pKillerOrange-C
Vector for fusing KillerOrange to the N-terminus of a partner protein.
Sequence Author: Evrogen
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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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Sticky ends from different BsrDI sites may not be compatible. |
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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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Efficient cleavage requires at least two copies of the NarI recognition sequence. |
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* Blocked by Dam methylation. |
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* Blocked by Dam methylation. |
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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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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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Prolonged incubation with NdeI may lead to removal of additional nucleotides. |
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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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Sticky ends from different BlpI sites may not be compatible. |
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Efficient cleavage requires at least two copies of the PaqCI recognition sequence. Sticky ends from different PaqCI sites may not be compatible.Cleavage can be improved with PaqCI Activator. |
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PaeR7I does not recognize the sequence CTCTCGAG. |
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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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Efficient cleavage requires at least two copies of the SacII 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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ApaI can be used between 25°C and 37°C. |
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SmaI can be used at 37°C for brief incubations. |
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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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* Blocked by Dam methylation. BclI is typically used at 50-55°C, but is 50% active at 37°C. |
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Sticky ends from different BtsαI 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. |
NeoR/KanR 2621 .. 3415 = 795 bp 264 amino acids = 29.0 kDa Product: aminoglycoside phosphotransferase from Tn5 confers resistance to neomycin, kanamycin, and G418 (Geneticin®) |
NeoR/KanR 2621 .. 3415 = 795 bp 264 amino acids = 29.0 kDa Product: aminoglycoside phosphotransferase from Tn5 confers resistance to neomycin, kanamycin, and G418 (Geneticin®) |
KillerOrange 613 .. 1323 = 711 bp 237 amino acids = 26.4 kDa Product: orange fluorescent protein KillerOrange, a genetically-encoded photosensitizer derived from KillerRed (Sarkisyan et al., 2015) mammalian codon-optimized |
KillerOrange 613 .. 1323 = 711 bp 237 amino acids = 26.4 kDa Product: orange fluorescent protein KillerOrange, a genetically-encoded photosensitizer derived from KillerRed (Sarkisyan et al., 2015) mammalian codon-optimized |
ori 4023 .. 4611 = 589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication |
ori 4023 .. 4611 = 589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication |
f1 ori 1641 .. 2096 = 456 bp f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis |
f1 ori 1641 .. 2096 = 456 bp f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis |
SV40 promoter 2229 .. 2586 = 358 bp SV40 enhancer and early promoter |
SV40 promoter 2229 .. 2586 = 358 bp SV40 enhancer and early promoter |
CMV enhancer 61 .. 364 = 304 bp human cytomegalovirus immediate early enhancer |
CMV enhancer 61 .. 364 = 304 bp human cytomegalovirus immediate early enhancer |
CMV promoter 365 .. 568 = 204 bp human cytomegalovirus (CMV) immediate early promoter |
CMV promoter 365 .. 568 = 204 bp human cytomegalovirus (CMV) immediate early promoter |
SV40 poly(A) signal 1513 .. 1634 = 122 bp SV40 polyadenylation signal |
SV40 poly(A) signal 1513 .. 1634 = 122 bp SV40 polyadenylation signal |
AmpR promoter 2123 .. 2227 = 105 bp |
AmpR promoter 2123 .. 2227 = 105 bp |
MCS 1324 .. 1389 = 66 bp multiple cloning site |
MCS 1324 .. 1389 = 66 bp multiple cloning site |
HSV TK poly(A) signal 3647 .. 3694 = 48 bp herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985) |
HSV TK poly(A) signal 3647 .. 3694 = 48 bp herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985) |
SV40 ori 2437 .. 2572 = 136 bp SV40 origin of replication |
SV40 ori 2437 .. 2572 = 136 bp SV40 origin of replication |
Kozak sequence 607 .. 616 = 10 bp vertebrate consensus sequence for strong initiation of translation (Kozak, 1987) |
Kozak sequence 607 .. 616 = 10 bp vertebrate consensus sequence for strong initiation of translation (Kozak, 1987) |
ORF: 613 .. 1404 = 792 bp ORF: 263 amino acids = 28.8 kDa |
ORF: 3436 .. 3885 = 450 bp ORF: 149 amino acids = 16.3 kDa |
ORF: 2621 .. 3415 = 795 bp ORF: 264 amino acids = 29.0 kDa |
ORF: 2793 .. 3179 = 387 bp ORF: 128 amino acids = 14.6 kDa |
ORF: 598 .. 1422 = 825 bp ORF: 274 amino acids = 26.7 kDa |
ORF: 447 .. 791 = 345 bp ORF: 114 amino acids = 12.4 kDa |
ORF: 939 .. 1307 = 369 bp ORF: 122 amino acids = 13.7 kDa |
ORF: 2930 .. 3466 = 537 bp ORF: 178 amino acids = 19.8 kDa |
ORF: 3641 .. 3874 = 234 bp ORF: 77 amino acids = 8.6 kDa |
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