p935sGFP B982
Ligation of NptGFP-Xho-Mlu* into B414p7o-LH*
- Promoter 1
- 35s
- Gene 1
- nptII-EGFP
- Features
- dicots
- Other markers
- NptIIo, Sm/Sp
- Size
- 10,390 bp, circular
- Record date
- 04-OCT-2017
Construct
- 35S–NptIIo–GFPm · E9 · payload cassette, 309–2,991
- MCS · 11 unique sites · at 3,001, 3,092
- SfiI ×2 · flanks the MCS · at 3,032, 3,096
- 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 35S |
| XmnI (PdmI) | GAANN^NNTTC | 594 | in 35S |
| XhoI (PaeR7I) | C^TCGAG | 727 | between 35S and NptIIo |
| SacI (Ecl136II) | GAGCT^C | 1,255 | in NptIIo |
| BstBI (Bsp119I) | TT^CGAA | 2,126 | in GFPm |
| MluI | A^CGCGT | 2,691 | between GFPm and E9 |
| PstI | CTGCA^G | 3,001 | between E9 and RB |
| SbfI (SdaI) | CCTGCA^GG | 3,001 | between E9 and RB |
| NcoI | C^CATGG | 3,027 | between E9 and RB |
| SpeI | A^CTAGT | 3,040 | between E9 and RB |
| BamHI | G^GATCC | 3,046 | between E9 and RB |
| SmaI (XmaI, TspMI) | CCC^GGG | 3,054 | between E9 and RB |
| NsiI (Mph1103I) | ATGCA^T | 3,062 | between E9 and RB |
| HindIII | A^AGCTT | 3,070 | between E9 and RB |
| SalI | G^TCGAC | 3,082 | between E9 and RB |
| StuI (Eco147I) | AGG^CCT | 3,090 | between E9 and RB |
| AflII (BspTI) | C^TTAAG | 3,092 | between E9 and RB |
| PmeI (MssI) | GTTT^AAAC | 3,151 | between E9 and RB |
| AatII (ZraI) | GACGT^C | 3,542 | between RB and ColE1 |
| BsiWI (Pfl23II) | C^GTACG | 7,756 | in pVS1 |
| NheI (BmtI) | G^CTAGC | 7,872 | in pVS1 |
Cut twice: EcoRI (1,780, 3,064), SphI (3,007, 3,276), NdeI (4,657, 6,416), BglII (1,769, 3,102), EcoRV (634, 3,663), ApaI (828, 3,113), SfiI (3,032, 3,096), AgeI (8,366, 8,674), BstEII (1,497, 9,237), BsmBI (4,835, 5,888), SapI (4,595, 8,492), AarI (1,218, 1,339).
No site: AscI, PacI, SwaI, AvrII, HpaI, BsrGI, SacII, 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 | + |
| 35S | Promoter misc_feature | 309 | 724 | 416 | + |
| NptIIo | Selection marker CDS | 745 | 1,043 | 299 | + |
| Int | Site CDS | 1,044 | 1,230 | 187 | + |
| NptIIo | Selection marker CDS | 1,231 | 1,734 | 504 | + |
| GFPm | Coding sequence CDS | 1,786 | 2,164 | 379 | + |
| Int | Site CDS | 2,165 | 2,353 | 189 | + |
| GFPm | Coding sequence CDS | 2,354 | 2,691 | 338 | + |
| E9 | Terminator misc_feature | 2,701 | 2,991 | 291 | + |
| RB | T-DNA border misc_feature | 3,167 | 3,190 | 24 | + |
| ColE1 | Origin / backbone misc_feature | 3,725 | 4,422 | 698 | + |
| bom | Origin / backbone misc_feature | 4,459 | 4,718 | 260 | + |
| pVS1 | Origin / backbone misc_feature | 4,971 | 8,676 | 3,706 | + |
| Sm/Sp | Selection marker CDS | 8,860 | 9,870 | 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.