IGEM:The Citadel/Notebook/Notes Brian/07-09-10

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  • Made glycerol stocks of the 5 parts that were minipreped on 06-08 and 06-30, since we had not prepared these already.
    • BBa_J23101
    • BBa_I13521
    • BBa_I20260
    • BBa_I0462
    • BBa_J45120

(The first two are from 06-08. The next three are from 06-30.)

NOTE: Cells should not be permitted to grow overnight for glycerol stock.

Notes from my construction of a Biobrick for PYY3-36:

mRNA sequence from Human PYY: 

       1 gcccctggag gaactgaacc cactatcggt catggggccg agactaaatg tggcgggttg
      61 tctttaatct gctgccaaga ggaaactcat tcaggcaagt tcagcccttt atgaggaatt
     121 cccctgtggt cacattccaa ttcctggacc tgctgccacc ctcagaactg catgctcctt
     181 cttcagactt tctaagaatg actcaggtca ttggtggagt gaagtcaaga tttccaactc
     241 agtcacctga agagatggag ataccattca tggagctgga ggtccctgga gatttgggaa
     301 ttcagataac aagctaagat aaggagtttg cctacctctg tcctagagcg aagcctgagc
     361 cttgggcgcg cagcacacca caagtatctg ttactgtgtt ttgcagaagc ttcaggcggg
     421 gatataagcc ccacaaggaa agcgctgagc agaggaggcc tcagcttgac ctgcggcagt
     481 gcagcccttg ggacttccct cgccttccac ctcctgctcg tctgcttcac aagctatcgc
     541 tatggtgttc gtgcgcaggc cgtggcccgc cttgaccaca gtgcttctgg ccctgctcgt
     601 ctgcctaggg gcgctggtcg acgcctaccc catcaaaccc gaggctcccg gcgaagacgc
     661 ctcgccggag gagctgaacc gctactacgc ctccctgcgc cactacctca acctggtcac
     721 ccggcagcgg tatgggaaaa gagacggccc ggacacgctt ctttccaaaa cgttcttccc
     781 cgacggcgag gaccgccccg tcaggtcgcg gtcggagggc ccagacctgt ggtgaggacc
     841 cctgaggcct cctgggagat ctgccaacca cgcccacgtc atttgcatac gcactcccga
     901 ccccagaaac ccggattctg cctcccgacg gcggcgtctg ggcagggttc gggtgcggcc
     961 ctccgcccgc gtctcggtgc ccccgccccc tgggctggag ggctgtgtgt ggtccttccc
    1021 tggtcccaaa ataaagagca aattccacag aaaaaaaaaa aaaaaaaaa


Start position = 542 on

Homo sapiens peptide YY (PYY), mRNA
NCBI Reference Sequence: NM_004160.4
(GeneID: 5697)
(PDB: 2DF0A)

Excerpt of PYY coding region: 

     541 ~atggtgttc gtgcgcaggc cgtggcccgc cttgaccaca gtgcttctgg ccctgctcgt
     601 ctgcctaggg gcgctggtcg acgcctaccc catcaaaccc gaggctcccg gcgaagacgc
     661 ctcgccggag gagctgaacc gctactacgc ctccctgcgc cactacctca acctggtcac
     721 ccggcagcgg tatgggaaaa gagacggccc ggacacgctt ctttccaaaa cgttcttccc
     781 cgacggcgag gaccgccccg tcaggtcgcg gtcggagggc ccagacctgt ggtgaggacc
     841 cctgaggcct cctgggagat ctgccaacca cgcccacgtc atttgcatac gcactcccga
     901 ccccagaaac ccggattctg cctcccgacg gcggcgtctg ggcagggttc gggtgcggcc
     961 ctccgcccgc gtctcggtgc ccccgccccc tgggctggag ggctgtgtgt ggtccttccc
    1021 tggtcccaaa ataaagagca aattccacag aaaaaaaaaa aaaaaaaaa


Complete PYY3-36:

atc aaa ccc gag gct ccc ggc gaa gac gcc
tcg ccg gag gag ctg aac cgc tac tac gcc tcc ctg cgc cac tac ctc aac ctg gtc acc
cgg cag cgg tat


Amino Acid Sequence:

IKPEAPGEDASPEELNRYYASLRHYLNLVTRQRY


mRNA Sequence:

atcaaaccc gaggctcc cggcgaa gacgcct cgccgg aggagct gaaccgc tactacg cctccct gcgccac tacctca acctggt cacccgg cagcgg tat


DNA Sequence:

tagtttgggctccgagggccgcttctgcggagcggcctcctcgacttggcgatgatgcggagggacgcggtgatggagttggaccagtgggccgtcgccata

DNA Sequence with Start Codon and Double Stop Codon Included:

atgtagtttgggctccgagggccgcttctgcggagcggcctcctcgacttggcgatgatgcggagggacgcggtgatggagttggaccagtgggccgtcgccatataataa

DNA Sequence w/ Start Codon, Double Stop Codon, Biobrick prefix and suffix Included:

gaattcgcggccgcttctagatgtagtttgggctccgagggccgcttctgcggagcggcctcctcgacttggcgatgatgcggagggacgcggtgatggagttggaccagtgggccgtcgccatataataatactagtagcggccgctgcag

DNA Sequence to have synthesized (sticky ends available):

aattcgcggccgcttctagatgtagtttgggctccgagggccgcttctgcggagcggcctcctcgacttggcgatgatgcggagggacgcggtgatggagttggaccagtgggccgtcgccatataataatactagtagcggccgctgca

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