pcDNA4 HisMax-TOPO (linearized)
Linearized mammalian vector with 3'-T overhangs and bound topoisomerase, for TOPO® TA cloning of PCR products and expression of N-terminally 6xHis-tagged proteins.
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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FseI gradually loses activity when stored at -20°C. |
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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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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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Sticky ends from different BlpI 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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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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ApaI can be used between 25°C and 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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| AmpR 3094 .. 3954 = 861 bp 286 amino acids = 31.6 kDa 2 segments Segment 2: 3094 .. 3885 = 792 bp 263 amino acids = 28.9 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
| AmpR 3094 .. 3954 = 861 bp 286 amino acids = 31.6 kDa 2 segments Segment 1: signal sequence 3886 .. 3954 = 69 bp 23 amino acids = 2.6 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
| AmpR 3094 .. 3954 = 861 bp 286 amino acids = 31.6 kDa 2 segments Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
| ori 2338 .. 2923 = 586 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication |
| ori 2338 .. 2923 = 586 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication |
| f1 ori 377 .. 805 = 429 bp f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis |
| f1 ori 377 .. 805 = 429 bp f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis |
| CMV enhancer 4325 .. 4704 = 380 bp human cytomegalovirus immediate early enhancer |
| CMV enhancer 4325 .. 4704 = 380 bp human cytomegalovirus immediate early enhancer |
| BleoR 1262 .. 1636 = 375 bp 124 amino acids = 13.8 kDa Product: antibiotic-binding protein confers resistance to bleomycin, phleomycin, and Zeocin™ |
| BleoR 1262 .. 1636 = 375 bp 124 amino acids = 13.8 kDa Product: antibiotic-binding protein confers resistance to bleomycin, phleomycin, and Zeocin™ |
| SV40 promoter 819 .. 1148 = 330 bp SV40 enhancer and early promoter |