p9oNoR-U10 D922
Ligation of B357p9ioR-35sCasWT* into C889p9oN-U10*
- Promoter 1
- Nos
- Gene 1
- nptII
- Promoter 2
- Ubq10
- Features
- dicots
- Other markers
- NptIIo, Sm/Sp
- Size
- 9,814 bp, circular
- Record date
- 26-AUG-2020
Construct
- PNOS–NptIIo · TE9 · selection cassette, 309–1,918
- Pro-Ubq10At–(empty)–OCS · empty expression cassette, 1,967–3,685
- MCS · 5 unique sites · at 3,443, 3,505
- SfiI ×2 · flanks the Pro-Ubq10At–(empty)–OCS cassette · at 1,959, 3,699
- T-DNA borders · 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 |
|---|---|---|---|
| SacII (Cfr42I) | CCGC^GG | 421 | in PNOS |
| XhoI (PaeR7I) | C^TCGAG | 607 | between PNOS and NptIIo |
| SbfI (SdaI) | CCTGCA^GG | 1,928 | between TE9 and Pro-Ubq10At |
| NcoI | C^CATGG | 1,954 | between TE9 and Pro-Ubq10At |
| SpeI | A^CTAGT | 3,443 | between Pro-Ubq10At and OCS |
| BamHI | G^GATCC | 3,449 | between Pro-Ubq10At and OCS |
| SmaI (XmaI, TspMI) | CCC^GGG | 3,457 | between Pro-Ubq10At and OCS |
| HindIII | A^AGCTT | 3,473 | between Pro-Ubq10At and OCS |
| BsmBI (Esp3I) | CGTCTC(1/5) | 3,505 | in OCS |
| StuI (Eco147I) | AGG^CCT | 3,693 | between OCS and RB |
| AflII (BspTI) | C^TTAAG | 3,695 | between OCS and RB |
| KpnI (Acc65I) | GGTAC^C | 3,722 | between OCS and RB |
| PmeI (MssI) | GTTT^AAAC | 3,754 | between OCS and RB |
| AatII (ZraI) | GACGT^C | 4,145 | between RB and ColE1 |
| BsiWI (Pfl23II) | C^GTACG | 7,180 | in pVS1 |
| ClaI (BspDI) | AT^CGAT | 7,376 | in pVS1 |
Cut twice: EcoRI (2,627, 3,467), SalI (2,805, 3,485), PstI (1,928, 3,465), SphI (1,934, 3,879), SacI (1,135, 3,060), NdeI (5,260, 5,840), NheI (503, 7,296), BglII (3,372, 3,705), ApaI (708, 3,716), MluI (1,618, 3,479), SfiI (1,959, 3,699), FseI (1,274, 6,952), MfeI (14, 1,028), AgeI (7,790, 8,098), XmnI (1,620, 2,228), BstEII (1,377, 8,661), AarI (1,098, 1,219).
No site: AscI, PacI, SwaI, AvrII, BstBI, HpaI, NsiI, BsrGI, 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 |
|---|---|---|---|---|---|
| BL | Site misc_feature | 1 | 24 | 24 | + |
| PNOS | Promoter misc_feature | 309 | 606 | 298 | + |
| NptIIo | Selection marker CDS | 625 | 923 | 299 | + |
| splice | Site misc_feature | 920 | 923 | 4 | + |
| int | Site CDS | 924 | 1,110 | 187 | + |
| splice | Site misc_feature | 1,109 | 1,112 | 4 | + |
| NptIIo | Selection marker CDS | 1,111 | 1,614 | 504 | + |
| TE9 | Terminator misc_feature | 1,628 | 1,918 | 291 | + |
| Pro-Ubq10At | Promoter misc_feature | 1,967 | 3,442 | 1,476 | + |
| TATA | Site misc_feature | 3,038 | 3,041 | 4 | + |
| Ubi cds 5 | Promoter misc_feature | 3,071 | 3,134 | 64 | + |
| íntron | Other misc_feature | 3,137 | 3,440 | 304 | + |
| OCS | Terminator misc_feature | 3,490 | 3,685 | 196 | + |
| RB | T-DNA border misc_feature | 3,770 | 3,793 | 24 | + |
| ColE1 | Origin / backbone misc_feature | 4,328 | 5,025 | 698 | + |
| pVS1 | Origin / backbone misc_feature | 5,313 | 8,100 | 2,788 | + |
| Sm/Sp | Selection marker CDS | 8,284 | 9,294 | 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.