pSpark Done (linearized)
Linearized high copy vector for very efficient cloning of blunt-ended PCR products, with an MCS containing EcoRI and NotI sites on each side of the insertion site.
Sequence Author: Canvax Biotech
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Efficient cleavage requires at least two copies of the SacII recognition sequence. |
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Sticky ends from different StyI sites may not be compatible. |
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ApaI can be used between 25°C and 37°C. |
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Efficient cleavage requires at least two copies of the NaeI recognition sequence. |
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Efficient cleavage requires at least two copies of the NgoMIV recognition sequence. |
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Sticky ends from different BtgZI sites may not be compatible.After cleavage, BtgZI can remain bound to DNA and alter its electrophoretic mobility.BtgZI is typically used at 60°C, but is 75% active at 37°C. |
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Sticky ends from different DraIII sites may not be compatible. |
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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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Efficient cleavage requires at least two copies of the BspMI recognition sequence. Sticky ends from different BspMI sites may not be compatible. |
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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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Prolonged incubation with NdeI may lead to removal of additional nucleotides. |
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Sticky ends from different BstXI 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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PciI is inhibited by nonionic detergents. |
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After cleavage, BseYI can remain bound to DNA and alter its electrophoretic mobility. |
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Sticky ends from different AlwNI sites may not be compatible. |
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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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Sticky ends from different BsaI sites may not be compatible.BsaI can be used between 37°C and 50°C. |
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Efficient cleavage requires at least two copies of the BpmI recognition sequence. Sticky ends from different BpmI sites may not be compatible.After cleavage, BpmI can remain bound to DNA and alter its electrophoretic mobility.BpmI quickly loses activity 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). |
AmpR 1278 .. 2138 = 861 bp 286 amino acids = 31.6 kDa 2 segments Segment 2: 1278 .. 2069 = 792 bp 263 amino acids = 28.9 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
AmpR 1278 .. 2138 = 861 bp 286 amino acids = 31.6 kDa 2 segments Segment 1: signal sequence 2070 .. 2138 = 69 bp 23 amino acids = 2.6 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
AmpR 1278 .. 2138 = 861 bp 286 amino acids = 31.6 kDa 2 segments Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
ori 519 .. 1107 = 589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication |
ori 519 .. 1107 = 589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication |
f1 ori 2321 .. 2776 = 456 bp f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis |
f1 ori 2321 .. 2776 = 456 bp f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis |
lacZα 1 .. 121 = 121 bp 40 amino acids = 4.3 kDa Product: LacZα fragment of β-galactosidase |
lacZα 1 .. 121 = 121 bp 40 amino acids = 4.3 kDa Product: LacZα fragment of β-galactosidase |
AmpR promoter 2139 .. 2243 = 105 bp |
AmpR promoter 2139 .. 2243 = 105 bp |
MCS 2966 .. 3016 = 51 bp multiple cloning site |
MCS 2966 .. 3016 = 51 bp multiple cloning site |
lac promoter 165 .. 195 = 31 bp 3 segments Segment 3: -10 165 .. 171 = 7 bp promoter for the E. coli lac operon |
lac promoter 165 .. 195 = 31 bp 3 segments Segment 2: 172 .. 189 = 18 bp promoter for the E. coli lac operon |
lac promoter 165 .. 195 = 31 bp 3 segments Segment 1: -35 190 .. 195 = 6 bp promoter for the E. coli lac operon |
lac promoter 165 .. 195 = 31 bp 3 segments promoter for the E. coli lac operon |
T7 promoter 2940 .. 2958 = 19 bp promoter for bacteriophage T7 RNA polymerase |
T7 promoter 2940 .. 2958 = 19 bp promoter for bacteriophage T7 RNA polymerase |
lac operator 141 .. 157 = 17 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 141 .. 157 = 17 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). |
M13 fwd 2917 .. 2933 = 17 bp common sequencing primer, one of multiple similar variants |
M13 fwd 2917 .. 2933 = 17 bp common sequencing primer, one of multiple similar variants |
lacZα 2753 .. 3015 = 263 bp 86 amino acids = 9.8 kDa Product: LacZα fragment of β-galactosidase |
lacZα 2753 .. 3015 = 263 bp 86 amino acids = 9.8 kDa Product: LacZα fragmen |