                         SEQUENCE LISTING

<110>  The Salk Institute for Biological Studies
       Wang, Lei
 
<120>  METHODS OF GENETICALLY ENCODING UNNATURAL AMINO ACIDS IN 
       EUKARYOTIC CELLS USING ORTHOGONAL tRNA/SYNTHETASE PAIRS

<130>  7159-78350-02

<150>  60/923,247
<151>  2007-04-13

<160>  34    

<170>  PatentIn version 3.3

<210>  1
<211>  48
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  forward primer sequences used to amplify the E. coli TyrRS gene

<400>  1
ccaccatgga actcgagatt ttgatggcaa gcagtaactt gattaaac                  48


<210>  2
<211>  43
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  reverse primer sequences used to amplify the E. coli TyrRS gene

<400>  2
acaagatctg ctagcttatt tccagcaaat cagacagtaa ttc                       43


<210>  3
<211>  69
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  forward  primer sequence used to amplify the gene for EctRNATyr 
       in construct tRNA2

<400>  3
gtgggatccc cggtggggtt cccgagcggc caaagggagc agactctaaa tctgccgtca     60

tcgacttcg                                                             69


<210>  4
<211>  69
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  reverse primer sequence used to amplify the gene for EctRNATyr in
       construct tRNA2

<400>  4
gataagcttt tccaaaaatg gtggtggggg aaggattcga accttcgaag tcgatgacgg     60

cagatttag                                                             69


<210>  5
<211>  26
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  forward primer sequence used to amplify the gene for the E. coli 
       LeuRS gene

<400>  5
gcctcgagaa gagcaatacc gcccgg                                          26


<210>  6
<211>  31
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  reverse primer sequence used to amplify the gene for the E. coli 
       LeuRS gene

<400>  6
cgctagctta gccaacgacc agattgagga g                                    31


<210>  7
<211>  37
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  forward primer sequence used to amplify the gene for 32P-labeled 
       DNA probes specific for the EctRNATyr

<400>  7
aaccttcgaa gtcgatgacg gcagatttac agtctgc                              37


<210>  8
<211>  33
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  reverse primer sequence used to amplify the gene for 32P-labeled 
       DNA probes specific for the EctRNATyr

<400>  8
ccgtctaaat gtcagacgag ggaaaccggc gag                                  33


<210>  9
<211>  26
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  forward sequence FW19 used to amplify a spacer sequence from 
       pCDNA3

<400>  9
atactagtgc gggcgctagg gcgctg                                          26


<210>  10
<211>  26
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  reverse primer sequence FW20 used to amplify a spacer sequence 
       from pCDNA3

<400>  10
atggtacccc tggaaggtgc cactcc                                          26


<210>  11
<211>  27
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  forward primer sequence FW21 used to amplify the E. coli TyrRS 
       gene from E. coli genomic DNA

<400>  11
caactagtat ggagattttg atggcaa                                         27


<210>  12
<211>  30
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  reverse primer sequence FW22 used to amplify the E. coli TyrRS 
       gene from E. coli genomic DNA

<400>  12
aactcgagtt atttccagca aatcagacag                                      30


<210>  13
<211>  35
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  forward primer sequence FW16 used to amplify the SNR52 promoter 
       from yeast genomic DNA

<400>  13
cactgcagtc tttgaaaaga taatgtatga ttatg                                35


<210>  14
<211>  44
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  reverse primer sequence FW17 used to amplify the SNR52 promoter 
       from yeast genomic DNA

<400>  14
ggccgctcgg gaaccccacc gatcatttat ctttcactgc ggag                      44


<210>  15
<211>  24
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  forward primer sequence FW14  used to amplify the EctRNATyr gene 
       followed by the 3'-flanking sequence of the SUP4 suppressor tRNA 
       from pEYCUA-YRS

<400>  15
ggtggggttc ccgagcggcc aaag                                            24


<210>  16
<211>  52
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  reverse primer sequence FW15 used to amplify the EctRNATyr gene 
       followed by the 3'-flanking sequence of the SUP4 suppressor tRNA 
       from pEYCUA-YRS

<400>  16
ggtcgacaga cataaaaaac aaaaaaatgg tgggggaagg attcgaacct tc             52


<210>  17
<211>  31
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  forward sequence FW12 used to amplify the RPR1 promoter from 
       yeast genomic DNA

<400>  17
cactgcagtc tgccaattga acataacatg g                                    31


<210>  18
<211>  45
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  reverse primer sequence FW13 used to amplify the RPR1 promoter 
       from yeast genomic DNA

