pRSITUR-U6Tet-sh-HTS3-UbiC-TetRep-2A-TagRFP

Lentiviral vector for inducible barcoded shRNA libraries, with an HTS3 cassette for high-throughput sequencing plus a TagRFP marker.

Sequence Author: Cellecta

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HomePlasmidsViral Expression & Packaging VectorspRSITUR-U6Tet-sh-HTS3-UbiC-TetRep-2A-TagRFP
MluI (1) SspI (7517) ScaI (7193) PciI (5820) SfiI (5398) NcoI (5352) AanI (5169) AanI (5113) KpnI (4778) Acc65I (4774) NaeI (4697) NgoMIV (4695) HincII (4179) SalI (4177) SphI (96) BssHII (490) NruI * (612) MfeI (964) BbvCI (1199) AleI (1352) PpuMI - SanDI (1709) BspDI - ClaI (1798) BsaAI (1928) BbsI - BpiI (2065) cloning site BbsI - BpiI (2091) EcoRI (2154) PaeR7I - XhoI (2165) BstBI (2356) AfeI (2416) XbaI (2760) BstXI (2856) BlpI (3314) BclI * (3344) BamHI (3391) BspEI (3451) BsrGI (3506) PasI (3687) PshAI (4096) pRSITUR-U6Tet-sh-HTS3-UbiC-TetRep-2A-TagRFP 8104 bp
MluI  (1)
1 site		 A C G C G T T G C G C A
SspI  (7517)
1 site		 A A T A T T T T A T A A
ScaI  (7193)
1 site		 A G T A C T T C A T G A
PciI  (5820)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
SfiI  (5398)
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.
NcoI  (5352)
1 site		 C C A T G G G G T A C C
AanI  (5169)
2 sites		 T T A T A A A A T A T T
AanI  (5113)
2 sites		 T T A T A A A A T A T T
KpnI  (4778)
1 site		 G G T A C C C C A T G G
Acc65I  (4774)
1 site		 G G T A C C C C A T G G
NaeI  (4697)
1 site		 G C C G G C C G G C C G
Efficient cleavage requires at least two copies of the NaeI recognition sequence.
NgoMIV  (4695)
1 site		 G C C G G C C G G C C G
Efficient cleavage requires at least two copies of the NgoMIV recognition sequence.
HincII  (4179)
1 site		 G T Y R A C C A R Y T G
SalI  (4177)
1 site		 G T C G A C C A G C T G
SphI  (96)
1 site		 G C A T G C C G T A C G
BssHII  (490)
1 site		 G C G C G C C G C G C G
BssHII is typically used at 50°C, but is 75% active at 37°C.
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
PpuMI  (1709)
1 site		 R G G W C C Y Y C C W G G R
Sticky ends from different PpuMI sites may not be compatible.
SanDI  (1709)
1 site		 G G G W C C C C C C W G G G
Sticky ends from different SanDI sites may not be compatible.
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
BsaAI  (1928)
1 site		 Y A C G T R R T G C A Y
BbsI  (2065)
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  (2065)
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  (2091)
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  (2091)
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.
EcoRI  (2154)
1 site		 G A A T T C C T T A A G
PaeR7I  (2165)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (2165)
1 site		 C T C G A G G A G C T C
BstBI  (2356)
1 site		 T T C G A A A A G C T T
AfeI  (2416)
1 site		 A G C G C T T C G C G A
XbaI  (2760)
1 site		 T C T A G A A G A T C T
BstXI  (2856)
1 site		 C C A N N N N N N T G G G G T N N N N N N A C C
Sticky ends from different BstXI sites may not be compatible.
BlpI  (3314)
1 site		 G C T N A G C C G A N T C G
Sticky ends from different BlpI sites may not be compatible.
BclI  (3344)
1 site		 T G A T C A A C T A G T
* Blocked by Dam methylation.
BclI is typically used at 50-55°C, but is 50% active at 37°C.
BamHI  (3391)
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.
BspEI  (3451)
1 site		 T C C G G A A G G C C T
BsrGI  (3506)
1 site		 T G T A C A A C A T G T
BsrGI is typically used at 37°C, but is even more active at 60°C.
PasI  (3687)
1 site		 C C C W G G G G G G W C C C
Sticky ends from different PasI sites may not be compatible.
PshAI  (4096)
1 site		 G A C N N N N G T C C T G N N N N C A G
PshAI quickly loses activity at 37°C, but can be used at 25°C for long incubations.
TetR
2770 .. 3390  =  621 bp
207 amino acids  =  23.4 kDa
Product: tetracycline repressor TetR
TetR binds to the tetracycline operator tetO to inhibit transcription. This inhibition can be relieved by adding tetracycline or doxycycline.
TetR
2770 .. 3390  =  621 bp
207 amino acids  =  23.4 kDa
Product: tetracycline repressor TetR
