pME-Cas9-T2A-GFP

Middle entry Gateway® vector encoding zebrafish codon-optimized S. pyogenes Cas9 with nuclear localization signals and with a C-terminal GFP.

Sequence Author: Zon Lab / Addgene #63155

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rrnB T2 terminator ATG pME-Cas9-T2A-GFP 7685 bp
ATG
756 .. 758  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
756 .. 758  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
SV40 NLS
768 .. 788  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
768 .. 788  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
Cas9
795 .. 4895  =  4101 bp
1367 amino acids  =  158.2 kDa
Product: Cas9 (Csn1) endonuclease from the Streptococcus pyogenes MGAS15252 Type II CRISPR/Cas system
generates RNA-guided double strand breaks in DNA
Cas9
795 .. 4895  =  4101 bp
1367 amino acids  =  158.2 kDa
Product: Cas9 (Csn1) endonuclease from the Streptococcus pyogenes MGAS15252 Type II CRISPR/Cas system
generates RNA-guided double strand breaks in DNA
SV40 NLS
4911 .. 4931  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
4911 .. 4931  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
T2A
4947 .. 5000  =  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
4947 .. 5000  =  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.
EGFP
5007 .. 5726  =  720 bp
239 amino acids  =  26.9 kDa
3 segments
   Segment 1:  
   5007 .. 5009  =  3 bp
   1 amino acid  =  149.2 Da
Product: enhanced GFP
mammalian codon-optimized
EGFP
5007 .. 5726  =  720 bp
239 amino acids  =  26.9 kDa
3 segments
   Segment 2:  1a  
   5010 .. 5012  =  3 bp
   1 amino acid  =  117.1 Da
Product: enhanced GFP
mammalian codon-optimized
EGFP
5007 .. 5726  =  720 bp
239 amino acids  =  26.9 kDa
3 segments
   Segment 3:  
   5013 .. 5726  =  714 bp
   237 amino acids  =  26.7 kDa
Product: enhanced GFP
mammalian codon-optimized
EGFP
5007 .. 5726  =  720 bp
239 amino acids  =  26.9 kDa
3 segments
Product: enhanced GFP
mammalian codon-optimized
KanR
6079 .. 6888  =  810 bp
269 amino acids  =  30.8 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes
KanR
6079 .. 6888  =  810 bp
269 amino acids  =  30.8 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes
ori
7035 .. 7623  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
7035 .. 7623  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
attL1
569 .. 668  =  100 bp
recombination site for the Gateway® LR reaction
attL1
569 .. 668  =  100 bp
recombination site for the Gateway® LR reaction
attL2
5810 .. 5909  =  100 bp
recombination site for the Gateway® LR reaction
attL2
5810 .. 5909  =  100 bp
recombination site for the Gateway® LR reaction
rrnB T1 terminator
387 .. 473  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
rrnB T1 terminator
387 .. 473  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
rrnB T2 terminator
268 .. 295  =  28 bp
transcription terminator T2 from the E. coli rrnB gene
rrnB T2 terminator
268 .. 295  =  28 bp
transcription terminator T2 from the E. coli rrnB gene
ORF:  628 .. 855  =  228 bp
ORF:  75 amino acids  =  9.0 kDa
ORF:  6079 .. 6888  =  810 bp
ORF:  269 amino acids  =  30.8 kDa
ORF:  756 .. 5726  =  4971 bp
ORF:  1656 amino acids  =  190.3 kDa
ORF:  2415 .. 2717  =  303 bp
ORF:  100 amino acids  =  11.3 kDa
ORF:  2817 .. 3311  =  495 bp
ORF:  164 amino acids  =  18.5 kDa
ORF:  3516 .. 3746  =  231 bp
ORF:  76 amino acids  =  8.6 kDa
ORF:  4107 .. 4838  =  732 bp
ORF:  243 amino acids  =  27.8 kDa
ORF:  581 .. 814  =  234 bp
ORF:  77 amino acids  =  8.7 kDa
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