p6o-LH-35s-LmEGFP D365
Ligation of D360p35s-LmEGFP* into B413p6o-LH*
- Promoter 1
- 35s
- Gene 1
- hptII
- Promoter 2
- 35s
- Gene 2
- link-mEGFP
- Features
- dicots
- Other markers
- Sm/Sp
- Size
- 11,254 bp, circular
- Record date
- 26-AUG-2020
Construct
- P35S–hptII · TE9 · selection cassette, 309–2,260
- 35S-Pro–mEGFP · OCS · payload cassette, 2,315–3,946
- MCS · 6 unique sites · at 2,736, 2,779
- SfiI ×2 · flanks the 35S-Pro–mEGFP cassette · at 2,301, 3,960
- 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 |
|---|---|---|---|
| XbaI | T^CTAGA | 303 | between LB and P35S |
| XhoI (PaeR7I) | C^TCGAG | 727 | between P35S and Kozak |
| SbfI (SdaI) | CCTGCA^GG | 2,270 | between TE9 and 35S-Pro |
| NcoI | C^CATGG | 2,296 | between TE9 and 35S-Pro |
| SpeI | A^CTAGT | 2,736 | in 35S-Pro |
| BamHI | G^GATCC | 2,743 | between 35S-Pro and Linker |
| SmaI (XmaI, TspMI) | CCC^GGG | 2,751 | between 35S-Pro and Linker |
| EcoRI | G^AATTC | 2,761 | between 35S-Pro and Linker |
| HindIII | A^AGCTT | 2,767 | between 35S-Pro and Linker |
| BsrGI (Bsp1407I) | T^GTACA | 2,779 | between 35S-Pro and Linker |
| BstBI (Bsp119I) | TT^CGAA | 3,176 | in mEGFP |
| SalI | G^TCGAC | 3,746 | between mEGFP and OCS |
| AflII (BspTI) | C^TTAAG | 3,956 | between OCS and RB |
| BglII | A^GATCT | 3,966 | between OCS and RB |
| ApaI (PspOMI) | GGGCC^C | 3,977 | between OCS and RB |
| KpnI (Acc65I) | GGTAC^C | 3,983 | between OCS and RB |
| PmeI (MssI) | GTTT^AAAC | 4,015 | between OCS and RB |
| AatII (ZraI) | GACGT^C | 4,406 | between RB and ColE1 |
| BsiWI (Pfl23II) | C^GTACG | 8,620 | in pVS1 |
| NheI (BmtI) | G^CTAGC | 8,736 | in pVS1 |
Cut twice: PstI (2,270, 2,759), SphI (2,276, 4,140), NdeI (5,521, 7,280), ClaI (3,407, 8,816), MluI (1,960, 2,773), SfiI (2,301, 3,960), FseI (6,287, 8,392), AgeI (9,230, 9,538), StuI (1,641, 3,954), SacII (1,115, 1,945), BstEII (1,035, 10,101), SapI (5,459, 9,356).
No site: SacI, AscI, PacI, SwaI, AvrII, HpaI, NsiI, PvuII, 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 |
|---|---|---|---|---|---|
| LB | T-DNA border misc_feature | 1 | 24 | 24 | + |
| P35S | Promoter misc_feature | 309 | 724 | 416 | + |
| Kozak | Site misc_feature | 740 | 744 | 5 | + |
| hptII | Selection marker CDS | 745 | 1,314 | 570 | + |
| int | Site misc_feature | 1,315 | 1,503 | 189 | + |
| hptII | Selection marker CDS | 1,504 | 1,959 | 456 | + |
| TE9 | Terminator misc_feature | 1,970 | 2,260 | 291 | + |
| 35S-Pro | Promoter misc_feature | 2,315 | 2,737 | 423 | + |
| TATA | Site CDS | 2,703 | 2,709 | 7 | + |
| Linker | Coding sequence CDS | 2,785 | 2,835 | 51 | + |
| mEGFP | Coding sequence CDS | 2,836 | 3,214 | 379 | + |
| F64L/S65T - enhanced fluorescence | Site misc_feature | 3,025 | 3,030 | 6 | + |
| ST-LS1 | Coding sequence CDS | 3,215 | 3,403 | 189 | + |
| mEGFP | Coding sequence CDS | 3,404 | 3,741 | 338 | + |
| A206K - monomeric protein | Site misc_feature | 3,640 | 3,642 | 3 | + |
| OCS | Terminator misc_feature | 3,751 | 3,946 | 196 | + |
| RB | T-DNA border misc_feature | 4,031 | 4,054 | 24 | + |
| ColE1 | Origin / backbone misc_feature | 4,589 | 5,286 | 698 | + |
| bom | Origin / backbone misc_feature | 5,323 | 5,582 | 260 | + |
| pVS1 | Origin / backbone misc_feature | 5,835 | 9,540 | 3,706 | + |
| Sm/Sp | Selection marker CDS | 9,724 | 10,734 | 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.