Friday, 28 Jun 2013

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~~=='''6/28/13 Researching the Sequence of kerA'''==~~

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*Planning subteams continued to order reagents and compile protocols.

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*The plasmid design team decided on gibson assembly to produce the kerA biobrick with geneblocks ordered from IDT.

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*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).

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~~CTCCTGCCAAGCTGAAGCGGTCTATTCATACTTTCGAACTGAACATTTTTCTAAAACAGTTATTAATAACCAAAAAATTT~~

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~~TAAATTGGCCCTCCAAAAAAATAGGCCTACCATATAATTCATTTTTTTTCTATAATAAATTAACAGAATAATTGGAATAG~~

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~~ATTATATTATCCTTCTATTTAAATTATTCTGAATAAAGAGGAGGAGAGTGAGTAATGATGAGGAAAAAGAGTTTTTGGCT~~

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~~TGGGATGCTGACGGCCTTCATGCTCGTGTTCACGATGGCATTCAGCGATTCCGCTTCTGCTGCTCAACCGGCGAAAAATG~~

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~~TTGAAAAGGATTATATTGTCGGATTTAAGTCAGGAGTGAAAACCGCATCTGTCAAAAAGGACATCATCAAAGAGAGCGGC~~

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~~GGAAAAGTGGACAAGCAGTTTAGAATCATCAACGCGGCAAAAGCGAAGCTAGACAAAGAAGCGCTTAAGGAAGTCAAAAA~~

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~~TGATCCGGATGTCGCTTATGTGGAAGAGGATCATGTGGCCCATGCCTTGGCGCAAACCGTTCCTTACGGCATTCCTCTCA~~

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~~TTAAAGCGGACAAAGTGCAGGCTCAAGGCTTTAAGGGAGCGAATGTAAAAGTAGCCGTCCTGGATACAGGAATCCAAGCT~~

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~~TCTCATCCGGACTTGAACGTAGTCGGCGGAGCAAGCTTTGTGGCTGGCGAAGCTTATAACACCGACGGCAACGGACACGG~~

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~~CACACATGTTGCCGGTACAGTAGCTGCGCTTGACAATACAACGGGTGTATTAGGCGTTGCGCCAAGCGTATCCTTGTACG~~

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~~CGGTTAAAGT ACTG AAT TC AAGCGGAAGCGGATCATACAGCGGCATTGTAAGCGGAATCGAGTGGGCGACAACAAACGGC~~

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~~ATGGATGTTATCAATATGAGCCTTGGGGGAGCATCAGGCTCGACAGCGATGAAACAGGCAGTCGACAATGCATATGCAAG~~

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~~AGGGGTTGTCGTTGT AGCTGCAGCAGGGAACAGCGGATCTTCAGGAAACACGAATACAATTGGCTATCCTGCGAAATACG~~

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~~ATTCTGTCATCGCTGTTGGTGCGGTAGACTCTAACAGCAACAGAGCTTCATTTTCCAGTGTGGGAGCAGAGCTTGAAGTC~~

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~~ATGGCTCCTGGCGCAGGCGTATACAGCACTTACCCAACGAACACTTATGCAACATTGAACGGAACGTCAATGGTTTCTCC~~

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~~TCATGTAGCGGGAGCAGCAGCTTTGATCTTGTCAAAACATCCGAACCTTTCAGCTTCACAAGTCCGCAACCGTCTCTCCA~~

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~~GCACGGCGACTTATTTGGGAAGCTCCTTCTACTATGGGAAAGGTCTGATCAATGTCGAAGCTGCCGCTCAATAACATATT~~

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~~CTAACAAATAGCATATAGAAAAAGCTAGTGTTTTTAGCACTAGCTTTTTCTTCATTCTGATGAAGGTTGTCCAATATTTT~~

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~~GAATCCGTTCCATGATC~~

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*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.

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*EcoRI (orange) and PstI (blue) sites within the gene are indicated and must be removed to produce the kerA biobrick.

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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.

From 2013.igem.org

Latest revision as of 08:15, 26 September 2013