                     SEQUENCE LISTING

<110>    North Carolina Central University
<120>    PREPARATION OF A FUNCTIONAL RAT LDL RECEPTOR MINIGENE
<130>    295200-9
<160>    NUMBER OF SEQ ID NOS: 29
<170>    KopatentIn 3.0 
<210>    1
<211>    816 
<212>    DNA
<213>    Artificial Sequence
<220>
<223>    genomic fragment #1 of minigene
<400>    1
gttttggtag cacttctggt tgattttact tacattcctc tctgagtgct atttatggtc       60
ggatccgaat aaccatctaa aacctgcaac tgtacttggg tcaaaaatct atgtgcccgt      120
tcctagaaag gagagtaaat gctgcaagga gtctgtgtgt atgtgcgcgc acacgagttg      180
ggggctccgg ggtgggggct ggggggtccc atacttctgt tttcctttgg aggtggaaag      240
gaaaacgtca gaaggcgacc cagggtgtgg aaggatgtgg gtcgtccttt gaacagtgag      300
agctgagtcc acagctaagc atctgggaac tggtgaaatt ctgaaggagg aactcgagga      360
acttcccaag gctaagggag cttcaggggt taaagatcct atgtcacatc ggccgttcca      420
agctcctccc cgcacagtga ggagtagatt tttgaaaatc accccactgc agactcctcc      480
ccctctggaa acctcgtccc tagggctgag aatgactctg gctctgcgcg tgtagtcagt      540
caccagttga cccagtgcgg cgtaggattg cagcccgcat accgtggggc ttgccaccca      600
ggttttgcag ctgagacacc gtgggacccg tgatcctgtg tttgcagcgg gaacatttcg      660
ggtctgtgat ccgagtgggg acgcgacgca gaggctgagg atgagcaccg cggatctgat      720
gctacgctgg gccatcgccc tgctcctggc tgctgctgga gttgcaggta aggcttattc      780
caggagcctg ccgagggcca gggccagccg ctctga                                816

<210>    2
<211>    830 
<212>    DNA
<213>    Artificial Sequence
<220>
<223>    genomic fragment #1 with primer ends (genomic fragment #1)
<400>    2

acgcgtgttt tggtagcact tctggttgat tttacttaca ttcctctctg agtgctattt       60
atggtcggat ccgaataacc atctaaaacc tgcaactgta cttgggtcaa aaatctatgt      120
gcccgttcct agaaaggaga gtaaatgctg caaggagtct gtgtgtatgt gcgcgcacac      180
gagttggggg ctccggggtg ggggctgggg ggtcccatac ttctgttttc ctttggaggt      240
ggaaaggaaa acgtcagaag gcgacccagg gtgtggaagg atgtgggtcg tcctttgaac      300
agtgagagct gagtccacag ctaagcatct gggaactggt gaaattctga aggaggaact      360
cgaggaactt cccaaggcta agggagcttc aggggttaaa gatcctatgt cacatcggcc      420
gttccaagct cctccccgca cagtgaggag tagatttttg aaaatcaccc cactgcagac      480
tcctccccct ctggaaacct cgtccctagg gctgagaatg actctggctc tgcgcgtgta      540
gtcagtcacc agttgaccca gtgcggcgta ggattgcagc ccgcataccg tggggcttgc      600
cacccaggtt ttgcagctga gacaccgtgg gacccgtgat cctgtgtttg cagcgggaac      660
atttcgggtc tgtgatccga gtggggacgc gacgcagagg ctgaggatga gcaccgcgga      720
tctgatgcta cgctgggcca tcgccctgct cctggctgct gctggagttg caggtaaggc      780
ttattccagg agcctgccga gggccagggc cagccgctct gagcggccgc                 830

<210>    3
<211>    28
<212>    DNA
<213>    Artificial Sequence
<220>
<223>    Primer P1
<400>    3
acgcgtgttt tggtagcact tctggttg                                          28

<210>    4
<211>    28
<212>    DNA
<213>    Artificial Sequence
<220>
<223>    Primer P2
<400>    4
gcggccgctc agagcggctg gccctggc                                          28

<210>    5
<211>    223
<212>    DNA
<213>    Artificial Sequence
<220>
<223>    genomic fragment #2 of minigene

<400>    5

catctttgct gtgctgatga ccacaccctt ccttcccttt cgctcttgca gcagaagatt       60
catgtggcaa gaacgagttc cagtgtagag acggaaaatg catcgtcagc aagtgggtgt      120
gtgacggcag ccgcgagtgc ccggatggct ccgatgagtc ccctgagaca tgcagtaagt      180 
cgcctctgat tgtgggtgtt tgtagagaca gaacccatgg ccc                        223

