pcDNA6.2 C-YFP-GW TOPO (linearized)
Linearized vector with 3'-T overhangs and bound topoisomerase, for TOPO® TA cloning to add a C-terminal YFP tag and create a Gateway® expression clone.
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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| * Blocked by Dam methylation. |
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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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SmaI can be used at 37°C for brief incubations. |
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Sticky ends from different AvaI sites may not be compatible. |
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Sticky ends from different BsoBI sites may not be compatible.BsoBI is typically used at 37°C, but can be used at temperatures up to 65°C. |
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Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present. |
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Prolonged incubation with NdeI may lead to removal of additional nucleotides. |
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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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| AmpR 3865 .. 4725 = 861 bp 286 amino acids = 31.6 kDa 2 segments Segment 2: 3865 .. 4656 = 792 bp 263 amino acids = 28.9 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
| AmpR 3865 .. 4725 = 861 bp 286 amino acids = 31.6 kDa 2 segments Segment 1: signal sequence 4657 .. 4725 = 69 bp 23 amino acids = 2.6 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
| AmpR 3865 .. 4725 = 861 bp 286 amino acids = 31.6 kDa 2 segments Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
| Topaz YFP 52 .. 771 = 720 bp 239 amino acids = 27.0 kDa 3 segments Segment 1: 52 .. 54 = 3 bp 1 amino acid = 149.2 Da Product: enhanced yellow variant of GFP mammalian codon-optimized |
| Topaz YFP 52 .. 771 = 720 bp 239 amino acids = 27.0 kDa 3 segments Segment 2: 1a 55 .. 57 = 3 bp 1 amino acid = 117.1 Da Product: enhanced yellow variant of GFP mammalian codon-optimized |
| Topaz YFP 52 .. 771 = 720 bp 239 amino acids = 27.0 kDa 3 segments Segment 3: 58 .. 771 = 714 bp 237 amino acids = 26.8 kDa Product: enhanced yellow variant of GFP mammalian codon-optimized |
| Topaz YFP 52 .. 771 = 720 bp 239 amino acids = 27.0 kDa 3 segments Product: enhanced yellow variant of GFP mammalian codon-optimized |
| ori 3106 .. 3694 = 589 bp high-copy-number colE1/pMB1/pBR322/pUC origin of replication |
| ori 3106 .. 3694 = 589 bp high-copy-number colE1/pMB1/pBR322/pUC origin of replication |
| f1 ori 1093 .. 1521 = 429 bp f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis |
| f1 ori 1093 .. 1521 = 429 bp f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis |
| BSD 1978 .. 2376 = 399 bp 132 amino acids = 13.7 kDa Product: blasticidin S deaminase confers resistance to blasticidin |
| BSD 1978 .. 2376 = 399 bp 132 amino acids = 13.7 kDa Product: blasticidin S deaminase confers resistance to blasticidin |
| CMV enhancer 5096 .. 5475 = 380 bp human cytomegalovirus immediate early enhancer |
| CMV enhancer 5096 .. 5475 = 380 bp human cytomegalovirus immediate early enhancer |
| SV40 promoter 1535 .. 1864 = 330 bp SV40 enhancer and early promoter |
| SV40 promoter 1535 .. 1864 = 330 bp SV40 enhancer and early promoter |
| CMV promoter 5476 .. 5679 = 204 bp human cytomegalovirus (CMV) immediate early promoter |
| CMV promoter 5476 .. 5679 = 204 bp human cytomegalovirus (CMV) immediate early promoter |
| SV40 poly(A) signal 2534 .. 2655 = 122 bp SV40 polyadenylation signal |
| SV40 poly(A) signal 2534 .. 2655 = 122 bp SV40 polyadenylation signal |
| AmpR promoter 4726 .. 4830 = 105 bp |
| AmpR promoter 4726 .. 4830 = 105 bp |
| HSV TK poly(A) signal 843 .. 891 = 49 bp herpesvirus thymidine kinase polyadenylation signal |
| HSV TK poly(A) signal 843 .. 891 = 49 bp herpesvirus thymidine kinase polyadenylation signal |
| EM7 promoter 1912 .. 1959 = 48 bp synthetic bacterial promoter |
| EM7 promoter 1912 .. 1959 = 48 bp synthetic bacterial promoter |
| attB2 14 .. 38 = 25 bp recombination site for the Gateway® BP reaction |
| attB2 14 .. 38 = 25 bp recombination site for the Gateway® BP reaction |
| attB1 5776 .. 5800 = 25 bp recombination site for the Gateway® BP reaction |
| attB1 5776 .. 5800 = 25 bp recombination site for the Gateway® BP reaction |
| T7 promoter 5724 .. 5742 = 19 bp promoter for bacteriophage T7 RNA polymerase |
| T7 promoter 5724 .. 5742 = 19 bp promoter for bacteriophage T7 RNA polymerase |
| SV40 ori 1715 .. 1850 = 136 bp SV40 origin of replication |
| SV40 ori 1715 .. 1850 = 136 bp SV40 origin of replication |
| ORF: 1 .. 771 = 771 bp ORF: 256 amino acids = 28.9 kDa (no start codon) |
| ORF: 1978 .. 2376 = 399 bp ORF: 132 amino acids = 13.7 kDa |
| ORF: 3995 .. 4261 = 267 bp ORF: 88 amino acids = 9.2 kDa |
| ORF: 1924 .. 2280 = 357 bp ORF: 118 amino acids = 12.8 kDa |
| ORF: 3865 .. 4725 = 861 bp ORF: 286 amino acids = 31.6 kDa |
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