Human ZDHHC6 ORF clone (NM_022494) (Cat. No.: V044508)

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Basic Information

Note: It expresses human ZDHHC6, a palmitoyltransferase for protein modification, with a 3xFLAG tag and CopGFP reporter driven by CMV promoter, and PuroR via T2A. It includes WPRE and viral elements (LTRs, HIV-1 Ψ, RRE, cPPT/CTS) for packaging, plus AmpR and ori for bacteria.

Name:: pLV3-CMV-ZDHHC6(human)-3xFLAG-CopGFP-Puro

Accession ID:: NM_022494.3

Antibiotic Resistance:: Ampicillin

Length:: 9531 bp

Type:: Protein expression

Replication origin:: ori

Host:: Mammalian cells, Lentivirus

Selection Marker:: Puro;CopGFP

Copy Number:: High

Promoter:: CMV

5' Primer:: CMV-F

Fusion Tag:: 3xFLAG

Growth Strain(s):: Stbl3

Growth Temperature:: 37℃

Expression Method:: Constiutive, Stable / Transient

Gene Synonyms:: DHHC-6; DHHC6; ZNF376

Transcript Definition:: Homo sapiens zinc finger DHHC-type palmitoyltransferase 6 (ZDHHC6), transcript variant 1, mRNA

Cat. No.: V044508 pLV3-CMV-ZDHHC6(human)-3xFLAG-CopGFP-Puro

$ 299.4

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

Resources

Transcript ID

NM_022494.3 NM_001351084.2

Fusion Tag

V5 tag HA 3xFLAG

Selection Marker

Neo/G418 Puro;CopGFP Puro;mCherry

Expression Method

Transient 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.