p4oM-LH B471
Ligation of AHAS-Xho-Bspe* into B474p7oM-LH*
- Promoter 1
- 35s
- Gene 1
- AHAS
- Features
- monocot
- Other markers
- Sm/Sp
- Size
- 10,653 bp, circular
- Record date
- 22-JAN-2016
Construct
- P35S–ahas · TE9 · selection cassette, 309–3,254
- MCS · 15 unique sites · at 3,264, 3,414
- SfiI ×2 · flanks the MCS · at 3,295, 3,359
- 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 |
| XmnI (PdmI) | GAANN^NNTTC | 594 | in P35S |
| XhoI (PaeR7I) | C^TCGAG | 727 | between P35S and ahas |
| PstI | CTGCA^G | 3,264 | between TE9 and RB |
| SbfI (SdaI) | CCTGCA^GG | 3,264 | between TE9 and RB |
| NcoI | C^CATGG | 3,290 | between TE9 and RB |
| SpeI | A^CTAGT | 3,303 | between TE9 and RB |
| BamHI | G^GATCC | 3,309 | between TE9 and RB |
| SmaI (XmaI, TspMI) | CCC^GGG | 3,317 | between TE9 and RB |
| NsiI (Mph1103I) | ATGCA^T | 3,325 | between TE9 and RB |
| EcoRI | G^AATTC | 3,327 | between TE9 and RB |
| HindIII | A^AGCTT | 3,333 | between TE9 and RB |
| SalI | G^TCGAC | 3,345 | between TE9 and RB |
| AflII (BspTI) | C^TTAAG | 3,355 | between TE9 and RB |
| BglII | A^GATCT | 3,365 | between TE9 and RB |
| ApaI (PspOMI) | GGGCC^C | 3,376 | between TE9 and RB |
| KpnI (Acc65I) | GGTAC^C | 3,382 | between TE9 and RB |
| PmeI (MssI) | GTTT^AAAC | 3,414 | between TE9 and RB |
| AatII (ZraI) | GACGT^C | 3,805 | between RB and ColE1 |
| BsiWI (Pfl23II) | C^GTACG | 8,019 | in pVS1 |
| NheI (BmtI) | G^CTAGC | 8,135 | in pVS1 |
| BstEII (Eco91I) | G^GTNACC | 9,500 | in Sm/Sp |
Cut twice: SphI (3,270, 3,539), NdeI (4,920, 6,679), ClaI (3,340, 8,215), EcoRV (634, 3,926), SfiI (3,295, 3,359), FseI (5,686, 7,791), MfeI (14, 1,490), AgeI (8,629, 8,937), StuI (1,645, 3,353), SapI (4,858, 8,755).
No site: SacI, MluI, AscI, PacI, SwaI, AvrII, BstBI, HpaI, BsrGI, SacII, 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 |
|---|---|---|---|---|---|
| LB | T-DNA border misc_feature | 1 | 24 | 24 | + |
| P35S | Promoter misc_feature | 309 | 724 | 416 | + |
| ahas | Coding sequence CDS | 745 | 1,383 | 639 | + |
| int | Site CDS | 1,384 | 1,572 | 189 | + |
| ahas | Coding sequence CDS | 1,573 | 2,946 | 1,374 | + |
| TE9 | Terminator misc_feature | 2,964 | 3,254 | 291 | + |
| RB | T-DNA border misc_feature | 3,430 | 3,453 | 24 | + |
| ColE1 | Origin / backbone misc_feature | 3,988 | 4,685 | 698 | + |
| pVS1 | Origin / backbone misc_feature | 5,234 | 8,939 | 3,706 | + |
| Sm/Sp | Selection marker CDS | 9,123 | 10,133 | 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.