<210>    6
<211>    237
<212>    DNA
<213>    Artificial Sequence
<220>
<223>    genomic fragment #2 with primer ends(genomic fragment #2)

<400>    6

gcggccgcca tctttgctgt gctgatgacc acacccttcc ttccctttcg ctcttgcagc       60
agaagattca tgtggcaaga acgagttcca gtgtagagac ggaaaatgca tcgtcagcaa      120
gtgggtgtgt gacggcagcc gcgagtgccc ggatggctcc gatgagtccc ctgagacatg      180
cagtaagtcg cctctgattg tgggtgtttg tagagacaga acccatggcc cgatatc         237

<210>    7
<211>    30
<212>    DNA
<213>    Artificial Sequence
<220>
<223>    Primer P3
<400>    7
gcggccgcca tctttgctgt gctgatgacc                                        30

<210>    8
<211>    25
<212>    DNA
<213>    Artificial Sequence
<220>
<223>    Primer P4
<400>    8
gatatcgggc catgggttct gtctc                                             25

<210>    9
<211>    224
<212>    DNA
<213>    Artificial Sequence
<220>
<223>    genomic fragment #3 of minigene

<400>    9
tggcctaact aaattgtgtt ccttttgagt gatggctcct ctttatccct agtgtctgtc       60
acctgtcgat ccggtgagtt cagctgtgga ggccgcgtca gccgatgcat tcctgactcc      120
tggagatgtg atgggcggac cgactgtgaa aatggctcgg atgaactaga ctgctgtaag      180
tgacgctcag accaccctgg atgtggcgtg tgtgtgcgcg cgcg                       224

<210>    10
<211>    236
<212>    DNA
<213>    Artificial Sequence
<220>
<223>    genomic fragment #3 with primer ends (genomic fragment #3)

<400>    10

gatatctggc ctaactaaat tgtgttcctt ttgagtgatg gctcctcttt atccctagtg       60
tctgtcacct gtcgatccgg tgagttcagc tgtggaggcc gcgtcagccg atgcattcct      120
gactcctgga gatgtgatgg gcggaccgac tgtgaaaatg gctcggatga actagactgc      180
tgtaagtgac gctcagacca ccctggatgt ggcgtgtgtg tgcgcgcgcg ctcgag          236

<210>    11
<211>    28
<212>    DNA
<213>    Artificial Sequence
<220>
<223>    Primer P5
<400>    11
gatatctggc ctaactaaat tgtgttcc                                          28

<210>    12
<211>    26
<212>    DNA
<213>    Artificial Sequence
<220>
<223>    Primer P6
<400>    12
ctcgagcgcg cgcgcacaca cacgcc                                            26

<210>    13
<211>    293
<212>    DNA
<213>    Artificial Sequence
<220>
<223>    genomic fragment #4 of minigene

<400>    13

ggtatgagcc atagcatgac agggcgctct cctctctgtg cacccccaca gcccccaaga       60
cgtgctccct ggatgagttc cgctgccagg atggcaagtg catctcccgg cagtttgtgt      120
gtgaccaaga ctgggattgc ctggatggct ctgacgaggc ccactgtgcg gccaccactt      180
gtggccctgc tcacttccgc tgcaactcct cttcctgcat ccccagcctg tgggcctgcg      240
acggggaccg ggactgtgac gatggctccg atgagtggcc gcagaactgc ggg             293

<210>    14
<211>    305
<212>    DNA
<213>    Artificial Sequence
<220>
<223>    genomic fragment #4 with primer ends(genomic fragment #4)

<400>    14

ctcgagggta tgagccatag catgacaggg cgctctcctc tctgtgcacc cccacagccc       60
ccaagacgtg ctccctggat gagttccgct gccaggatgg caagtgcatc tcccggcagt      120
ttgtgtgtga ccaagactgg gattgcctgg atggctctga cgaggcccac tgtgcggcca      180
ccacttgtgg ccctgctcac ttccgctgca actcctcttc ctgcatcccc agcctgtggg      240
cctgcgacgg ggaccgggac tgtgacgatg gctccgatga gtggccgcag aactgcgggg      300
aattc                                                                  305

<210>    15
<211>    29
<212>    DNA
<213>    Artificial Sequence
<220>
<223>    Primer P7
<400>    15
ctcgagggta tgagccatag catgacagg                                         29

