pAAV-Guide-it-1 Vector (Linear)

Linearized CRISPR/Cas9 adeno-associated virus (AAV) vector with S. aureus Cas9, for cloning and expression of a single guide RNA (sgRNA).

Sequence Author: Clontech (TaKaRa)

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Guide-itâ„¢ Sequencing Primer 1 (6950 .. 6973) SV40 NLS polIII terminator CMV enhancer ATG pAAV-Guide-it-1 Vector (Linear) 7028 bp
Guide-itâ„¢ Sequencing Primer 1
24-mer  /  38% GC
1 binding site
6950 .. 6973  =  24 annealed bases
Tm  =  56°C
ATG
3314 .. 3316  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
3314 .. 3316  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
SV40 NLS
3323 .. 3343  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
3323 .. 3343  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SaCas9
3344 .. 6499  =  3156 bp
1052 amino acids  =  123.8 kDa
Product: Cas9 (Csn1) endonuclease from the Staphylococcus aureus Type II CRISPR/Cas system
generates RNA-guided double strand breaks in DNA
SaCas9
3344 .. 6499  =  3156 bp
1052 amino acids  =  123.8 kDa
Product: Cas9 (Csn1) endonuclease from the Staphylococcus aureus Type II CRISPR/Cas system
generates RNA-guided double strand breaks in DNA
SV40 NLS
6515 .. 6535  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
6515 .. 6535  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
FLAG
6536 .. 6559  =  24 bp
8 amino acids  =  1.0 kDa
Product: FLAG® epitope tag, followed by an enterokinase cleavage site
FLAG
6536 .. 6559  =  24 bp
8 amino acids  =  1.0 kDa
Product: FLAG® epitope tag, followed by an enterokinase cleavage site
AmpR
603 .. 1463  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   603 .. 671  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
603 .. 1463  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   672 .. 1463  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
603 .. 1463  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
1634 .. 2222  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
1634 .. 2222  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
CMV enhancer
2776 .. 3079  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
2776 .. 3079  =  304 bp
human cytomegalovirus immediate early enhancer
U6 promoter
6783 .. 7031  =  249 bp
RNA polymerase III promoter for human U6 snRNA (Domitrovich & Kunkel, 2003)
U6 promoter
6783 .. 7031  =  249 bp
RNA polymerase III promoter for human U6 snRNA (Domitrovich & Kunkel, 2003)
bGH poly(A) signal
6569 .. 6776  =  208 bp
bovine growth hormone polyadenylation signal
bGH poly(A) signal
6569 .. 6776  =  208 bp
bovine growth hormone polyadenylation signal
CMV promoter
3080 .. 3283  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
3080 .. 3283  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
AAV2 ITR
96 .. 236  =  141 bp
inverted terminal repeat of adeno-associated virus serotype 2
AAV2 ITR
96 .. 236  =  141 bp
inverted terminal repeat of adeno-associated virus serotype 2
AAV2 ITR
2567 .. 2707  =  141 bp
inverted terminal repeat of adeno-associated virus serotype 2
AAV2 ITR
2567 .. 2707  =  141 bp
inverted terminal repeat of adeno-associated virus serotype 2
AmpR promoter
498 .. 602  =  105 bp
AmpR promoter
498 .. 602  =  105 bp
Sa gRNA scaffold
1 .. 76  =  76 bp
guide RNA scaffold for the Staphylococcus aureus CRISPR/Cas9 system
Sa gRNA scaffold
1 .. 76  =  76 bp
guide RNA scaffold for the Staphylococcus aureus CRISPR/Cas9 system
polIII terminator
77 .. 81  =  5 bp
RNA polymerase III transcription terminator
polIII terminator
77 .. 81  =  5 bp
RNA polymerase III transcription terminator
Kozak sequence
3308 .. 3317  =  10 bp
vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)
Kozak sequence
3308 .. 3317  =  10 bp
vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)
ORF:  110 .. 391  =  282 bp
ORF:  93 amino acids  =  9.9 kDa
ORF:  3239 .. 6562  =  3324 bp
ORF:  1107 amino acids  =  130.0 kDa
ORF:  603 .. 1463  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  3604 .. 3843  =  240 bp
ORF:  79 amino acids  =  8.5 kDa
ORF:  4705 .. 4962  =  258 bp
ORF:  85 amino acids  =  9.8 kDa
ORF:  6538 .. 6810  =  273 bp
ORF:  90 amino acids  =  10.0 kDa
ORF:  1067 .. 1333  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  3287 .. 6478  =  3192 bp
ORF:  1063 amino acids  =  114.4 kDa
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