Long poly(a) plasmids and methods for introduction of long poly(a) sequences into the plasmid

A polynucleotide comprising:

• a first nucleic acid sequence that comprises at least one endonuclease recognition site;
• and
• a second nucleic acid sequence that comprises a plurality of thymine nucleotides, wherein said plurality of thymine nucleotides comprises at least, greater than, equal to, or any number in between 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500 or 550 covalently linked thymine nucleotides, wherein the second nucleic acid comprises an endonuclease cleavage site that is within the plurality of thymine nucleotides or within 1, 2, 3, 4, 5, 6, 7, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 nucleotides from the 5′ terminal thymine in the plurality of thymine nucleotides;
• and
• wherein said first nucleic acid is covalently linked to said second nucleic acid at one end of said second nucleic acid and, wherein said endonuclease recognition site is within 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54 or 55 nucleotides of the endonuclease cleavage site and said endonuclease cleavage site is within 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or 40 nucleotides of the 5′ terminal thymine in the plurality of thymine nucleotides.