pmGFPD-nos B445
Ligation of GFPdi-Hind-Sal* into pNos-BM*
- Promoter 2
- MCS
- Gene 2
- GFP
- Features
- dicots
- Other markers
- Amp
- Size
- 4,150 bp, circular
- Record date
- 22-JAN-2016
Construct
- MCS · 12 unique sites · at 103, 162
- SfiI ×2 · flanks the MCS · at 118, 1,374
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 | 103 | between ColE1 and gfp |
| XbaI | T^CTAGA | 105 | between ColE1 and gfp |
| NcoI | C^CATGG | 113 | between ColE1 and gfp |
| NotI | GC^GGCCGC | 119 | between ColE1 and gfp |
| EagI (Eco52I) | C^GGCCG | 119 | between ColE1 and gfp |
| SpeI | A^CTAGT | 126 | between ColE1 and gfp |
| BamHI | G^GATCC | 132 | between ColE1 and gfp |
| SmaI (XmaI, TspMI) | CCC^GGG | 140 | between ColE1 and gfp |
| PstI | CTGCA^G | 148 | between ColE1 and gfp |
| EcoRI | G^AATTC | 150 | between ColE1 and gfp |
| HindIII | A^AGCTT | 156 | between ColE1 and gfp |
| MluI | A^CGCGT | 162 | between ColE1 and gfp |
| BstBI (Bsp119I) | TT^CGAA | 519 | in gfp |
| MfeI (MunI) | C^AATTG | 664 | in int |
| ClaI (BspDI) | AT^CGAT | 750 | in gfp |
| SalI | G^TCGAC | 1,088 | between gfp and Tnos |
| NsiI (Mph1103I) | ATGCA^T | 1,226 | in Tnos |
| StuI (Eco147I) | AGG^CCT | 1,368 | between Tnos and Amp |
| BglII | A^GATCT | 1,380 | between Tnos and Amp |
| ApaI (PspOMI) | GGGCC^C | 1,391 | between Tnos and Amp |
| XmnI (PdmI) | GAANN^NNTTC | 2,363 | in Amp |
| BsaI (Eco31I) | GGTCTC(1/5) | 2,897 | in Amp |
| SapI (BspQI, LguI) | GCTCTTC(1/4) | 3,968 | between ColE1 and gfp |
Cut twice: KpnI (400, 1,397), SfiI (118, 1,374), AflII (1,135, 1,370), PvuII (1,518, 4,031).
No site: SphI, XhoI, NdeI, NheI, EcoRV, AscI, PacI, SbfI, FseI, PmeI, SwaI, AvrII, BsiWI, AgeI, HpaI, BsrGI, SacII, AatII, BstEII, BbsI, BsmBI, 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 |
|---|---|---|---|---|---|
| gfp | Coding sequence CDS | 179 | 557 | 379 | + |
| int | Site CDS | 558 | 746 | 189 | + |
| gfp | Coding sequence CDS | 747 | 1,084 | 338 | + |
| Tnos | Terminator misc_feature | 1,106 | 1,359 | 254 | + |
| Amp | Selection marker misc_feature | 2,176 | 3,036 | 861 | + |
| ColE1 | Origin / backbone CDS | 3,079 | 3,795 | 717 | + |
PromoterCoding sequenceTerminatorSelection markerOrigin / backboneT-DNA borderSite
Feature types are inferred from the labels in the GenBank file; the file itself is the reference.