pcDNA3.1 CT-GFP
Vector for expressing C-terminally GFP-tagged proteins in mammalian cells.
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 BsmI sites may not be compatible. |
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Sticky ends from different PfoI 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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BssHII is typically used at 50°C, but is 75% active at 37°C. |
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The 1-base overhangs produced by PflFI may be hard to ligate.Sticky ends from different PflFI sites may not be compatible. |
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The 1-base overhangs produced by Tth111I may be hard to ligate.Sticky ends from different Tth111I sites may not be compatible. |
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Efficient cleavage requires at least two copies of the PluTI 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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EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage. |
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| * Blocked by Dam methylation. |
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BsrGI is typically used at 37°C, but is even more active at 60°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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Sticky ends from different DraIII 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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Sticky ends from different BseRI sites may not be compatible.BseRI quickly loses activity at 37°C.Prolonged incubation with BseRI may lead to degradation of the DNA. |
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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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| * Blocked by Dam methylation. |
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Efficient cleavage requires at least two copies of the NarI recognition sequence. |
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| AmpR 5145 .. 6005 = 861 bp 286 amino acids = 31.6 kDa 2 segments Segment 2: 5145 .. 5936 = 792 bp 263 amino acids = 28.9 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
| AmpR 5145 .. 6005 = 861 bp 286 amino acids = 31.6 kDa 2 segments Segment 1: signal sequence 5937 .. 6005 = 69 bp 23 amino acids = 2.6 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
| AmpR 5145 .. 6005 = 861 bp 286 amino acids = 31.6 kDa 2 segments Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
| NeoR/KanR 2844 .. 3638 = 795 bp 264 amino acids = 29.0 kDa Product: aminoglycoside phosphotransferase from Tn5 confers resistance to neomycin, kanamycin, and G418 (Geneticin®) |
| NeoR/KanR 2844 .. 3638 = 795 bp 264 amino acids = 29.0 kDa Product: aminoglycoside phosphotransferase from Tn5 confers resistance to neomycin, kanamycin, and G418 (Geneticin®) |
| Cycle 3 GFP 988 .. 1707 = 720 bp 239 amino acids = 26.9 kDa 3 segments Segment 1: 988 .. 990 = 3 bp 1 amino acid = 149.2 Da Product: Cycle 3 GFP (Crameri et al., 1996) mammalian codon-optimized |
| Cycle 3 GFP 988 .. 1707 = 720 bp 239 amino acids = 26.9 kDa 3 segments Segment 2: 1a 991 .. 993 = 3 bp 1 amino acid = 89.1 Da Product: Cycle 3 GFP (Crameri et al., 1996) mammalian codon-optimized |
| Cycle 3 GFP 988 .. 1707 = 720 bp 239 amino acids = 26.9 kDa 3 segments Segment 3: 994 .. 1707 = 714 bp 237 amino acids = 26.7 kDa Product: Cycle 3 GFP (Crameri et al., 1996) mammalian codon-optimized |
| Cycle 3 GFP 988 .. 1707 = 720 bp 239 amino acids = 26.9 kDa 3 segments Product: Cycle 3 GFP (Crameri et al., 1996) mammalian codon-optimized |
| ori 4386 .. 4974 = 589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication |
| ori 4386 .. 4974 = 589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication |
| f1 ori 2005 .. 2433 = 429 bp f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis |
| f1 ori 2005 .. 2433 = 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 |
| SV40 promoter 2447 .. 2777 = 331 bp SV40 enhancer and early promoter |
| SV40 promoter 2447 .. 2777 = 331 bp SV40 enhancer and early promoter |
| bGH poly(A) signal 1735 .. 1959 = 225 bp bovine growth hormone polyadenylation signal |
| bGH poly(A) signal 1735 .. 1959 = 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 3814 .. 3935 = 122 bp SV40 polyadenylation signal |
| SV40 poly(A) signal 3814 .. 3935 = 122 bp SV40 polyadenylation signal |
| AmpR promoter 6006 .. 6110 = 105 bp |
| AmpR promoter 6006 .. 6110 = 105 bp |
| MCS 908 .. 987 = 80 bp multiple cloning site |
| MCS 908 .. 987 = 80 bp multiple cloning site |
| 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 2628 .. 2763 = 136 bp SV40 origin of replication |
| SV40 ori 2628 .. 2763 = 136 bp SV40 origin of replication |
| ORF: 988 .. 1707 = 720 bp ORF: 239 amino acids = 26.9 kDa |
| ORF: 3016 .. 3402 = 387 bp ORF: 128 amino acids = 14.6 kDa |
| ORF: 5275 .. 5541 = 267 bp ORF: 88 amino acids = 9.2 kDa |
| ORF: 1956 .. 2222 = 267 bp ORF: 88 amino acids = 9.3 kDa |
| ORF: 2844 .. 3638 = 795 bp ORF: 264 amino acids = 29.0 kDa |
| ORF: 3153 .. 3407 = 255 bp ORF: 84 amino acids = 9.6 kDa |
| ORF: 5145 .. 6005 = 861 bp ORF: 286 amino acids = 31.6 kDa |
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