pBABE-ɑ-Syn-nYFP vector (Cat. No.: V038495)

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Basic Information

Name:: pBABE-ɑ-Syn-nYFP

Antibiotic Resistance:: Ampicillin

Length:: 7541 bp

Type:: BiFC

Replication origin:: ori

Host:: Mammalian cells, Retroviral

Selection Marker:: Neo/G418;VN155(I152L)

Copy Number:: High

Promoter:: CMV

Growth Strain(s):: Stable

Growth Temperature:: 37℃

Expression Method:: Transient

$ 199.0

In stock, 1 week for quality controls

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

Resources

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.

pBABE-ɑ-Syn-nYFP vector (Cat. No.: V038495) Sequence

LOCUS       pBABE-ɑ-Syn-nYFP        7541 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V038495
VERSION     V038495
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             22..491
                     /note="long terminal repeat from Moloney murine leukemia
                     virus"
                     /label="LTR"
     misc_feature    556..755
                     /note="packaging signal of Moloney murine leukemia virus
                     (MMLV)"
                     /label="MMLV Ψ"
     CDS             956..1372
                     /codon_start=1
                     /note="truncated Moloney murine leukemia virus
                     (MMLV) gag gene lacking the start codon"
                     /transl_table=1
                     /label="gag (truncated)"
     enhancer        1600..1979
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        1980..2183
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        2414..2432
                     /note="promoter for bacteriophage SP6 RNA polymerase"
                     /label="SP6 promoter"
     protein_bind    2510..2534
                     /bound_moiety="BP Clonase™"
                     /gene="mutant version of attB"
                     /note="recombination site for the Gateway® BP reaction"
                     /label="attB1"
     protein_bind    complement(2991..3015)
                     /bound_moiety="BP Clonase™"
                     /gene="mutant version of attB"
                     /note="recombination site for the Gateway® BP reaction"
                     /label="attB2"
     CDS             3077..3538
                     /codon_start=1
                     /product="improved N-terminal fragment of mVenus for
                     use in bimolecular fluorescence complementation (BiFC)
                     (Kodama and Hu, 2010)"
                     /transl_table=1
                     /translation="V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITA"
                     /label="VN155(I152L)"
     CDS             4050..4853
                     /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="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     LTR             5049..5518
                     /note="long terminal repeat from Moloney murine leukemia
                     virus"
                     /label="LTR"
     rep_origin      complement(5819..6407)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6567..7427)
                     /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"
     promoter        complement(7428..7532)
                     /gene="bla"
                     /label="AmpR promoter"