Basic Vector Information
- Vector Name:
- pEF1hCASP-9-His
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7393 bp
- Type:
- Mammalian expression vector
- Replication origin:
- ori
- Source/Author:
- De Schamphelaire W, Olbrechts A, Meert J, Verhelst K, Roggeman Fonseca M, Vanhoucke M, Beyaert R.
- Promoter:
- EF-1α
pEF1hCASP-9-His 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 ℃.
pEF1hCASP-9-His vector Sequence
LOCUS V007626 7393 bp DNA circular SYN 17-DEC-2018 DEFINITION Exported. ACCESSION V007626 VERSION V007626 KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct . REFERENCE 1 (bases 1 to 7393) AUTHORS De Schamphelaire W, Olbrechts A, Meert J, Verhelst K, Roggeman Fonseca M, Vanhoucke M, Beyaert R. TITLE BCCM/LMBP Plasmid collection JOURNAL Unpublished REFERENCE 2 (bases 1 to 7393) AUTHORS De Schamphelaire W. TITLE Direct Submission JOURNAL Submitted (02-FEB-2017) BCCM/LMBP, Universiteit Gent, Technologiepark 927, 9052, BELGIUM REFERENCE 3 (bases 1 to 7393) TITLE Direct Submission REFERENCE 4 (bases 1 to 7393) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Unpublished" SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (02-FEB-2017) BCCM/LMBP, Universiteit Gent, Technologiepark 927, 9052, BELGIUM" SGRef: number: 3; type: "Journal Article" FEATURES Location/Qualifiers source 1..7393 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 473..1651 /label="EF-1-alpha promoter" /note="strong constitutive promoter for human elongation factor EF-1-alpha" promoter 1668..1686 /label="T7 promoter" /note="promoter for bacteriophage T7 RNA polymerase" CDS 1758..3005 /gene="CASP9" /label="Caspase-9" /note="Caspase-9 from Homo sapiens. Accession#: P55211" CDS 3024..3065 /label="V5 tag" /note="epitope tag from simian virus 5" CDS 3075..3092 /label="6xHis" /note="6xHis affinity tag" polyA_signal 3121..3345 /label="bGH poly(A) signal" /note="bovine growth hormone polyadenylation signal" rep_origin 3391..3819 /label="f1 ori" /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 3833..4162 /label="SV40 promoter" /note="SV40 enhancer and early promoter" CDS 4229..5020 /label="NeoR/KanR" /note="aminoglycoside phosphotransferase" polyA_signal 5199..5332 /label="SV40 poly(A) signal" /note="SV40 polyadenylation signal" primer_bind complement(5369..5385) /label="M13 rev" /note="common sequencing primer, one of multiple similar variants" protein_bind complement(5393..5409) /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(5417..5447) /label="lac promoter" /note="promoter for the E. coli lac operon" protein_bind complement(5462..5483) /label="CAP binding site" /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(5760..6348) /direction=LEFT /label="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(6522..7379) /label="AmpR" /note="beta-lactamase" promoter complement(join(7380..7393,1..91)) /label="AmpR promoter"
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