p9o-LH-35s-ocs B425
Ligation of p35-OCS-BM* into B415p9o-LH*
- Promoter 1
- 35s
- Gene 1
- nptII
- Promoter 2
- 35s
- Gene 2
- MCS
- Features
- dicots
- Other markers
- nptIIo, Sm/Sp
- Size
- 10,065 bp, circular
- Record date
- 22-JAN-2016
Construct
- P35S–nptIIo · TE9 · selection cassette, 309–2,038
- P35S–(empty)–Tocs · empty expression cassette, 2,093–2,757
- MCS · 6 unique sites · at 2,514, 2,557
- SfiI ×2 · flanks the P35S–(empty)–Tocs cassette · at 2,079, 2,771
- T-DNA borders · LB, RB
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 LB and P35S |
| XhoI (PaeR7I) | C^TCGAG | 727 | between P35S and nptIIo |
| SacI (Ecl136II) | GAGCT^C | 1,255 | in nptIIo |
| SbfI (SdaI) | CCTGCA^GG | 2,048 | between TE9 and P35S |
| NcoI | C^CATGG | 2,074 | between TE9 and P35S |
| SpeI | A^CTAGT | 2,514 | in P35S |
| BamHI | G^GATCC | 2,521 | between P35S and Tocs |
| SmaI (XmaI, TspMI) | CCC^GGG | 2,529 | between P35S and Tocs |
| EcoRI | G^AATTC | 2,539 | between P35S and Tocs |
| HindIII | A^AGCTT | 2,545 | between P35S and Tocs |
| SalI | G^TCGAC | 2,557 | between P35S and Tocs |
| StuI (Eco147I) | AGG^CCT | 2,765 | between Tocs and RB |
| AflII (BspTI) | C^TTAAG | 2,767 | between Tocs and RB |
| BglII | A^GATCT | 2,777 | between Tocs and RB |
| KpnI (Acc65I) | GGTAC^C | 2,794 | between Tocs and RB |
| PmeI (MssI) | GTTT^AAAC | 2,826 | between Tocs and RB |
| AatII (ZraI) | GACGT^C | 3,217 | between RB and ColE1 |
| BsiWI (Pfl23II) | C^GTACG | 7,431 | in pVS1 |
| NheI (BmtI) | G^CTAGC | 7,547 | in pVS1 |
| ClaI (BspDI) | AT^CGAT | 7,627 | in pVS1 |
Cut twice: PstI (2,048, 2,537), SphI (2,054, 2,951), NdeI (4,332, 6,091), ApaI (828, 2,788), MluI (1,738, 2,551), SfiI (2,079, 2,771), MfeI (14, 1,148), AgeI (8,041, 8,349), BstEII (1,497, 8,912), SapI (4,270, 8,167), AarI (1,218, 1,339).
No site: AscI, PacI, SwaI, AvrII, BstBI, HpaI, NsiI, BsrGI, SacII, PvuII.
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 |
|---|---|---|---|---|---|
| LB | T-DNA border misc_feature | 1 | 24 | 24 | + |
| P35S | Promoter misc_feature | 309 | 724 | 416 | + |
| nptIIo | Selection marker CDS | 745 | 1,043 | 299 | + |
| splice | Site misc_signal | 1,040 | 1,043 | 4 | + |
| int | Site CDS | 1,044 | 1,230 | 187 | + |
| splice | Site misc_signal | 1,229 | 1,232 | 4 | + |
| nptIIo | Selection marker CDS | 1,231 | 1,734 | 504 | + |
| TE9 | Terminator misc_feature | 1,748 | 2,038 | 291 | + |
| P35S | Promoter misc_feature | 2,093 | 2,515 | 423 | + |
| Tocs | Terminator misc_feature | 2,562 | 2,757 | 196 | + |
| RB | T-DNA border misc_feature | 2,842 | 2,865 | 24 | + |
| ColE1 | Origin / backbone misc_feature | 3,400 | 4,097 | 698 | + |
| bom | Origin / backbone misc_feature | 4,134 | 4,393 | 260 | + |
| pVS1 | Origin / backbone misc_feature | 4,646 | 8,351 | 3,706 | + |
| Sm/Sp | Selection marker CDS | 8,535 | 9,545 | 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.
Used in
- Efficient and versatile rapeseed transformation for new breeding technologies – Ille K, Melzer S, The Plant Journal 2025; 123:e70330