pSPORT 1
Vector for cloning, sequencing, and bacterial expression of cDNAs.
Sequence Author: Thermo Fisher (Invitrogen)
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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 DraIII 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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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 AlwNI sites may not be compatible. |
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BsiWI is typically used at 55°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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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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Sticky ends from different AvaI sites may not be compatible. |
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Sticky ends from different BsoBI sites may not be compatible.BsoBI is typically used at 37°C, but can be used at temperatures up to 65°C. |
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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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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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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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| * Blocked by Dcm methylation. 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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Sticky ends from different BsmBI sites may not be compatible.BsmBI-v2 is an improved version of BsmBI. |
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Sticky ends from different Esp3I 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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ApaI can be used between 25°C and 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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| * 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 PflMI sites may not be compatible. |
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PciI is inhibited by nonionic detergents. |
| lacI 464 .. 1546 = 1083 bp 360 amino acids = 38.5 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 464 .. 1546 = 1083 bp 360 amino acids = 38.5 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). |
| AmpR 2661 .. 3521 = 861 bp 286 amino acids = 31.6 kDa 2 segments Segment 2: 2661 .. 3452 = 792 bp 263 amino acids = 28.9 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
| AmpR 2661 .. 3521 = 861 bp 286 amino acids = 31.6 kDa 2 segments Segment 1: signal sequence 3453 .. 3521 = 69 bp 23 amino acids = 2.6 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
| AmpR 2661 .. 3521 = 861 bp 286 amino acids = 31.6 kDa 2 segments Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
| ori 1902 .. 2490 = 589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication |
| ori 1902 .. 2490 = 589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication |
| f1 ori 3653 .. 4108 = 456 bp f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis |
| f1 ori 3653 .. 4108 = 456 bp f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis |
| AmpR promoter 3522 .. 3626 = 105 bp |
| AmpR promoter 3522 .. 3626 = 105 bp |
| MCS 191 .. 286 = 96 bp multiple cloning site |
| MCS 191 .. 286 = 96 bp multiple cloning site |
| lacI promoter 1547 .. 1624 = 78 bp |
| lacI promoter 1547 .. 1624 = 78 bp |
| lac promoter 385 .. 415 = 31 bp 3 segments Segment 3: -10 385 .. 391 = 7 bp promoter for the E. coli lac operon |
| lac promoter 385 .. 415 = 31 bp 3 segments Segment 2: 392 .. 409 = 18 bp promoter for the E. coli lac operon |
| lac promoter 385 .. 415 = 31 bp 3 segments Segment 1: -35 410 .. 415 = 6 bp promoter for the E. coli lac operon |
| lac promoter 385 .. 415 = 31 bp 3 segments promoter for the E. coli lac operon |
| SP6 promoter 164 .. 182 = 19 bp promoter for bacteriophage SP6 RNA polymerase |
| SP6 promoter 164 .. 182 = 19 bp promoter for bacteriophage SP6 RNA polymerase |
| T7 promoter 300 .. 318 = 19 bp promoter for bacteriophage T7 RNA polymerase |
| T7 promoter 300 .. 318 = 19 bp promoter for bacteriophage T7 RNA polymerase |
| M13 fwd 140 .. 156 = 17 bp common sequencing primer, one of multiple similar variants |
| M13 fwd 140 .. 156 = 17 bp common sequencing primer, one of multiple similar variants |
| M13 rev 337 .. 353 = 17 bp common sequencing primer, one of multiple similar variants |
| M13 rev 337 .. 353 = 17 bp common sequencing primer, one of multiple similar variants |
| lac operator 361 .. 377 = 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 361 .. 377 = 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). |
| lacZα 4085 .. 341 = 366 bp 121 amino acids = 13.7 kDa Product: LacZα fragment of β-galactosidase |
| lacZα 4085 .. 341 = 366 bp 121 amino acids = 13.7 kDa Product: LacZα fragment of β-galactosidase |
| ORF: 430 .. 681 = 252 bp ORF: 83 amino acids = 9.1 kDa |
| ORF: 2791 .. 3057 = 267 bp ORF: 88 amino acids = 9.2 kDa |
| ORF: 4085 .. 341 = 366 bp ORF: 121 amino acids = 13.7 kDa |
| ORF: 444 .. 707 = 264 bp ORF: 87 amino acids = 8.9 kDa |
| ORF: 1296 .. 1613 = 318 bp ORF: 105 amino acids = 11.2 kDa |
| ORF: 2661 .. 3521 = 861 bp ORF: 286 amino acids = 31.6 kDa |
| ORF: 464 .. 1423 = 960 bp ORF: 319 amino acids = 34.1 kDa |
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