<400>  18
ggccgctcgg gaaccccacc tgccaatcgc agctcccaga gtttc                     45


<210>  19
<211>  34
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  forward primer sequence used to amplify a gene cassette 
       containing the 5' flanking sequence of the SUP4 suppressor tRNA, 
       the EctRNATyr, and the 3' flanking sequence of the SUP4 
       suppressor tRNA from plasmid  pEYCUA-YRS-tRNA-5

<400>  19
cactgcagct ctttttcaat tgtaatgtgt tatg                                 34


<210>  20
<211>  30
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  forward primer sequence FW27 used to amplify a gene cassette 
       containing the 5' flanking sequence of the SUP4 suppressor tRNA, 
       the EctRNALeu, and the 3' flanking sequence of the SUP4 
       suppressor tRNA from plasmid pLeuRSB8T252A

<400>  20
caaagcttct ctttttcaat tgtatatgtg                                      30


<210>  21
<211>  31
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  reverse primer sequenc3 FW28 used to amplify a gene cassette 
       containing the 5' flanking sequence of the SUP4 suppressor tRNA, 
       the EctLeu, and the 3' flanking sequence of the SUP4 suppressor 
       tRNA from plasmid pLeuRSB8T252A

<400>  21
gagtcgacag acataaaaaa caaaaaaata c                                    31


<210>  22
<211>  29
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  forward primer sequence FW29 used to amplify the E. coli LeuRS 
       gene from E. coli genomic DNA

<400>  22
agctcgagtt agccaacgac cagattgag                                       29


<210>  23
<211>  29
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  reverse primer sequence FW30 used to amplify the E. coli LeuRS 
       gene from E. coli genomic DNA

<400>  23
agactagtat gcaagagcaa taccgcccg                                       29


<210>  24
<211>  44
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  reverse primer sequence FW31 used to amplify the SNR52 promoter 
       from pSNR-TyrRS

<400>  24
ctaccgattc caccatccgg gcgatcattt atctttcact gcgg                      44


<210>  25
<211>  22
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  forward primer sequence FW32 used to amplify the EctRNALeu -3' 
       flanking sequence fragment from pLeuRSB8T252A

<400>  25
gcccggatgg tggaatcggt ag                                              22


<210>  26
<211>  53
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  forward  primer sequence JT171 used to amplify a mutant GFP-TAG 
       gene

<400>  26
tagtcggatc ctcagtgatg gtgatggtga tgcttgtaca gctcgtccat gcc            53


<210>  27
<211>  56
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  reverse primer sequence JT172 used to amplify a mutant GFP-TAG 
       gene

<400>  27
tagtcgtcga catggattac aaagatgatg atgataaagt gagcaagggc gaggag         56


<210>  28
<211>  29
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  sequence of a biotinylated probe FW39 which is specific for the 
       E. coli tRNATyr and the EctRNATyr

<400>  28
tctgctccct ttggccgctc gggaacccc                                       29


<210>  29
<211>  26
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  forward primer sequence FW5 used to amplify a gene cassette 
       containing ~200 bp upstream of UPF1, the Kan-MX6, and ~200 bp 
       downstream of UPF1

<400>  29
aatgaaaagc ttaccagaaa cttacg                                          26


<210>  30
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  reverse primer sequence FW6 used to amplify a gene cassette 
       containing ~200 bp upstream of UPF1, the Kan-MX6, and ~200 bp 
       downstream of UPF1

<400>  30
ggctaggata tcaagtccat gcc                                             23


<210>  31
<211>  26
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  forward primer sequence used to amplify genomic DNA ~300 bp away 
       from the UPF1 gene

<400>  31
gatttgggag ggacaccttt atacgc                                          26


<210>  32
<211>  25
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  reverse primer sequence used to amplify genomic DNA ~300 bp away 
       from the UPF1 gene

<400>  32
ttcattagaa gtacaatggt agccc                                           25


<210>  33
<211>  85
<212>  DNA
<213>  E. coli

<400>  33
ggtggggttc ccgagcggcc aaagggagca gactctaaat ctgccgtcat cgacttcgaa     60

ggttcgaatc cttcccccac cacca                                           85


<210>  34
<211>  87
<212>  DNA
<213>  E. coli

<400>  34
gcccggatgg tggaatcggt agacacaagg gattctaaat ccctcggcgt tcgcgctgtg     60

cgggttcaag tcccgctccg ggtacca                                         87


