Basic Vector Information
HBV 1.3-mer WT replicon 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 ℃.
HBV 1.3-mer WT replicon vector Sequence
LOCUS 40924_1284 6821 bp DNA circular SYN 13-MAY-2021 DEFINITION containing 1.3 units of the HBV genome (subtype ayw). ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6821) AUTHORS Wang H, Kim S, Ryu WS TITLE DDX3 DEAD-Box RNA helicase inhibits hepatitis B virus reverse transcription by incorporation into nucleocapsids. JOURNAL J Virol. 2009 Jun;83(11):5815-24. doi: 10.1128/JVI.00011-09. Epub 2009 Mar 18. PUBMED 19297497 REFERENCE 2 (bases 1 to 6821) TITLE Direct Submission REFERENCE 3 (bases 1 to 6821) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10.1128/JVI.00011-09"; journalName: "J Virol"; date: "2009-06"; volume: "83"; issue: "11"; pages: "5815-24" COMMENT SGRef: number: 2; type: "Journal Article" FEATURES Location/Qualifiers source 1..6821 /mol_type="other DNA" /organism="synthetic DNA construct" CDS 843..1685 /codon_start=1 /label=HBsAg (pre-S2/S) /note="middle (pre-S2/S) envelope protein of hepatitis B virus" /translation="MQWNSTTFHQTLQDPRVRGLYFPAGGSSSGTVNPVLTTASPLSSI FSRIGDPALNMENITSGFLGPLLVLQAGFFLLTRILTIPQSLDSWWTSLNFLGGTTVCL GQNSQSPTSNHSPTSCPPTCPGYRWMCLRRFIIFLFILLLCLIFLLVLLDYQGMLPVCP LIPGSSTTSTGPCRTCMTTAQGTSMYPSCCCTKPSDGNCTCIPIPSSWAFGKFLWEWAS ARFSWLSLLVPFVQWFVGLSPTVWLSVIWMMWYWGPSLYSILSPFLPLLPIFFCLWVYI " promoter complement(2866..2884) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2903..2919) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind complement(2927..2943) /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(2951..2981) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(2996..3017) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3134..3151) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3305..3893) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(4067..4924) /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(4925..5029) /label=AmpR promoter primer_bind 5097..5115 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(5153..5175) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 5275..5294 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 5503..5519 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 5527..5545 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase"
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