p6oAct-LH-nos C138
Ligation of pNos-BM* into B488p6oAct-LH* corrected sequence
- Promoter 1
- Act
- Gene 1
- hptII
- Gene 2
- MCS
- Features
- monocots
- Other markers
- Sm/Sp
- Size
- 10,930 bp, circular
- Record date
- 04-OCT-2017
Construct
- pAct–hptII · E9 · selection cassette, 313–3,260
- MCS · 7 unique sites · at 3,270, 3,351
- SfiI ×2 · flanks the MCS · at 3,301, 3,636
- 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 pAct |
| AarI | CACCTGC(4/8) | 1,002 | in pAct |
| SacI (Ecl136II) | GAGCT^C | 1,212 | in pAct |
| XhoI (PaeR7I) | C^TCGAG | 1,720 | between pAct and hptII |
| XmnI (PdmI) | GAANN^NNTTC | 2,620 | in hptII |
| SbfI (SdaI) | CCTGCA^GG | 3,270 | between E9 and NOS-T |
| NcoI | C^CATGG | 3,296 | between E9 and NOS-T |
| SpeI | A^CTAGT | 3,309 | between E9 and NOS-T |
| SmaI (XmaI, TspMI) | CCC^GGG | 3,323 | between E9 and NOS-T |
| HindIII | A^AGCTT | 3,339 | between E9 and NOS-T |
| MluI | A^CGCGT | 3,345 | between E9 and NOS-T |
| SalI | G^TCGAC | 3,351 | between E9 and NOS-T |
| ApaI (PspOMI) | GGGCC^C | 3,653 | between NOS-T and RB |
| KpnI (Acc65I) | GGTAC^C | 3,659 | between NOS-T and RB |
| PmeI (MssI) | GTTT^AAAC | 3,691 | between NOS-T and RB |
| AatII (ZraI) | GACGT^C | 4,082 | between RB and ColE1 |
| EcoRV (Eco32I) | GAT^ATC | 4,203 | between RB and ColE1 |
| BbsI (BpiI) | GAAGAC(2/6) | 5,492 | between ColE1 and pVS1 |
| BsiWI (Pfl23II) | C^GTACG | 8,296 | in pVS1 |
| ClaI (BspDI) | AT^CGAT | 8,492 | in pVS1 |
Cut twice: EcoRI (691, 3,333), BamHI (1,408, 3,315), PstI (3,270, 3,331), SphI (3,276, 3,816), NheI (1,715, 8,412), SfiI (3,301, 3,636), MfeI (14, 2,414), AgeI (8,906, 9,214), StuI (2,634, 3,630), NsiI (561, 3,489), AflII (3,398, 3,632), SacII (2,108, 2,938), SapI (5,135, 9,032).
No site: AscI, PacI, SwaI, AvrII, BstBI, HpaI, BsrGI, PvuII.
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 | + |
| pAct | Promoter misc_feature | 313 | 1,710 | 1,398 | + |
| splice | Site misc_feature | 1,244 | 1,247 | 4 | + |
| splice | Site misc_feature | 1,706 | 1,709 | 4 | + |
| hptII | Selection marker CDS | 1,738 | 2,307 | 570 | + |
| int | Site misc_feature | 2,308 | 2,496 | 189 | + |
| hptII | Selection marker CDS | 2,497 | 2,952 | 456 | + |
| E9 | Terminator misc_feature | 2,970 | 3,260 | 291 | + |
| NOS-T | Terminator misc_feature | 3,369 | 3,621 | 253 | + |
| RB | T-DNA border misc_feature | 3,707 | 3,730 | 24 | + |
| ColE1 | Origin / backbone misc_feature | 4,265 | 4,962 | 698 | + |
| pVS1 | Origin / backbone misc_feature | 5,511 | 9,216 | 3,706 | + |
| Sm/Sp | Selection marker CDS | 9,400 | 10,410 | 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.