Basic Vector Information
- Vector Name:
- HITI-EF1a
- Antibiotic Resistance:
- Ampicillin
- Length:
- 3732 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Gunadi A, Orchard N, Qu F, Finer J.
HITI-EF1a 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-EF1a vector Sequence
LOCUS 62056_1181 3732 bp DNA circular SYN 01-FEB-2020
DEFINITION Cloning vector HITI-EF1a, complete sequence.
ACCESSION MN043931
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3732)
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JOURNAL Unpublished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 3732)
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 3732)
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"Unpublished"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..3732
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 379..395
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
CDS 429..1145
/codon_start=1
/label=EGFP
/note="enhanced GFP"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITHGMDELYK"
terminator 1181..1433
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
primer_bind complement(1511..1527)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(1535..1551)
/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(1559..1589)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(1604..1625)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(1913..2501)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2675..3532)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(3533..3637)
/label=AmpR promoter
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