Basic Vector Information
- Vector Name:
- pEF1mtBIDHis
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6561 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
pEF1mtBIDHis 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 ℃.
pEF1mtBIDHis vector Sequence
LOCUS 40924_16989 6561 bp DNA circular SYN 17-DEC-2018 DEFINITION Mammalian expression vector pEF1mtBIDHis, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6561) 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A, Meert J, Verhelst K, Roggeman Fonseca M, Vanhoucke M, Beyaert R. TITLE BCCM/LMBP Plasmid collection JOURNAL Unpublished REFERENCE 2 (bases 1 to 6561) AUTHORS De Schamphelaire W. TITLE Direct Submission JOURNAL Submitted (02-FEB-2017) BCCM/LMBP, Universiteit Gent, Technologiepark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 6561) TITLE Direct Submission REFERENCE 4 (bases 1 to 6561) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Unpublished" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (02-FEB-2017) BCCM/LMBP, Universiteit Gent, Technologiepark 927, 9052, BELGIUM" COMMENT SGRef: number: 3; type: "Journal Article" FEATURES Location/Qualifiers source 1..6561 /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 2192..2233 /codon_start=1 /label=V5 tag /note="epitope tag from simian virus 5" /translation="GKPIPNPLLGLDST" CDS 2243..2260 /codon_start=1 /label=6xHis /note="6xHis affinity tag" /translation="HHHHHH" polyA_signal 2289..2513 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2559..2987 /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 3001..3330 /label=SV40 promoter /note="SV40 enhancer and early promoter" CDS 3397..4188 /codon_start=1 /label=NeoR/KanR /note="aminoglycoside phosphotransferase" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" polyA_signal 4367..4500 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4537..4553) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind complement(4561..4577) /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(4585..4615) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(4630..4651) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(4928..5516) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(5690..6547) /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(join(6548..6561,1..91)) /label=AmpR promoter
This page is informational only.