pcDNA6.2 V5-pL-DEST
Promoterless Gateway® destination vector for cloning of mammalian promoters and genes.
Sequence Author: Thermo Fisher (Invitrogen)
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Sticky ends from different BglI 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 PfoI sites may not be compatible. |
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Sticky ends from different BlpI sites may not be compatible. |
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| * Blocked by Dam methylation. |
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Efficient cleavage requires at least two copies of the BsgI recognition sequence. Sticky ends from different BsgI sites may not be compatible.For full activity, add fresh S-adenosylmethionine (SAM). |
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Sticky ends from different PpuMI sites may not be compatible. |
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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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| * Blocked by Dam methylation. BclI is typically used at 50-55°C, but is 50% active at 37°C. |
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Efficient cleavage requires at least two copies of the BfuAI recognition sequence. Sticky ends from different BfuAI sites may not be compatible.BfuAI is typically used at 50°C, but is 50% active at 37°C. |
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Efficient cleavage requires at least two copies of the BspMI recognition sequence. Sticky ends from different BspMI sites may not be compatible. |
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Sticky ends from different BstXI sites may not be compatible. |
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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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BssHII is typically used at 50°C, but is 75% active at 37°C. |
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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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| * Blocked by Dam methylation. |
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ApaI can be used between 25°C and 37°C. |
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Efficient cleavage requires at least two copies of the SacII recognition sequence. |
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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. |
| AmpR 5583 .. 6443 = 861 bp 286 amino acids = 31.6 kDa 2 segments Segment 2: 5583 .. 6374 = 792 bp 263 amino acids = 28.9 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
| AmpR 5583 .. 6443 = 861 bp 286 amino acids = 31.6 kDa 2 segments Segment 1: signal sequence 6375 .. 6443 = 69 bp 23 amino acids = 2.6 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
| AmpR 5583 .. 6443 = 861 bp 286 amino acids = 31.6 kDa 2 segments Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
| CmR 1352 .. 2005 = 654 bp 218 amino acids = 25.6 kDa Product: chloramphenicol acetyltransferase confers resistance to chloramphenicol |
| CmR 1352 .. 2005 = 654 bp 218 amino acids = 25.6 kDa Product: chloramphenicol acetyltransferase confers resistance to chloramphenicol |
| ori 4824 .. 5412 = 589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication |
| ori 4824 .. 5412 = 589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication |
| f1 ori 2811 .. 3239 = 429 bp f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis |
| f1 ori 2811 .. 3239 = 429 bp f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis |
| BSD 3696 .. 4094 = 399 bp 132 amino acids = 13.7 kDa Product: blasticidin S deaminase confers resistance to blasticidin |
| BSD 3696 .. 4094 = 399 bp 132 amino acids = 13.7 kDa Product: blasticidin S deaminase confers resistance to blasticidin |
| SV40 promoter 3253 .. 3582 = 330 bp SV40 enhancer and early promoter |
| SV40 promoter 3253 .. 3582 = 330 bp SV40 enhancer and early promoter |
| ccdB 697 .. 1002 = 306 bp 101 amino acids = 11.7 kDa Product: CcdB, a bacterial toxin that poisons DNA gyrase Plasmids containing the ccdB gene cannot be propagated in standard E. coli strains. |
| ccdB 697 .. 1002 = 306 bp 101 amino acids = 11.7 kDa Product: CcdB, a bacterial toxin that poisons DNA gyrase Plasmids containing the ccdB gene cannot be propagated in standard E. coli strains. |
| attR1 144 .. 268 = 125 bp recombination site for the Gateway® LR reaction |
| attR1 144 .. 268 = 125 bp recombination site for the Gateway® LR reaction |
| attR2 2286 .. 2410 = 125 bp recombination site for the Gateway® LR reaction |
| attR2 2286 .. 2410 = 125 bp recombination site for the Gateway® LR reaction |
| SV40 poly(A) signal 4252 .. 4373 = 122 bp SV40 polyadenylation signal |
| SV40 poly(A) signal 4252 .. 4373 = 122 bp SV40 polyadenylation signal |
| AmpR promoter 6444 .. 6548 = 105 bp |
| AmpR promoter 6444 .. 6548 = 105 bp |
| cat promoter 2006 .. 2108 = 103 bp promoter of the E. coli cat gene |
| cat promoter 2006 .. 2108 = 103 bp promoter of the E. coli cat gene |
| HSV TK poly(A) signal 2561 .. 2609 = 49 bp herpesvirus thymidine kinase polyadenylation signal |
| HSV TK poly(A) signal 2561 .. 2609 = 49 bp herpesvirus thymidine kinase polyadenylation signal |
| EM7 promoter 3630 .. 3677 = 48 bp synthetic bacterial promoter |
| EM7 promoter 3630 .. 3677 = 48 bp synthetic bacterial promoter |
| V5 tag 2436 .. 2477 = 42 bp 14 amino acids = 1.4 kDa Product: epitope tag from simian virus 5 |
| V5 tag 2436 .. 2477 = 42 bp 14 amino acids = 1.4 kDa Product: epitope tag from simian virus 5 |
| T7 promoter 96 .. 114 = 19 bp promoter for bacteriophage T7 RNA polymerase |
| T7 promoter 96 .. 114 = 19 bp promoter for bacteriophage T7 RNA polymerase |
| SV40 ori 3433 .. 3568 = 136 bp SV40 origin of replication |
| SV40 ori 3433 .. 3568 = 136 bp SV40 origin of replication |
| ORF: 5713 .. 5979 = 267 bp ORF: 88 amino acids = 9.2 kDa |
| ORF: 3696 .. 4094 = 399 bp ORF: 132 amino acids = 13.7 kDa |
| ORF: 697 .. 1002 = 306 bp ORF: 101 amino acids = 11.7 kDa |
| ORF: 3642 .. 3998 = 357 bp ORF: 118 amino acids = 12.8 kDa |
| ORF: 5583 .. 6443 = 861 bp ORF: 286 amino acids = 31.6 kDa |
| ORF: 1322 .. 2005 = 684 bp ORF: 227 amino acids = 26.7 kDa |
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