p7N-35S-Ubi10-RNAi
Ligation of inverted 670pD1-35s-Ubi10-RNAi* into p7N*
- Promoter 1
- Nos
- Gene 1
- bar
- Promoter 2
- 35S
- Gene 2
- gus-spacer
- Features
- double promoter construct for use in dicots (Arabidopsis+Tobacco)
- Other markers
- Sm/Sp
- Size
- 11,474 bp, circular
- Record date
- 12-JUL-2013
Construct
- Pnos–bar · T35S · selection cassette, 7–1,093
- P35S–(empty)–T35S · empty expression cassette, 1,152–2,364
- SfiI ×2 · flanks the P35S–(empty)–T35S cassette · at 1,131, 3,878
- 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 |
|---|---|---|---|
| XhoI (PaeR7I) | C^TCGAG | 305 | between Pnos and bar |
| AarI | CACCTGC(4/8) | 653 | in bar |
| PvuII | CAG^CTG | 813 | in bar |
| PstI | CTGCA^G | 1,100 | between T35S and P35S |
| SbfI (SdaI) | CCTGCA^GG | 1,100 | between T35S and P35S |
| SwaI (SmiI) | ATTT^AAAT | 1,141 | between T35S and P35S |
| BsrGI (Bsp1407I) | T^GTACA | 1,733 | between T35s and Gus |
| HindIII | A^AGCTT | 1,740 | between T35s and Gus |
| AflII (BspTI) | C^TTAAG | 2,296 | between Gus and T35S |
| SmaI (XmaI, TspMI) | CCC^GGG | 2,305 | between Gus and T35S |
| BamHI | G^GATCC | 2,310 | between Gus and T35S |
| SpeI | A^CTAGT | 2,372 | between T35S and Ubq10At |
| EcoRI | G^AATTC | 3,188 | in Ubq10At |
| PacI | TTAAT^TAA | 3,867 | between Ubq10At and RB |
| PmeI (MssI) | GTTT^AAAC | 3,933 | between Ubq10At and RB |
| AatII (ZraI) | GACGT^C | 4,324 | between RB and ColE1 |
| BsiWI (Pfl23II) | C^GTACG | 8,538 | in pVS1 ORI |
| ClaI (BspDI) | AT^CGAT | 8,734 | in pVS1 ORI |
| BstEII (Eco91I) | G^GTNACC | 10,019 | in Sm/Sp |
Cut twice: SacI (1,676, 2,763), NcoI (317, 1,126), NdeI (5,439, 7,198), NheI (201, 8,654), MluI (875, 1,726), SfiI (1,131, 3,878), FseI (6,205, 8,310), AgeI (9,148, 9,456), SacII (119, 753), BsaI (9,057, 9,477).
No site: AscI, AvrII, BstBI, HpaI, StuI, NsiI.
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 |
|---|---|---|---|---|---|
| Pnos | Promoter misc_feature | 7 | 304 | 298 | + |
| bar | Selection marker misc_feature | 319 | 870 | 552 | + |
| T35S | Terminator misc_feature | 893 | 1,093 | 201 | + |
| P35S | Promoter misc_feature | 1,152 | 1,664 | 513 | + |
| T35s | Terminator misc_feature | 1,678 | 1,726 | 49 | − |
| unnamed | Coding sequence CDS | 1,746 | 2,295 | 550 | + |
| Gus | Coding sequence CDS | 1,746 | 2,295 | 550 | − |
| T35S | Terminator misc_feature | 2,316 | 2,364 | 49 | + |
| Ubq10At | Other misc_feature | 2,378 | 3,853 | 1,476 | − |
| íntron | Other misc_feature | 2,380 | 2,683 | 304 | − |
| RB | T-DNA border misc_feature | 3,949 | 3,972 | 24 | + |
| ColE1 | Origin / backbone misc_feature | 4,507 | 5,204 | 698 | + |
| pVS1 ORI | Origin / backbone misc_feature | 5,753 | 9,458 | 3,706 | + |
| Sm/Sp | Selection marker CDS | 9,642 | 10,652 | 1,011 | + |
| LB | T-DNA border misc_feature | 11,173 | 11,196 | 24 | + |
PromoterCoding sequenceTerminatorSelection markerOrigin / backboneT-DNA borderSite
Feature types are inferred from the labels in the GenBank file; the file itself is the reference.