p7U10-RNAi
Ligation of inverted pD1-35s-RNAi* into p7U10*
- Promoter 1
- UBQ10AT
- Gene 1
- bar
- Promoter 2
- 35S
- Gene 2
- gus-spacer
- Features
- double promoter construct for dicots
- Other markers
- Sm/Sp
- Size
- 11,610 bp, circular
- Record date
- 09-OCT-2009
Construct
- UBQ10–bar · T35S · selection cassette, 7–2,278
- Pro-35S–(empty)–T35S · empty expression cassette, 2,337–3,549
- Pro-35S–Gus · T35s · payload cassette, 2,863–3,983 (− strand)
- SfiI ×2 · flanks 2 cassettes · at 2,316, 4,014
- 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 |
|---|---|---|---|
| EcoRI | G^AATTC | 669 | in UBQ10 |
| XhoI (PaeR7I) | C^TCGAG | 1,490 | between UBQ10 and bar |
| AarI | CACCTGC(4/8) | 1,838 | in bar |
| SacII (Cfr42I) | CCGC^GG | 1,938 | in bar |
| PvuII | CAG^CTG | 1,998 | in bar |
| PstI | CTGCA^G | 2,285 | between T35S and Pro-35S |
| SbfI (SdaI) | CCTGCA^GG | 2,285 | between T35S and Pro-35S |
| SwaI (SmiI) | ATTT^AAAT | 2,326 | between T35S and Pro-35S |
| BsrGI (Bsp1407I) | T^GTACA | 2,918 | between T35s and (null) |
| HindIII | A^AGCTT | 2,925 | between T35s and (null) |
| AflII (BspTI) | C^TTAAG | 3,481 | between (null) and T35S |
| SmaI (XmaI, TspMI) | CCC^GGG | 3,490 | between (null) and T35S |
| BamHI | G^GATCC | 3,495 | between (null) and T35S |
| SpeI | A^CTAGT | 3,557 | between T35S and Pro-35S |
| PacI | TTAAT^TAA | 4,003 | between Pro-35S and RB |
| PmeI (MssI) | GTTT^AAAC | 4,069 | between Pro-35S and RB |
| AatII (ZraI) | GACGT^C | 4,460 | between RB and ColE1 |
| BsiWI (Pfl23II) | C^GTACG | 8,674 | in pVS1 ORI |
| NheI (BmtI) | G^CTAGC | 8,790 | in pVS1 ORI |
| ClaI (BspDI) | AT^CGAT | 8,870 | in pVS1 ORI |
| BstEII (Eco91I) | G^GTNACC | 10,155 | in Sm/Sp |
Cut twice: XbaI (673, 1,411), SalI (847, 2,850), SacI (1,102, 2,861), NcoI (1,502, 2,311), NdeI (5,575, 7,334), MluI (2,060, 2,911), SfiI (2,316, 4,014), FseI (6,341, 8,446), AgeI (9,284, 9,592), BsaI (9,193, 9,613).
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 |
|---|---|---|---|---|---|
| UBQ10 | Other misc_feature | 7 | 1,484 | 1,478 | + |
| íntron | Other misc_feature | 1,179 | 1,482 | 304 | + |
| bar | Selection marker CDS | 1,504 | 2,055 | 552 | + |
| T35S | Terminator misc_feature | 2,078 | 2,278 | 201 | + |
| Pro-35S | Promoter misc_feature | 2,337 | 2,849 | 513 | + |
| T35s | Terminator misc_feature | 2,863 | 2,911 | 49 | − |
| (null) | Coding sequence CDS | 2,931 | 3,480 | 550 | + |
| Gus | Coding sequence CDS | 2,931 | 3,480 | 550 | − |
| T35S | Terminator misc_feature | 3,501 | 3,549 | 49 | + |
| Pro-35S | Promoter misc_feature | 3,562 | 3,983 | 422 | − |
| RB | T-DNA border misc_feature | 4,085 | 4,108 | 24 | + |
| ColE1 | Origin / backbone misc_feature | 4,643 | 5,340 | 698 | + |
| pVS1 ORI | Origin / backbone misc_feature | 5,889 | 9,594 | 3,706 | + |
| Sm/Sp | Selection marker CDS | 9,778 | 10,788 | 1,011 | + |
| LB | T-DNA border misc_feature | 11,309 | 11,332 | 24 | + |
PromoterCoding sequenceTerminatorSelection markerOrigin / backboneT-DNA borderSite
Feature types are inferred from the labels in the GenBank file; the file itself is the reference.