Basic Vector Information
SARS-CoV-2 S HexaPro 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 ℃.
SARS-CoV-2 S HexaPro vector Sequence
LOCUS 40924_48748 8376 bp DNA circular SYN 13-MAY-2021
DEFINITION Mammalian expression vector for expression of the SARS-CoV-2 spike
HexaPro variant.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8376)
AUTHORS Hsieh CL, Goldsmith JA, Schaub JM, DiVenere AM, Kuo HC, Javanmardi
K, Le KC, Wrapp D, Lee AG, Liu Y, Chou CW, Byrne PO, Hjorth CK,
Johnson NV, Ludes-Meyers J, Nguyen AW, Park J, Wang N, Amengor D,
Lavinder JJ, Ippolito GC, Maynard JA, Finkelstein IJ, McLellan JS
TITLE Structure-based design of prefusion-stabilized SARS-CoV-2 spikes.
JOURNAL Science. 2020 Sep 18;369(6510):1501-1505. doi:
10.1126/science.abd0826. Epub 2020 Jul 23.
PUBMED 32703906
REFERENCE 2 (bases 1 to 8376)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 8376)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1126/science.abd0826"; journalName: "Science"; date:
"2020-09-18- 18"; volume: "369"; issue: "6510"; pages: "1501-1505"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..8376
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 5..384
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
intron 662..1678
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
CDS 5486..5509
/codon_start=1
/label=HRV 3C site
/note="recognition and cleavage site for human rhinovirus
3C and PreScission proteases"
/translation="LEVLFQGP"
CDS 5513..5536
/codon_start=1
/label=8xHis
/note="8xHis affinity tag"
/translation="HHHHHHHH"
CDS 5543..5566
/codon_start=1
/label=Strep-Tag II
/note="peptide that binds Strep-Tactin(R), an engineered
form
of streptavidin"
/translation="WSHPQFEK"
CDS 5606..5629
/codon_start=1
/label=Strep-Tag II
/note="peptide that binds Strep-Tactin(R), an engineered
form
of streptavidin"
/translation="WSHPQFEK"
primer_bind complement(5731..5750)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_signal 5796..5851
/label=beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(5850..5869)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
primer_bind complement(6212..6228)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(6236..6252)
/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(6260..6290)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(6305..6326)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(6455..6472)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(6626..7214)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(7388..8245)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(8246..8350)
/label=AmpR promoter
promoter 8376
/label=CAG
/note="CMV early enhancer fused to modified chicken
beta-actin promoter"
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