pBABE-hLin28B vector (V001654)

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Price Information

Cat No.	Plasmid Name	Availability	Add to cart
V001654	pBABE-hLin28B	In stock, 1 week for quality controls	Buy one, get one free!

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

Basic Vector Information

Vector Name:: pBABE-hLin28B

Antibiotic Resistance:: Ampicillin

Length:: 5875 bp

Type:: Mammalian Expression, Retroviral

Replication origin:: ori

Selection Marker:: Puromycin

Copy Number:: High Copy

Promoter:: SV40

Cloning Method:: Restriction Enzyme

5' Primer:: pBABE 5' primer

pBABE-hLin28B vector Map

Copy Sequence View Map

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-hLin28B vector Sequence

LOCUS       V001654                 5875 bp    DNA     circular SYN 13-MAY-2021
DEFINITION  Exported.
ACCESSION   V001654
VERSION     V001654
KEYWORDS    pBABE-hLin28B
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 5875)
  AUTHORS   Viswanathan SR, Powers JT, Einhorn W, Hoshida Y, Ng TL, Toffanin S,
            O'Sullivan M, Lu J, Phillips LA, Lockhart VL, Shah SP, Tanwar PS,
            Mermel CH, Beroukhim R, Azam M, Teixeira J, Meyerson M, Hughes TP,
            Llovet JM, Radich J, Mullighan CG, Golub TR, Sorensen PH, Daley GQ
  TITLE     Lin28 promotes transformation and is associated with advanced human
            malignancies.
  JOURNAL   Nat Genet. 2009 Jul . 41(7):843-8.
   PUBMED   19483683
REFERENCE   2  (bases 1 to 5875)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 5875)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Genet.
            2009 Jul . 41(7):843-8."
            SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5875
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             complement(120..711)
                     /label="LTR"
                     /note="long terminal repeat from Moloney murine leukemia
                     virus"
     CDS             complement(813..1409)
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     promoter        complement(1419..1748)
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             complement(1768..2517)
                     /gene="LIN28B"
                     /label="Protein lin-28 homolog B"
                     /note="Protein lin-28 homolog B from Homo sapiens.
                     Accession#: Q6ZN17"
     CDS             complement(2560..2583)
                     /label="FLAG"
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             complement(2600..3016)
                     /label="gag (truncated)"
                     /note="truncated Moloney murine leukemia virus (MMLV) gag
                     gene lacking the start codon"
     misc_feature    complement(3217..3416)
                     /label="MMLV Psi"
                     /note="packaging signal of Moloney murine leukemia virus
                     (MMLV)"
     LTR             complement(3481..3950)
                     /label="LTR"
                     /note="long terminal repeat from Moloney murine leukemia
                     virus"
     promoter        3981..4085
                     /label="AmpR promoter"
     CDS             4086..4943
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      5106..5694
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     primer_bind     5848..5865
                     /label="L4440"
                     /note="L4440 vector, forward primer"