pRSGC2-U6-sg-HTS6-UbiC-Cas9-2A-Puro

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HomePlasmidsViral Expression & Packaging VectorspRSGC2-U6-sg-HTS6-UbiC-Cas9-2A-Puro
AmpR promoter SspI (10,966) PciI (9269) SfiI (8847) AanI (8618) AanI (8562) KpnI (8227) Acc65I (8223) HincII (7628) SalI (7626) SexAI * (7587) BstEII (7154) RsrII (7136) BsiWI (7076) PflFI - Tth111I (7062) BamHI (6954) NotI (6946) FseI (6925) nucleoplasmin NLS BsaBI * (6291) MluI (1) NruI * (612) MfeI (964) BbvCI (1199) AleI (1352) BspDI - ClaI (1798) BbsI - BpiI (2064) BbsI - BpiI (2090) PaeR7I - PspXI - XhoI (2113) EcoRI (2280) AgeI (2289) BstBI (2298) AfeI (2358) SV40 NLS SV40 NLS SV40 NLS PspOMI (4148) ApaI (4152) EcoRV (4646) PmlI (5582) pRSGC2-U6-sg-HTS6-UbiC-Cas9-2A-Puro 11,553 bp
SspI  (10,966)
1 site		 A A T A T T T T A T A A
PciI  (9269)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
SfiI  (8847)
1 site		 G G C C N N N N N G G C C C C G G N N N N N C C G G
Efficient cleavage requires at least two copies of the SfiI recognition sequence.
Sticky ends from different SfiI sites may not be compatible.
AanI  (8618)
2 sites		 T T A T A A A A T A T T
AanI  (8562)
2 sites		 T T A T A A A A T A T T
KpnI  (8227)
1 site		 G G T A C C C C A T G G
Acc65I  (8223)
1 site		 G G T A C C C C A T G G
HincII  (7628)
1 site		 G T Y R A C C A R Y T G
SalI  (7626)
1 site		 G T C G A C C A G C T G
SexAI  (7587)
1 site		 A C C W G G T T G G W C C A
* Blocked by Dcm methylation.
Sticky ends from different SexAI sites may not be compatible.
BstEII  (7154)
1 site		 G G T N A C C C C A N T G G
Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
RsrII  (7136)
1 site		 C G G W C C G G C C W G G C
Efficient cleavage requires at least two copies of the RsrII recognition sequence.
Sticky ends from different RsrII sites may not be compatible.
For full activity, add fresh DTT.
BsiWI  (7076)
1 site		 C G T A C G G C A T G C
BsiWI is typically used at 55°C, but is 50% active at 37°C.
PflFI  (7062)
1 site		 G A C N N N G T C C T G N N N C A G
The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (7062)
1 site		 G A C N N N G T C C T G N N N C A G
The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
BamHI  (6954)
1 site		 G G A T C C C C T A G G
After cleavage, BamHI-HF® (but not the original BamHI) can remain bound to DNA and alter its electrophoretic mobility.
NotI  (6946)
1 site		 G C G G C C G C C G C C G G C G
FseI  (6925)
1 site		 G G C C G G C C C C G G C C G G
FseI gradually loses activity when stored at -20°C.
BsaBI  (6291)
1 site		 G A T N N N N A T C C T A N N N N T A G
* Blocked by Dam methylation.
MluI  (1)
1 site		 A C G C G T T G C G C A
NruI  (612)
1 site		 T C G C G A A G C G C T
* Blocked by Dam methylation.
MfeI  (964)
1 site		 C A A T T G G T T A A C
BbvCI  (1199)
1 site		 C C T C A G C G G A G T C G
AleI  (1352)
1 site		 C A C N N N N G T G G T G N N N N C A C
BspDI  (1798)
1 site		 A T C G A T T A G C T A
ClaI  (1798)
1 site		 A T C G A T T A G C T A
BbsI  (2064)
2 sites		 G A A G A C N N C T T C T G N N ( N ) 4
Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
BpiI  (2064)
2 sites		 G A A G A C N N C T T C T G N N ( N ) 4
Sticky ends from different BpiI sites may not be compatible.
BbsI  (2090)
2 sites		 G A A G A C N N C T T C T G N N ( N ) 4
Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
BpiI  (2090)
2 sites		 G A A G A C N N C T T C T G N N ( N ) 4
Sticky ends from different BpiI sites may not be compatible.