<210>    16
<211>    27
<212>    DNA
<213>    Artificial Sequence
<220>
<223>    Primer P8
<400>    16
gaattccccg cagttctgcg gccactc                                           27

<210>    17
<211>    36
<212>    DNA
<213>    Artificial Sequence
<220>
<223>    Oligonucleotides used to insert Mlu I site into the pSG5 vector 
          
<400>    17
gaaccagctg tggaacgcgt gtcagttagg gtgtgg                                 36

<210>    18
<211>    42
<212>    DNA
<213>    Artificial Sequence
<220>
<223>    Oligonucleotides used to mutate the LDL receptor promoter
          within the assembled-minigene for motif -156
<400>    18
ctgcgcgtgt gatcagtcac cgtcctgccc agcgcggcgt ag                          42

<210>    19
<211>    37
<212>    DNA
<213>    Artificial Sequence
<220>
<223>    Oligonucleotides used to mutate the LDL receptor promoter 
         within the assembled-minigene for motif -612
<400>    19
cctgcaactg tacttaaatt aaaaatctat gtgcccg                                37

<210>    20
<211>    20
<212>    DNA
<213>    Artificial Sequence
<220>
<223>    sequencing primer(1)
<400>    20
gcgattaagt tgggtaacgc                                                   20
 
<210>    21
<211>    20
<212>    DNA
<213>    Artificial Sequence
<220>
<223>    sense primer (2)
<400>    21
tgagcaccgc ggatctgatg                                                   20

<210>    22
<211>    18
<212>    DNA
<213>    Artificial Sequence
<220>
<223>    antisense primer (3)
<400>    22
gtaaccatta taagctgc                                                     18

<210>    23
<211>    15
<212>    DNA
<213>    Artificial Sequence
<220>
<223>    antisense primer (5)
<400>    23
accattataa gctgc                                                        15

<210>    24
<211>    20
<212>    DNA
<213>    Artificial Sequence
<220>
<223>    new sense primer (4)
<400>    24
ctagactgct cccccaagac                                                   20

<210>    25
<211>    18
<212>    DNA
<213>    Artificial Sequence
<220>
<223>    primer
<400>    25
gggacaagtg gcgttcag                                                     18

<210>    26
<211>    20
<212>    DNA
<213>    Artificial Sequence
<220>
<223>    primer
<400>    26
cgctgagcca gtcagtgtag                                                   20

<210>    27
<211>    868
<212>    DNA
<213>    Artificial Sequence
<220>
<223>     mRNA sequence for Rat LDL receptor minigene
<400>    27

ttgacccagt gcggcgtagg attgcagccc gcataccgtg gggcttgcca cccaggtttt       60
gcagctgaga caccgtggga cccgtgatcc tgtgtttgca gcgggaacat ttcgggtctg      120
tgatccgagt ggggacgcga cgcagaggct gagatgagca ccgcggatct gatgctacgc      180
tgggccatcg ccctgctcct ggctgctgct ggagttgcag cagaagattc atgtggcaag      240
aacgagttcc agtgtagaga cggaaaatgc atcgtcagca agtgggtgtg tgacggcagc      300
cgcgagtgcc cggatggctc cgatgagtcc cctgagacat gcatgtctgt cacctgtcga      360
tccggtgagt tcagctgtgg aggccgcgtc agccgatgca ttcctgactc ctggagatgt      420
gatgggcgga ccgactgtga aaatggctcg gatgaactag actgctcccc caagacgtgc      480
tccctggatg agttccgctg ccaggatggc aagtgcatct cccggcagtt tgtgtgtgac      540
caagactggg attgcctgga tggctctgac gaggcccact gtgcggccac cacttgtggc      600
cctgctcact tccgctgcaa ctcctcttcc tgcatcccca gcctgtgggc ctgcgacggg      660
gaccgggact gtgacgatgg ctccgatgag tggccgcaga actgcgggga attcggatcc      720
agatcttatt aaagcagaac ttgtttattg cagcttataa tggttacaaa taaagcaata      780
gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca      840
aactcatcaa tgtatcttat catgtctg                                         868


