p7M2x35s-UZmC9i H027
Ligation of F653p6M2x35s-UbiZmCas9i* into E760p7M2x35s*
- Promoter 1
- 2x35s
- Gene 1
- bar
- Promoter 2
- UbiZm
- Gene 2
- Cas9
- Features
- monocots
- Other markers
- strep/spec
- Size
- 13,437 bp, circular
- Record date
- 26-NOV-2025
Construct
- P2x35s–Bar · E9 · selection cassette, 174–2,024
- Ubi-int–Cas9WT · NOS-T · payload cassette, 2,056–8,137
- PU6C4–gRNA · gRNA cassette, 8,147–8,709
- SfiI ×2 · flanks 2 cassettes · at 2,042, 8,724
- BsmBI Golden Gate · overhangs AGCT, GTTT · at 8,594, 8,626
- 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 |
|---|---|---|---|
| AarI | CACCTGC(4/8) | 1,499 | in Bar |
| SbfI (SdaI) | CCTGCA^GG | 2,034 | between E9 and Ubi-int |
| NotI | GC^GGCCGC | 2,043 | between E9 and Ubi-int |
| SpeI | A^CTAGT | 2,050 | between E9 and Ubi-int |
| NheI (BmtI) | G^CTAGC | 3,175 | in Intron |
| ClaI (BspDI) | AT^CGAT | 3,288 | in Intron |
| SacI (Ecl136II) | GAGCT^C | 3,630 | between NLS and int |
| SapI (BspQI, LguI) | GCTCTTC(1/4) | 4,877 | in Cas9WT |
| SmaI (XmaI, TspMI) | CCC^GGG | 7,881 | between Cas9WT and NOS-T |
| EcoRI | G^AATTC | 8,141 | between NOS-T and PU6C4 |
| BstBI (Bsp119I) | TT^CGAA | 8,607 | between PU6C4 and targetRNA |
| HindIII | A^AGCTT | 8,610 | between PU6C4 and targetRNA |
| SalI | G^TCGAC | 8,710 | between gRNA scaffold and Right Border Region |
| KpnI (Acc65I) | GGTAC^C | 8,752 | between gRNA scaffold and Right Border Region |
| PmeI (MssI) | GTTT^AAAC | 8,784 | in Right Border Region |
| BstEII (Eco91I) | G^GTNACC | 12,587 | in strep/spec |
Cut twice: SphI (6,990, 8,909), XhoI (958, 2,755), NcoI (3,102, 5,146), SfiI (2,042, 8,724), EagI (2,043, 9,166), AatII (4,265, 9,175), BsmBI (8,594, 8,626).
No site: BamHI, MluI, AscI, PacI, FseI, SwaI, AvrII, BsiWI, AgeI, HpaI, BsrGI, BsaI.
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 | + |
| Bar | Selection marker CDS | 976 | 1,209 | 234 | + |
| STLS1 | Other misc_feature | 1,210 | 1,398 | 189 | + |
| Bar | Selection marker CDS | 1,399 | 1,716 | 318 | + |
| 'E9 | Terminator misc_feature | 1,734 | 2,024 | 291 | + |
| Ubi-int | Promoter misc_feature | 2,056 | 3,564 | 1,509 | + |
| Intron | Site misc_feature | 3,043 | 3,562 | 520 | + |
| NLS | Site misc_feature | 3,595 | 3,615 | 21 | + |
| int | Site misc_feature | 4,006 | 4,098 | 93 | + |
| Cas9WT | Coding sequence CDS | 4,204 | 7,878 | 3,675 | + |
| NLS | Site misc_feature | 7,828 | 7,875 | 48 | + |
| NOS-T | Terminator misc_feature | 7,884 | 8,137 | 254 | + |
| PU6C4 | Promoter misc_feature | 8,147 | 8,599 | 453 | + |
| targetRNA | Site CDS | 8,611 | 8,626 | 16 | + |
| hairpin | Site CDS | 8,627 | 8,656 | 30 | + |
| gRNA scaffold | Site misc_feature | 8,627 | 8,709 | 83 | + |
| Right Border Region | Other misc_feature | 8,755 | 9,357 | 603 | + |
| RB | T-DNA border misc_feature | 8,800 | 8,823 | 24 | + |
| 'ColE1'' | Origin / backbone misc_feature | 9,371 | 10,029 | 659 | + |
| ''pVS1 ORI''' | Origin / backbone misc_feature | 10,058 | 12,021 | 1,964 | + |
| ParB | Coding sequence CDS | 10,384 | 11,559 | 1,176 | + |
| strep/spec | Selection marker CDS | 12,210 | 13,220 | 1,011 | + |
| Left Border Region | T-DNA border misc_feature | 13,288 | 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.