HBV 1.3-mer WT replicon vector (V009181)

Price Information

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V009181 HBV 1.3-mer WT replicon In stock, instant shipping

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Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.

Basic Vector Information

Vector Name:
HBV 1.3-mer WT replicon
Antibiotic Resistance:
Ampicillin
Length:
6821 bp
Type:
Mammalian Expression
Replication origin:
ori
Growth Strain(s):
DH10B
Growth Temperature:
37℃

HBV 1.3-mer WT replicon vector Map

HBV 1.3-mer WT replicon6821 bp3006009001200150018002100240027003000330036003900420045004800510054005700600063006600HBsAg (pre-S2/S)T7 promoterM13 revlac operatorlac promoterCAP binding siteL4440oriAmpRAmpR promoterpBRforEcopGEX 3'pRS-markerM13 fwdSP6 promoter

Plasmid 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 ℃.

6. The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it

General Plasmid Transform Protocol

1. Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.

2. Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.

3. Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.

4. Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.

5. Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.

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"