pMJ806

pET-derived bacterial vector for expressing 6xHis- and MBP-tagged S. pyogenes Cas9.

Sequence Author: Doudna Lab / Addgene #39312

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T7 promoter RBS ATG 6xHis pMJ806 10,577 bp
ATG
2803 .. 2805  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
2803 .. 2805  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
6xHis
2806 .. 2823  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
2806 .. 2823  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
MBP
2827 .. 3924  =  1098 bp
366 amino acids  =  40.2 kDa
Product: maltose binding protein from E. coli
This version of the gene does not encode a signal sequence, so MBP will remain in the cytosol.
MBP
2827 .. 3924  =  1098 bp
366 amino acids  =  40.2 kDa
Product: maltose binding protein from E. coli
This version of the gene does not encode a signal sequence, so MBP will remain in the cytosol.
TEV site
3955 .. 3975  =  21 bp
7 amino acids  =  869.9 Da
Product: tobacco etch virus (TEV) protease recognition and cleavage site
TEV site
3955 .. 3975  =  21 bp
7 amino acids  =  869.9 Da
Product: tobacco etch virus (TEV) protease recognition and cleavage site
Cas9
3985 .. 8091  =  4107 bp
1368 amino acids  =  158.4 kDa
Product: Cas9 (Csn1) endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system
generates RNA-guided double strand breaks in DNA
Cas9
3985 .. 8091  =  4107 bp
1368 amino acids  =  158.4 kDa
Product: Cas9 (Csn1) endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system
generates RNA-guided double strand breaks in DNA
lacI
1246 .. 2328  =  1083 bp
360 amino acids  =  38.6 kDa
Product: lac repressor
The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).
lacI
1246 .. 2328  =  1083 bp
360 amino acids  =  38.6 kDa
Product: lac repressor
The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).
KanR
8871 .. 9686  =  816 bp
271 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes
KanR
8871 .. 9686  =  816 bp
271 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes
ori
9808 .. 10,396  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
9808 .. 10,396  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
8324 .. 8779  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
8324 .. 8779  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
rop
246 .. 437  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at low copy number
rop
246 .. 437  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein, which maintains plasmids at low copy number
lacI promoter
2329 .. 2406  =  78 bp
lacI promoter
2329 .. 2406  =  78 bp
T7 terminator
8240 .. 8287  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
8240 .. 8287  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
lac operator
2734 .. 2758  =  25 bp
The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).
lac operator
2734 .. 2758  =  25 bp
The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).
RBS
2773 .. 2795  =  23 bp
efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)
RBS
2773 .. 2795  =  23 bp
efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)
T7 promoter
2715 .. 2733  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
2715 .. 2733  =  19 bp
promoter for bacteriophage T7 RNA polymerase
ORF:  469 .. 837  =  369 bp
ORF:  122 amino acids  =  14.2 kDa
ORF:  2803 .. 8091  =  5289 bp
ORF:  1762 amino acids  =  201.7 kDa
ORF:  2504 .. 2743  =  240 bp
ORF:  79 amino acids  =  8.0 kDa
ORF:  1212 .. 1463  =  252 bp
ORF:  83 amino acids  =  9.1 kDa
ORF:  2367 .. 2630  =  264 bp
ORF:  87 amino acids  =  9.5 kDa
ORF:  246 .. 470  =  225 bp
ORF:  74 amino acids  =  8.5 kDa
ORF:  834 .. 1190  =  357 bp
ORF:  118 amino acids  =  13.0 kDa
ORF:  8871 .. 9686  =  816 bp
ORF:  271 amino acids  =  31.0 kDa
ORF:  1226 .. 1489  =  264 bp
ORF:  87 amino acids  =  8.9 kDa
ORF:  2078 .. 2578  =  501 bp
ORF:  166 amino acids  =  17.5 kDa
ORF:  1246 .. 2205  =  960 bp
ORF:  319 amino acids  =  34.1 kDa
ORF:  2575 .. 2943  =  369 bp
ORF:  122 amino acids  =  13.3 kDa
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