Friday, 28 Jun 2013

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6/28/13 Researching the Sequence of kerA

Planning subteams continued to order reagents and compile protocols.
The plasmid design team decided on gibson assembly to produce the kerA biobrick with geneblocks ordered from IDT.
kerA sequence was obtained from “Nucleotide Sequence and Expression of kerA, the Gene Encoding a Keratinolytic Protease of Bacillus licheniformis PWD-1” (Lin et al. 1995).

CTCCTGCCAAGCTGAAGCGGTCTATTCATACTTTCGAACTGAACATTTTTCTAAAACAGTTATTAATAACCAAAAAATTT TAAATTGGCCCTCCAAAAAAATAGGCCTACCATATAATTCATTTTTTTTCTATAATAAATTAACAGAATAATTGGAATAG ATTATATTATCCTTCTATTTAAATTATTCTGAATAAAGAGGAGGAGAGTGAGTAATGATGAGGAAAAAGAGTTTTTGGCT TGGGATGCTGACGGCCTTCATGCTCGTGTTCACGATGGCATTCAGCGATTCCGCTTCTGCTGCTCAACCGGCGAAAAATG TTGAAAAGGATTATATTGTCGGATTTAAGTCAGGAGTGAAAACCGCATCTGTCAAAAAGGACATCATCAAAGAGAGCGGC GGAAAAGTGGACAAGCAGTTTAGAATCATCAACGCGGCAAAAGCGAAGCTAGACAAAGAAGCGCTTAAGGAAGTCAAAAA TGATCCGGATGTCGCTTATGTGGAAGAGGATCATGTGGCCCATGCCTTGGCGCAAACCGTTCCTTACGGCATTCCTCTCA TTAAAGCGGACAAAGTGCAGGCTCAAGGCTTTAAGGGAGCGAATGTAAAAGTAGCCGTCCTGGATACAGGAATCCAAGCT TCTCATCCGGACTTGAACGTAGTCGGCGGAGCAAGCTTTGTGGCTGGCGAAGCTTATAACACCGACGGCAACGGACACGG CACACATGTTGCCGGTACAGTAGCTGCGCTTGACAATACAACGGGTGTATTAGGCGTTGCGCCAAGCGTATCCTTGTACG CGGTTAAAGT ACTG AAT TC AAGCGGAAGCGGATCATACAGCGGCATTGTAAGCGGAATCGAGTGGGCGACAACAAACGGC ATGGATGTTATCAATATGAGCCTTGGGGGAGCATCAGGCTCGACAGCGATGAAACAGGCAGTCGACAATGCATATGCAAG AGGGGTTGTCGTTGT AGCTGCAGCAGGGAACAGCGGATCTTCAGGAAACACGAATACAATTGGCTATCCTGCGAAATACG ATTCTGTCATCGCTGTTGGTGCGGTAGACTCTAACAGCAACAGAGCTTCATTTTCCAGTGTGGGAGCAGAGCTTGAAGTC ATGGCTCCTGGCGCAGGCGTATACAGCACTTACCCAACGAACACTTATGCAACATTGAACGGAACGTCAATGGTTTCTCC TCATGTAGCGGGAGCAGCAGCTTTGATCTTGTCAAAACATCCGAACCTTTCAGCTTCACAAGTCCGCAACCGTCTCTCCA GCACGGCGACTTATTTGGGAAGCTCCTTCTACTATGGGAAAGGTCTGATCAATGTCGAAGCTGCCGCTCAATAACATATT CTAACAAATAGCATATAGAAAAAGCTAGTGTTTTTAGCACTAGCTTTTTCTTCATTCTGATGAAGGTTGTCCAATATTTT GAATCCGTTCCATGATC

Two putative promoters (red), a possible ribosomal binding site (green), and a transcriptional terminator sequence (light blue, single underline) are indicated. The putative starting residues of the preprotein (pre = blue), proprotein (pro = pink), and mature protein (mature = purple) are indicated.
EcoRI (orange) and PstI (blue) sites within the gene are indicated and must be removed to produce the kerA biobrick.