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Human LYPLA1 ORF clone (NM_006330) (Cat. No.: V021545)

pCMV-Neo-LYPLA1(human)-EGFP5430 bp60012001800240030003600420048005400CMV enhancerCMV promoterEGFPLYPLA1(NM_006330)SV40 poly(A) signalf1 oriAmpR promoterSV40 promoterNeoR/KanRHSV TK poly(A) signalori
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Basic Information

Name:
pCMV-Neo-LYPLA1(human)-EGFP
Accession ID:
NM_006330.4
Antibiotic Resistance:
Kanamycin
Length:
5430 bp
Type:
Protein expression
Replication origin:
ori
Host:
Mammalian cells
Selection Marker:
Neo/G418;EGFP
Promoter:
CMV
Fusion Tag:
EGFP
Expression Method:
Transient
Gene Synonyms:
APT-1; APT1; hAPT1; LPL-I; LPL1
Transcript Definition:
Homo sapiens lysophospholipase 1 (LYPLA1), transcript variant 1, mRNA
Cat. No.: V021545 pCMV-Neo-LYPLA1(human)-EGFP
$ 299.0
In stock, 1-2 weeks for quality controls
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Two tubes of transfection grade plasmid, each tube is about 100 µg (lyophilized).
Transcript ID
NM_001279356.2 NM_006330.4
Fusion Tag
3xFLAG EGFP HA
Selection Marker
Neo/G418 Neo/G418;EGFP Puro;CopGFP
Expression Method
Transient Constiutive, Stable / Transient

Resources

Plasmid Protocol

Plasmid Reconstitution 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.

Human LYPLA1 ORF clone (NM_006330) (Cat. No.: V021545) Annotation

LOCUS       V021545                 5430 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021545
VERSION     V021545
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        61..364
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        365..568
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             613..1329
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK"
                     /label="EGFP"
     CDS             1396..2085
                     /label="LYPLA1(NM_006330)"
                     /note="LYPLA1(NM_006330)"
                     /gene="LYPLA1"
     polyA_signal    2218..2339
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      complement(2346..2801)
                     /direction=LEFT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2828..2932
                     /gene="bla"
                     /label="AmpR promoter"
     promoter        2934..3291
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3326..4120
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    4352..4399
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4728..5316
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"