pcDNA4 V5-His C
Mammalian vector for expressing C-terminally V5- and 6xHis-tagged proteins. For other reading frames, use pcDNA™4/V5-His A or pcDNA™4/V5-His B.
Sequence Author: Thermo Fisher (Invitrogen)
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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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PciI is inhibited by nonionic detergents. |
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Sticky ends from different BspQI sites may not be compatible. |
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Sticky ends from different SapI sites may not be compatible.SapI gradually settles in solution, so a tube of SapI should be mixed before removing an aliquot. |
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Sticky ends from different BsmI sites may not be compatible. |
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Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present. This recognition sequence is asymmetric, so ligating sticky ends generated by Bpu10I will not always regenerate a Bpu10I site.Sticky ends from different Bpu10I sites may not be compatible. |
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Prolonged incubation with NdeI may lead to removal of additional nucleotides. |
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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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EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage. |
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PaeR7I does not recognize the sequence CTCTCGAG. |
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Sticky ends from different BstEII sites may not be compatible.BstEII is typically used at 60°C, but is 50% active at 37°C. |
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Sticky ends from different BbsI sites may not be compatible.BbsI gradually loses activity when stored at -20°C. |
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BssHII is typically used at 50°C, but is 75% active at 37°C. |
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Efficient cleavage requires at least two copies of the SgrAI recognition sequence. |
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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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FseI gradually loses activity when stored at -20°C. |
| AmpR 4092 .. 4952 = 861 bp 286 amino acids = 31.6 kDa 2 segments Segment 2: 4092 .. 4883 = 792 bp 263 amino acids = 28.9 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
| AmpR 4092 .. 4952 = 861 bp 286 amino acids = 31.6 kDa 2 segments Segment 1: signal sequence 4884 .. 4952 = 69 bp 23 amino acids = 2.6 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
| AmpR 4092 .. 4952 = 861 bp 286 amino acids = 31.6 kDa 2 segments Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
| ori 3333 .. 3921 = 589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication |
| ori 3333 .. 3921 = 589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication |
| f1 ori 1372 .. 1800 = 429 bp f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis |
| f1 ori 1372 .. 1800 = 429 bp f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis |
| CMV enhancer 235 .. 614 = 380 bp human cytomegalovirus immediate early enhancer |
| CMV enhancer 235 .. 614 = 380 bp human cytomegalovirus immediate early enhancer |
| BleoR 2257 .. 2631 = 375 bp 124 amino acids = 13.8 kDa Product: antibiotic-binding protein confers resistance to bleomycin, phleomycin, and Zeocin™ |
| BleoR 2257 .. 2631 = 375 bp 124 amino acids = 13.8 kDa Product: antibiotic-binding protein confers resistance to bleomycin, phleomycin, and Zeocin™ |
| SV40 promoter 1814 .. 2143 = 330 bp SV40 enhancer and early promoter |
| SV40 promoter 1814 .. 2143 = 330 bp SV40 enhancer and early promoter |
| bGH poly(A) signal 1102 .. 1326 = 225 bp bovine growth hormone polyadenylation signal |
| bGH poly(A) signal 1102 .. 1326 = 225 bp bovine growth hormone polyadenylation signal |
| CMV promoter 615 .. 818 = 204 bp human cytomegalovirus (CMV) immediate early promoter |
| CMV promoter 615 .. 818 = 204 bp human cytomegalovirus (CMV) immediate early promoter |
| SV40 poly(A) signal 2761 .. 2882 = 122 bp SV40 polyadenylation signal |
| SV40 poly(A) signal 2761 .. 2882 = 122 bp SV40 polyadenylation signal |
| MCS 895 .. 1004 = 110 bp multiple cloning site |
| MCS 895 .. 1004 = 110 bp multiple cloning site |
| AmpR promoter 4953 .. 5057 = 105 bp |
| AmpR promoter 4953 .. 5057 = 105 bp |
| V5 tag 1005 .. 1046 = 42 bp 14 amino acids = 1.4 kDa Product: epitope tag from simian virus 5 |
| V5 tag 1005 .. 1046 = 42 bp 14 amino acids = 1.4 kDa Product: epitope tag from simian virus 5 |
| 6xHis 1056 .. 1073 = 18 bp 6 amino acids = 840.9 Da Product: 6xHis affinity tag |
| 6xHis 1056 .. 1073 = 18 bp 6 amino acids = 840.9 Da Product: 6xHis affinity tag |
| EM7 promoter 2191 .. 2238 = 48 bp synthetic bacterial promoter |
| EM7 promoter 2191 .. 2238 = 48 bp synthetic bacterial promoter |
| T7 promoter 863 .. 881 = 19 bp promoter for bacteriophage T7 RNA polymerase |
| T7 promoter 863 .. 881 = 19 bp promoter for bacteriophage T7 RNA polymerase |
| SV40 ori 1994 .. 2129 = 136 bp SV40 origin of replication |
| SV40 ori 1994 .. 2129 = 136 bp SV40 origin of replication |
| ORF: 2257 .. 2631 = 375 bp ORF: 124 amino acids = 13.8 kDa |
| ORF: 4222 .. 4488 = 267 bp ORF: 88 amino acids = 9.2 kDa |
| ORF: 1323 .. 1589 = 267 bp ORF: 88 amino acids = 9.3 kDa |
| ORF: 2250 .. 2537 = 288 bp ORF: 95 amino acids = 10.1 kDa |
| ORF: 4092 .. 4952 = 861 bp ORF: 286 amino acids = 31.6 kDa |