>NcoI Restriction site 1 (5’)
ccatgg
> backbone1
ttaaagttatcggtcgtcgttccctcggagtgcaaagaatatttgatattggtcttccccaagaccataattttctgctagccaatggggcgatcg
> mutation1
g
>backbone2
ccacaat
>Variable Area	-	111-134 (may vary depending on actual size of peptide insert)
tccggtggatatctgccgccgctg
>backbone3
tgcttaagttttggcaccgaaattttaaccgttgagtacggcccattgcccattggcaaaattgtgagtgaagaaattaattgttctg
>BsrGI Restriction site 2 (3’)
tgtaca
>backbone4
gtgttgatccagaagggagagtttacacccaggcgatcgcccaatggcatgaccggggagagcaggaagtattggaatatgaattggaagatggctcagtaatccgagctacctctgaccaccgctttttaaccaccgattatcaactgttggcgatcgaagaaatttttgctaggcaactggacttgttgactttagaaaatattaagcaaactgaagaagctcttgacaaccatcgtcttccctttccattacttgacgctggtaccattaaaacgacaaatcctggtgtatccgcttggcaggtcaacacagcttatactgcgggacaattggtcacatataacggcaagacgtataaatgtttgcagccccacacctccctggcaggatgggaaccatccaacgttcctgccttgtggcagcttcaatgaaagcttatcgatgataagctgtcaaacatgagaattacaacttatatcgtatggggctgacttcaggtgctacatttgaagagataaattgcactgaaatctagaaatattttatctgattaataagatgatcttcttgagatcgttttggtctgcgcgtaatctcttgctctgaaaacgaaaaaaccgccttgcagggcggtttttcgaaggttctctgagctaccaactctttgaaccgaggtaactggcttggaggagcgcagtcaccaaaacttgtcctttcagtttagccttaaccggcgcatgacttcaagactaactcctctaaatcaattaccagtggctgctgccagtggtgcttttgcatgtctttccgggttggactcaagacgatagttaccggataaggcgcagcggtcggactgaacggggggttcgtgcatacagtccagcttggagcgaactgcctacccggaactgagtgtcaggcgtggaatgagacaaacgcggccataacagcggaatgacaccggtaaaccgaaaggcaggaacaggagagcgcacgagggagccgccagggggaaacgcctggtatctttatagtcctgtcgggtttcgccaccactgatttgagcgtcagatttcgtgatgcttgtcaggggggcggagcctatggaaaaacggctttgccgcggccctctcacttccctgttaagtatcttcctggcatcttccaggaaatctccgccccgttcgtaagccatttccgctcgccgcagtcgaacgaccgagcgtagcgagtcagtgagcgaggaagcggaatatatcctgtatcacatattctgctgacgcaccggtgcagccttttttctcctgccacatgaagcacttcactgacaccctcatcagtgccaacatagtaagccagtatacactccgctagcgctgatgtccggcggtgcttttgccgttacgcaccaccccgtcagtagctgaacaggagggacagctgatagaaacagaagccactggagcacctcaaaaacaccatcatacactaaatcagtaagttggcagcatcacccgacgcactttgcgccgaataaatacctgtgacggaagatcacttcgcagaataaataaatcctggtgtccctgttgataccgggaagccctgggccaacttttggcgaaaatgagacgttgatcggcacgtaagaggttccaactttcaccataatgaaataagatcactaccgggcgtattttttgagttatcgagattttcaggagctaaggaagctaaaatggagaaaaaaatcactggatataccaccgttgatatatcccaatggcatcgtaaagaacattttgaggcatttcagtcagttgctcaatgtacctataaccagaccgttcagctggatattacggcctttttaaagaccgtaaagaaaaataagcacaagttttatccggcctttattcacattcttgcccgcctgatgaatgctcatccggaattccgtatggcaatgaaagacggtgagctggtgatatgggatagtgttcacccttgttacaccgttttccatgagcaaactgaaacgttttcatcgctctggagtgaataccacgacgatttccggcagtttctacacatatattcgcaagatgtggcgtgttacggtgaaaacctggcctatttccctaaagggtttattgagaatatgtttttcgtctcagccaatccctgggtgagtttcaccagttttgatttaaacgtggccaatatggacaacttcttcgcccccgttttcacaatgggcaaatattatacgcaaggcgacaaggtgctgatgccgctggcgattcaggttcatcatgccgtctgtgatggcttccatgtcggcagaatgcttaatgaattacaacagtactgcgatgagtggcagggcggggcgtaatttttttaaggcagttattggtgcccttaaacgcctggtgctacgcctgaataagtgataataagcggatgaatggcagaaattcgaatagttacggcttatgacatctttgtggacacatcattcactttttattcacatccggccctgaactcgctaggacttgccccggtgcattttttaaatacccgcgaaaaatagagctgatcgtcaaatccaacattgcgcccaacggtcgctatcggcattcgcgtagtgctaagcagaagtttcgcctggctgatacgctgatcttcgcgccagctcaatacgctaatgcctaactgctggcggaacagatgtgataaccgggagggcgacaggcagacatgctgggcgacgctggcgatatcaaaatggctgtccgccagatggtcgctgatatactggcaggcatcgcgcacacggctatccatcggcgggtgcaacgactcattaattaccgccatacgtctgagcaacaactgctccagcagattgatcgccagtagctcagaatagcgaccttccccttgcccggcgctgatgatctgcccgaacagttcgctgaaatgcggctggcgcgcctcgtccgggcggaaaaatcctgtctgggcaaagattgtcggccaggtcagccactcctgccagtaggcgcgaggccggaaataaacccactggtgataccactcgctggcgtccggatgccgtccatagtgatgaatctcgcccggcggaaacaataatatatcgccaggccgacagacaaactgctcgccattattattaatgacgccctctccgcggatggtcaggttaagaatatatcccttcatgcccaacggacgatcgataaaaaaatccagatatccattcgcttcaattggcgtcagcccggcgaccagatgggcattaaatgaatatcccggcaatagcggatcattttgcgtttcagccatgatttctctaccccccgatgttcagagaagaaacaaattgtccatatcgaccaggacgacagagcttccgtctccgcaagactttgcgcttgatgaaagcacgtatcaaccccgcttgtgaaaagcgctttgtaacaaaagcgtacagttcaggcgataaaattaagtaacagaagtgtctataactatggctggaatgtccacattgaatatttgcacagcgtcacactttgcaaagcattagcatttttgtccataagattagcggatcctgcctgacggtttttgccgcgactctctataatttctccatacctgtttttctggatggagtaaga