p6M2x35s-UbiZmGFP C242
Ligation of B595pUbiZm-mGFPM-ocs* into C241p6M2x35s*
- Promoter 1
- 2x35s
- Gene 1
- hptII
- Promoter 2
- UbiZm
- Gene 2
- EGFP
- Features
- monocots
- Other markers
- hptII, strep/spec
- Size
- 9,915 bp, circular
- Record date
- 10-APR-2017
Construct
- P2x35s–hptII · E9 · selection cassette, 174–2,498
- Ubi-int–GFP · OCS · payload cassette, 2,546–5,188
- MCS · 5 unique sites · at 2,508, 2,538
- SfiI ×2 · flanks the Ubi-int–GFP cassette · at 2,516, 5,202
- BsaI Golden Gate · overhangs GCGA, CCGC · at 1,488, 1,943
- 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 |
|---|---|---|---|
| SbfI (SdaI) | CCTGCA^GG | 2,508 | between E9 and Ubi-int |
| NotI | GC^GGCCGC | 2,517 | between E9 and Ubi-int |
| SpeI | A^CTAGT | 2,524 | between E9 and Ubi-int |
| BamHI | G^GATCC | 2,530 | between E9 and Ubi-int |
| SmaI (XmaI, TspMI) | CCC^GGG | 2,538 | between E9 and Ubi-int |
| NcoI | C^CATGG | 3,591 | in Intron |
| NheI (BmtI) | G^CTAGC | 3,664 | in Intron |
| NsiI (Mph1103I) | ATGCA^T | 3,822 | in Intron |
| HindIII | A^AGCTT | 4,056 | between Ubi-int and GFP |
| MluI | A^CGCGT | 4,062 | between Ubi-int and GFP |
| BstBI (Bsp119I) | TT^CGAA | 4,419 | in GFP |
| SalI | G^TCGAC | 4,988 | between GFP and OCS |
| BsmBI (Esp3I) | CGTCTC(1/5) | 5,008 | in OCS |
| PmeI (MssI) | GTTT^AAAC | 5,262 | in Right Border Region |
| SphI (PaeI) | GCATG^C | 5,387 | in Right Border Region |
| AatII (ZraI) | GACGT^C | 5,653 | in Right Border Region |
Cut twice: PstI (2,508, 4,050), KpnI (4,300, 5,230), XhoI (958, 3,244), BglII (3,491, 5,208), ClaI (3,777, 4,650), EcoRV (826, 5,774), ApaI (3,604, 5,221), SfiI (2,516, 5,202), EagI (2,517, 5,644), SacII (1,346, 2,176), BstEII (1,266, 9,065), BsaI (1,488, 1,943).
No site: EcoRI, SacI, AscI, PacI, FseI, SwaI, AvrII, BsiWI, AgeI, HpaI, AflII, BsrGI, PvuII, SapI, 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 |
|---|---|---|---|---|---|
| LB' | T-DNA border misc_feature | 1 | 24 | 24 | + |
| P2x35s' | Promoter misc_feature | 174 | 958 | 785 | + |
| Kozak | Site misc_feature | 971 | 975 | 5 | + |
| hptII | Selection marker CDS | 976 | 1,545 | 570 | + |
| int | Site misc_feature | 1,546 | 1,734 | 189 | + |
| 'hptII | Selection marker CDS | 1,735 | 2,190 | 456 | + |
| 'E9 | Terminator misc_feature | 2,208 | 2,498 | 291 | + |
| 'Ubi-int | Promoter misc_feature | 2,546 | 4,053 | 1,508 | + |
| Intron | Site misc_feature | 3,532 | 4,051 | 520 | + |
| GFP | Coding sequence CDS | 4,079 | 4,457 | 379 | + |
| ST-LS1 | Other misc_feature | 4,458 | 4,646 | 189 | + |
| GFP | Coding sequence CDS | 4,647 | 4,984 | 338 | + |
| OCS | Terminator misc_feature | 4,993 | 5,188 | 196 | + |
| Right Border Region | Other misc_feature | 5,233 | 5,835 | 603 | + |
| RB | T-DNA border misc_feature | 5,278 | 5,301 | 24 | + |
| 'ColE1'' | Origin / backbone misc_feature | 5,849 | 6,507 | 659 | + |
| ''pVS1 ORI''' | Origin / backbone misc_feature | 6,536 | 8,499 | 1,964 | + |
| strep/spec | Selection marker CDS | 8,688 | 9,698 | 1,011 | + |
| Left Border Region | T-DNA border misc_feature | 9,766 | 163 | 0 | + |
PromoterCoding sequenceTerminatorSelection markerOrigin / backboneT-DNA borderSite
Feature types are inferred from the labels in the GenBank file; the file itself is the reference.