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Price Information

Cat No. Plasmid Name Availability Buy one, get one free! (?)
V018140 Human H-IgM WT In stock, 1 week for quality controls

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

Basic Vector Information

High-level human immunoglobulin expression in mammalian cells, especially HEK293 derivatives (like 293-6E), by leveraging the Epstein-Barr virus (EBV) origin of replication (OriP) and a strong CMV promoter.

Vector Name:
Human H-IgM WT
Antibiotic Resistance:
Ampicillin
Length:
5814 bp
Type:
Mammalian Expression Vectors
Copy Number:
High copy number
Promoter:
CMV
Growth Strain(s):
DH5a
Growth Temperature:
37℃
Expression Method:
Transient

Human H-IgM WT vector Map

Copy Sequence View Map
Human H-IgM WT5814 bp60012001800240030003600420048005400CMV enhancerCMV promoterAdenovirus TPL and MLPhuman interleukin 10 signal peptidehuIgMbeta-globin poly(A) signalOriPAmpR promoterAmpRori

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.

Human H-IgM WT vector Sequence

LOCUS       Exported                5814 bp DNA     circular SYN 08-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5814)
  AUTHORS   11111111
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5814
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        42..421
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        422..625
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        688..1172
                     /label=Adenovirus TPL and MLP
                     /label=Adenovirus Tripartite Leader and Major Late
                     Promoter
                     /note="Regulatory DNA sequence from adenovirus containing
                     both the tripartite leader and major late promoter 
                     elements, functioning to initiate and enhance transcription
                     of late viral genes.; id: 3685243"
     sig_peptide     1236..1289
                     /product="human interleukin 10 signal peptide"
     CDS             1302..2663
                     /codon_start=1
                     /product="immunoglobulin mu heavy chain"
                     /label=huIgM
                     /translation="GSASAPTLFPLVSCENSPSDTSSVAVGCLAQDFLPDSITFSWKYK
                     NNSDISSTRGFPSVLRGGKYAATSQVLLPSKDVMQGTDEHVVCKVQHPNGNKEKNVPLP
                     VIAELPPKVSVFVPPRDGFFGNPRKSKLICQATGFSPRQIQVSWLREGKQVGSGVTTDQ
                     VQAEAKESGPTTYKVTSTLTIKESDWLGQSMFTCRVDHRGLTFQQNASSMCVPDQDTAI
                     RVFAIPPSFASIFLTKSTKLTCLVTDLTTYDSVTISWTRQNGEAVKTHTNISESHPNAT
                     FSAVGEASICEDDWNSGERFTCTVTHTDLPSPLKQTISRPKGVALHRPDVYLLPPAREQ
                     LNLRESATITCLVTGFSPADVFVQWMQRGQPLSPEKYVTSAPMPEPQAPGRYFAHSILT
                     VSEEEWNTGETYTCVVAHEALPNRVTERTVDKSTGKPTLYNVSLVMSDTAGTCY"
     polyA_site      2690..2745
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal"
     misc_feature    3058..3785
                     /label=OriP
                     /note="Epstein-Barr virus (EBV) origin of replication"
     promoter        3914..4018
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4019..4879
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5050..5638
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"