pMirTarget

Reporter construct with TurboRFP and neomycin resistance markers, for cloning 3' UTR sequences downstream of firefly luciferase to validate miRNA targets.

Sequence Author: OriGene

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HomePlasmidsLuciferase VectorspMirTarget
SpeI (7507) AseI (7501) NdeI (7274) SnaBI (7170) T7 promoter AccI (6881) SalI (6880) BamHI (6873) BglII (6857) PshAI (6214) PaeR7I - PspXI - XhoI (6136) EcoRV (6096) PmeI (6062) FseI (6056) SmaI (6020) TspMI - XmaI (6018) AhdI (5907) BsmBI (5668) BstXI (5589) BstBI (4065) XbaI (4058) NotI (4047) MluI (4038) RsrII (4030) AscI - BssHII (4022) AsiSI - SgfI (4015) AanI - PsiI (72) StuI (884) BspDI * - ClaI * (903) PflFI - Tth111I (1181) AclI (1766) PspOMI (1856) ApaI (1860) XmnI (1955) HindIII (1967) PmlI (2059) AarI (2082) Acc65I (2184) KpnI (2188) BmgBI (2286) MreI - SgrAI (2397) HpaI (3539) EcoRI (4003) pMirTarget 7868 bp
SpeI  (7507)
1 site		 A C T A G T T G A T C A
AseI  (7501)
1 site		 A T T A A T T A A T T A
NdeI  (7274)
1 site		 C A T A T G G T A T A C
Prolonged incubation with NdeI may lead to removal of additional nucleotides.
SnaBI  (7170)
1 site		 T A C G T A A T G C A T
AccI  (6881)
1 site		 G T M K A C C A K M T G
Efficient cleavage with AccI requires ≥13 bp on each side of the recognition sequence.
Sticky ends from different AccI sites may not be compatible.
SalI  (6880)
1 site		 G T C G A C C A G C T G
BamHI  (6873)
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.
BglII  (6857)
1 site		 A G A T C T T C T A G A
PshAI  (6214)
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.
PaeR7I  (6136)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (6136)
1 site		 V C T C G A G B B G A G C T C V
XhoI  (6136)
1 site		 C T C G A G G A G C T C
EcoRV  (6096)
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.
PmeI  (6062)
1 site		 G T T T A A A C C A A A T T T G
FseI  (6056)
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.
SmaI  (6020)
1 site		 C C C G G G G G G C C C
SmaI can be used at 37°C for brief incubations.
TspMI  (6018)
1 site		 C C C G G G G G G C C C
XmaI  (6018)
1 site		 C C C G G G G G G C C C
Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
AhdI  (5907)
1 site		 G A C N N N N N G T C C T G N N N N N C A G
The 1-base overhangs produced by AhdI may be hard to ligate.
Sticky ends from different AhdI sites may not be compatible.
BsmBI  (5668)
1 site		 C G T C T C N G C A G A G N ( N ) 4
Sticky ends from different BsmBI sites may not be compatible.
BsmBI-v2 is an improved version of BsmBI.
BstXI  (5589)
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.
BstBI  (4065)
1 site		 T T C G A A A A G C T T
XbaI  (4058)
1 site		 T C T A G A A G A T C T
NotI  (4047)
1 site		 G C G G C C G C C G C C G G C G
MluI  (4038)
1 site		 A C G C G T T G C G C A
RsrII  (4030)
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.
AscI  (4022)
1 site		 G G C G C G C C C C G C G C G G
BssHII  (4022)
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.
AsiSI  (4015)
1 site		 G C G A T C G C C G C T A G C G
SgfI  (4015)
1 site		 G C G A T C G C C G C T A G C G
AanI  (72)
1 site		 T T A T A A A A T A T T
PsiI  (72)
1 site		 T T A T A A A A T A T T
StuI  (884)
1 site		 A G G C C T T C C G G A
BspDI  (903)
1 site		 A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (903)
1 site		 A T C G A T T A G C T A
* Blocked by Dam methylation.
PflFI  (1181)
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  (1181)
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.
AclI  (1766)
1 site		 A A C G T T T T G C A A
PspOMI  (1856)
1 site		 G G G C C C C C C G G G
ApaI  (1860)
1 site		 G G G C C C C C C G G G
ApaI can be used between 25°C and 37°C.
XmnI  (1955)
1 site		 G A A N N N N T T C C T T N N N N A A G
HindIII  (1967)
1 site		 A A G C T T T T C G A A
PmlI  (2059)
1 site		 C A C G T G G T G C A C
AarI  (2082)
1 site		 C A C C T G C ( N ) 4 G T G G A C G ( N ) 4 ( N ) 4
