p6M2x35s-UbiZmGusXi C906
Ligation of C673pC119p6oAct-P4-oGusXi* into C242p6M2x35s-UbiZmGFP*
- Promoter 1
- 2x35s
- Gene 1
- hptII
- Promoter 2
- UbiZm
- Gene 2
- oGus
- Features
- monocots
- Other markers
- hptII, strep/spec
- Size
- 11,007 bp, circular
- Record date
- 15-DEC-2022
Construct
- P2x35s–hptII · E9 · selection cassette, 174–2,498
- Ubi-int–GusX · OCS · payload cassette, 2,546–6,280
- MCS · 5 unique sites · at 2,508, 2,538
- SfiI ×2 · flanks the Ubi-int–GusX cassette · at 2,516, 6,294
- 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 |
|---|---|---|---|
| SbfI (SdaI) | CCTGCA^GG | 2,508 | between E9 and Ubi-int |
| NotI | GC^GGCCGC | 2,517 | between E9 and Ubi-int |
| SpeI | A^CTAGT | 2,524 | between E9 and Ubi-int |
| BamHI | G^GATCC | 2,530 | between E9 and Ubi-int |
| SmaI (XmaI, TspMI) | CCC^GGG | 2,538 | between E9 and Ubi-int |
| NcoI | C^CATGG | 3,591 | in Intron |
| NheI (BmtI) | G^CTAGC | 3,664 | in Intron |
| ClaI (BspDI) | AT^CGAT | 3,777 | in Intron |
| NsiI (Mph1103I) | ATGCA^T | 3,822 | in Intron |
| HindIII | A^AGCTT | 4,056 | between Ubi-int and GusX' |
| MluI | A^CGCGT | 4,062 | between Ubi-int and GusX' |
| HpaI (KspAI) | GTT^AAC | 4,375 | in GusX' |
| BstBI (Bsp119I) | TT^CGAA | 4,419 | in GusX' |
| AarI | CACCTGC(4/8) | 4,870 | in 'Intron |
| SalI | G^TCGAC | 6,080 | between 'GusX and OCS |
| BsmBI (Esp3I) | CGTCTC(1/5) | 6,100 | in OCS |
| KpnI (Acc65I) | GGTAC^C | 6,322 | between OCS and Right Border Region |
| PmeI (MssI) | GTTT^AAAC | 6,354 | in Right Border Region |
| SphI (PaeI) | GCATG^C | 6,479 | in Right Border Region |
Cut twice: PstI (2,508, 4,050), XhoI (958, 3,244), BglII (3,491, 6,300), ApaI (3,604, 6,313), SfiI (2,516, 6,294), SacII (1,346, 2,176).
No site: EcoRI, SacI, AscI, PacI, FseI, SwaI, AvrII, BsiWI, AgeI, AflII, BsrGI, PvuII, SapI.
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 | + |
| P2x35s' | Promoter misc_feature | 174 | 958 | 785 | + |
| Kozak | Site misc_feature | 971 | 975 | 5 | + |
| hptII | Selection marker CDS | 976 | 1,545 | 570 | + |
| int | Site misc_feature | 1,546 | 1,734 | 189 | + |
| 'hptII | Selection marker CDS | 1,735 | 2,190 | 456 | + |
| 'E9 | Terminator misc_feature | 2,208 | 2,498 | 291 | + |
| 'Ubi-int | Promoter misc_feature | 2,546 | 4,053 | 1,508 | + |
| Intron | Site misc_feature | 3,532 | 4,051 | 520 | + |
| GusX' | Coding sequence CDS | 4,079 | 4,693 | 615 | + |
| ''Intron | Site misc_feature | 4,694 | 4,882 | 189 | + |
| ''GusX | Coding sequence CDS | 4,883 | 6,079 | 1,197 | + |
| OCS | Terminator misc_feature | 6,085 | 6,280 | 196 | + |
| Right Border Region | Other misc_feature | 6,325 | 6,927 | 603 | + |
| RB | T-DNA border misc_feature | 6,370 | 6,393 | 24 | + |
| 'ColE1'' | Origin / backbone misc_feature | 6,941 | 7,599 | 659 | + |
| ''pVS1 ORI''' | Origin / backbone misc_feature | 7,628 | 9,591 | 1,964 | + |
| strep/spec | Selection marker CDS | 9,780 | 10,790 | 1,011 | + |
| Left Border Region | T-DNA border misc_feature | 10,858 | 163 | 0 | + |
PromoterCoding sequenceTerminatorSelection markerOrigin / backboneT-DNA borderSite
Feature types are inferred from the labels in the GenBank file; the file itself is the reference.