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Plasmid Files

pHSN401

Plant vector for expressing a single guide RNA (sgRNA) together with maize codon-optimized Cas9.

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pHSN401 Sequence and MappHSN401.dna
Map and Sequence File   
Sequence Author:  Chen Lab / Addgene #50588
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 CaMV 35S promoter (enhanced) BsaI (527) BsaI (1752) polIII terminator ATG 3xFLAG SV40 NLS pHSN401 12,729 bp
BsaI  (527)
2 sites
G G T C T C N C C A G A G N ( N ) 4

Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
BsaI  (1752)
2 sites
G G T C T C N C C A G A G N ( N ) 4

Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
Cas9
3086 .. 7186  =  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
3086 .. 7186  =  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
HygR
8398 .. 9423  =  1026 bp
341 amino acids  =  38.0 kDa
Product: hygromycin B phosphotransferase
confers resistance to hygromycin
HygR
8398 .. 9423  =  1026 bp
341 amino acids  =  38.0 kDa
Product: hygromycin B phosphotransferase
confers resistance to hygromycin
KanR
11,692 .. 12,501  =  810 bp
269 amino acids  =  30.7 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin in bacteria or G418
(Geneticin®) in eukaryotes
KanR
11,692 .. 12,501  =  810 bp
269 amino acids  =  30.7 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin in bacteria or G418
(Geneticin®) in eukaryotes
SmR
810 .. 1601  =  792 bp
263 amino acids  =  29.2 kDa
Product: aminoglycoside adenylyltransferase
(Murphy, 1985)
confers resistance to spectinomycin and
streptomycin
SmR
810 .. 1601  =  792 bp
263 amino acids  =  29.2 kDa
Product: aminoglycoside adenylyltransferase
(Murphy, 1985)
confers resistance to spectinomycin and
streptomycin
CaMV 35S promoter (enhanced)
7653 .. 8330  =  678 bp
cauliflower mosaic virus 35S promoter with a
duplicated enhancer region
CaMV 35S promoter (enhanced)
7653 .. 8330  =  678 bp
cauliflower mosaic virus 35S promoter with a
duplicated enhancer region
ori
11,014 .. 11,602  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
11,014 .. 11,602  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
pSa ori
10,464 .. 10,899  =  436 bp
origin of replication from bacterial plasmid pSa
pSa ori
10,464 .. 10,899  =  436 bp
origin of replication from bacterial plasmid pSa
AtU6-26 promoter
106 .. 531  =  426 bp
promoter for the Arabidopsis thaliana U6-26 snRNA
gene (Waibel and Filipowicz, 1990)
AtU6-26 promoter
106 .. 531  =  426 bp
promoter for the Arabidopsis thaliana U6-26 snRNA
gene (Waibel and Filipowicz, 1990)
CaMV 35S promoter
2442 .. 2786  =  345 bp
strong constitutive promoter from cauliflower
mosaic virus
CaMV 35S promoter
2442 .. 2786  =  345 bp
strong constitutive promoter from cauliflower
mosaic virus
NOS terminator
7285 .. 7537  =  253 bp
nopaline synthase terminator and poly(A) signal
NOS terminator
7285 .. 7537  =  253 bp
nopaline synthase terminator and poly(A) signal
CaMV poly(A) signal
9463 .. 9637  =  175 bp
cauliflower mosaic virus polyadenylation signal
CaMV poly(A) signal
9463 .. 9637  =  175 bp
cauliflower mosaic virus polyadenylation signal
rrnB T1 terminator
10,209 .. 10,295  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
10,209 .. 10,295  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
gRNA scaffold
1753 .. 1828  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system
gRNA scaffold
1753 .. 1828  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system
3xFLAG
2969 .. 3034  =  66 bp
22 amino acids  =  2.7 kDa
Product: three tandem FLAG® epitope tags,
followed by an enterokinase cleavage site
3xFLAG
2969 .. 3034  =  66 bp
22 amino acids  =  2.7 kDa
Product: three tandem FLAG® epitope tags,
followed by an enterokinase cleavage site
nucleoplasmin NLS
7187 .. 7234  =  48 bp
16 amino acids  =  1.7 kDa
Product: bipartite nuclear localization signal from
nucleoplasmin
nucleoplasmin NLS
7187 .. 7234  =  48 bp
16 amino acids  =  1.7 kDa
Product: bipartite nuclear localization signal from
nucleoplasmin
rrnB T2 terminator
10,090 .. 10,117  =  28 bp
transcription terminator T2 from the E. coli rrnB
gene
rrnB T2 terminator
10,090 .. 10,117  =  28 bp
transcription terminator T2 from the E. coli rrnB
gene
RB T-DNA repeat
1 .. 25  =  25 bp
right border repeat from nopaline C58 T-DNA
RB T-DNA repeat
1 .. 25  =  25 bp
right border repeat from nopaline C58 T-DNA
LB T-DNA repeat
9787 .. 9811  =  25 bp
left border repeat from nopaline C58 T-DNA
LB T-DNA repeat
9787 .. 9811  =  25 bp
left border repeat from nopaline C58 T-DNA
SV40 NLS
3041 .. 3061  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T
antigen
SV40 NLS
3041 .. 3061  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T
antigen
polIII terminator
1829 .. 1834  =  6 bp
RNA polymerase III transcription terminator
polIII terminator
1829 .. 1834  =  6 bp
RNA polymerase III transcription terminator
ATG
2966 .. 2968  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
2966 .. 2968  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
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