Cleavage may be enhanced when more than one copy of the AarI recognition sequence is present.
Sticky ends from different AarI sites may not be compatible.
After cleavage, AarI can remain bound to DNA and alter its electrophoretic mobility.
Acc65I  (2184)
1 site		 G G T A C C C C A T G G
KpnI  (2188)
1 site		 G G T A C C C C A T G G
BmgBI  (2286)
1 site		 C A C G T C G T G C A G
This recognition sequence is asymmetric, so ligating blunt ends generated by BmgBI will not always regenerate a BmgBI site.
MreI  (2397)
1 site		 C G C C G G C G G C G G C C G C
SgrAI  (2397)
1 site		 C R C C G G Y G G Y G G C C R C
Efficient cleavage requires at least two copies of the SgrAI recognition sequence.
HpaI  (3539)
1 site		 G T T A A C C A A T T G
EcoRI  (4003)
1 site		 G A A T T C C T T A A G
luciferase
2334 .. 3986  =  1653 bp
550 amino acids  =  60.6 kDa
Product: firefly luciferase
synthetic version of the luciferase gene
luciferase
2334 .. 3986  =  1653 bp
550 amino acids  =  60.6 kDa
Product: firefly luciferase
synthetic version of the luciferase gene
NeoR/KanR
935 .. 1729  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR/KanR
935 .. 1729  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
TurboRFP
6150 .. 6842  =  693 bp
231 amino acids  =  26.1 kDa
Product: red fluorescent protein from Entacmaea quadricolor
mammalian codon-optimized
TurboRFP
6150 .. 6842  =  693 bp
231 amino acids  =  26.1 kDa
Product: red fluorescent protein from Entacmaea quadricolor
mammalian codon-optimized
hGH poly(A) signal
5398 .. 6020  =  623 bp
human growth hormone polyadenylation signal
hGH poly(A) signal
5398 .. 6020  =  623 bp
human growth hormone polyadenylation signal
ori
4661 .. 5249  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
4661 .. 5249  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
IRES
1766 .. 2318  =  553 bp
internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)
IRES
1766 .. 2318  =  553 bp
internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)
f1 ori
7844 .. 431  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
7844 .. 431  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
CMV enhancer
7147 .. 7526  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
7147 .. 7526  =  380 bp
human cytomegalovirus immediate early enhancer
CMV promoter
6943 .. 7146  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
6943 .. 7146  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
SV40 ori
752 .. 886  =  135 bp
SV40 origin of replication
SV40 ori
752 .. 886  =  135 bp
SV40 origin of replication
AmpR promoter
458 .. 562  =  105 bp
AmpR promoter
458 .. 562  =  105 bp
MCS
4003 .. 4069  =  67 bp
multiple cloning site
MCS
4003 .. 4069  =  67 bp
multiple cloning site
HSV TK poly(A) signal
4285 .. 4332  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
HSV TK poly(A) signal
4285 .. 4332  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
T7 promoter
6899 .. 6917  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
6899 .. 6917  =  19 bp
promoter for bacteriophage T7 RNA polymerase
ORF:  6496 .. 6741  =  246 bp
ORF:  81 amino acids  =  9.1 kDa
ORF:  935 .. 1729  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  2318 .. 2587  =  270 bp
ORF:  89 amino acids  =  10.7 kDa
ORF:  5432 .. 5656  =  225 bp
ORF:  74 amino acids  =  8.1 kDa
ORF:  1107 .. 1493  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  2226 .. 3986  =  1761 bp
ORF:  586 amino acids  =  64.9 kDa
ORF:  4074 .. 4523  =  450 bp
ORF:  149 amino acids  =  16.3 kDa
ORF:  2241 .. 2561  =  321 bp
ORF:  106 amino acids  =  12.1 kDa
ORF:  3330 .. 4058  =  729 bp
ORF:  242 amino acids  =  25.2 kDa
ORF:  6030 .. 6842  =  813 bp
ORF:  270 amino acids  =  30.6 kDa
ORF:  1244 .. 1498  =  255 bp
ORF:  84 amino acids  =  9.6 kDa
ORF:  4279 .. 4512  =  234 bp
ORF:  77 amino acids  =  8.6 kDa
ORF:  6601 .. 6987  =  387 bp
ORF:  128 amino acids  =  15.0 kDa
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Download pMirTarget.dna file

