pCas

Bacterial plasmid with a temperature-sensitive replicon encoding Cas9, an inducible single guide RNA (sgRNA), and the λ Red recombination system.

Sequence Author: Yang Lab / Addgene #62225

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lac operator pCas 12,545 bp
Cas9
4917 .. 9023  =  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
4917 .. 9023  =  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
404 .. 1486  =  1083 bp
360 amino acids  =  38.5 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
404 .. 1486  =  1083 bp
360 amino acids  =  38.5 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).
Rep101(Ts)
1698 .. 2648  =  951 bp
316 amino acids  =  37.4 kDa
Product: temperature-sensitive version of the RepA protein needed for replication with the pSC101 origin (Armstrong et al., 1984)
Rep101(Ts)
1698 .. 2648  =  951 bp
316 amino acids  =  37.4 kDa
Product: temperature-sensitive version of the RepA protein needed for replication with the pSC101 origin (Armstrong et al., 1984)
araC
9153 .. 10,031  =  879 bp
292 amino acids  =  33.4 kDa
Product: L-arabinose regulatory protein
araC
9153 .. 10,031  =  879 bp
292 amino acids  =  33.4 kDa
Product: L-arabinose regulatory protein
KanR
3572 .. 4387  =  816 bp
271 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes
KanR
3572 .. 4387  =  816 bp
271 amino acids  =  31.0 kDa
Product: aminoglycoside phosphotransferase
confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes
Beta
10,808 .. 11,593  =  786 bp
261 amino acids  =  29.7 kDa
Product: single-stranded DNA binding recombinase in the λ Red system
mediates pairing of a single-stranded DNA with a complementary target
Beta
10,808 .. 11,593  =  786 bp
261 amino acids  =  29.7 kDa
Product: single-stranded DNA binding recombinase in the λ Red system
mediates pairing of a single-stranded DNA with a complementary target
Gam
10,386 .. 10,802  =  417 bp
138 amino acids  =  16.3 kDa
Product: inhibitor of the host RecBCD nuclease in the λ Red system
protects the double-stranded DNA substrate for recombination
Gam
10,386 .. 10,802  =  417 bp
138 amino acids  =  16.3 kDa
Product: inhibitor of the host RecBCD nuclease in the λ Red system
protects the double-stranded DNA substrate for recombination
araBAD promoter
10,058 .. 10,342  =  285 bp
promoter of the L-arabinose operon of E. coli; the araC regulatory gene is transcribed in the opposite direction (Guzman et al., 1995)
araBAD promoter
10,058 .. 10,342  =  285 bp
promoter of the L-arabinose operon of E. coli; the araC regulatory gene is transcribed in the opposite direction (Guzman et al., 1995)
λ tL3 terminator
12,271 .. 12,515  =  245 bp
transcription terminator tL3 from phage λ
λ tL3 terminator
12,271 .. 12,515  =  245 bp
transcription terminator tL3 from phage λ
pSC101 ori
2696 .. 2918  =  223 bp
low-copy replication origin that requires the Rep101 protein
pSC101 ori
2696 .. 2918  =  223 bp
low-copy replication origin that requires the Rep101 protein
tracrRNA
4543 .. 4621  =  79 bp
trans-activating CRISPR RNA for the Streptococcus pyogenes CRISPR/Cas9 system
tracrRNA
4543 .. 4621  =  79 bp
trans-activating CRISPR RNA for the Streptococcus pyogenes CRISPR/Cas9 system
lacIq promoter
1487 .. 1564  =  78 bp
In the lacIq allele, a single base change in the promoter boosts expression of the lacI gene about 10-fold.
lacIq promoter
1487 .. 1564  =  78 bp
In the lacIq allele, a single base change in the promoter boosts expression of the lacI gene about 10-fold.
gRNA scaffold
14 .. 89  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system
gRNA scaffold
14 .. 89  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system
trc promoter
143 .. 172  =  30 bp
3 segments
   Segment 3:  -10  
   143 .. 149  =  7 bp
strong E. coli promoter; hybrid between the trp and lac UV5 promoters
trc promoter
143 .. 172  =  30 bp
3 segments
   Segment 2:  
   150 .. 166  =  17 bp
strong E. coli promoter; hybrid between the trp and lac UV5 promoters
trc promoter
143 .. 172  =  30 bp
3 segments
   Segment 1:  -35  
   167 .. 172  =  6 bp
strong E. coli promoter; hybrid between the trp and lac UV5 promoters
trc promoter
143 .. 172  =  30 bp
3 segments
strong E. coli promoter; hybrid between the trp and lac UV5 promoters
lac operator
119 .. 135  =  17 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
119 .. 135  =  17 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).
Exo
11,590 .. 12,270  =  681 bp
226 amino acids  =  25.9 kDa
Product: 5' to 3' double-stranded DNA exonuclease in the λ Red system
creates 3' single-stranded DNA tails
Exo
11,590 .. 12,270  =  681 bp
226 amino acids  =  25.9 kDa
Product: 5' to 3' double-stranded DNA exonuclease in the λ Red system
creates 3' single-stranded DNA tails
ORF:  11,590 .. 12,270  =  681 bp
ORF:  226 amino acids  =  25.9 kDa
ORF:  10,808 .. 11,593  =  786 bp
ORF:  261 amino acids  =  29.7 kDa
ORF:  4917 .. 9023  =  4107 bp
ORF:  1368 amino acids  =  158.4 kDa
ORF:  9717 .. 9965  =  249 bp
ORF:  82 amino acids  =  9.3 kDa
ORF:  10,386 .. 10,802  =  417 bp
ORF:  138 amino acids  =  16.3 kDa
ORF:  11,019 .. 11,276  =  258 bp
ORF:  85 amino acids  =  9.3 kDa
ORF:  384 .. 647  =  264 bp
ORF:  87 amino acids  =  8.9 kDa
ORF:  1236 .. 1553  =  318 bp
ORF:  105 amino acids  =  11.2 kDa
ORF:  1698 .. 2648  =  951 bp
ORF:  316 amino acids  =  37.4 kDa
ORF:  9153 .. 10,082  =  930 bp
ORF:  309 amino acids  =  35.2 kDa
ORF:  11,421 .. 11,684  =  264 bp
ORF:  87 amino acids  =  9.4 kDa
ORF:  12,144 .. 12,434  =  291 bp
ORF:  96 amino acids  =  10.6 kDa
ORF:  404 .. 1363  =  960 bp
ORF:  319 amino acids  =  34.1 kDa
ORF:  3572 .. 4387  =  816 bp
ORF:  271 amino acids  =  31.0 kDa
ORF:  10,946 .. 11,245  =  300 bp
ORF:  99 amino acids  =  11.1 kDa
ORF:  10,690 .. 10,941  =  252 bp
ORF:  83 amino acids  =  9.0 kDa
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