pActOs-LGFP B635
Ligation of B629p35s-LGFP* into B627pAct-ocs*
- Promoter 2
- ActOs
- Gene 2
- Link-GFP
- Features
- monocots
- Other markers
- Amp
- Size
- 5,526 bp, circular
- Record date
- 08-FEB-2016
Construct
- ACT–gfpm · OCS · payload cassette, 103–2,710
- MCS · 6 unique sites · at 1,500, 1,543
- SfiI ×2 · flanks the ACT–gfpm cassette · at 92, 2,724
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 | 79 | between ColE1 and ACT |
| NcoI | C^CATGG | 87 | between ColE1 and ACT |
| NotI | GC^GGCCGC | 93 | between ColE1 and ACT |
| EagI (Eco52I) | C^GGCCG | 93 | between ColE1 and ACT |
| NsiI (Mph1103I) | ATGCA^T | 351 | in ACT |
| AarI | CACCTGC(4/8) | 792 | in ACT |
| FseI | GGCCGG^CC | 865 | in ACT |
| BstEII (Eco91I) | G^GTNACC | 1,035 | in mRNA |
| SpeI | A^CTAGT | 1,500 | between ACT and link |
| SmaI (XmaI, TspMI) | CCC^GGG | 1,515 | between ACT and link |
| PstI | CTGCA^G | 1,523 | between ACT and link |
| HindIII | A^AGCTT | 1,531 | between ACT and link |
| MluI | A^CGCGT | 1,537 | between ACT and link |
| BsrGI (Bsp1407I) | T^GTACA | 1,543 | between ACT and link |
| BstBI (Bsp119I) | TT^CGAA | 1,940 | in gfpm |
| MfeI (MunI) | C^AATTG | 2,085 | in int |
| ClaI (BspDI) | AT^CGAT | 2,171 | in gfpm |
| SalI | G^TCGAC | 2,510 | between gfpm and OCS |
| StuI (Eco147I) | AGG^CCT | 2,718 | between OCS and Amp |
| AflII (BspTI) | C^TTAAG | 2,720 | between OCS and Amp |
| ApaI (PspOMI) | GGGCC^C | 2,741 | between OCS and Amp |
| KpnI (Acc65I) | GGTAC^C | 2,747 | between OCS and Amp |
| XmnI (PdmI) | GAANN^NNTTC | 3,713 | in Amp |
| SapI (BspQI, LguI) | GCTCTTC(1/4) | 5,318 | between ColE1 and ACT |
Cut twice: EcoRI (481, 1,525), BamHI (1,198, 1,507), SacI (77, 1,002), NdeI (115, 436), SfiI (92, 2,724), PvuI (2,899, 3,944), PvuII (2,868, 5,381), BsaI (1,094, 4,247).
No site: SphI, XhoI, NheI, EcoRV, AscI, PacI, SbfI, PmeI, SwaI, AvrII, BsiWI, AgeI, HpaI, SacII, AatII, BbsI.
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 |
|---|---|---|---|---|---|
| ACT | Promoter misc_feature | 103 | 1,500 | 1,398 | + |
| mRNA | Coding sequence CDS | 946 | 1,500 | 555 | + |
| int | Site misc_feature | 1,036 | 1,497 | 462 | + |
| link | Other misc_feature | 1,549 | 1,599 | 51 | + |
| gfpm | Coding sequence CDS | 1,600 | 1,978 | 379 | + |
| int | Site CDS | 1,979 | 2,167 | 189 | + |
| gfpm | Coding sequence CDS | 2,168 | 2,505 | 338 | + |
| OCS | Terminator misc_feature | 2,515 | 2,710 | 196 | + |
| Amp | Selection marker misc_feature | 3,526 | 4,386 | 861 | + |
| ColE1 | Origin / backbone misc_feature | 4,429 | 5,145 | 717 | + |
PromoterCoding sequenceTerminatorSelection markerOrigin / backboneT-DNA borderSite
Feature types are inferred from the labels in the GenBank file; the file itself is the reference.