Basic Vector Information
- Vector Name:
- GST-HA-GFP11-N1
- Antibiotic Resistance:
- Kanamycin
- Length:
- 4759 bp
- Type:
- Mammalian Expression
- Replication origin:
- ori
- Selection Marker:
- Kan/Neo
- Copy Number:
- High Copy
- Promoter:
- CMV
- Cloning Method:
- Restriction Enzyme
GST-HA-GFP11-N1 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 ℃.
GST-HA-GFP11-N1 vector Sequence
LOCUS 40924_1239 4759 bp DNA circular SYN 13-MAY-2021
DEFINITION Expresses GST-HA-GFP11 in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4759)
AUTHORS Ruan L, Zhou C, Jin E, Kucharavy A, Zhang Y, Wen Z, Florens L, Li R
TITLE Cytosolic proteostasis through importing of misfolded proteins into
mitochondria.
JOURNAL Nature. 2017 Mar 16;543(7645):443-446. doi: 10.1038/nature21695.
Epub 2017 Mar 1.
PUBMED 28241148
REFERENCE 2 (bases 1 to 4759)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 4759)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1038/nature21695"; journalName: "Nature"; date: "2017-03-16-
16"; volume: "543"; issue: "7645"; pages: "443-446"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..4759
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 124..427
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 428..631
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
CDS 691..1344
/codon_start=1
/label=GST
/note="glutathione S-transferase from Schistosoma
japonicum"
/translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
CDS 1372..1398
/codon_start=1
/label=HA
/note="HA (human influenza hemagglutinin) epitope tag"
/translation="YPYDVPDYA"
CDS 1438..1485
/codon_start=1
/label=GFP11
/note="11th beta-strand of superfolder GFP"
/translation="RDHMVLHEYVNAAGIT"
polyA_signal 1610..1731
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
rep_origin complement(1738..2193)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 2220..2324
/label=AmpR promoter
promoter 2326..2683
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
CDS 2718..3509
/codon_start=1
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(3700..3719)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 3744..3791
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4120..4708
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
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