pHTN HaloTag CMV-neo
Mammalian expression vector with a traditional MCS and ampicillin and G418 resistance markers, encoding a cleavable N-terminal HaloTag®.
Sequence Author: Promega
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PciI is inhibited by nonionic detergents. |
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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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After cleavage, BamHI-HF® (but not the original BamHI) can remain bound to DNA and alter its electrophoretic mobility. |
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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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Efficient cleavage requires at least two copies of the NaeI recognition sequence. |
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Efficient cleavage requires at least two copies of the NgoMIV recognition sequence. |
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Efficient cleavage requires at least two copies of the PluTI recognition sequence. |
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BsrGI is typically used at 37°C, but is even more active at 60°C. |
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Prolonged incubation with NdeI may lead to removal of additional nucleotides. |
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Sticky ends from different PflMI sites may not be compatible. |
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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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This recognition sequence is asymmetric, so ligating blunt ends generated by BmgBI will not always regenerate a BmgBI site. |
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Sticky ends from different PasI sites may not be compatible. |
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| * Blocked by Dam methylation. BclI is typically used at 50-55°C, but is 50% active at 37°C. |
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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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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 XmaI recognition sequence is present. |
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SmaI can be used at 37°C for brief incubations. |
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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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PaeR7I does not recognize the sequence CTCTCGAG. |
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Efficient cleavage requires at least two copies of the SacII recognition sequence. |
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EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage. |
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ApaI can be used between 25°C and 37°C. |
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Sticky ends from different BlpI sites may not be compatible. |
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Sticky ends from different CsiI sites may not be compatible. |
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| * Blocked by Dcm methylation. Sticky ends from different SexAI sites may not be compatible. |
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Efficient cleavage requires at least two copies of the SfiI recognition sequence. Sticky ends from different SfiI sites may not be compatible. |
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Efficient cleavage requires at least two copies of the NarI recognition sequence. |
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| HaloTag® 1067 .. 1957 = 891 bp 297 amino acids = 33.6 kDa Product: modified bacterial dehalogenase that forms covalent bonds with chloroalkane derivatives |
| HaloTag® 1067 .. 1957 = 891 bp 297 amino acids = 33.6 kDa Product: modified bacterial dehalogenase that forms covalent bonds with chloroalkane derivatives |
| TEV site 1970 .. 1990 = 21 bp 7 amino acids = 900.9 Da Product: tobacco etch virus (TEV) protease recognition and cleavage site |
| TEV site 1970 .. 1990 = 21 bp 7 amino acids = 900.9 Da Product: tobacco etch virus (TEV) protease recognition and cleavage site |
| AmpR 4085 .. 4945 = 861 bp 286 amino acids = 31.6 kDa 2 segments Segment 1: signal sequence 4085 .. 4153 = 69 bp 23 amino acids = 2.6 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
| AmpR 4085 .. 4945 = 861 bp 286 amino acids = 31.6 kDa 2 segments Segment 2: 4154 .. 4945 = 792 bp 263 amino acids = 28.9 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
| AmpR 4085 .. 4945 = 861 bp 286 amino acids = 31.6 kDa 2 segments Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
| NeoR/KanR 2918 .. 3712 = 795 bp 264 amino acids = 29.0 kDa Product: aminoglycoside phosphotransferase from Tn5 confers resistance to neomycin, kanamycin, and G418 (Geneticin®) |
| NeoR/KanR 2918 .. 3712 = 795 bp 264 amino acids = 29.0 kDa Product: aminoglycoside phosphotransferase from Tn5 confers resistance to neomycin, kanamycin, and G418 (Geneticin®) |
| ori 5103 .. 5691 = 589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication |
| ori 5103 .. 5691 = 589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication |
| CMV enhancer 138 .. 517 = 380 bp human cytomegalovirus immediate early enhancer |
| CMV enhancer 138 .. 517 = 380 bp human cytomegalovirus immediate early enhancer |
| SV40 promoter 2510 .. 2867 = 358 bp SV40 enhancer and early promoter |
| SV40 promoter 2510 .. 2867 = 358 bp SV40 enhancer and early promoter |
| cer region 5807 .. 6090 = 284 bp ColE1-derived recombination site that helps to maintain plasmids as monomers |
| cer region 5807 .. 6090 = 284 bp ColE1-derived recombination site that helps to maintain plasmids as monomers |
| CMV promoter 518 .. 721 = 204 bp human cytomegalovirus (CMV) immediate early promoter |
| CMV promoter 518 .. 721 = 204 bp human cytomegalovirus (CMV) immediate early promoter |
| chimeric intron 857 .. 989 = 133 bp chimera between introns from human β-globin and immunoglobulin heavy chain genes |
| chimeric intron 857 .. 989 = 133 bp chimera between introns from human β-globin and immunoglobulin heavy chain genes |
| SV40 poly(A) signal 2144 .. 2265 = 122 bp SV40 polyadenylation signal |
| SV40 poly(A) signal 2144 .. 2265 = 122 bp SV40 polyadenylation signal |
| AmpR promoter 3980 .. 4084 = 105 bp |
| AmpR promoter 3980 .. 4084 = 105 bp |
| MCS 1997 .. 2072 = 76 bp multiple cloning site |
| MCS 1997 .. 2072 = 76 bp multiple cloning site |
| poly(A) signal 3776 .. 3824 = 49 bp synthetic polyadenylation signal |
| poly(A) signal 3776 .. 3824 = 49 bp synthetic polyadenylation signal |
| T7 promoter 1033 .. 1051 = 19 bp promoter for bacteriophage T7 RNA polymerase |
| T7 promoter 1033 .. 1051 = 19 bp promoter for bacteriophage T7 RNA polymerase |
| SV40 ori 2718 .. 2853 = 136 bp SV40 origin of replication |
| SV40 ori 2718 .. 2853 = 136 bp SV40 origin of replication |
| ORF: 1067 .. 2107 = 1041 bp ORF: 346 amino acids = 39.0 kDa |
| ORF: 2918 .. 3712 = 795 bp ORF: 264 amino acids = 29.0 kDa |
| ORF: 4085 .. 4945 = 861 bp ORF: 286 amino acids = 31.6 kDa |
| ORF: 3090 .. 3476 = 387 bp ORF: 128 amino acids = 14.6 kDa |
| ORF: 2034 .. 2282 = 249 bp ORF: 82 amino acids = 9.2 kDa |
| ORF: 3588 .. 3893 = 306 bp ORF: 101 amino acids = 11.5 kDa |
| ORF: 821 .. 1630 = 810 bp ORF: 269 amino acids = 29.4 kDa |
| ORF: 3227 .. 3763 = 537 bp ORF: 178 amino acids = 19.8 kDa |
| ORF: 1630 .. 1866 = 237 bp ORF: 78 amino acids = 8.0 kDa |
| ORF: 4549 .. 4815 = 267 bp ORF: 88 amino acids = 9.2 kDa |
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