p7D35s-UbiAt-mEGFP E039
Ligation of E038pUbiAt-mEGFP* into D011p7D35s*
- Promoter 1
- 35s
- Gene 1
- bar
- Promoter 2
- Ubq10
- Gene 2
- mEGFP
- Features
- dicots
- Other markers
- strep/spec
- Size
- 9,028 bp, circular
- Record date
- 26-AUG-2020
Construct
- P35S–Bar · TE9 · selection cassette, 169–1,646
- Pro-Ubq10At–mEGFP · OCS · payload cassette, 1,690–4,301
- MCS · 6 unique sites · at 1,656, 1,686
- SfiI ×2 · flanks the Pro-Ubq10At–mEGFP cassette · at 1,664, 4,315
- 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 |
|---|---|---|---|
| XhoI (PaeR7I) | C^TCGAG | 587 | between P35S and Bar |
| AarI | CACCTGC(4/8) | 1,128 | in Bar |
| PstI | CTGCA^G | 1,656 | between TE9 and Pro-Ubq10At |
| SbfI (SdaI) | CCTGCA^GG | 1,656 | between TE9 and Pro-Ubq10At |
| NotI | GC^GGCCGC | 1,665 | between TE9 and Pro-Ubq10At |
| SpeI | A^CTAGT | 1,672 | between TE9 and Pro-Ubq10At |
| BamHI | G^GATCC | 1,678 | between TE9 and Pro-Ubq10At |
| SmaI (XmaI, TspMI) | CCC^GGG | 1,686 | between TE9 and Pro-Ubq10At |
| EcoRI | G^AATTC | 2,350 | in Pro-Ubq10At |
| SacI (Ecl136II) | GAGCT^C | 2,783 | in Pro-Ubq10At |
| SapI (BspQI, LguI) | GCTCTTC(1/4) | 2,788 | in Pro-Ubq10At |
| HindIII | A^AGCTT | 3,168 | between Pro-Ubq10At and mEGFP |
| BstBI (Bsp119I) | TT^CGAA | 3,531 | in mEGFP |
| ClaI (BspDI) | AT^CGAT | 3,762 | in mEGFP |
| BsmBI (Esp3I) | CGTCTC(1/5) | 4,121 | in OCS |
| PmeI (MssI) | GTTT^AAAC | 4,375 | in Right Border Region |
| SphI (PaeI) | GCATG^C | 4,500 | in Right Border Region |
| AatII (ZraI) | GACGT^C | 4,766 | in Right Border Region |
| BstEII (Eco91I) | G^GTNACC | 8,178 | in strep/spec |
Cut twice: SalI (2,528, 4,101), KpnI (3,412, 4,343), BglII (3,095, 4,321), MluI (1,346, 3,174), SfiI (1,664, 4,315), StuI (1,658, 4,309), EagI (1,665, 4,757), SacII (1,228, 1,245).
No site: NcoI, NdeI, NheI, AscI, PacI, FseI, SwaI, AvrII, BsiWI, AgeI, HpaI, NsiI, AflII, BsrGI, PvuII, BsaI.
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 | 169 | 584 | 416 | + |
| Bar | Selection marker CDS | 605 | 838 | 234 | + |
| STLS1 | Other misc_feature | 839 | 1,027 | 189 | + |
| Bar | Selection marker CDS | 1,028 | 1,343 | 316 | + |
| TE9 | Terminator misc_feature | 1,356 | 1,646 | 291 | + |
| Pro-Ubq10At | Promoter misc_feature | 1,690 | 3,165 | 1,476 | + |
| TATA | Site misc_feature | 2,761 | 2,764 | 4 | + |
| Ubi cds 5 | Promoter misc_feature | 2,794 | 2,859 | 66 | + |
| íntron | Other misc_feature | 2,860 | 3,163 | 304 | + |
| mEGFP | Coding sequence CDS | 3,191 | 3,569 | 379 | + |
| ST-LS1 | Coding sequence CDS | 3,570 | 3,758 | 189 | + |
| mEGFP | Coding sequence CDS | 3,759 | 4,096 | 338 | + |
| A206K - monomeric protein | Site misc_feature | 3,995 | 3,997 | 3 | + |
| OCS | Terminator misc_feature | 4,106 | 4,301 | 196 | + |
| Right Border Region | Other misc_feature | 4,346 | 4,948 | 603 | + |
| RB | T-DNA border misc_feature | 4,391 | 4,414 | 24 | + |
| ColE1 | Origin / backbone misc_feature | 4,962 | 5,620 | 659 | + |
| ''pVS1 ORI''' | Origin / backbone misc_feature | 5,649 | 7,612 | 1,964 | + |
| ParB | Coding sequence CDS | 5,975 | 7,150 | 1,176 | + |
| strep/spec | Selection marker CDS | 7,801 | 8,811 | 1,011 | + |
| Left Border Region | T-DNA border misc_feature | 8,879 | 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.