TetR binds to the tetracycline operator tetO to inhibit transcription. This inhibition can be relieved by adding tetracycline or doxycycline.
T2A
3397 .. 3450  =  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
3397 .. 3450  =  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.
TagRFP
3457 .. 4170  =  714 bp
237 amino acids  =  26.7 kDa
Product: monomeric derivative of red fluorescent protein from Entacmaea quadricolor (Merzlyak et al., 2007)
mammalian codon-optimized
TagRFP
3457 .. 4170  =  714 bp
237 amino acids  =  26.7 kDa
Product: monomeric derivative of red fluorescent protein from Entacmaea quadricolor (Merzlyak et al., 2007)
mammalian codon-optimized
AmpR
6640 .. 7500  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   6640 .. 7431  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6640 .. 7500  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   7432 .. 7500  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6640 .. 7500  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
WPRE
4183 .. 4771  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
WPRE
4183 .. 4771  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
ori
5881 .. 6469  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
5881 .. 6469  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
UbiC promoter
2361 .. 2760  =  400 bp
human ubiquitin C promoter
UbiC promoter
2361 .. 2760  =  400 bp
human ubiquitin C promoter
U6-Tet promoter
1819 .. 2064  =  246 bp
RNA polymerase III promoter for human U6 snRNA, modified to contain two tetracycline operator (tetO) sites
U6-Tet promoter
1819 .. 2064  =  246 bp
RNA polymerase III promoter for human U6 snRNA, modified to contain two tetracycline operator (tetO) sites
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.
3' LTR (ΔU3)
4845 .. 5078  =  234 bp
self-inactivating 3' long terminal repeat (LTR) from HIV-1
3' LTR (ΔU3)
4845 .. 5078  =  234 bp
self-inactivating 3' long terminal repeat (LTR) from HIV-1
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
5311 .. 5446  =  136 bp
SV40 origin of replication
SV40 ori
5311 .. 5446  =  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
5150 .. 5271  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
5150 .. 5271  =  122 bp
SV40 polyadenylation signal
cPPT/CTS
2195 .. 2312  =  118 bp
central polypurine tract and central termination sequence of HIV-1
cPPT/CTS
2195 .. 2312  =  118 bp
central polypurine tract and central termination sequence of HIV-1
AmpR promoter
7501 .. 7605  =  105 bp
AmpR promoter
7501 .. 7605  =  105 bp
cloning site
2066 .. 2095  =  30 bp
shRNA cloning site
cloning site
2066 .. 2095  =  30 bp
shRNA cloning site
tet operator
2046 .. 2064  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2046 .. 2064  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2020 .. 2037  =  18 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2020 .. 2037  =  18 bp
bacterial operator O2 for the tetR and tetA genes
ORF:  2770 .. 4170  =  1401 bp
ORF:  466 amino acids  =  52.2 kDa
ORF:  4273 .. 4818  =  546 bp
ORF:  181 amino acids  =  19.7 kDa
ORF:  956 .. 1837  =  882 bp
ORF:  293 amino acids  =  33.8 kDa
ORF:  3014 .. 3346  =  333 bp
ORF:  110 amino acids  =  12.4 kDa
ORF:  4286 .. 4876  =  591 bp
ORF:  196 amino acids  =  20.9 kDa
ORF:  6770 .. 7036  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  7886 .. 180  =  399 bp
ORF:  132 amino acids  =  15.1 kDa
ORF:  2175 .. 2510  =  336 bp
ORF:  111 amino acids  =  12.5 kDa
ORF:  5172 .. 5408  =  237 bp
ORF:  78 amino acids  =  8.6 kDa
ORF:  2039 .. 2266  =  228 bp
ORF:  75 amino acids  =  8.4 kDa
ORF:  871 .. 1251  =  381 bp
ORF:  126 amino acids  =  13.3 kDa
ORF:  6640 .. 7500  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  7846 .. 161  =  420 bp
ORF:  139 amino acids  =  15.8 kDa
ORF:  2559 .. 2939  =  381 bp
ORF:  126 amino acids  =  14.1 kDa
ORF:  3066 .. 3512  =  447 bp
ORF:  148 amino acids  =  16.4 kDa
ORF:  3570 .. 3815  =  246 bp
ORF:  81 amino acids  =  9.1 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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