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The translocations occur within an 8.3 kb breakpoint cluster region of the gene and result in the replacement of ALL-1-coding sequence 3' to the breakpoint by the coding sequence of the translocation partner (Gu et al, 1992; Tkachuk et al, 1992).
Our molecular characterization (Figure 2 and File S1) confirmed that the coding sequence of the ADGF-A gene had been accurately replaced by the coding sequence of dGFP.
To test the importance of this motif, we constructed a reporter containing 2.5 kb of the Pfk promoter, followed by the coding sequence for GFP.
The DNA was subcloned into a pcDNA3.1 vector (Invitrogen); the sequence coding for the neomycin resistance gene was replaced by the coding sequence for either citrine or EGFP.
The +1 reading frame is determined by the coding sequence of the reference ORF for each transcript (independently of the gene or transcript).
We made reporter constructs containing different lengths of the promoter region of Ldh, followed by the coding sequence for green fluorescent protein (GFP).
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Therefore the "knock in" cassette (generated in an intermediate vector) offers a modular structure that allows the simple replacement of the promoter (by XhoI/EcoRV), the coding sequence (by EcoRV/BglII) and the terminator DNA sequences (by BglII/SacI).
The pMS plasmid encoding EGb-H22 scFv) EGb-H22 scFvconstruct EGb-Ki4(scFv) by replacing the coding sequence for the single chain variable fragment H22(scFv) by Ki4(scFv).
To generate dominant negative constructs, cytoplasmic kinase domains were removed by truncating the coding sequence after the transmembrane and extracellular domains.
A prokaryotic RBM24 expression plasmid pET33b-EK-RBM24 pET33b-EK-RBM24 pET33b-EK-RBM24ding sequence of RBM24 into pET33b.
The product was ligated into a similarly digested pET28-M13-mEos2 plasmid, which was constructed by swapping the coding sequence of rsFastlime with mEos2.
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