PaeR7I  (2113)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (2113)
1 site		 V C T C G A G B B G A G C T C V
XhoI  (2113)
1 site		 C T C G A G G A G C T C
EcoRI  (2280)
1 site		 G A A T T C C T T A A G
AgeI  (2289)
1 site		 A C C G G T T G G C C A
BstBI  (2298)
1 site		 T T C G A A A A G C T T
AfeI  (2358)
1 site		 A G C G C T T C G C G A
PspOMI  (4148)
1 site		 G G G C C C C C C G G G
ApaI  (4152)
1 site		 G G G C C C C C C G G G
ApaI can be used between 25°C and 37°C.
EcoRV  (4646)
1 site		 G A T A T C C T A T A G
EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage.
PmlI  (5582)
1 site		 C A C G T G G T G C A C
SV40 NLS
2721 .. 2741  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
2721 .. 2741  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
2745 .. 2765  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
2745 .. 2765  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
2769 .. 2789  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
2769 .. 2789  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
Cas9
2796 .. 6896  =  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
2796 .. 6896  =  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
nucleoplasmin NLS
6897 .. 6944  =  48 bp
16 amino acids  =  1.7 kDa
Product: bipartite nuclear localization signal from nucleoplasmin
nucleoplasmin NLS
6897 .. 6944  =  48 bp
16 amino acids  =  1.7 kDa
Product: bipartite nuclear localization signal from nucleoplasmin
T2A
6960 .. 7013  =  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
6960 .. 7013  =  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.
PuroR
7020 .. 7619  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
7020 .. 7619  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
AmpR
10,089 .. 10,949  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   10,089 .. 10,880  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
10,089 .. 10,949  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   10,881 .. 10,949  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
10,089 .. 10,949  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
9330 .. 9918  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
9330 .. 9918  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
WPRE
7632 .. 8220  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
WPRE
7632 .. 8220  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
UbiC promoter
2303 .. 2702  =  400 bp
human ubiquitin C promoter
UbiC promoter
2303 .. 2702  =  400 bp
human ubiquitin C promoter
U6 promoter
1819 .. 2067  =  249 bp
RNA polymerase III promoter for human U6 snRNA (Domitrovich & Kunkel, 2003)
U6 promoter
1819 .. 2067  =  249 bp
RNA polymerase III promoter for human U6 snRNA (Domitrovich & Kunkel, 2003)
3' LTR (ΔU3)
8294 .. 8527  =  234 bp
self-inactivating 3' long terminal repeat (LTR) from HIV-1
3' LTR (ΔU3)
8294 .. 8527  =  234 bp
self-inactivating 3' long terminal repeat (LTR) from HIV-1
RRE
1078 .. 1311  =  234 bp
The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm.
