pNIC-CH
Bacterial vector encoding a C-terminal 6xHis-tag plus a SacB negative selection marker, for purification of recombinant proteins.
Sequence Author: Structural Genomics Consortium
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Sticky ends from different BstAPI sites may not be compatible. |
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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 BstEII sites may not be compatible.BstEII is typically used at 60°C, but is 50% active at 37°C. |
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Efficient cleavage requires at least two copies of the NmeAIII recognition sequence. Sticky ends from different NmeAIII sites may not be compatible.For full activity, add fresh S-adenosylmethionine (SAM). |
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ApaI can be used between 25°C and 37°C. |
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BssHII is typically used at 50°C, but is 75% active at 37°C. |
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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 BglI sites may not be compatible. |
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Sticky ends from different PpuMI 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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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 AlwNI sites may not be compatible. |
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SmaI can be used at 37°C for brief incubations. |
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Efficient cleavage requires at least two copies of the SgrAI recognition sequence. |
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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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| * Blocked by Dcm methylation. |
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BsrGI is typically used at 37°C, but is even more active at 60°C. |
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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 AarI recognition sequence is present. Sticky ends from different AarI sites may not be compatible.After cleavage, AarI can remain bound to DNA and alter its electrophoretic mobility. |
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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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After cleavage, BamHI-HF® (but not the original BamHI) can remain bound to DNA and alter its electrophoretic mobility. |
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PaeR7I does not recognize the sequence CTCTCGAG. |
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Sticky ends from different BlpI sites may not be compatible. |
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Sticky ends from different DraIII sites may not be compatible. |
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Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present. |
| SacB 781 .. 2202 = 1422 bp 473 amino acids = 53.0 kDa 2 segments Segment 1: signal peptide 781 .. 867 = 87 bp 29 amino acids = 3.0 kDa Product: secreted levansucrase that renders bacterial growth sensitive to sucrose negative selection marker |
| SacB 781 .. 2202 = 1422 bp 473 amino acids = 53.0 kDa 2 segments Segment 2: 868 .. 2202 = 1335 bp 444 amino acids = 50.0 kDa Product: secreted levansucrase that renders bacterial growth sensitive to sucrose negative selection marker |
| SacB 781 .. 2202 = 1422 bp 473 amino acids = 53.0 kDa 2 segments Product: secreted levansucrase that renders bacterial growth sensitive to sucrose negative selection marker |
| lacI 6002 .. 7084 = 1083 bp 360 amino acids = 38.6 kDa Product: lac repressor The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG). |
| lacI 6002 .. 7084 = 1083 bp 360 amino acids = 38.6 kDa Product: lac repressor The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG). |
| KanR 3047 .. 3862 = 816 bp 271 amino acids = 31.0 kDa Product: aminoglycoside phosphotransferase confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes |
| KanR 3047 .. 3862 = 816 bp 271 amino acids = 31.0 kDa Product: aminoglycoside phosphotransferase confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes |
| ori 3984 .. 4572 = 589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication |
| ori 3984 .. 4572 = 589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication |
| f1 ori 2499 .. 2954 = 456 bp f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis |
| f1 ori 2499 .. 2954 = 456 bp f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis |
| sacB promoter 348 .. 780 = 433 bp sacB promoter and control region |
| sacB promoter 348 .. 780 = 433 bp sacB promoter and control region |
| rop 5002 .. 5193 = 192 bp 63 amino acids = 7.2 kDa Product: Rop protein, which maintains plasmids at low copy number |
| rop 5002 .. 5193 = 192 bp 63 amino acids = 7.2 kDa Product: Rop protein, which maintains plasmids at low copy number |
| lacI promoter 7085 .. 7162 = 78 bp |
| lacI promoter 7085 .. 7162 = 78 bp |
| T7 terminator 2415 .. 2462 = 48 bp transcription terminator for bacteriophage T7 RNA polymerase |
| T7 terminator 2415 .. 2462 = 48 bp transcription terminator for bacteriophage T7 RNA polymerase |
| lac operator 272 .. 296 = 25 bp The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG). |
| lac operator 272 .. 296 = 25 bp The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG). |
| T7 promoter 253 .. 271 = 19 bp promoter for bacteriophage T7 RNA polymerase |
| T7 promoter 253 .. 271 = 19 bp promoter for bacteriophage T7 RNA polymerase |
| 6xHis 2264 .. 2281 = 18 bp 6 amino acids = 840.9 Da Product: 6xHis affinity tag |
| 6xHis 2264 .. 2281 = 18 bp 6 amino acids = 840.9 Da Product: 6xHis affinity tag |
| 6xHis 2331 .. 2348 = 18 bp 6 amino acids = 840.9 Da Product: 6xHis affinity tag |
| 6xHis 2331 .. 2348 = 18 bp 6 amino acids = 840.9 Da Product: 6xHis affinity tag |
| RBS 327 .. 332 = 6 bp ribosome binding site |
| RBS 327 .. 332 = 6 bp ribosome binding site |
| ORF: 781 .. 2202 = 1422 bp ORF: 473 amino acids = 53.0 kDa |
| ORF: 5968 .. 6219 = 252 bp ORF: 83 amino acids = 9.1 kDa |
| ORF: 7123 .. 168 = 264 bp ORF: 87 amino acids = 9.5 kDa |
| ORF: 5225 .. 5593 = 369 bp ORF: 122 amino acids = 14.2 kDa |
| ORF: 42 .. 281 = 240 bp ORF: 79 amino acids = 8.0 kDa |
| ORF: 5002 .. 5226 = 225 bp ORF: 74 amino acids = 8.5 kDa |
| ORF: 5590 .. 5946 = 357 bp ORF: 118 amino acids = 13.0 kDa |
| ORF: 6834 .. 116 = 501 bp ORF: 166 amino acids = 17.5 kDa |
| ORF: 5982 .. 6245 = 264 bp ORF: 87 amino acids = 8.9 kDa |
| ORF: 3047 .. 3862 = 816 bp ORF: 271 amino acids = 31.0 kDa |
| ORF: 6002 .. 6961 = 960 bp ORF: 319 amino acids = 34.1 kDa |
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