p9oU10-35s-LGFP C859
Ligation of B629p35s-LGFP* into B435p9oU10-LH*
- Promoter 1
- Ubq10
- Gene 1
- nptII
- Promoter 2
- 35s
- Gene 2
- EGFP
- Features
- dicots
- Other markers
- NptIIo, Sm/Sp
- Size
- 12,091 bp, circular
- Record date
- 26-AUG-2020
Construct
- Pro-Ubq10At–NptIIo · TE9 · selection cassette, 309–3,097
- 35S-Pro–EGFP · OCS · payload cassette, 3,152–4,783
- MCS · 5 unique sites · at 3,573, 3,616
- SfiI ×2 · flanks the 35S-Pro–EGFP cassette · at 3,138, 4,797
- 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 | 1,786 | between Pro-Ubq10At and NptIIo |
| SbfI (SdaI) | CCTGCA^GG | 3,107 | between TE9 and 35S-Pro |
| NcoI | C^CATGG | 3,133 | between TE9 and 35S-Pro |
| SpeI | A^CTAGT | 3,573 | in 35S-Pro |
| BamHI | G^GATCC | 3,580 | between 35S-Pro and link |
| SmaI (XmaI, TspMI) | CCC^GGG | 3,588 | between 35S-Pro and link |
| HindIII | A^AGCTT | 3,604 | between 35S-Pro and link |
| BsrGI (Bsp1407I) | T^GTACA | 3,616 | between 35S-Pro and link |
| BstBI (Bsp119I) | TT^CGAA | 4,013 | in EGFP |
| StuI (Eco147I) | AGG^CCT | 4,791 | between OCS and RB |
| AflII (BspTI) | C^TTAAG | 4,793 | between OCS and RB |
| KpnI (Acc65I) | GGTAC^C | 4,820 | between OCS and RB |
| PmeI (MssI) | GTTT^AAAC | 4,852 | between OCS and RB |
| AatII (ZraI) | GACGT^C | 5,243 | between RB and ColE1 |
| BsiWI (Pfl23II) | C^GTACG | 9,457 | in pVS1 |
| NheI (BmtI) | G^CTAGC | 9,573 | in pVS1 |
Cut twice: EcoRI (969, 3,598), SalI (1,147, 4,583), PstI (3,107, 3,596), SphI (3,113, 4,977), SacI (1,402, 2,314), NdeI (6,358, 8,117), BglII (1,714, 4,803), ClaI (4,244, 9,653), ApaI (1,887, 4,814), MluI (2,797, 3,610), SfiI (3,138, 4,797), AgeI (10,067, 10,375), BstEII (2,556, 10,938), AarI (2,277, 2,398).
No site: AscI, PacI, SwaI, AvrII, HpaI, NsiI, 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 | + |
| Pro-Ubq10At | Promoter misc_feature | 309 | 1,784 | 1,476 | + |
| Ubi cds 5 | Promoter misc_feature | 1,413 | 1,476 | 64 | + |
| íntron | Other misc_feature | 1,479 | 1,782 | 304 | + |
| NptIIo | Selection marker CDS | 1,804 | 2,102 | 299 | + |
| splice | Site misc_feature | 2,099 | 2,102 | 4 | + |
| int | Site CDS | 2,103 | 2,289 | 187 | + |
| splice | Site misc_feature | 2,288 | 2,291 | 4 | + |
| NptIIo | Selection marker CDS | 2,290 | 2,793 | 504 | + |
| TE9 | Terminator misc_feature | 2,807 | 3,097 | 291 | + |
| 35S-Pro | Promoter misc_feature | 3,152 | 3,574 | 423 | + |
| TATA | Site CDS | 3,540 | 3,546 | 7 | + |
| link | Site misc_feature | 3,622 | 3,672 | 51 | + |
| EGFP | Coding sequence CDS | 3,673 | 4,051 | 379 | + |
| ST-LS1 | Coding sequence CDS | 4,052 | 4,240 | 189 | + |
| EGFP | Coding sequence CDS | 4,241 | 4,578 | 338 | + |
| OCS | Terminator misc_feature | 4,588 | 4,783 | 196 | + |
| RB | T-DNA border misc_feature | 4,868 | 4,891 | 24 | + |
| ColE1 | Origin / backbone misc_feature | 5,426 | 6,123 | 698 | + |
| bom | Origin / backbone misc_feature | 6,160 | 6,419 | 260 | + |
| pVS1 | Origin / backbone misc_feature | 6,672 | 10,377 | 3,706 | + |
| Sm/Sp | Selection marker CDS | 10,561 | 11,571 | 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.