RRE
1078 .. 1311  =  234 bp
The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm.
RSV promoter
6 .. 232  =  227 bp
Rous sarcoma virus enhancer/promoter
RSV promoter
6 .. 232  =  227 bp
Rous sarcoma virus enhancer/promoter
5' LTR (truncated)
233 .. 413  =  181 bp
truncated 5' long terminal repeat (LTR) from HIV-1
5' LTR (truncated)
233 .. 413  =  181 bp
truncated 5' long terminal repeat (LTR) from HIV-1
SV40 ori
8760 .. 8895  =  136 bp
SV40 origin of replication
SV40 ori
8760 .. 8895  =  136 bp
SV40 origin of replication
HIV-1 Ψ
460 .. 585  =  126 bp
packaging signal of human immunodeficiency virus type 1
HIV-1 Ψ
460 .. 585  =  126 bp
packaging signal of human immunodeficiency virus type 1
SV40 poly(A) signal
8599 .. 8720  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
8599 .. 8720  =  122 bp
SV40 polyadenylation signal
cPPT/CTS
2122 .. 2237  =  116 bp
central polypurine tract and central termination sequence of HIV-1 (lacking the first T)
cPPT/CTS
2122 .. 2237  =  116 bp
central polypurine tract and central termination sequence of HIV-1 (lacking the first T)
AmpR promoter
10,950 .. 11,054  =  105 bp
AmpR promoter
10,950 .. 11,054  =  105 bp
cloning site
2065 .. 2094  =  30 bp
sgRNA cloning site
cloning site
2065 .. 2094  =  30 bp
sgRNA cloning site
ORF:  7735 .. 8325  =  591 bp
ORF:  196 amino acids  =  20.9 kDa
ORF:  10,219 .. 10,485  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  11,335 .. 180  =  399 bp
ORF:  132 amino acids  =  15.1 kDa
ORF:  956 .. 1837  =  882 bp
ORF:  293 amino acids  =  33.8 kDa
ORF:  8621 .. 8857  =  237 bp
ORF:  78 amino acids  =  8.6 kDa
ORF:  2715 .. 7619  =  4905 bp
ORF:  1634 amino acids  =  187.0 kDa
ORF:  7722 .. 8267  =  546 bp
ORF:  181 amino acids  =  19.7 kDa
ORF:  871 .. 1251  =  381 bp
ORF:  126 amino acids  =  13.3 kDa
ORF:  11,295 .. 161  =  420 bp
ORF:  139 amino acids  =  15.8 kDa
ORF:  2703 .. 5312  =  2610 bp
ORF:  869 amino acids  =  91.7 kDa
ORF:  10,089 .. 10,949  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  2501 .. 2920  =  420 bp
ORF:  139 amino acids  =  15.7 kDa
ORF:  4142 .. 4435  =  294 bp
ORF:  97 amino acids  =  10.8 kDa
ORF:  4706 .. 5155  =  450 bp
ORF:  149 amino acids  =  17.3 kDa
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Individual Sequences & Maps

pAAV-AC-GFP
pLenti6.2 N-Lumio V5-DEST
pLVX-rHom-1
pRetroX-Tet3G
pAAV-AC-Myc-DDK
pLenti6.2 V5-DEST
pLVX-rHom-Nuc1
pRetroX-TetOne
pAAV-AC-RFP
pLenti6.3 TO V5-DEST
pLVX-rHom-Sec1
pRetroX-TetOne-Puro
pAAV2-EF1a-tGFP-WPRE
pLenti6.3 V5-DEST
pLVX-Tet-Off Advanced
pRetroX-Tight-Hyg
pAd BLOCK-iT-DEST
pLenti6.4 R4R2 V5-DEST
pLVX-Tet-On Advanced
pRetroX-Tight-Pur
pAd CMV V5-DEST
pLenti6 BLOCK-iT-DEST
pLVX-Tet3G
pRetroX-TRE3G
pAd PL-DEST
pLenti6 capTEV-CT-DEST
pLVX-TetOne
pRFP-C-RS
pAdenoX-CMV Linear
pLenti6 capTEV-NT-DEST1
pLVX-TetOne-Puro
pRFP-CB-shLenti
pAdenoX-DsRed-Express Linear
pLenti6 TR
pLVX-Tight-Puro
pRS
pAdenoX-PRLS Linear
pLenti6 UbC V5-DEST
pLVX-TRE3G
pRSGC1-U6-sg-HTS6-CMV-Cas9-2A-Puro
pAdenoX-PRLS-DsRed-Express Linear