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Individual Sequences & Maps

pBI-CMV5
pGL4.22[luc2CP Puro]
pGL4.81[hRlucCP Neo]
pRL-null
pBIND-ER(alpha)
pGL4.23[luc2 minP]
pGL4.82[hRluc Puro]
pRL-SV40
pBIND-GR
pGL4.24[luc2P minP]
pGL4.83[hRlucP Puro]
pRL-TK
pCBG68-Basic
pGL4.25[luc2CP minP]
pGL4.84[hRlucCP Puro]
pSELECT-zeo-Lucia
pCBG68-Control
pGL4.26[luc2 minP Hygro]
pGLuc Mini-TK 2
pSF-CMV-EMCV-FLuc
pCBG99-Basic
pGL4.27[luc2P minP Hygro]
pGLuc-Basic 2
pSF-CMV-EMCV-RLuc
pCBG99-Control
pGL4.28[luc2CP minP Hygro]
phRG-B
pSF-CMV-FLuc
pCBR-Basic
pGL4.29[luc2P CRE Hygro]
phRG-TK
pSF-CMV-FMDV-FLuc
pCBR-Control
pGL4.30[luc2P NFAT-RE Hygro]
phRL-CMV
pSF-CMV-FMDV-RLuc
pCLuc Mini-TK 2
pGL4.31[luc2P GAL4UAS Hygro]
phRL-null
pSF-CMV-PGK-FLuc
pCLuc-Basic 2
pGL4.32[luc2P NF-kB-RE Hygro]
phRL-SV40
pSF-CMV-PGK-RLuc
pCMV-CLuc 2
pGL4.33[luc2P SRE Hygro]
phRL-TK
pSF-CMV-RLuc
pCMV-Cypridina Luc
pGL4.34[luc2P SRF-RE Hygro]
phRL-TK(Int-)
pSF-CMV-RSV-FLuc AscI
pCMV-Gaussia Luc
pGL4.35[luc2P 9XGAL4UAS Hygro]
pLightSwitch_Prom
pSF-CMV-RSV-RLuc AscI
pCMV-Gaussia-Dura Luc
pGL4.36[luc2P MMTV Hygro]
pLVX-MetLuc Control
pSF-CMV-Ub-FLuc AscI
pCMV-GLuc 2
pGL4.37[luc2P ARE Hygro]
pLVX-MetLuc Reporter
pSF-CMV-Ub-RLuc AscI
pCMV-Green Renilla Luc
pGL4.38[luc2P p53 RE Hygro]
pMCS-Cypridina Luc
pSF-CMVe-FLuc
pCMV-Red Firefly Luc
pGL4.39[luc2P ATF6 RE Hygro]
pMCS-Gaussia Luc
pSF-CMVe-RLuc
pCRE-MetLuc2-Reporter
pGL4.40[luc2P MRE Hygro]
pMCS-Gaussia-Dura Luc
pSF-MinProm-FLuc
pFN26A (BIND) hRluc-neo Flexi Vector
pGL4.41[luc2P HSE Hygro]
pMCS-Green Renilla Luc
pSF-MinProm-RLuc
pGEM-luc
pGL4.42[luc2P HRE Hygro]
pMCS-Red Firefly Luc
pSF-pA-CMVe-FLuc
pGL2-Basic
pGL4.43[luc2P XRE Hygro]
pMetLuc-Mem Control
pSF-pA-CMVe-RLuc
pGL2-Control
pGL4.44[luc2P AP1 RE Hygro]
pMetLuc-Mem Reporter
pSF-pA-MinProm-FLuc
pGL2-Enhancer
pGL4.45[luc2P ISRE Hygro]
pMetLuc2-Control
pSF-pA-MinProm-RLuc
pGL2-Promoter
pGL4.47[luc2P SIE Hygro]
pMetLuc2-Reporter
pSF-pA-PromMCS-FLuc
pGL3-Basic
pGL4.48[luc2P SBE Hygro]
pmirGLO
pSF-pA-PromMCS-RLuc
pGL3-Control
pGL4.49[luc2P TCF-LEF RE Hygro]
pMirTarget
pSF-PromMCS-FLuc
pGL3-Enhancer
pGL4.50[luc2 CMV Hygro]
pNFkB-MetLuc2-Reporter
pSF-PromMCS-RLuc
pGL3-Promoter
pGL4.51[luc2 CMV Neo]
pNL1.1.CMV[Nluc CMV]
pSF-RecA-FLuc
pGL4.10[luc2]
pGL4.52[luc2P STAT5 RE Hygro]
pNL1.1[Nluc]
pSP-luc+NF
pGL4.11[luc2P]
pGL4.70[hRluc]
pNL1.2[NlucP]
pSV40-CLuc
pGL4.12[luc2CP]
pGL4.71[hRlucP]
pNL1.3.CMV[secNluc CMV]
pSV40-GLuc
pGL4.13[luc2 SV40]
pGL4.72[hRlucCP]
pNL1.3[secNluc]
pTK-CLuc
pGL4.14[luc2 Hygro]
pGL4.73[hRluc SV40]
pNL2.1[Nluc Hygro]
pTK-Cypridina Luc
pGL4.15[luc2P Hygro]
pGL4.74[hRluc TK]
pNL2.2[NlucP Hygro]
pTK-Gaussia Luc
pGL4.16[luc2CP Hygro]
pGL4.75[hRluc CMV]
pNL2.3[secNluc Hygro]
pTK-Gaussia-Dura Luc
pGL4.17[luc2 Neo]
pGL4.76[hRluc Hygro]
pNL3.1[Nluc minP]
pTK-GLuc
pGL4.18[luc2P Neo]
pGL4.77[hRlucP Hygro]
pNL3.2.NF-kB-RE[NlucP NF-kB-RE Hygro]
pTK-Green Renilla Luc
pGL4.19[luc2CP Neo]
pGL4.78[hRlucCP Hygro]
pNL3.2[NlucP minP]
pTK-Red Firefly Luc
pGL4.20[luc2 Puro]
pGL4.79[hRluc Neo]
pNL3.3[secNluc minP]
pXPG
pGL4.21[luc2P Puro]
pGL4.80[hRlucP Neo]
pRL-CMV

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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