Transgenic selection methods and compositions

A eukaryotic cell, comprising

• (a) a first vector comprising (i) a nucleotide sequence encoding an N-terminal fragment of a selectable marker protein, which is upstream from a nucleotide sequence encoding an N-terminal fragment of an intein and (ii) a nucleotide sequence encoding a first molecule of interest;
• and
• (b) a second vector comprising (i) a nucleotide sequence encoding a C-terminal fragment of the intein, which is upstream from a C-terminal fragment of the selectable marker protein and (ii) a nucleotide sequence encoding a second molecule of interest,
• wherein the N-terminal fragment and the C-terminal fragment of the intein catalyze joining of the N-terminal fragment and the C-terminal fragment of the selectable marker protein to produce a full-length antibiotic resistance protein.

A kit, comprising

• (a) a first vector comprising a nucleotide sequence encoding an N-terminal fragment of a selectable marker protein, which is upstream from a nucleotide sequence encoding an N-terminal fragment of an intein;
• and
• (b) a second vector comprising a nucleotide sequence encoding a C-terminal fragment of the intein, which is upstream from a C-terminal fragment of the selectable marker protein,
• wherein the N-terminal fragment and the C-terminal fragment of the intein catalyze joining of the N-terminal fragment and the C-terminal fragment of the selectable marker protein to produce a full-length selectable marker protein.

A method comprising delivering to eukaryotic cells

• (a) a first vector comprising (i) a nucleotide sequence encoding an N-terminal fragment of a selectable marker protein, which is upstream from a nucleotide sequence encoding an N-terminal fragment of a first intein and (ii) a nucleotide sequence encoding a first molecule of interest,
• (b) a second vector comprising (i) a nucleotide sequence encoding a C-terminal fragment of the first intein, which is upstream from a nucleotide sequence encoding a central fragment of the selectable marker protein, which is upstream from a nucleotide sequence encoding an N-terminal fragment of a second intein and (ii) a nucleotide sequence encoding a second molecule of interest, and
• (c) a third vector comprising (i) a nucleotide sequence encoding a C-terminal fragment of the second intein, which is upstream from a nucleotide sequence encoding a C-terminal fragment of the selectable marker protein and (ii) a nucleotide sequence encoding a third molecule of interest,
• wherein the N-terminal fragment and the C-terminal fragment of the first intein catalyze joining of N-terminal fragment of the selectable marker protein to the central fragment of the selectable marker protein, and the N-terminal fragment and the C-terminal fragment of the second intein catalyze joining of central fragment of the selectable marker protein to the C-terminal fragment of the selectable marker protein, to produce a full-length selectable marker protein.

A eukaryotic cell comprising:

• (a) a first vector comprising (i) a nucleotide sequence encoding an N-terminal fragment of a selectable marker protein, which is upstream from a nucleotide sequence encoding an N-terminal fragment of a first intein and (ii) a nucleotide sequence encoding a first molecule of interest,
• (b) a second vector comprising (i) a nucleotide sequence encoding a C-terminal fragment of the first intein, which is upstream from a nucleotide sequence encoding a central fragment of the selectable marker protein, which is upstream from a nucleotide sequence encoding an N-terminal fragment of a second intein and (ii) a nucleotide sequence encoding a second molecule of interest, and
• (c) a third vector comprising (i) a nucleotide sequence encoding a C-terminal fragment of the second intein, which is upstream from a nucleotide sequence encoding a C-terminal fragment of the selectable marker protein and (ii) a nucleotide sequence encoding a third molecule of interest,
• wherein the N-terminal fragment and the C-terminal fragment of the first intein catalyze joining of N-terminal fragment of the selectable marker protein to the central fragment of the selectable marker protein, and the N-terminal fragment and the C-terminal fragment of the second intein catalyze joining of central fragment of the selectable marker protein to the C-terminal fragment of the selectable marker protein, to produce a full-length selectable marker protein.

A kit comprising:

• (a) a first vector comprising a nucleotide sequence encoding an N-terminal fragment of a selectable marker protein, which is upstream from a nucleotide sequence encoding an N-terminal fragment of a first intein,
• (b) a second vector comprising a nucleotide sequence encoding a C-terminal fragment of the first intein, which is upstream from a nucleotide sequence encoding a central fragment of the selectable marker protein, which is upstream from a nucleotide sequence encoding an N-terminal fragment of a second intein, and
• (c) a third vector comprising a nucleotide sequence encoding a C-terminal fragment of the second intein, which is upstream from a nucleotide sequence encoding a C-terminal fragment of the selectable marker protein,
• wherein the N-terminal fragment and the C-terminal fragment of the first intein catalyze joining of N-terminal fragment of the selectable marker protein to the central fragment of the selectable marker protein, and the N-terminal fragment and the C-terminal fragment of the second intein catalyze joining of central fragment of the selectable marker protein to the C-terminal fragment of the selectable marker protein, to produce a full-length selectable marker protein.