p9i-UbiH-2xStp-RNAi
Ligation of inverted A252pD1-UbiH-2xStp-RNAi* into p9i*
- Promoter 1
- 35S
- Gene 1
- npt-intron
- Promoter 2
- GUBB1HA
- Gene 2
- gus-spacer
- Features
- double promoter construct GUBB1HA with improved termination signals
- Other markers
- Sm/Sp
- Size
- 10,741 bp, circular
- Record date
- 20-JUN-2013
Construct
- 35S–NptII · selection cassette, 7–1,431
- UbiH–(empty)–T35S · empty expression cassette, 1,704–2,756
- UbiH–Gus · T35S · payload cassette, 2,062–3,114 (− strand)
- MCS · 4 unique sites · at 3,134, 3,200
- SfiI ×2 · flanks 2 cassettes · at 1,683, 3,145
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 |
|---|---|---|---|
| XmnI (PdmI) | GAANN^NNTTC | 292 | in 35S |
| XhoI (PaeR7I) | C^TCGAG | 425 | between 35S and NptII |
| PvuII | CAG^CTG | 871 | in NptII |
| SbfI (SdaI) | CCTGCA^GG | 1,652 | between T-35S and UbiH |
| SwaI (SmiI) | ATTT^AAAT | 1,693 | between T-35S and UbiH |
| SacI (Ecl136II) | GAGCT^C | 2,060 | between UbiH and T35S |
| MluI | A^CGCGT | 2,114 | between T35S and Gus |
| BsrGI (Bsp1407I) | T^GTACA | 2,121 | between T35S and Gus |
| HindIII | A^AGCTT | 2,128 | between T35S and Gus |
| SmaI (XmaI, TspMI) | CCC^GGG | 2,693 | between Gus and T35S |
| BamHI | G^GATCC | 2,698 | between Gus and T35S |
| SpeI | A^CTAGT | 2,764 | between T35S and UbiH |
| PacI | TTAAT^TAA | 3,134 | between UbiH and BR |
| BglII | A^GATCT | 3,151 | between UbiH and BR |
| ApaI (PspOMI) | GGGCC^C | 3,162 | between UbiH and BR |
| PmeI (MssI) | GTTT^AAAC | 3,200 | between UbiH and BR |
| AatII (ZraI) | GACGT^C | 3,591 | between BR and ColE1 |
| BsiWI (Pfl23II) | C^GTACG | 7,805 | in pVS1 ORI |
| NheI (BmtI) | G^CTAGC | 7,921 | in pVS1 ORI |
| ClaI (BspDI) | AT^CGAT | 8,001 | in pVS1 ORI |
| BstEII (Eco91I) | G^GTNACC | 9,286 | in Sm/Sp |
Cut twice: PstI (818, 1,652), KpnI (1,702, 3,168), NdeI (4,706, 6,465), EcoRV (332, 3,712), SfiI (1,683, 3,145), FseI (5,472, 7,577), AgeI (8,415, 8,723), BsmBI (4,884, 5,937).
No site: EcoRI, AscI, AvrII, BstBI, HpaI, StuI, NsiI, SacII, 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 |
|---|---|---|---|---|---|
| 35S | Promoter misc_feature | 7 | 422 | 416 | + |
| NptII | Site CDS | 439 | 475 | 37 | + |
| int | Site misc_feature | 476 | 664 | 189 | + |
| NptII | Selection marker CDS | 665 | 1,431 | 767 | + |
| T-35S | Promoter misc_feature | 1,444 | 1,645 | 202 | + |
| UbiH | Promoter misc_feature | 1,704 | 2,047 | 344 | + |
| T35S | Terminator misc_feature | 2,062 | 2,114 | 53 | − |
| unnamed | Coding sequence CDS | 2,134 | 2,683 | 550 | + |
| Gus | Coding sequence CDS | 2,134 | 2,683 | 550 | − |
| T35S | Terminator misc_feature | 2,704 | 2,756 | 53 | + |
| UbiH | Promoter misc_feature | 2,771 | 3,114 | 344 | − |
| BR | Site misc_feature | 3,216 | 3,239 | 24 | + |
| ColE1 | Origin / backbone misc_feature | 3,774 | 4,471 | 698 | + |
| pVS1 ORI | Origin / backbone misc_feature | 5,020 | 8,725 | 3,706 | + |
| Sm/Sp | Selection marker CDS | 8,909 | 9,919 | 1,011 | + |
| BL | Site misc_feature | 10,440 | 10,463 | 24 | + |
PromoterCoding sequenceTerminatorSelection markerOrigin / backboneT-DNA borderSite
Feature types are inferred from the labels in the GenBank file; the file itself is the reference.