pet28-MBP-NLS-Mad7-NLS F460
Ligation of PCR Mad7 TEV* into F455pet28-MBP-HisOpti*
- Gene 2
- MAD7
- Size
- 10,403 bp, circular
- Record date
- 20-APR-2023
Files
Order
Vectors from the list cost 250 €, Cas9 vectors 400 €, net. Send the vector name to info@dna-cloning.com and we ship on invoice.
Restriction sites
| Unique cutter | Site | Cuts at | Location |
|---|---|---|---|
| XbaI | T^CTAGA | 67 | between lacI and ORF-1 |
| NdeI | CA^TATG | 160 | in ORF-1 |
| BsaI (Eco31I) | GGTCTC(1/5) | 225 | in malE (mutated) |
| BsiWI (Pfl23II) | C^GTACG | 452 | in malE (mutated) |
| BamHI | G^GATCC | 1,326 | in ORF-1 |
| SalI | G^TCGAC | 1,345 | between ORF-1 and TEV |
| KpnI (Acc65I) | GGTAC^C | 1,408 | in Mad7 |
| HindIII | A^AGCTT | 2,743 | in Mad7 |
| NheI (BmtI) | G^CTAGC | 2,972 | in Mad7 |
| PstI | CTGCA^G | 5,241 | between Mad7 and NLS |
| XhoI (PaeR7I) | C^TCGAG | 5,278 | in ORF-1 |
| SmaI (XmaI, TspMI) | CCC^GGG | 6,509 | in aph(3')-Ia |
| SapI (BspQI, LguI) | GCTCTTC(1/4) | 7,698 | between aph(3')-Ia and bom |
| ApaI (PspOMI) | GGGCC^C | 9,479 | in lacI |
| SphI (PaeI) | GCATG^C | 10,215 | between lacI and ORF-1 |
Cut twice: SacI (1,272, 1,342), NcoI (106, 1,123), MluI (2,782, 9,682), NsiI (6,271, 6,537).
No site: NotI, SpeI, AscI, PacI, SfiI, SbfI, FseI, PmeI, SwaI, AvrII, MfeI, AgeI, StuI, AflII, EagI, SacII, AatII, AarI.
Computed from the GenBank sequence for 50 common enzymes; positions are the cut in the top strand, without Dam/Dcm methylation. Compare vectors in the restriction site finder.
Annotated features
| Feature | Type | Start | End | Length | Strand |
|---|---|---|---|---|---|
| ORF-1 | Coding sequence CDS | 137 | 1,345 | 1,209 | + |
| ORF-1' | Coding sequence CDS | 137 | 1,345 | 1,209 | + |
| thrombin recognition and cleavage site | Site CDS | 141 | 158 | 18 | + |
| malE (mutated) | Coding sequence CDS | 162 | 1,262 | 1,101 | + |
| TEV | Site CDS | 1,351 | 1,373 | 23 | + |
| start | Site misc_feature | 1,374 | 1,376 | 3 | + |
| Mad7 | Coding sequence CDS | 1,374 | 5,231 | 3,858 | + |
| 'NLS | Site misc_feature | 5,242 | 5,277 | 36 | + |
| ORF-1 | Coding sequence CDS | 5,242 | 5,304 | 63 | + |
| 6xHis affinity tag | Site CDS | 5,284 | 5,301 | 18 | + |
| aph(3')-Ia | Coding sequence CDS | 6,000 | 6,815 | 816 | − |
| bom | Origin / backbone misc_feature | 7,711 | 7,853 | 143 | − |
| rop | Coding sequence CDS | 7,955 | 8,146 | 192 | − |
| lacI | Coding sequence CDS | 8,955 | 10,037 | 1,083 | − |
PromoterCoding sequenceTerminatorSelection markerOrigin / backboneT-DNA borderSite
Feature types are inferred from the labels in the GenBank file; the file itself is the reference.