pB-Nos-DoGus-ocs D749
Ligation of B492pNos-DoGusi-ocs* into pB-BA*
- Promoter 1
- Nos
- Gene 1
- oGus
- Features
- dicots
- Other markers
- Sm/Sp
- Size
- 10,176 bp, circular
- Record date
- 19-AUG-2020
Construct
- MCS · 5 unique sites · at 303, 338
- SfiI ×2 · flanks the MCS · at 316, 2,882
- 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 P-NOS |
| NcoI | C^CATGG | 311 | between LB and P-NOS |
| SpeI | A^CTAGT | 324 | between LB and P-NOS |
| BamHI | G^GATCC | 330 | between LB and P-NOS |
| SmaI (XmaI, TspMI) | CCC^GGG | 338 | between LB and P-NOS |
| SacII (Cfr42I) | CCGC^GG | 455 | in P-NOS |
| HindIII | A^AGCTT | 644 | between P-NOS and ogus |
| MluI | A^CGCGT | 650 | between P-NOS and ogus |
| AarI | CACCTGC(4/8) | 1,458 | in Intron |
| HpaI (KspAI) | GTT^AAC | 1,566 | in ogus |
| XmnI (PdmI) | GAANN^NNTTC | 2,303 | in ogus |
| SalI | G^TCGAC | 2,668 | between ogus and OCS |
| AflII (BspTI) | C^TTAAG | 2,878 | between OCS and RB |
| ApaI (PspOMI) | GGGCC^C | 2,899 | between OCS and RB |
| KpnI (Acc65I) | GGTAC^C | 2,905 | between OCS and RB |
| PmeI (MssI) | GTTT^AAAC | 2,937 | between OCS and RB |
| SphI (PaeI) | GCATG^C | 3,062 | between RB and ColE1 |
| AatII (ZraI) | GACGT^C | 3,328 | between RB and ColE1 |
| BbsI (BpiI) | GAAGAC(2/6) | 4,738 | between BOM and pVS1 ORI |
| BsiWI (Pfl23II) | C^GTACG | 7,542 | in pVS1 ORI |
| ClaI (BspDI) | AT^CGAT | 7,738 | in pVS1 ORI |
| BstEII (Eco91I) | G^GTNACC | 9,023 | in Sm/Sp |
Cut twice: NdeI (4,443, 6,202), NheI (537, 7,658), BglII (1,669, 2,888), EcoRV (1,644, 3,449), SfiI (316, 2,882), FseI (5,209, 7,314), MfeI (14, 1,388), AgeI (8,152, 8,460), StuI (2,172, 2,876), SapI (4,381, 8,278).
No site: EcoRI, PstI, SacI, XhoI, AscI, PacI, SbfI, SwaI, AvrII, BstBI, NsiI, 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 | + |
| P-NOS | Other misc_feature | 343 | 640 | 298 | + |
| ogus | Coding sequence CDS | 667 | 1,281 | 615 | + |
| 'Intron | Site misc_feature | 1,282 | 1,470 | 189 | + |
| ogus | Coding sequence CDS | 1,471 | 2,667 | 1,197 | + |
| OCS | Terminator misc_feature | 2,673 | 2,868 | 196 | + |
| RB | T-DNA border misc_feature | 2,953 | 2,976 | 24 | + |
| ColE1 | Origin / backbone misc_feature | 3,511 | 4,208 | 698 | + |
| BOM | Origin / backbone misc_feature | 4,245 | 4,504 | 260 | + |
| pVS1 ORI | Origin / backbone misc_feature | 4,757 | 8,462 | 3,706 | + |
| Sm/Sp | Selection marker misc_feature | 8,646 | 9,656 | 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.