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


 * 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