p35sTnos-35sTocs A744
Ligation of Nos-35s-Spe-Sal* into p35-OCS-BM*
- Promoter 1
- 35s
- Other markers
- Amp
- Size
- 4,293 bp, circular
- Record date
- 18-DEC-2013
Construct
- 35S-Pro–(empty)–NOS-T · empty expression cassette, 106–807
- 35S-Pro–(empty)–OCS · empty expression cassette, 834–1,477
- MCS · 5 unique sites · at 77, 93
- SfiI ×2 · flanks 2 cassettes · at 92, 1,491
- BsmBI Golden Gate · overhangs CCCG, TATG · at 812, 1,297
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 |
|---|---|---|---|
| SacI (Ecl136II) | GAGCT^C | 77 | between ColE1 and 35S-Pro' |
| XbaI | T^CTAGA | 79 | between ColE1 and 35S-Pro' |
| NcoI | C^CATGG | 87 | between ColE1 and 35S-Pro' |
| NotI | GC^GGCCGC | 93 | between ColE1 and 35S-Pro' |
| EagI (Eco52I) | C^GGCCG | 93 | between ColE1 and 35S-Pro' |
| SpeI | A^CTAGT | 527 | between 35S-Pro' and NOS-T |
| BamHI | G^GATCC | 536 | between 35S-Pro' and NOS-T |
| NsiI (Mph1103I) | ATGCA^T | 674 | in NOS-T |
| SmaI (XmaI, TspMI) | CCC^GGG | 815 | between NOS-T and 35S-Pro |
| EcoRI | G^AATTC | 1,259 | between 35S-Pro and OCS |
| AvrII (XmaJI) | C^CTAGG | 1,268 | between 35S-Pro and OCS |
| SalI | G^TCGAC | 1,277 | between 35S-Pro and OCS |
| StuI (Eco147I) | AGG^CCT | 1,485 | between OCS and Amp |
| BglII | A^GATCT | 1,497 | between OCS and Amp |
| ApaI (PspOMI) | GGGCC^C | 1,508 | between OCS and Amp |
| KpnI (Acc65I) | GGTAC^C | 1,514 | between OCS and Amp |
| BsaI (Eco31I) | GGTCTC(1/5) | 3,014 | in Amp |
| SapI (BspQI, LguI) | GCTCTTC(1/4) | 4,085 | between ColE1 and 35S-Pro' |
Cut twice: EcoRV (434, 1,162), SfiI (92, 1,491), AflII (583, 1,487), PvuII (1,635, 4,148), BsmBI (812, 1,297).
No site: HindIII, PstI, SphI, XhoI, NdeI, NheI, ClaI, MluI, AscI, PacI, SbfI, FseI, PmeI, SwaI, BsiWI, MfeI, AgeI, BstBI, HpaI, BsrGI, SacII, AatII, BstEII, 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-Pro' | Promoter misc_feature | 106 | 527 | 422 | + |
| TATA | Site CDS | 494 | 500 | 7 | + |
| NOS-T | Terminator misc_feature | 554 | 807 | 254 | + |
| 35S-Pro | Promoter misc_feature | 834 | 1,256 | 423 | + |
| TATA | Site CDS | 1,222 | 1,228 | 7 | + |
| OCS | Terminator misc_feature | 1,282 | 1,477 | 196 | + |
| Amp | Selection marker misc_feature | 2,293 | 3,153 | 861 | + |
| ColE1 | Origin / backbone misc_feature | 3,196 | 3,912 | 717 | + |
PromoterCoding sequenceTerminatorSelection markerOrigin / backboneT-DNA borderSite
Feature types are inferred from the labels in the GenBank file; the file itself is the reference.