Basic Vector Information
- Vector Name:
- pEGFP-C1-AR
- Antibiotic Resistance:
- Kanamycin
- Length:
- 7942 bp
- Type:
- Mammalian Expression
- Replication origin:
- ori
- Selection Marker:
- Neomycin (select with G418)
- Copy Number:
- High Copy
- Promoter:
- CMV
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- EGFP-C
- 3' Primer:
- SV40pA-R
pEGFP-C1-AR 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 ℃.
pEGFP-C1-AR vector Sequence
LOCUS 40924_17139 7942 bp DNA circular SYN 13-MAY-2021
DEFINITION Mammalian expression of Androgen receptor fused to EGFP.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7942)
AUTHORS Stenoien DL, Cummings CJ, Adams HP, Mancini MG, Patel K, DeMartino
GN, Marcelli M, Weigel NL, Mancini MA
TITLE Polyglutamine-expanded androgen receptors form aggregates that
sequester heat shock proteins, proteasome components and SRC-1, and
are suppressed by the HDJ-2 chaperone.
JOURNAL Hum Mol Genet. 1999 May . 8(5):731-41.
PUBMED 10196362
REFERENCE 2 (bases 1 to 7942)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 7942)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Hum Mol
Genet. 1999 May . 8(5):731-41."
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..7942
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 86..389
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 390..593
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
CDS 638..1354
/codon_start=1
/label=EGFP
/note="enhanced GFP"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
polyA_signal 4755..4876
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
rep_origin complement(4883..5338)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 5365..5469
/label=AmpR promoter
promoter 5471..5828
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
CDS 5863..6654
/codon_start=1
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(6845..6864)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 6889..6936
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 7265..7853
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
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