p9o2x35s-UbiZm-GFP C872
Ligation of B595pUbiZm-mGFPM-ocs* into B485p9o2x35s-LH*
- Promoter 1
- 35s
- Gene 1
- nptII
- Promoter 2
- UbiZm
- Gene 2
- EGFP
- Features
- monocots
- Other markers
- NptIIo, Sm/Sp
- Size
- 12,437 bp, circular
- Record date
- 26-AUG-2020
Construct
- P35S–NptIIo · TE9 · selection cassette, 314–2,416
- PUbi–EGFP · OCS · payload cassette, 2,487–5,129
- MCS · 4 unique sites · at 2,426, 2,479
- SfiI ×2 · flanks the PUbi–EGFP cassette · at 2,457, 5,143
- 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 |
|---|---|---|---|
| SacI (Ecl136II) | GAGCT^C | 1,626 | in NptIIo |
| SbfI (SdaI) | CCTGCA^GG | 2,426 | between TE9 and PUbi |
| SpeI | A^CTAGT | 2,465 | between TE9 and PUbi |
| BamHI | G^GATCC | 2,471 | between TE9 and PUbi |
| SmaI (XmaI, TspMI) | CCC^GGG | 2,479 | between TE9 and PUbi |
| NsiI (Mph1103I) | ATGCA^T | 3,763 | in Intron |
| HindIII | A^AGCTT | 3,997 | between PUbi and EGFP |
| MluI | A^CGCGT | 4,003 | between PUbi and EGFP |
| BstBI (Bsp119I) | TT^CGAA | 4,360 | in EGFP |
| SalI | G^TCGAC | 4,929 | between EGFP and OCS |
| StuI (Eco147I) | AGG^CCT | 5,137 | between OCS and RB |
| AflII (BspTI) | C^TTAAG | 5,139 | between OCS and RB |
| PmeI (MssI) | GTTT^AAAC | 5,198 | between OCS and RB |
| AatII (ZraI) | GACGT^C | 5,589 | between RB and ColE1 |
| BsiWI (Pfl23II) | C^GTACG | 9,803 | in pVS1 |
Cut twice: PstI (2,426, 3,991), SphI (2,432, 5,323), KpnI (4,241, 5,166), XhoI (1,098, 3,185), NcoI (2,452, 3,532), NheI (3,605, 9,919), BglII (3,432, 5,149), EcoRV (966, 5,710), SfiI (2,457, 5,143), AgeI (10,413, 10,721), XmnI (601, 926), BstEII (1,868, 11,284), SapI (6,642, 10,539), AarI (1,589, 1,710).
No site: EcoRI, AscI, PacI, SwaI, AvrII, HpaI, BsrGI, 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 | + |
| P35S | Promoter misc_feature | 314 | 1,098 | 785 | + |
| NptIIo | Selection marker CDS | 1,116 | 1,414 | 299 | + |
| int | Site CDS | 1,415 | 1,601 | 187 | + |
| NptIIo | Selection marker CDS | 1,602 | 2,108 | 507 | + |
| TE9 | Terminator misc_feature | 2,126 | 2,416 | 291 | + |
| PUbi | Promoter misc_feature | 2,487 | 3,994 | 1,508 | + |
| Intron | Site misc_feature | 3,473 | 3,992 | 520 | + |
| EGFP | Coding sequence CDS | 4,020 | 4,398 | 379 | + |
| ST-LS1 | Coding sequence CDS | 4,399 | 4,587 | 189 | + |
| EGFP | Coding sequence CDS | 4,588 | 4,925 | 338 | + |
| OCS | Terminator misc_feature | 4,934 | 5,129 | 196 | + |
| RB | T-DNA border misc_feature | 5,214 | 5,237 | 24 | + |
| ColE1 | Origin / backbone misc_feature | 5,772 | 6,469 | 698 | + |
| pVS1 | Origin / backbone misc_feature | 7,018 | 10,723 | 3,706 | + |
| Sm/Sp | Selection marker CDS | 10,907 | 11,917 | 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.