p2x35s-LGFP B632
Ligation of B629p35s-LGFP* into 400pD35-ocs*
- Promoter 2
- 2x35s
- Gene 2
- Link-GFP
- Features
- monocots
- Other markers
- Amp
- Size
- 4,924 bp, circular
- Record date
- 08-FEB-2016
Construct
- P2x35s–gfpm · Tocs · payload cassette, 110–2,108
- MCS · 8 unique sites · at 898, 941
- SfiI ×2 · flanks the P2x35s–gfpm cassette · at 92, 2,122
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 | 77 | between ColE1 and P2x35s |
| XbaI | T^CTAGA | 79 | between ColE1 and P2x35s |
| NcoI | C^CATGG | 87 | between ColE1 and P2x35s |
| NotI | GC^GGCCGC | 93 | between ColE1 and P2x35s |
| EagI (Eco52I) | C^GGCCG | 93 | between ColE1 and P2x35s |
| EcoRV (Eco32I) | GAT^ATC | 762 | in P2x35s |
| SpeI | A^CTAGT | 898 | between P2x35s and link |
| BamHI | G^GATCC | 905 | between P2x35s and link |
| SmaI (XmaI, TspMI) | CCC^GGG | 913 | between P2x35s and link |
| PstI | CTGCA^G | 921 | between P2x35s and link |
| EcoRI | G^AATTC | 923 | between P2x35s and link |
| HindIII | A^AGCTT | 929 | between P2x35s and link |
| MluI | A^CGCGT | 935 | between P2x35s and link |
| BsrGI (Bsp1407I) | T^GTACA | 941 | between P2x35s and link |
| BstBI (Bsp119I) | TT^CGAA | 1,338 | in gfpm |
| MfeI (MunI) | C^AATTG | 1,483 | in int |
| ClaI (BspDI) | AT^CGAT | 1,569 | in gfpm |
| SalI | G^TCGAC | 1,908 | between gfpm and Tocs |
| BsmBI (Esp3I) | CGTCTC(1/5) | 1,928 | in Tocs |
| StuI (Eco147I) | AGG^CCT | 2,116 | between Tocs and Amp |
| AflII (BspTI) | C^TTAAG | 2,118 | between Tocs and Amp |
| BglII | A^GATCT | 2,128 | between Tocs and Amp |
| ApaI (PspOMI) | GGGCC^C | 2,139 | between Tocs and Amp |
| KpnI (Acc65I) | GGTAC^C | 2,145 | between Tocs and Amp |
| BsaI (Eco31I) | GGTCTC(1/5) | 3,645 | in Amp |
| SapI (BspQI, LguI) | GCTCTTC(1/4) | 4,716 | between ColE1 and P2x35s |
Cut twice: SfiI (92, 2,122), PvuI (2,297, 3,342), PvuII (2,266, 4,779).
No site: SphI, XhoI, NdeI, NheI, AscI, PacI, SbfI, FseI, PmeI, SwaI, AvrII, BsiWI, AgeI, HpaI, NsiI, SacII, AatII, BstEII, 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 |
|---|---|---|---|---|---|
| P2x35s | Promoter misc_feature | 110 | 894 | 785 | + |
| mut | Site misc_signal | 724 | 724 | 1 | + |
| TATA | Site misc_signal | 822 | 828 | 7 | + |
| link | Other misc_feature | 947 | 997 | 51 | + |
| gfpm | Coding sequence CDS | 998 | 1,376 | 379 | + |
| int | Site CDS | 1,377 | 1,565 | 189 | + |
| gfpm | Coding sequence CDS | 1,566 | 1,903 | 338 | + |
| Tocs | Terminator misc_feature | 1,913 | 2,108 | 196 | + |
| Amp | Selection marker misc_feature | 2,924 | 3,784 | 861 | + |
| ColE1 | Origin / backbone CDS | 3,827 | 4,543 | 717 | + |
PromoterCoding sequenceTerminatorSelection markerOrigin / backboneT-DNA borderSite
Feature types are inferred from the labels in the GenBank file; the file itself is the reference.