Basic Vector Information
- Vector Name:
- HygR-5' EGFP reporter
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6083 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Konishi Y, Karnan S, Takahashi M, Ota A, Damdindorj L, Hosokawa Y, Konishi H.
- Promoter:
- CMV
HygR-5' EGFP reporter 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 ℃.
HygR-5' EGFP reporter vector Sequence
LOCUS 40924_1389 6083 bp DNA circular SYN 17-DEC-2018
DEFINITION Cloning vector HygR-5' EGFP reporter, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6083)
AUTHORS Konishi Y, Karnan S, Takahashi M, Ota A, Damdindorj L, Hosokawa Y,
Konishi H.
TITLE A system for the measurement of gene targeting efficiency in human
cell lines using an antibiotic resistance-GFP fusion gene
JOURNAL BioTechniques 53 (3), 141-152 (2012)
PUBMED 22963476
REFERENCE 2 (bases 1 to 6083)
AUTHORS Konishi H.
TITLE Direct Submission
JOURNAL Submitted (07-JAN-2012) Aichi Medical University School of Medicine,
1-1 Yazako-Karimta, Nagakute, Aichi 480-1195, Japan
REFERENCE 3 (bases 1 to 6083)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 6083)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"BioTechniques"; date: "2012"; volume: "53"; issue: "3"; pages:
"141-152"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(07-JAN-2012) Aichi Medical University School of Medicine, 1-1
Yazako-Karimta, Nagakute, Aichi 480-1195, Japan"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6083
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 235..614
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 615..818
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 863..881
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory 936..945
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 957..1973
/label=HygR
/note="aminoglycoside phosphotransferase from E. coli"
CDS 2025..2531
/label=VN173
/note="N-terminal fragment of mVenus for use in bimolecular
fluorescence complementation (BiFC) (Kodama and Hu, 2010)"
polyA_signal 2673..2794
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
misc_feature 2865..3883
/note="partial SV40 largeT; an internal portion of SV40
largeT exon 2"
primer_bind complement(3926..3942)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(3950..3966)
/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(3974..4004)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(4019..4040)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(4328..4916)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5090..5947)
/label=AmpR
/note="beta-lactamase"
promoter complement(5948..6052)
/label=AmpR promoter
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