Basic Vector Information
- Vector Name:
- pEF1crmA
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7646 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
pEF1crmA 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 ℃.
pEF1crmA vector Sequence
LOCUS V007628 7646 bp DNA circular SYN 17-DEC-2018 DEFINITION Exported. ACCESSION V007628 VERSION V007628 KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct . REFERENCE 1 (bases 1 to 7646) 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A, Meert J, Verhelst K, Roggeman Fonseca M, Vanhoucke M, Beyaert R. TITLE BCCM/LMBP Plasmid collection JOURNAL Unpublished REFERENCE 2 (bases 1 to 7646) AUTHORS De Schamphelaire W. TITLE Direct Submission JOURNAL Submitted (02-FEB-2017) BCCM/LMBP, Universiteit Gent, Technologiepark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 7646) TITLE Direct Submission REFERENCE 4 (bases 1 to 7646) 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 TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Unpublished" SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (02-FEB-2017) BCCM/LMBP, Universiteit Gent, Technologiepark 927, 9052, BELGIUM" SGRef: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..7646 /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 2047..3069 /gene="OPG199" /label="Serine proteinase inhibitor 2" /note="Serine proteinase inhibitor 2 from Cowpox virus (strain Brighton Red). Accession#: P07385" CDS 3277..3318 /label="V5 tag" /note="epitope tag from simian virus 5" CDS 3328..3345 /label="6xHis" /note="6xHis affinity tag" polyA_signal 3374..3598 /label="bGH poly(A) signal" /note="bovine growth hormone polyadenylation signal" rep_origin 3644..4072 /label="f1 ori" /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 4086..4415 /label="SV40 promoter" /note="SV40 enhancer and early promoter" CDS 4482..5273 /label="NeoR/KanR" /note="aminoglycoside phosphotransferase" polyA_signal 5452..5585 /label="SV40 poly(A) signal" /note="SV40 polyadenylation signal" primer_bind complement(5622..5638) /label="M13 rev" /note="common sequencing primer, one of multiple similar variants" protein_bind complement(5646..5662) /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(5670..5700) /label="lac promoter" /note="promoter for the E. coli lac operon" protein_bind complement(5715..5736) /label="CAP binding site" /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(6013..6601) /direction=LEFT /label="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(6775..7632) /label="AmpR" /note="beta-lactamase" promoter complement(join(7633..7646,1..91)) /label="AmpR promoter"
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