pLenti6 V5-DEST
pLVX-TRE3G-Hom1
pRSGC2-U6-sg-HTS6-UbiC-Cas9-2A-Puro
pAdenoX-PRLS-ZsGreen1 Linear
pLenti7.3 V5-DEST
pLVX-TRE3G-IRES
pRSI12-U6-sh-HTS4-UbiC-TagRFP-2A-Puro
pAdenoX-Tet3G Linear
pLEX-MCS
pLVX-TRE3G-mCherry
pRSI16cb-U6-sh-13kCB18-HTS6-UbiC-TagRFP-2A-Puro
pAdenoX-ZsGreen1 Linear
pLHCX
pLVX-TRE3G-ZsGreen1
pRSI16-U6-sh-HTS6-UbiC-TagRFP-2A-Puro
pB-RS
pLKO.1
pLX301
pRSI17cb-U6-sh-13kCB18-HTS6-UbiC-TagGFP2-2A-Puro
pBABE-Hygro
pLKO.1 puro
pLX302
pRSI17-U6-sh-HTS6-UbiC-TagGFP2-2A-Puro
pBABE-Neo
pLMN-ZsGreen-Neomycin
pLX303
pRSIT12-U6Tet-sh-HTS4-CMV-TetRep-2A-TagRFP-2A-Puro
pBABE-Puro
pLMPd-Ametrine
pLX304
pRSIT16-U6Tet-sh-HTS6-CMV-TetRep-2A-TagRFP-2A-Puro
pBABE-Zeo
pLNCX2
pLX304-V5-Blast-Empty
pRSIT17-U6Tet-sh-HTS6-CMV-TetRep-2A-TagGFP2-2A-Puro
pCMV-VSV-G
pLP VSVG
pLXIN
pRSITEP-U6Tet-sh-noHTS-EF1-TetRep-2A-Puro
pGFP-B-RS
pLP1
pLXSN
pRSITUR-U6Tet-sh-HTS3-UbiC-TetRep-2A-TagRFP
pGFP-C-shLenti
pLP2
pMD2.G
pShuttle2
pGFP-V-RS
pLPCX
pMSCV16URP-U6-sh-HTS6-UbiC-TagRFP-2A-Puro
pSIREN-RetroQ
pGIPZ
pLVX-EF1alpha-IRES-mCherry
pMSCVhyg
pSIREN-RetroQ-DsRed-Express
pLenti-C-GFP
pLVX-EF1alpha-IRES-Puro
pMSCVneo
pSIREN-RetroQ-TetH
pLenti-C-mGFP
pLVX-EF1alpha-IRES-ZsGreen1
pMSCVpuro
pSIREN-RetroQ-TetP
pLenti-C-Myc-DDK
pLVX-EF1alpha-Tet3G
pQC-tTS-IN
pSIREN-RetroQ-ZsGreen1
pLenti-C-Myc-DDK-IRES-BSD
pLVX-Het-1
pQCXIH
psPAX2
pLenti-C-Myc-DDK-IRES-GFP
pLVX-Het-2
pQCXIN
pTRE3G-Hom1
pLenti-C-Myc-DDK-IRES-Neo
pLVX-Het-Mem1
pQCXIP
pTRIPZ
pLenti-C-Myc-DDK-IRES-Puro
pLVX-Het-Nuc1
pQCXIX
ptTS-Neo
pLenti-C-Myc-DDK-IRES-tRFP
pLVX-Hom-1
pRetro-Lib
pZIP-hCMV-RFP-Puro
pLenti-C-tRFP
pLVX-Hom-Mem1
pRetroQ-AcGFP1-C1
pZIP-hCMV-ZSGreen-Puro
pLenti-C-tYFP
pLVX-Hom-Nuc1
pRetroQ-AcGFP1-N1
pZIP-hEF1-alpha-RFP-Puro
pLenti-EF1a-C-mGFP
pLVX-IRES-Hyg
pRetroQ-DsRed-Monomer-C1
pZIP-hEF1-alpha-ZsGreen-Puro
pLenti-EF1a-C-Myc-DDK-IRES-Puro
pLVX-IRES-mCherry
pRetroQ-DsRed-Monomer-N1
pZIP-mCMV-RFP-Puro
pLenti-EF1a-C-tGFP
pLVX-IRES-Neo
pRetroQ-mCherry-C1
pZIP-mCMV-ZsGreen-Puro
pLenti-N-tGFP
pLVX-IRES-Puro
pRetroQ-mCherry-N1
pZIP-mEF1-alpha-RFP-Puro
pLenti-N-tRFP
pLVX-IRES-tdTomato
pRetroX-IRES-DsRedExpress
pZIP-mEF1-alpha-ZsGreen-Puro
pLenti-N-tYFP
pLVX-IRES-ZsGreen1
pRetroX-IRES-ZsGreen1
pZIP-SFFV-RFP-Puro
pLenti3.3 TR
pLVX-PTuner
pRetroX-PTuner
pZIP-SFFV-ZsGreen-Puro
pLenti4 BLOCK-iT-DEST
pLVX-PTuner Green
pRetroX-PTuner IRES
pZIP-TRE3G-ZsGreen-Puro
pLenti4 V5-DEST
pLVX-PTunerC
pRetroX-Tet-Off Advanced
pSIREN-Shuttle (linearized)
pLenti6.2 C-Lumio V5-DEST
pLVX-Puro
pRetroX-Tet-On Advanced
scramble shRNA

Plasmid Sets

Basic Cloning Vectors
I.M.A.G.E. Consortium Plasmids
pET & Duet Vectors (Novagen)
TA and GC Cloning Vectors
Coronavirus Resources
Insect Cell Vectors
pGEX Vectors (GE Healthcare)
TOPO Cloning Vectors
CRISPR Plasmids
Luciferase Vectors
Plant Vectors
Viral Expression & Packaging Vectors
Fluorescent Protein Genes & Plasmids
Lucigen Vectors
Qiagen Vectors
Yeast Plasmids
Gateway® Cloning Vectors
Mammalian Expression Vectors
Structural Genomics Vectors

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