Friday, 28 Jun 2013

From 2013.igem.org

(Difference between revisions)
(Created page with "=='''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 p...")
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*The plasmid design team decided on gibson assembly to produce the kerA biobrick with geneblocks ordered from IDT.
*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).  
*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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[[File:UChicago 2013 kerA plasmid design.png]]
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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
+
*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.
*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.
*EcoRI (orange) and PstI (blue) sites within the gene are indicated and must be removed to produce the kerA biobrick.

Revision as of 07:17, 26 September 2013

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).

UChicago 2013 kerA plasmid design.png

  • 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.
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