Human GOLIM4 ORF clone (NM_014498) (Cat. No.: V042410)

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Basic Information

Note: This is a 10937 bp lentiviral vector, pLV2-CMV-Puro-GOLIM4(human)-EGFP. It is designed to express the human GOLIM4 gene fused with EGFP, driven by a CMV promoter. The construct includes a puromycin resistance gene (PuroR) for selection and essential lentiviral elements such as the HIV-1 Psi packaging signal, RRE, cPPT/CTS, and WPRE for efficient viral transduction and RNA stability. It also contains an IRES2 sequence and bacterial elements for propagation.

Name:: pLV2-CMV-Puro-GOLIM4(human)-EGFP

Accession ID:: NM_014498.5

Antibiotic Resistance:: Ampicillin

Length:: 10937 bp

Type:: Protein expression

Replication origin:: ori

Host:: Mammalian cells, Lentivirus

Selection Marker:: Puro;EGFP

Copy Number:: Low

Promoter:: CMV

5' Primer:: CMV-F

Fusion Tag:: EGFP

Growth Strain(s):: EPI400

Growth Temperature:: 30℃

Expression Method:: Constiutive, Stable / Transient

Gene Synonyms:: GIMPC; GOLPH4; GPP130; P138

Transcript Definition:: Homo sapiens golgi integral membrane protein 4 (GOLIM4), transcript variant 1, mRNA

Cat. No.: V042410 pLV2-CMV-Puro-GOLIM4(human)-EGFP

$ 298.2

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Two tubes of transfection grade plasmid, each tube is about 100 µg (lyophilized).

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Transcript ID

NM_014498.5

Fusion Tag

EGFP 3xFLAG

Selection Marker

Puro;EGFP Puro;CopGFP

Expression Method

Constiutive, Stable / Transient

Plasmid Protocol

1. Centrifuge at 5,000×g for 5 min.

2. Carefully open the tube and add sterile water to dissolve the DNA: add 20 μl for 5 μg plasmid, and 100 μl for 100 μg plasmid.

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.