Basic Vector Information
- Vector Name:
- HITI-GmScreaM8-recipient
- Antibiotic Resistance:
- Ampicillin
- Length:
- 4238 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Gunadi A, Orchard N, Qu F, Finer J.
HITI-GmScreaM8-recipient 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 ℃.
HITI-GmScreaM8-recipient vector Sequence
LOCUS 62056_1186 4238 bp DNA circular SYN 01-FEB-2020
DEFINITION Cloning vector HITI-GmScreaM8-recipient, complete sequence.
ACCESSION MN043944
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4238)
AUTHORS Gunadi A, Orchard N, Qu F, Finer J.
TITLE Enhanced CRISPR/Cas9 mediated homology-independent targeted
integration in plant cotyledonary tissues
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 4238)
AUTHORS Gunadi A, Orchard N, Qu F, Finer J.
TITLE Direct Submission
JOURNAL Submitted (06-JUN-2019) Horticulture and Crop Science, The Ohio
State University, 216 Williams Hall, 1680 Madison Ave., Wooster, OH
44691, USA
REFERENCE 3 (bases 1 to 4238)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(06-JUN-2019) Horticulture and Crop Science, The Ohio State
University, 216 Williams Hall, 1680 Madison Ave., Wooster, OH 44691,
USA"
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..4238
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 379..395
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
regulatory 405..1978
/label=GmScreamM8 promoter from soybean
/note="GmScreamM8 promoter from soybean"
/regulatory_class="promoter"
misc_feature 1979..2001
/note="directional target for CRISPR/Cas9-mediated
double-strand break"
primer_bind complement(2017..2033)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(2041..2057)
/label=lac operator
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(2065..2095)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(2110..2131)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(2419..3007)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(3181..4038)
/label=AmpR
/note="beta-lactamase"
promoter complement(4039..4143)
/label=AmpR promoter
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