 7844  =  122 bp
SV40 polyadenylation signal
cPPT/CTS
41 .. 158  =  118 bp
central polypurine tract and central termination sequence of HIV-1
cPPT/CTS
41 .. 158  =  118 bp
central polypurine tract and central termination sequence of HIV-1
AmpR promoter
8021 .. 8090  =  70 bp
AmpR promoter
8021 .. 8090  =  70 bp
tet operator
220 .. 238  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
220 .. 238  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
256 .. 274  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
256 .. 274  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
291 .. 309  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
291 .. 309  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
327 .. 345  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
327 .. 345  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
363 .. 381  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
363 .. 381  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
398 .. 416  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
398 .. 416  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
434 .. 452  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
434 .. 452  =  19 bp
bacterial operator O2 for the tetR and tetA genes
ORF:  5119 .. 6744  =  1626 bp
ORF:  541 amino acids  =  59.0 kDa
ORF:  7141 .. 7497  =  357 bp
ORF:  118 amino acids  =  12.5 kDa
ORF:  10,108 .. 10,353  =  246 bp
ORF:  81 amino acids  =  8.9 kDa
ORF:  278 .. 541  =  264 bp
ORF:  87 amino acids  =  10.1 kDa
ORF:  6155 .. 6457  =  303 bp
ORF:  100 amino acids  =  11.1 kDa
ORF:  6821 .. 7345  =  525 bp
ORF:  174 amino acids  =  19.1 kDa
ORF:  7745 .. 7981  =  237 bp
ORF:  78 amino acids  =  8.6 kDa
ORF:  11,129 .. 410  =  1167 bp
ORF:  388 amino acids  =  44.8 kDa
ORF:  549 .. 4820  =  4272 bp
ORF:  1423 amino acids  =  164.6 kDa
ORF:  6834 .. 7130  =  297 bp
ORF:  98 amino acids  =  10.2 kDa
ORF:  8091 .. 8951  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  369 .. 5348  =  4980 bp
ORF:  1659 amino acids  =  176.3 kDa
ORF:  6450 .. 6731  =  282 bp
ORF:  93 amino acids  =  9.5 kDa
ORF:  10,035 .. 10,334  =  300 bp
ORF:  99 amino acids  =  10.9 kDa
ORF:  2015 .. 2308  =  294 bp
ORF:  97 amino acids  =  10.8 kDa
ORF:  2579 .. 3028  =  450 bp
ORF:  149 amino acids  =  17.3 kDa
ORF:  8555 .. 8821  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  11,044 .. 11,424  =  381 bp
ORF:  126 amino acids  =  13.3 kDa
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Download pCW-Cas9.dna file

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Individual Sequences & Maps

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