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While the bacterial ribosome binds to a well-conserved, internal sequence some bases before the start codon to initiate translation, the eukaryotic one is loaded on the 5′ terminus with the help of certain eukaryotic initiation factors.
To subclone each gene into the vector, a Kozak cassette was introduced at the 5' end before the start codon.
For the LEU2-MAT interval, we used the S. cerevisiae sequences from the nucleotide after the start codon of LEU2 to the nucleotide before the start codon of MAT-alpha (chromosomal co-ordinates 92419 198667).
The program searched 80 nucleotides before the start codon and 20 nucleotides after the start codon.
In fact, C. exaltata is somewhat contracted relative to Nicotiana and ends slightly before the start codon of ycf1.
The forward primer has four additional bases right before the start codon for TOPO cloning (in bold).
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The genes lob1, lob2AB, and lob2D contain SSRs either just before the start codons or within the open reading frame [ 47].
Moreover, we also found that a significant number of genes contain their main, or unique, splice site very close (sometimes immediately before and sometimes after) the start codon (AUG).
In another case the start codon was before the first amino acid resulting in the sequence MVVAITG….
NA nucleotide sequences were excluded from the analysis if the stalk region was not fully sequenced, i.e. if the first sequenced nucleotide position was after position 90 or the last sequenced position was before 270, counting adenine (A) in the start codon (ATG) as position 1.
With gfp inserted immediately before the termination codon or after the start codon of pqn-21 in WRM0622dE07, nuclear-localized GFP was observed in apparently all somatic cells (WBID Expr9751/9754).
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com