pOSIP-KT
Prokaryotic one-step cloning and chromosomal integration vector encoding a kanamycin resistance marker and the phage φ21 integrase.
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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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Sticky ends from different BglI sites may not be compatible. |
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* Blocked by Dcm methylation. Sticky ends from different PpuMI sites may not be compatible. |
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The 1-base overhangs produced by EcoNI may be hard to ligate.Sticky ends from different EcoNI sites may not be compatible. |
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Sticky ends from different DraIII 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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BssHII is typically used at 50°C, but is 75% active at 37°C. |
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BsrGI is typically used at 37°C, but is even more active at 60°C. |
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PaeR7I does not recognize the sequence CTCTCGAG. |
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Efficient cleavage requires at least two copies of the NarI recognition sequence. |
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Efficient cleavage requires at least two copies of the PluTI recognition sequence. |
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ApaI can be used between 25°C and 37°C. |
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Cleavage may be enhanced when more than one copy of the EarI recognition sequence is present. Sticky ends from different EarI sites may not be compatible. |
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Sticky ends from different BstAPI sites may not be compatible. |
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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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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. |
phage φ21 integrase 2299 .. 3441 = 1143 bp 380 amino acids = 43.9 kDa Product: integrase from phage φ21 |
phage φ21 integrase 2299 .. 3441 = 1143 bp 380 amino acids = 43.9 kDa Product: integrase from phage φ21 |
KanR 3676 .. 4491 = 816 bp 271 amino acids = 31.0 kDa Product: aminoglycoside phosphotransferase confers resistance to kanamycin |
KanR 3676 .. 4491 = 816 bp 271 amino acids = 31.0 kDa Product: aminoglycoside phosphotransferase confers resistance to kanamycin |
λ repressor (ts) 1424 .. 2137 = 714 bp 237 amino acids = 26.2 kDa Product: temperature-sensitive variant of the phage λ repressor thermosensitivity is conferred by the A67T mutation |
λ repressor (ts) 1424 .. 2137 = 714 bp 237 amino acids = 26.2 kDa Product: temperature-sensitive variant of the phage λ repressor thermosensitivity is conferred by the A67T mutation |
ori 477 .. 1065 = 589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication |
ori 477 .. 1065 = 589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication |
phage φ21 attP 5139 .. 5638 = 500 bp attachment site of phage φ21 |
phage φ21 attP 5139 .. 5638 = 500 bp attachment site of phage φ21 |
R6K γ ori 4613 .. 5001 = 389 bp γ replication origin from E. coli plasmid R6K; requires the R6K initiator protein pi for replication |
R6K γ ori 4613 .. 5001 = 389 bp γ replication origin from E. coli plasmid R6K; requires the R6K initiator protein pi for replication |
ccdB 142 .. 447 = 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 142 .. 447 = 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. |
λ tL3 terminator 5659 .. 5905 = 247 bp transcription terminator tL3 from phage λ |
λ tL3 terminator 5659 .. 5905 = 247 bp transcription terminator tL3 from phage λ |
rrnB T1 terminator 6225 .. 6311 = 87 bp transcription terminator T1 from the E. coli rrnB gene |