Basic Vector Information
- Vector Name:
- 1-UBI_Cas9_PinII
- Antibiotic Resistance:
- Kanamycin
- Length:
- 9396 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Gao H.
- Promoter:
- Ubi
1-UBI_Cas9_PinII vector Vector Map
Plasmid Resuspension Protocol:
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5.Store the plasmid at -20 ℃.
1-UBI_Cas9_PinII vector Sequence
LOCUS 62056_66 9396 bp DNA circular SYN 19-MAR-2020
DEFINITION Cloning vector 1-UBI_Cas9_PinII, complete sequence.
ACCESSION MN294713
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9396)
AUTHORS Gao H.
TITLE Direct Submission
JOURNAL Submitted (08-AUG-2019) Molecular Engineering in AST, Corteva
Agriscience, 8305 NW 62nd Ave., Johnston, IA 50131, USA
REFERENCE 2 (bases 1 to 9396)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Submitted
(08-AUG-2019) Molecular Engineering in AST, Corteva Agriscience,
8305 NW 62nd Ave., Johnston, IA 50131, USA"
COMMENT ##Assembly-Data-START##
Assembly Method :: Vector NTI v. 10
Sequencing Technology :: Illumina
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..9396
/mol_type="other DNA"
/organism="synthetic DNA construct"
regulatory 13..322
/regulatory_class="terminator"
protein_bind complement(378..477)
/label=attL2
/note="recombination site for the Gateway(R) LR reaction"
promoter complement(495..513)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(518..534)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
CDS 647..1453
/label=KanR
/note="aminoglycoside phosphotransferase"
rep_origin 1603..2191
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
terminator complement(2521..2548)
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator complement(2640..2726)
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
primer_bind 2790..2806
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 2822..2921
/label=attL1
/note="recombination site for the Gateway(R) LR reaction"
promoter 3052..5046
/label=Ubi promoter
/note="maize polyubiquitin gene promoter"
CDS 5073..5093
/label=SV40 NLS
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
intron 5472..5660
/label=IV2 intron
/note="modified second intron of the potato ST-LS1 gene
(Vancanneyt et al., 1990)"
CDS 6891..9389
/label=Cas9(C)
/note="C-terminal portion of Streptococcus pyogenes Cas9
(Zetsche et al., 2015)"
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