p6oN-ocs-LH B422
Ligation of pOCS-BM* into B399p6oN-LH*
- Promoter 1
- Nos
- Gene 1
- hptII
- Gene 2
- MCS
- Features
- dicots
- Other markers
- Sm/Sp
- Size
- 9,739 bp, circular
- Record date
- 21-JAN-2016
Construct
- Pnos–hptII · TE9 · selection cassette, 309–2,140
- MCS · 8 unique sites · at 2,150, 2,231
- SfiI ×2 · flanks the MCS · at 2,181, 2,445
Read out of the GenBank file; compare every vector in the multiple expression cassettes and cloning sites table.
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 | 303 | between BL and Pnos |
| XhoI (PaeR7I) | C^TCGAG | 607 | between Pnos and hptII |
| SbfI (SdaI) | CCTGCA^GG | 2,150 | between TE9 and Tocs |
| NcoI | C^CATGG | 2,176 | between TE9 and Tocs |
| SpeI | A^CTAGT | 2,189 | between TE9 and Tocs |
| BamHI | G^GATCC | 2,195 | between TE9 and Tocs |
| SmaI (XmaI, TspMI) | CCC^GGG | 2,203 | between TE9 and Tocs |
| EcoRI | G^AATTC | 2,213 | between TE9 and Tocs |
| HindIII | A^AGCTT | 2,219 | between TE9 and Tocs |
| SalI | G^TCGAC | 2,231 | between TE9 and Tocs |
| AflII (BspTI) | C^TTAAG | 2,441 | between Tocs and BR |
| BglII | A^GATCT | 2,451 | between Tocs and BR |
| ApaI (PspOMI) | GGGCC^C | 2,462 | between Tocs and BR |
| KpnI (Acc65I) | GGTAC^C | 2,468 | between Tocs and BR |
| PmeI (MssI) | GTTT^AAAC | 2,500 | between Tocs and BR |
| AatII (ZraI) | GACGT^C | 2,891 | between BR and ColE1 |
| EcoRV (Eco32I) | GAT^ATC | 3,012 | between BR and ColE1 |
| BbsI (BpiI) | GAAGAC(2/6) | 4,301 | between ColE1 and pVS1 ORI |
| BsiWI (Pfl23II) | C^GTACG | 7,105 | in pVS1 ORI |
| ClaI (BspDI) | AT^CGAT | 7,301 | in pVS1 ORI |
Cut twice: PstI (2,150, 2,211), SphI (2,156, 2,625), NdeI (4,006, 5,765), NheI (503, 7,221), MluI (1,840, 2,225), SfiI (2,181, 2,445), FseI (4,772, 6,877), MfeI (14, 1,301), AgeI (7,715, 8,023), StuI (1,521, 2,439), XmnI (1,507, 1,842), BstEII (915, 8,586), SapI (3,944, 7,841).
No site: SacI, AscI, PacI, SwaI, AvrII, BstBI, HpaI, NsiI, BsrGI, PvuII, 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 |
|---|---|---|---|---|---|
| BL | Site misc_feature | 1 | 24 | 24 | + |
| Pnos | Promoter misc_feature | 309 | 606 | 298 | + |
| hptII | Selection marker CDS | 625 | 1,194 | 570 | + |
| int | Site misc_feature | 1,195 | 1,383 | 189 | + |
| hptII | Selection marker CDS | 1,384 | 1,839 | 456 | + |
| TE9 | Terminator misc_feature | 1,850 | 2,140 | 291 | + |
| Tocs | Terminator misc_feature | 2,236 | 2,431 | 196 | + |
| BR | Site misc_feature | 2,516 | 2,539 | 24 | + |
| ColE1 | Origin / backbone misc_feature | 3,074 | 3,771 | 698 | + |
| pVS1 ORI | Origin / backbone misc_feature | 4,320 | 8,025 | 3,706 | + |
| Sm/Sp | Selection marker CDS | 8,209 | 9,219 | 1,011 | + |
PromoterCoding sequenceTerminatorSelection markerOrigin / backboneT-DNA borderSite
Feature types are inferred from the labels in the GenBank file; the file itself is the reference.