3.1. Choose your protein.
Gene: GFP
MSKGEELFTGVVPILVELDGDVNGQKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFYKD DGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKMEYNYNSHNVYIMADKPKNG IKVNFKIRHNIKDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHM ILLEFVTAAGITHGMDELYK
3.2. Reverse Translate: Protein (amino acid) sequence to DNA (nucleotide) sequence.
DNA:
atgagcaaaggcgaagaactgtttaccggcgtggtgccgattctggtggaactggatggc gatgtgaacggccagaaatttagcgtgagcggcgaaggcgaaggcgatgcgacctatggc aaactgaccctgaaatttatttgcaccaccggcaaactgccggtgccgtggccgaccctg gtgaccacctttagctatggcgtgcagtgctttagccgctatccggatcatatgaaacag catgatttttttaaaagcgcgatgccggaaggctatgtgcaggaacgcaccattttttat aaagatgatggcaactataaaacccgcgcggaagtgaaatttgaaggcgataccctggtg aaccgcattgaactgaaaggcattgattttaaagaagatggcaacattctgggccataaa atggaatataactataacagccataacgtgtatattatggcggataaaccgaaaaacggc attaaagtgaactttaaaattcgccataacattaaagatggcagcgtgcagctggcggat cattatcagcagaacaccccgattggcgatggcccggtgctgctgccggataaccattat ctgagcacccagagcgcgctgagcaaagatccgaacgaaaaacgcgatcatatgattctg ctggaatttgtgaccgcggcgggcattacccatggcatggatgaactgtataaa
3.3. Codon optimization.
Optimized for E. Coli
atgagcaaaggcgaagaactgtttaccggcgtggtgccgattctggtggaactggatggc
gatgtgaacggccagaaatttagcgtgagcggcgaaggcgaaggcgatgcgacctatggc
aaactgaccctgaaatttatttgcaccaccggcaaactgccggtgccgtggccgaccctg
gtgaccacctttagctatggcgtgcagtgctttagccgctatccggatcatatgaaacag
catgatttttttaaaagcgcgatgccggaaggctatgtgcaggaacgcaccattttttat
aaagatgatggcaactataaaacccgcgcggaagtgaaatttgaaggcgataccctggtg
aaccgcattgaactgaaaggcattgattttaaagaagatggcaacattctgggccataaa
atggaatataactataacagccataacgtgtatattatggcggataaaccgaaaaacggc
attaaagtgaactttaaaattcgccataacattaaagatggcagcgtgcagctggcggat
cattatcagcagaacaccccgattggcgatggcccggtgctgctgccggataaccattat
ctgagcacccagagcgcgctgagcaaagatccgaacgaaaaacgcgatcatatgattctg