ALDH3A1

Aldehyde dehydrogenase 3 family, member A1 [ Homo sapiens ]

Location: 17p11.2

Gene report:

Source: http://www.ncbi.nlm.nih.gov/gene/218

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Defining the region where the promoter is likely to be found

Proposed Promoter Sequence: (position -19649444 to -19648446) 998bp

TATAGGCGTGAGCCACCGCGCCCGGCCTGTAACCTGTTTTCTTTCTTATCCCCAGTGCCT GAGACACCCCTGGTGCAGAGAGAGCCCCTACAAACATGCTGAATAGCCTGCTCCACCT AATCCCTAAATACTGCACAGTAAACCCTTACGTGGGTGTGACAGGAGTCATTTAAACTG TCTCCTTTCTTTCGGATCTTTGGGATGTTTACAATACTTCCCTGTTATAGAAAACAGTGA TGACTTTCTTATTCGTGAATGTTTAGGAAAGGCTGTAGGACCTTTTCGTCACAGAGAAG TATGTTTGAAAAACTTTTGCCACATATTGCCAAATTGCCCCCAGCAGGGCCATCCCGAT TTGCCACTATACTTTCGTGTATTGGTAGCAGGAATCGCCTCCACACCTCTTTGTGGAAAT CTGCAGCTCTCTGAATCCTTGGCGGGCATGGGCGTGCGTCTCCCAGGTGTGGCACGCGT GGTTTACACACCAAGCCTGGTTCAGCTCTGCCAAGAAACCAGCGCTTTTTAGAACAATG GCTTAGCCAGGAGGGTGGTGCTGGGTTTTTCTTTATCGGCTTGGGTGTGGAATCGCGGA CTGCGCCCCTCCAATCTGATCTGGAGAGTGGCTGACTGCGTGTGCAGAGCTCTGGGGGT GTTGGTCTGGGATTGTGCCATCAGCTTTGCAAGCAAGTAAGGGAGCGGAAAAGGCCGG GAAAGGCCCTGCCGCGAGCACGCTGCCAAGAGCCCCCAGCAGCAGTTCGGCTTAGGAC TCGGGTTGCGGCGGGTGTCACCTTCTCAGGGGCTAGCAAGGCAGCCAGGGCCCAGGCG TCTGAGTGAGGGGCGGGAGAGGAGGCGAGGCAGAAAGTGGACCTTCCAGCGGAAAGG CCATTTTCCCCAAGGCCGAGCCCAGGGAAGTCCCTTCCTATAGAATTCAGGCAGGGTGG GAGGCAGGGCGCGCTCGTGCCCCTCAGCCAGCTGCAGGTGCTCTCTGTCCCCAGGCG

Source: http://www.ncbi.nlm.nih.gov/gene/218

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Finding restriction sites

The proposed promoter sequence was checked for restriction sites using NEBcutter 2.0. The following enzymes cut the sequence:

NOTE: There is a recognition site for EcoRI within the sequence. Source: http://tools.neb.com/NEBcutter2/cutshow.php?name=6d7a0d36-ALDH3A1_Promoter

Defining the best pair of primers

The following primers were designed to amplify the selected area. Forward-Primer: TGAACTTCTGACCTCAGGTGA Reverse-Primer:

CGGCCTCGCTGATCTTGC

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These primers were selected with www.yeastgenome.org

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The sequence (1080 bp) of DNA that will be amplified by these primers is:
TGAACTTCTG 1 51 GCTAGGATTA 101 TTCTTATCCC 151 AACATGCTGA 201 CCCTTACGTG 251 ATCTTTGGGA 301 TTCTTATTCG 351 GAAGTATGTT 401 GGGCCATCCC 451 CCTCCACACC 501 ATGGGCGTGC 551 GGTTCAGCTC 601 GGAGGGTGGT 651 CTGCGCCCCT 701 TCTGGGGGTG 751 GGAGCGGAAA 801 CCCCAGCAGC 851 TCAGGGGCTA 901 AGAGGAGGCG 951 CAAGGCCGAG 1001 AGGCAGGGCG 1051 CAGGCGCCAT ACCTCAGGTG TAGGCGTGAG CAGTGCCTGA ATAGCCTGCT GGTGTGACAG TGTTTACAAT TGAATGTTTA TGAAAAACTT GATTTGCCAC TCTTTGTGGA GTCTCCCAGG TGCCAAGAAA GCTGGGTTTT CCAATCTGAT TTGGTCTGGG AGGCCGGGAA AGTTCGGCTT GCAAGGCAGC AGGCAGAAAG CCCAGGGAAG CGCTCGTGCC GAGCAAGATC ACCCGCCCAC CCACCGCGCC GACACCCCTG CCACCTAATC GAGTCATTTA ACTTCCCTGT GGAAAGGCTG TTGCCACATA TATACTTTCG AATCTGCAGC TGTGGCACGC CCAGCGCTTT TCTTTATCGG CTGGAGAGTG ATTGTGCCAT AGGCCCTGCC AGGACTCGGG CAGGGCCCAG TGGACCTTCC TCCCTTCCTA CCTCAGCCAG AGCGAGGCCG CTCAGCCTCC CGGCCTGTAA GTGCAGAGAG CCTAAATACT AACTGTCTCC TATAGAAAAC TAGGACCTTT TTGCCAAATT TGTATTGGTA TCTCTGAATC GTGGTTTACA TTAGAACAAT CTTGGGTGTG GCTGACTGCG CAGCTTTGCA GCGAGCACGC TTGCGGCGGG GCGTCTGAGT AGCGGAAAGG TAGAATTCAG CTGCAGGTGC CAAAGTGTTT CCTGTTTTCT AGCCCCTACA GCACAGTAAA TTTCTTTCGG AGTGATGACT TCGTCACAGA GCCCCCAGCA GCAGGAATCG CTTGGCGGGC CACCAAGCCT GGCTTAGCCA GAATCGCGGA TGTGCAGAGC AGCAAGTAAG TGCCAAGAGC TGTCACCTTC GAGGGGCGGG CCATTTTCCC GCAGGGTGGG TCTCTGTCCC

Standard Part Fabrication

The iGEM prefix and suffix were added to the primer sequence. PREFIX PRIMER SUFFIX

5' GAATTC GCGGCCGC T TCTAGA G TGAACTTCTGACCTCAGGTGA T ACTAGT A GCGGCCG CTGCAG 3'

3' CTGCAG CGGCCGC T ACTAGT A CGGCCTCGCTGATCTTGC C TCTAGA A GCGGCCGC GAATTC 5'

Source: Alignment of various methods

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