Human MBNL3 ORF clone (NM_001386896) (Cat. No.: V026075)

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Basic Information

Name:: pT3-EF1a-MBNL3(human)-Myc

Accession ID:: NM_001386896.1

Antibiotic Resistance:: Ampicillin

Length:: 6664 bp

Type:: Protein expression

Replication origin:: ori

Host:: Mammalian cells

Promoter:: EF1a

Fusion Tag:: Myc

Growth Strain(s):: Stbl3

Growth Temperature:: 37℃

Expression Method:: Transient

Gene Synonyms:: CHCR; MBLX; MBLX39; MBXL

Transcript Definition:: Homo sapiens muscleblind like splicing regulator 3 (MBNL3), transcript variant 11, mRNA

Cat. No.: V026075 pT3-EF1a-MBNL3(human)-Myc

$ 299.0

In stock, 1-2 weeks for quality controls

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

Resources

Transcript ID

NM_001386889.1 NM_001386896.1

Fusion Tag

3xFLAG Myc

Selection Marker

Puro

Expression Method

Constiutive, Stable / Transient Transient

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 MBNL3 ORF clone (NM_001386896) (Cat. No.: V026075) Sequence

LOCUS       V026075                 6664 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V026075
VERSION     V026075
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        90..1268
                     /note="strong constitutive promoter for human elongation
                     factor EF-1α"
                     /label="EF-1α promoter"
     promoter        1285..1303
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     protein_bind    1382..1406
                     /bound_moiety="BP Clonase™"
                     /gene="mutant version of attB"
                     /note="recombination site for the Gateway® BP reaction"
                     /label="attB1"
     CDS             1417..1446
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             1453..2514
                     /label="MBNL3(NM_001386896)"
                     /note="MBNL3(NM_001386896)"
                     /gene="MBNL3"
     protein_bind    complement(2519..2543)
                     /bound_moiety="BP Clonase™"
                     /gene="mutant version of attB"
                     /note="recombination site for the Gateway® BP reaction"
                     /label="attB2"
     CDS             2596..2637
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /transl_table=1
                     /translation="GKPIPNPLLGLDST"
                     /label="V5 tag"
     polyA_signal    2681..2905
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     protein_bind    2975..3008
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTATGC) (Shaw et al., 2021)."
                     /label="loxP"
     primer_bind     complement(3466..3482)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     promoter        3956..4060
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             4061..4921
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     rep_origin      5092..5680
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     protein_bind    5968..5989
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        6004..6034
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6042..6058
                     /bound_moiety="lac repressor encoded by lacI"
                     /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-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     primer_bind     6066..6082
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6580..6613
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTATGC) (Shaw et al., 2021)."
                     /label="loxP"