<210>    28
<211>    579
<212>    DNA
<213>    Artificial Sequence
<220>
<223>    Coding sequence
<223>    
<400>    28

atgagcaccg cggatctgat gctacgctgg gccatcgccc tgctcctggc tgctgctgga       60
gttgcagcag aagattcatg tggcaagaac gagttccagt gtagagacgg aaaatgcatc      120
gtcagcaagt gggtgtgtga cggcagccgc gagtgcccgg atggctccga tgagtcccct      180
gagacatgca tgtctgtcac ctgtcgatcc ggtgagttca gctgtggagg ccgcgtcagc      240
cgatgcattc ctgactcctg gagatgtgat gggcggaccg actgtgaaaa tggctcggat      300
gaactagact gctcccccaa gacgtgctcc ctggatgagt tccgctgcca ggatggcaag      360
tgcatctccc ggcagtttgt gtgtgaccaa gactgggatt gcctggatgg ctctgacgag      420
gcccactgtg cggccaccac ttgtggccct gctcacttcc gctgcaactc ctcttcctgc      480
atccccagcc tgtgggcctg cgacggggac cgggactgtg acgatggctc cgatgagtgg      540
ccgcagaact gcggggaatt cggatccaga tcttattaa                             579


<210>    29
<211>    187
<212>    PRT
<213>    Synthetic Peptide
<220>
<223>    WT minigene sequence
<400>    29
			      
Met Ser Thr Ala Asp Leu Met Leu Arg Trp Ala Ile Ala Leu Leu Leu 
  1               5                  10                  15
Ala Ala Ala Gly Val Ala Ala Glu Asp Ser Cys Gly Lys Asn Glu Phe 
             20                  25                  30
Gln Cys Arg Asp Gly Lys Cys Ile Val Ser Lys Trp Val Cys Asp Gly 
         35                  40                  45
Ser Arg Glu Cys Pro Asp Gly Ser Asp Glu Ser Pro Glu Thr Cys Met 
     50                  55                  60 
Ser Val Thr Cys Arg Ser Gly Glu Phe Ser Cys Gly Gly Arg Val Ser 
65                  70                  75                  80
Arg Cys Ile Pro Asp Ser Trp Arg Cys Asp Gly Arg Thr Asp Cys Glu 
                 85                  90                  95
Asn Gly Ser Asp Glu Leu Asp Cys Ser Pro Lys Thr Cys Ser Leu Asp 
            100                 105                 110 
Glu Phe Arg Cys Gln Asp Gly Lys Cys Ile Ser Arg Gln Phe Val Cys 
        115                 120                 125
Asp Gln Asp Trp Asp Cys Leu Asp Gly Ser Asp Glu Ala His Cys Ala 
    130                 135                 140 
Ala Thr Thr Cys Gly Pro Ala His Phe Arg Cys Asn Ser Ser Ser Cys 
145                 150                 155                 160
Ile Pro Ser Leu Trp Ala Cys Asp Gly Asp Arg Asp Cys Asp Asp Gly 
                165                 170                 175 
Ser Asp Glu Trp Pro Gln Asn Cys Gly Glu Phe  
            180                 185      

<210>    30
<211>    192
<212>    PRT
<213>    Synthetic Peptide
<220>
<223>    WT minigene sequence
<400>    30
			      
Met Ser Thr Ala Asp Leu Met Leu Arg Trp Ala Ile Ala Leu Leu Leu 
  1               5                  10                  15
Ala Ala Ala Gly Val Ala Ala Glu Asp Ser Cys Gly Lys Asn Glu Phe 
             20                  25                  30
Gln Cys Arg Asp Gly Lys Cys Ile Val Ser Lys Trp Val Cys Asp Gly 
         35                  40                  45
Ser Arg Glu Cys Pro Asp Gly Ser Asp Glu Ser Pro Glu Thr Cys Met 
     50                  55                  60 
Ser Val Thr Cys Arg Ser Gly Glu Phe Ser Cys Gly Gly Arg Val Ser 
65                  70                  75                  80
Arg Cys Ile Pro Asp Ser Trp Arg Cys Asp Gly Arg Thr Asp Cys Glu 
                 85                  90                  95
Asn Gly Ser Asp Glu Leu Asp Cys Ser Pro Lys Thr Cys Ser Leu Asp 
            100                 105                 110 
Glu Phe Arg Cys Gln Asp Gly Lys Cys Ile Ser Arg Gln Phe Val Cys 
        115                 120                 125
Asp Gln Asp Trp Asp Cys Leu Asp Gly Ser Asp Glu Ala His Cys Ala 
    130                 135                 140 
Ala Thr Thr Cys Gly Pro Ala His Phe Arg Cys Asn Ser Ser Ser Cys 
145                 150                 155                 160
Ile Pro Ser Leu Trp Ala Cys Asp Gly Asp Arg Asp Cys Asp Asp Gly 
                165                 170                 175 
Ser Asp Glu Trp Pro Gln Asn Cys Gly Glu Phe Gly Ser Arg Ser Tyr
            180                 185                 190

