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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Prolonged incubation with NdeI may lead to removal of additional nucleotides. |
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Efficient cleavage with AccI requires ≥13 bp on each side of the recognition sequence.Sticky ends from different AccI sites may not be compatible. |
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After cleavage, BamHI-HF® (but not the original BamHI) can remain bound to DNA and alter its electrophoretic mobility. |
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PshAI quickly loses activity at 37°C, but can be used at 25°C for long incubations. |
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PaeR7I does not recognize the sequence CTCTCGAG. |
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EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage. |
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FseI gradually loses activity when stored at -20°C. |
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SmaI can be used at 37°C for brief incubations. |
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Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present. |
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The 1-base overhangs produced by AhdI may be hard to ligate. Sticky ends from different AhdI sites may not be compatible. |
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Sticky ends from different BsmBI sites may not be compatible.BsmBI-v2 is an improved version of BsmBI. |
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Sticky ends from different BstXI sites may not be compatible. |
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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. |
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BssHII is typically used at 50°C, but is 75% active at 37°C. |
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* Blocked by Dam methylation. |
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* Blocked by Dam methylation. |
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The 1-base overhangs produced by PflFI may be hard to ligate.Sticky ends from different PflFI sites may not be compatible. |
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The 1-base overhangs produced by Tth111I may be hard to ligate.Sticky ends from different Tth111I sites may not be compatible. |
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ApaI can be used between 25°C and 37°C. |
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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. |
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This recognition sequence is asymmetric, so ligating blunt ends generated by BmgBI will not always regenerate a BmgBI site. |
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Efficient cleavage requires at least two copies of the SgrAI recognition sequence. |
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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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