Table 1. Twenty overlapping oligonucleotides utilized to construct the synthetic huEPO gene. The overlapping complementary sequences are underlined.
(a) Oligonucleotide sets for synthetic huEPO gene A - 531 bases
(b) Oligonucleotide sets for synthetic huEPO gene B - 609 bases.


a) huEPO gene

Oligo

Sequence

F-1A

R-1A

5’-AAAAAAGAATTCATGGCCCCACCACGCCTCATCTGTGACAGCCGAGTC-3’

3’-CGGGGTGGTGCGGAGTAGACACTGTCGGCTCAGGACCTCTCCATG-5’

 

F-2A

R-2A

5’-AGGTACCTCTTGGAGGCCAAGGAGGCCGAGAATATCACGACGGGC-3’

3’-AACCTCCGGTTCCTCCGGCTCTTATAGTGCTGCCCGACACGACTT-5’

 

F-3A

R-3A

5’-GAACACTGCAGCTTGAATGAGAATATCACTGTCCCAGACACCAAA-3’

3’-TCGAACTTACTCTTATAGTGACAGGGTCTGTGGTTTCAATTAAAGATA-5’

 

F-4A

R-4A

5’-TTCTATGCCTGGAAGAGGATGGAGGTCGGGCAGCAGGCCGTAGAAGTC-3’

3’-TTCTCCTACCTCCAGCCCGTCGTCCGGCATCTTCAGACCGTCCCGGAC-5’

 

F-5A

R-5A

5’-GGCCTGGCCCTGCTGTCGGAAGCTGTCCTGCGGGGCCAGGCCCTGTTG-3’

3’-GACGACAGCCTTCGACAGGACGCCCCGGTCCGGGACAACCAGTTGAGAAGG-5’

 

F-6A

R-6A

5’-TCCCAGCCGTGGGAGCCCCTGCAGCTGCATGTGGATAAAGCCGTCAGTGGC-3’

3’-ACCCTCGGGGACGTCGACGTACACCTATTTCGGCAGTCACCGGAAGCGTCG-5’

 

F-7A

R-7A

5’-AGCCTCACCACTCTGCTTCGGGCTCTGGGAGCCCAGAAGGAAGCC-3’

3’-TGGTGAGACGAAGCCCGAGACCCTCGGGTCTTCCTTCGGTAGAGGGGA-5’

 

F-8A

R-8A

5’-CCTCCAGATGCGGCCTCAGCTGCTCCACTCCGAACAATCACTGCT-3’

3’-CGCCGGAGTCGACGAGGTGAGGCTTGTTAGTGACGACTGTGAAAG-5’

 

F-9A

R-9A

5’-TTCCGCAAACTCTTCCGAGTCTACTCCAATTTCCTCCGGGGAAAG-3’

3’-GAGAAGGCTCAGATGAGGTTAAAGGAGGCCCCTTTCGACTTCGAC-5’

 

F-10A

R-10A

5’-AAGCTGTACACAGGGGAGGCCTGCAGGACAGGGGACAGATAATTT-3’

3’-CCCCTCCGGACGTCCTGTCCCCTGTCTATTAAAGGATCCAAAAAA-5’

 

b) huEPO B

Oligo

Sequence

F-1B

R-1B

5’-AAAAAAGAATTCATGGGGGTGCACGAATGTCCTGCCTGG-3’

3’-TACCCCCACGTGCTTACAGGACGGACCGACACCGAAGAGGACAGGGACGACAGC-5’

 

F-2B R-2B

5’-CTTCTCCTGTCCCTGCTGTCGCTCCCTCTGGGCCTCCCAGTCCTGGGCGCC-3’

3’-CCGGAGGGTGACGACCCGCGGGGTGGTGCGGAGTAGACACTGTCGGCTCAGGAC-5’

 

F-3B

R-3B

5’-CTCATCTGTGACAGCCGAGTCCTGGAGAGGTACCTCTTGGAGGCCAAGGAGGCC-3’

3’-GAGAAGCTCCGGTTCCTCCGGCTCTTATAGTGCTGCCCGACACGACTTGTGACGTTA-5’

 

F-4B

R-4B

5’-ACGACGGGCTGTGCTGAACACTGCAGCTTGAATGAGAATATCACTGTCCCA-3’

3’-CTCTTATAGTGACAGGGTCTGTGGTTTCAATTAAAGATAGCCACCTTCTCCCCC-5’

 

F-5B

R-5B

5’-GTTAATTTCTATGCCTGGAAGAGGATGGAGGTCGGGCAGCAGGCCGTAGAAGTC-3’

3’-GTCGTCCGGCATCTTCAGACCGACCCGGAGCGGGACGACAGCCTTCGA-5’

 

F-6B

R-6B

5’-GGCCTGGCCCTGCTGTCGGAAGCTGTCCTGCGGGGCCAGGCCCTGTTG -3’

3’-GCCCCGGTCCGGGACAACCAGTGGAGAAGGGTCGGCACCCTCGGGGACGTCGAC-5’

 

F-7B

R-7B

5’-CAGCCGTGGGAGCCCCTGCAGCTGCATGTGGATAAAGCCGTCAGTGGCCTT-3’

3’-CTATTTCGGCAGTCACCGGAAGCGTCGGAGTGGTGAGACGAAGCCCGAGAC-5’

 

F-8B

R-8B

5’-CTCACCACTCTGCTTCGGGCTCTGGGAGCCCAGAAGGAAGCCATCTCCCCTCCAGAT-3’

3’-TTCCTTCGGTAGAGGGGAGGTCTACGCCGGAGTCGACGAGGTGAGGCTTGTTAG-5’

 

F-9B

R-9B

5’-GCTGCTCCACTCCGAACAATCACTGCTGACACTTTCCGCAAACTCTTCCGAGTC-3’

3’-AAGGCGTTTGAGAAGGCTCAGATGAGGTTAAAGGAGGCCCCTTTCGACTTCGAC-5’

 

F-10B

R-10B

5’-CGGGGAAAGCTGAAGCTGTACACAGGGGAGGCCTGCAGGACAGGGGACAGATAA-3’

3’-CCCCTCCGGACGTCCTGTCCCCTGTCTATTAAAGGATCCAAA AAA-5’

 

 


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